METHODS OF PRODUCING REUTERIN AND PROPENOIC ACID USING AN ISOLATED STRAIN OF LACTOBACILLUS REUTERI

Cross-Reference to Related Applications

[0001] This application claims priority to U.S. provisional application No. 63/413,950, filed on October 6, 2022, the contents of which is hereby incorporated by reference in its entirety.

Field

[0002] The present disclosure relates in some aspects to isolated strains of Lactobacillus reuteri, and their use in methods of producing 3-hydroxypropanal (reuterin) and/or propenoic acid, along with kits and compositions thereof.

Background

[0003] Propenoic (acrylic) acid is a simple carboxylic acid that is used to make thousands of specialized chemicals. It is the source of vinyls, polishes, paints, clear plastics, coatings, adhesives, and elastomers, which are found in everything from cleaning products to mobile phones. The most infamous derivatives driving consumer demand for green biomanufacturing are polyacrylic acids — the superabsorbent polymers used in diapers, feminine hygiene products, and absorbent pads found under packaged meat products, such as chicken and ground beef. Polyacrylic acid (polypropenoic acid) may account for up to 30% of the manufacturing price of diapers, for instance (Journal of Cleaner Production, Disposable baby diapers: life cycle costs, eco-efficiency and circular economy. November 2018). However, existing methods of producing propenoic acid can be environmentally unfriendly and prohibitively expensive. Accordingly, there is a need in the field of producing propenoic acid in an economical and environmentally friendly way. The methods, isolated strains, kits, and compositions described herein satisfy this need.

Summary

[0004] Provided herein is a method of producing 3-hydroxypropanal (reuterin), which includes: (a) culturing an isolated strain of Lactobacillus reuteri in a growth medium, thereby resulting in a population of the isolated strain of Lactobacillus reuteri; and (b) culturing the population in a production medium containing a glycerol source to produce 3-hydroxypropanal. In some embodiments, the method further includes separating the production medium containing 3-hydroxypropanal from the population, thereby resulting in a separated production medium containing 3-hydroxypropanal.

[0005] Provided herein is a method of producing propenoic acid, which includes: (a) culturing an isolated strain of Lactobacillus reuteri in a growth medium, thereby resulting in a population of the isolated strain of Lactobacillus reuteri; (b) culturing the population in a production medium containing a glycerol source to produce 3-hydroxypropanal, thereby resulting in a production medium containing 3- hydroxypropanal; (c) converting the 3-hydroxypropanal into propenoic acid. In some embodiments, the method further includes separating the production medium containing 3-hydroxypropanal from the population prior to the converting, thereby resulting in a separated production medium containing the 3- hy droxypropanal .

[0006] In some of any embodiments, the isolated strain of Lactobacillus reuteri was isolated from feces of an animal. In some embodiments, the animal is selected from the group consisting of humans, porcine, poultry, rodents, and cattle. In some embodiments, the animal is a livestock animal. In some embodiments, the livestock animal is selected from the group consisting of porcine, poultry, and cattle. In some of any embodiments, the animal is a porcine animal. In some embodiments, the porcine animal is a pig, hog, or boar. In some of any embodiments, the animal is a poultry animal. In some embodiments, the poultry animal is a chicken, turkey, duck, goose, fowl, or pheasant. In some of any embodiments, the animal is a cattle animal. In some embodiments, the cattle animal is a cow, steer, or bull. In some of any embodiments, the animal is selected from the group consisting of a human, a mouse, a rat, a pig, a hog, a boar, a chicken, a turkey, a duck, a goose, a fowl, a pheasant, a cow, a steer, a bull, a buffalo, and a bison.

[0007] In some of any embodiments, the isolated strain of Lactobacillus reuteri was isolated by a method which includes: a) suspending the feces in buffer, thereby resulting in a fecal suspension, and b) culturing the fecal suspension in an isolation medium that promotes the growth of Lactobacillus reuteri. In some embodiments, the isolation medium contains glucose, dipotassium hydrogen phosphate, magnesium sulfate heptahydrate, manganous sulfate tetrahydrate, peptone, yeast extract, meat extract, triammonium citrate, and sodium acetate trihydrate. In some embodiments, the isolation medium further contains glycerol and vancomycin.

[0008] In some embodiments, the isolated strain may be genetically engineered to alter the DNA sequence of the isolated organism. In some embodiments, the isolated strain is a strain that is genetically engineered for any of many reasons, including but not limited to: a) decreasing the cost of the media used for growth of the isolated strain or production of 3-hydroxypropanal, b) increasing the production of 3- hydroxypropanal by the isolated strain, and/or c) enabling growth of the isolated strain in a media containing a carbon or nitrogen source the isolated strain is natively unable to grow on.

[0009] In some of any embodiments, the growth medium contains components that promote the growth of Lactobacillus reuteri. In some of any embodiments, the growth medium contains vitamins, amino acids, nucleotides, metal salts, acid whey, glycerol, sodium acetate, and polysorbate 80. In some of any embodiments, the growth medium does not contain glucose.

[0010] In some of any embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium includes incubating the isolated strain in the growth medium at a temperature of between or between about 28 degrees C and 50 degrees C. In some of any embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium includes incubating the isolated strain in the growth medium at a temperature of: (i) between or between about 28° C, 29° C, 31° C, 32° C, 33° C, or 34° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C; or (ii) between or between about 35° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C; or (iii) between or between about 36° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C; or (iv) between or between about 37° C and 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C; or (v) between or between about 37° C and 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, or 45° C; or (vi) at or about 28° C, 29° C, 30° C, 31° C, 32° C, 33° C, 34° C, 35° C, 36° C, 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C; or (vii) at or about 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, or 45° C; or (viii) at or about 37° C; or (ix) at or about 45° C.

[0011] In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium occurs for a duration of time sufficient to result in an optical density (OD) within the range of or of about 10-50. In some of any embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium occurs for a duration of time sufficient to result in an optical density (OD) within the range of or of about 10-12. In some of any embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium occurs for a duration of time that is: (i) at least or at least about 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 22 hours, 24 hours, 30 hours, 36 hours, 42 hours, or 48 hours; or (ii) is or is about 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, or 20 hours; or (iii) is between or between about 4 hours and 10 hours, 4 hours and 9 hours, 4 hours and 8 hours, 4 hours and 7 hours, 4 hours and 6 hours, 5 hours and 10 hours, 5 hours and 9 hours, 5 hours and 8 hours, or 5 hours and 7 hours; or (iv) is or is about 5 hours, 6 hours, 7 hours, or 8 hours; or (v) is between or between about 8 hours and 14 hours, 8 hours and 13 hours, 8 hours and 12 hours, 9 hours and 14 hours, 9 hours and 13 hours, 9 hours and 12 hours, 10 hours and 14 hours, 10 hours and 13 hours, or 10 hours and 12 hours; or (vi) is or is about 9 hours, 10 hours, 11 hours, 12 hours, or 13 hours. In some of any embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium includes incubating the isolated strain in the growth medium at a temperature at or about 37 degrees C for a duration of time that is or is about 10 hours, 11, or 12 hours. In some of any embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium includes incubating the isolated strain in the growth medium at a temperature at or about 45 degrees C for a duration of time that is or is about 5 hours, 6 hours, 7 hours, or 8 hours.

[0012] In some of any embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium includes incubating the isolated strain in the growth medium until CO2 generation has reduced to between or between about 1/3 to 1/2 of peak CO2 generation. In some of any embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium includes incubating the isolated strain in the growth medium until CO2 generation has reduced to about 1/2 of peak CO2 generation.

[0013] In some of any embodiments, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an optical density (OD) between or between about 9 and 50. In some of any embodiments, the OD is between or between about 9 and 45, 10 and 45, 11 and 45, 12 and 45, 13 and 45, 14 and 45, 9 and 40, 10 and 40, 11 and 40, 12 and 40, 13 and 40, 14 and 40, 9 and 35, 10 and 35, 11 and 35, 12 and 35, 13 and 35, 14 and 35, 9 and 30, 10 and 30, 11 and 30, 12 and 30, 13 and 30, 14 and 30, 9 and 25, 10 and 25, 11 and 25, 12 and 25, 13 and 25, 14 and 25, 9 and 20, 10 and 20, 11 and 20, 12 and 20, 13 and 20, 14 and 20, 15 and 50, 20 and 50, 25 and 50, 30 and 50, 35 and 50, 40 and 50, 15 and 45, 20 and 45, 25 and 45, 30 and 45, 35 and 45, 40 and 45, 15 and 40, 20 and 40, 25 and 40, 30 and 40, 35 and 40, 15 and 35, 20 and 35, 25 and 35, 30 and 35, 15 and 30, 20 and 30, or 25 and 30. In some of any embodiments, the OD is between or between about 9 and 15, 10 and 14, 12 and 20, 18 and 26, 20 and 30, or 22 and 50.

[0014] In some of any embodiments, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an optical density (OD) between or between about 9 and 15 or 10 and 14. In some embodiments, the OD is between or between about 10 and 14. In some of any embodiments, the OD is or is about 10, 11, 12, 13, or 14. In some of any embodiments, the OD is between or between about 9 and 15. In some of any embodiments, the OD is between or between about 18 and 26. In some of any embodiments, the OD is between or between about 22 and 50. In some of any embodiments, the OD is between or between about 25 and 50. [0015] In some of any embodiments, the method includes separating the population of the isolated strain of Lactobacillus reuteri from the growth media. In some embodiments, the separating includes the use of tangential flow filtration, followed by centrifuging the population, and followed by removing the growth media.

[0016] In some of any embodiments, the glycerol source is glycerol. In some of any embodiments, the glycerol source is from an organic waste source. In some embodiments, the organic waste source has been pasteurized.

[0017] In some of any embodiments, the production medium comprises glycerol, a carbohydrate source, and a buffering agent.

[0018] In some of any embodiments, the production medium comprises between or between about 50 rnM and 800 mM glycerol, between or between about 0.1 and 60 mM of a carbohydrate source, and between or between about 20 rnM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 25 mM of a carbohydrate source, and between or between about 20 mM and 125 rnM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 rnM and 800 mM glycerol, between or between about 0.1 and 20 mM of a carbohydrate source, and between or between about 20 rnM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 15 mM of a carbohydrate source, and between or between about 20 mM and 125 rnM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 rnM and 800 mM glycerol, between or between about 0.1 and 10 mM of a carbohydrate source, and between or between about 20 rnM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 5 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.5 and 10 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 rnM and 800 mM glycerol, between or between about 0.5 and 5 rnM of a carbohydrate source, and between or between about 20 mM and 125 rnM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.5 and 3 rnM of a carbohydrate source, and between or between about 20 rnM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.5 and 2 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 1 and 10 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 1 and 5 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 1 and 3 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 1 and 2 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent.

[0019] In some of any embodiments, the production medium comprises glycerol, a carbohydrate source, a buffering agent, sodium chloride, potassium chloride, metals, and iron(II) sulfate. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 60 mM of a carbohydrate source, between or between about 20 mM and 125 mM of a buffering agent, between or between about 10 and 60 mM sodium chloride, between or between about 1 mM and 8 mM potassium chloride, one or more metals, and between or between about 2 pM and 40 pM iron(II) sulfate.

[0020] In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 60 mM lactose, between or between about 20 mM and 125 mM of a buffering agent, optionally wherein the buffering agent is MOPS or phosphate buffer, between or between about 10 and 60 mM sodium chloride, between or between about 1 mM and 8 mM potassium chloride, metals comprising Al, B, Ca, Co, Cu, Fe, Mg, Mn, Mo, Ni, Se, V, W, and Zn, and between or between about 2 pM and 40 pM iron(II) sulfate.

[0021] In some of any embodiments, the carbohydrate source comprises a carbohydrate selected from the group consisting of lactose, galactose, sucrose, glucose, maltose, arabinose, and fructose, or any combination thereof. In some of any embodiments, the carbohydrate source is lactose.

[0022] In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 60 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 25 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 20 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 15 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 10 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 5 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.5 and 10 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.5 and 5 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.5 and 3 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.5 and 2 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 1 and 10 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 1 and 5 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 1 and 3 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 1 and 2 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent.

[0023] In some of any embodiments, the buffering agent is MOPS or phosphate buffer. In some of any embodiments, the buffering agent is MOPS pH 7. [0024] In some of any embodiments, the production medium comprises glycerol, lactose, a buffering agent, sodium chloride, potassium chloride, metals, and iron(II) sulfate. In some embodiments, the buffering agent is MOPS, e.g., MOPS pH 7. In some of any embodiments, the production medium comprises glycerol, lactose, MOPS pH 7, sodium chloride, potassium chloride, metals, and iron(II) sulfate.

[0025] In some of any embodiments, the production medium contains glycerol, lactose, a buffering agent, optionally wherein the buffering agent is MOPS or phosphate buffer, sodium chloride, potassium chloride, metals, and iron(II) sulfate. In some embodiments, the production medium contains 200 mM glycerol, 1.5 mM lactose, 50 mM of a buffering agent, optionally wherein the buffering agent is MOPS or phosphate buffer, 25 mM sodium chloride, 3.125 mM potassium chloride, metals, and 10 pM iron(II) sulfate. In some of any embodiments, the production medium contains glycerol, lactose, a buffering agent, optionally wherein the buffering agent is MOPS or phosphate buffer, polysorbate 80, sodium chloride, potassium chloride, metals, and iron(II) sulfate. In some embodiments, the production medium contains 200 mM glycerol, 1.5 mM lactose, 50 mM of a buffering agent, optionally wherein the buffering agent is MOPS or phosphate buffer, 1 g/L polysorbate 80, 25 mM sodium chloride, 3.125 mM potassium chloride, metals, and 10 pM iron(II) sulfate.

[0026] In some of any embodiments, the production medium contains glycerol, lactose, MOPS pH 7, sodium chloride, potassium chloride, metals, and iron(II) sulfate. In some embodiments, the production medium contains 200 mM glycerol, 1.5 mM lactose, 50 mM MOPS pH 7, 25 mM sodium chloride, 3.125 mM potassium chloride, metals, and 10 pM iron(II) sulfate. In some of any embodiments, the production medium contains glycerol, lactose, MOPS pH 7, polysorbate 80, sodium chloride, potassium chloride, metals, and iron(II) sulfate. In some embodiments, the production medium contains 200 mM glycerol, 1.5 mM lactose, 50 mM MOPS pH 7, 1 g/L polysorbate 80, 25 mM sodium chloride, 3.125 mM potassium chloride, metals, and 10 pM iron(II) sulfate.

[0027] In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 10 and 60 mM lactose, between or between about 20 mM and 125 mM of a buffering agent, optionally wherein the buffering agent is MOPS or phosphate buffer, between or between about 10 and 60 mM sodium chloride, between or between about 1 mM and 8 mM potassium chloride, one or more metals, and between or between about 2 pM and 40 pM iron(II) sulfate. In some of any embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 10 and 60 mM lactose, between or between about 20 mM and 125 mM of a buffering agent, optionally wherein the buffering agent is MOPS or phosphate buffer, between or between about 10 and 60 mM sodium chloride, between or between about 1 mM and 8 mM potassium chloride, metals comprising Al, B, Ca, Co, Cu, Fe, Mg, Mn, Mo, Ni, Se, V, W, and Zn, and between or between about 2 uM and 40 uM iron(II) sulfate. In some of any embodiments, the production medium comprises 200 mM glycerol, 25 mM lactose, 50 mM of a buffering agent, optionally wherein the buffering agent is MOPS or phosphate buffer, 25 mM sodium chloride, 3.125 mM potassium chloride, metals, and 10 pM iron(II) sulfate.

[0028] In some of any embodiments, the metals include one or more metals selected from the group consisting of Mg, Mn, Ca, Fe, Zn, Co, Cu, Al, B, Mo, Ni, W, V, and Se. In some of any embodiments, the metals include Mg, Mn, Ca, Fe, Zn, Co, Cu, Al, B, Mo, Ni, W, V, and Se.

[0029] In some of any embodiments, the metals comprise between or between about 250 pM and 2000 pM Mg, between or between about 0.02 pM and 5 pM Mn, between or between about 50 pM and 600 pM Ca, between or between about 1 pM and 30 pM Fe, between or between about 5 pM and 45 pM Zn, between or between about 0.02 pM and 50 pM Co, between or between about 0.05 pM and 2 pM Cu, between or between about 1 pM and 20 pM Al, between or between about 1 pM and 20 pM B, between or between about 0.02 pM and 2 pM Mo, between or between about 0.02 pM and 2 pM Ni, between or between about 0.001 pM and 0.5 pM W, between or between about 0.5 pM and 10 pM V, and between or between about 0.02 pM and 2 pM Se. In some of any embodiments, wherein the metals include between or between about 250 pM and 2000 pM Mg, between or between about 0.02 pM and 5 pM Mn, between or between about 50 pM and 600 pM Ca, between or between about 1 pM and 30 pM Fe, between or between about 5 pM and 45 pM Zn, between or between about 0.02 pM and 1 pM Co, between or between about 0.05 pM and 2 pM Cu, between or between about 1 pM and 20 pM Al, between or between about 1 pM and 20 pM B, between or between about 0.02 pM and 2 pM Mo, between or between about 0.02 pM and 2 pM Ni, between or between about 0.001 pM and 0.5 pM W, between or between about 0.5 pM and 10 pM V, and between or between about 0.02 pM and 2 pM Se.

[0030] In some of any embodiments, culturing the population in the production medium occurs for a duration of time that results in converting at least or at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the glycerol source to 3- hydroxypropanal. In some embodiments, culturing the population in the production medium occurs for a duration of time that results in converting at least or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the glycerol source to 3-hydroxypropanal. In some of any embodiments, culturing the population in the production medium occurs for a duration of time that results in converting at or about 100% of the glycerol source to 3-hydroxypropanal. [0031] In some of any embodiments, culturing the population in the production medium occurs for a duration of time that is or is at least 30 minutes. In some of any embodiments, culturing the population in the production medium occurs for a duration of time that is between or between about 30 minutes and 48 hours. In some embodiments, culturing the population in the production medium occurs for a duration of time that is between or between about 30 minutes and 5 hours. In some of any embodiments, culturing the population in the production medium occurs for a duration of time that is between or between about 1 hour and 4 hours. In some of any embodiments, culturing the population in the production medium occurs for a duration of time that is between or between about 2 and 3 hours.

[0032] In some embodiments, culturing the population in the production medium occurs for a duration of time that is or is at least 30 minutes, 45 minutes, 1 hour, 1.25 hours, 1.5 hours, 1.75 hours, 2 hours, 2.25 hours, 2.5 hours, 2.75 hours, 3 hours, 3.25 hours, 3.5 hours, or 4 hours.

[0033] In some embodiments, culturing the population in the production medium occurs for a duration of time that is or is between 3 and 48 hours. In some embodiments, culturing the population in the production medium occurs for a duration of time that is between or between about 1 and 48 hours, 1 and 42 hours, 1 and 36 hours, 1 and 30 hours, 1 and 24 hours, 1 and 18 hours, 1 and 12 hours, 1 and 6 hours, 2 and 48 hours, 2 and 42 hours, 2 and 36 hours, 2 and 30 hours, 2 and 24 hours, 2 and 18 hours, 2 and 12 hours, 2 and 6 hours, 4 and 48 hours, 4 and 42 hours, 4 and 36 hours, 4 and 30 hours, 4 and 24 hours, 4 and 18 hours, 4 and 12 hours, 8 and 48 hours, 8 and 42 hours, 8 and 36 hours, 8 and 30 hours, 8 and 24 hours, 8 and 18 hours, 8 and 12 hours, 12 and 48 hours, 12 and 42 hours, 12 and 36 hours, 12 and 30 hours, 12 and 24 hours, 12 and 18 hours, 16 and 48 hours, 16 and 42 hours, 16 and 36 hours, 16 and 30 hours, 16 and 24 hours, 20 and 48 hours, 20 and 42 hours, 20 and 36 hours, 20 and 30 hours, 20 and 24 hours, 24 and 48 hours, 24 and 42 hours, 24 and 36 hours, or 24 and 30 hours. In some embodiments, culturing the population in the production medium occurs for a duration of time that is or is between 6 and 48 hours. In some embodiments, culturing the population in the production medium occurs for a duration of time that is or is between 6 and 36 hours. In some embodiments, culturing the population in the production medium occurs for a duration of time that is or is between 12 and 48 hours. In some embodiments, culturing the population in the production medium occurs for a duration of time that is or is between 12 and 36 hours. In some embodiments, culturing the population in the production medium occurs for a duration of time that is or is between 18 and 48 hours. In some embodiments, culturing the population in the production medium occurs for a duration of time that is or is between 18 and 30 hours.

[0034] In some embodiments, culturing the population in the production medium occurs for a duration of time that is or is at least 4 hours, 5 hours, 6 hours, 8 hours, 10 hours, 12 hours, 16 hours, 20 hours, 25 hours, 30 hours, 36 hours, 42 hours, or 48 hours. In some embodiments, culturing the population in the production medium occurs for a duration of time that is or is about 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 8 hours, 10 hours, 12 hours, 16 hours, 20 hours, 25 hours, 30 hours, 36 hours, 42 hours, or 48 hours.

[0035] In some of any embodiments, culturing the population in the production medium includes incubating the population in the production medium at a temperature at or about 37 degrees C.

[0036] In some of any embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22 g/L/hour. In some of any embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or about 3, 4, 5, 6, 7, 8, 9, or 10 g/L/hour. In some of any embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22 g/L/hour. In some of any embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or at least 6 g/L/hour. In some of any embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of between or between about 3 and 10 g/L/hour, 3 and 9 g/L/hour, 3 and 8 g/L/hour, 3 and 7 g/L/hour, 4 and 10 g/L/hour, 4 and 9 g/L/hour, 4 and 8 g/L/hour, 4 and 7 g/L/hour, 5 and 10 g/L/hour, 5 and 9 g/L/hour, 5 and 8 g/L/hour, 5 and 7 g/L/hour, 6 and 10 g/L/hour, 6 and 9 g/L/hour, 6 and 8 g/L/hour, or 6 and 7 g/L/hour. In some of any embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or at least 10 g/L/hour. In some of any embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or at least 15-22 g/L/hour. In some of any embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or at least 15 g/L/hour. In some of any embodiments, during the culturing in the production medium, 3- hydroxypropanal is produced at a rate of between or between about 3 and 30 g/L/hour, 3 and 25 g/L/hour, 3 and 20 g/L/hour, 3 and 15 g/L/hour, 5 and 30 g/L/hour, 5 and 25 g/L/hour, 5 and 20 g/L/hour, 5 and 15 g/L/hour, 7 and 30 g/L/hour, 7 and 25 g/L/hour, 7 and 20 g/L/hour, 7 and 15 g/L/hour, 10 and 30 g/L/hour, 10 and 25 g/L/hour, 10 and 20 g/L/hour, 10 and 15 g/L/hour, 15 and 30 g/L/hour, 15 and 25 g/L/hour, 15 and 20 g/L/hour, 20 and 30 g/L/hour, or 20 and 25 g/L/hour.

[0037] In some of any embodiments, separating the production medium containing 3-hydroxypropanal from the population includes centrifuging the population and removing the production medium containing 3- hy droxypropanal .

[0038] In some of any embodiments, the separated production medium comprising 3-hydroxypropanal comprises at or at least 17 g/L, 18 g/L, 19 g/L, 20 g/L, 21 g/L, 22 g/L, 23 g/L, 24 g/L, 25 g/L, 30 g/L, 40 g/L, 50 g/L, 75 g/L, 100 g/L, 125 g/L, 150 g/L, 175 g/L, or 200 g/L of 3-hydroxypropanal.

[0039] In some of any embodiments, the separated production medium containing 3-hydroxypropanal contains at or at least 6 g/L, 7 g/L, 8 g/L, 9 g/L, 10 g/L, 11 g/L, 12 g/L, 13 g/L, 14 g/L, 15 g/L, 16 g/L, 17 g/L, 18 g/L, 19 g/L, 20 g/L, 21 g/L, 22 g/L, 23 g/L, or 24 g/L of 3-hydroxypropanal. In some of any embodiments, the separated production medium containing 3-hydroxypropanal contains between or between about 6 g/L and 40 g/L, 6 g/L and 36 g/L, 6 g/L and 32 g/L, 6 g/L and 28 g/L, 6 g/L and 24 g/L, 6 g/L and 20 g/L, 9 g/L and 40 g/L, 9 g/L and 36 g/L, 9 g/L and 32 g/L, 9 g/L and 28 g/L, 9 g/L and 24 g/L, 9 g/L and 20 g/L, 12 g/L and 40 g/L, 12 g/L and 36 g/L, 12 g/L and 32 g/L, 12 g/L and 28 g/L, 12 g/L and 24 g/L, 12 g/L and 20 g/L, 15 g/L and 40 g/L, 15 g/L and 36 g/L, 15 g/L and 32 g/L, 15 g/L and 28 g/L, 15 g/L and 24 g/L, 15 g/L and 20 g/L, 18 g/L and 40 g/L, 18 g/L and 36 g/L, 18 g/L and 32 g/L, 18 g/L and 28 g/L, 18 g/L and 24 g/L, 18 g/L and 20 g/L, 15 g/L to 30 g/L, 20 g/L to 35 g/L, 30 g/L to 45 g/L, 40 g/L and 80 g/L, or 40 g/L and 200 g/L of 3-hydroxypropanal. In some of any embodiments, the separated production medium comprising 3-hydroxypropanal comprises between or between about 6 and 200 g/L, 6 and 150 g/L, 6 and 100 g/L, 6 and 80 g/L, 6 and 60 g/L, 6 and 50 g/L, 6 and 30 g/L, 6 and 20 g/L, 6 and 15 g/L, 10 and 200 g/L, 10 and 150 g/L, 10 and 100 g/L, 10 and 80 g/L, 10 and 60 g/L, 10 and 50 g/L, 10 and 30 g/L, 10 and 20 g/L, 10 and 15 g/L, 15 and 200 g/L, 15 and 150 g/L, 15 and 100 g/L, 15 and 80 g/L, 15 and 60 g/L, 15 and 50 g/L, 15 and 30 g/L, 15 and 20 g/L, 20 and 200 g/L, 20 and 150 g/L, 20 and 100 g/L, 20 and 80 g/L, 20 and 60 g/L, 20 and 50 g/L, 20 and 30 g/L, 30 and 200 g/L, 30 and 150 g/L, 30 and 100 g/L, 30 and 80 g/L, 30 and 60 g/L, 30 and 50 g/L, 40 and 200 g/L, 40 and 150 g/L, 40 and 100 g/L, 40 and 80 g/L, 40 and 60 g/L, 40 and 50 g/L, 50 and 200 g/L, 50 and 150 g/L, 50 and 100 g/L, 50 and 80 g/L, 50 and 60 g/L, 60 and 200 g/L, 60 and 150 g/L, 60 and 100 g/L, or 60 and 80 g/L. In some of any embodiments, the separated production medium containing 3-hydroxypropanal contains between or between about 12 g/L and 18 g/L of 3-hydroxypropanal.

[0040] In some of any embodiments, converting the 3-hydroxypropanal into propenoic acid includes converting the 3-hydroxypropanal into prop-2-enal and converting the prop-2 -enal into propenoic acid. In some embodiments, converting the 3-hydroxypropanal into prop-2-enal includes acidifying the production medium containing 3-hydroxypropanal, thereby resulting in an acidified production medium; and converting the 3-hydroxypropanal in the acidified production medium into gaseous prop-2-enal. In some embodiments, converting the 3-hydroxypropanal into prop-2-enal includes acidifying the production medium containing 3- hydroxypropanal or the separated production medium containing 3-hydroxypropanal, thereby resulting in an acidified production medium; and converting the 3-hydroxypropanal in the acidified production medium into gaseous prop-2-enal. In some of any embodiments, converting prop-2 -enal into propenoic acid includes oxidizing the prop-2-enal in the presence of a catalyst to produce the propenoic acid.

[0041] In some of any embodiments, converting the 3-hydroxypropanal into propenoic acid includes: (i) acidifying the production medium containing 3-hydroxypropanal, thereby resulting in an acidified production medium; (ii) converting the 3-hydroxypropanal in the acidified production medium into gaseous prop-2 -enal; and (iii) oxidizing the prop-2 -enal in the presence of a catalyst to produce the propenoic acid.

[0042] In some of any embodiments, converting the 3-hydroxypropanal into propenoic acid includes: (i) acidifying the production medium containing 3-hydroxypropanal or the separated production medium containing 3-hydroxypropanal, thereby resulting in an acidified production medium; (ii) converting the 3- hydroxypropanal in the acidified production medium into gaseous prop-2-enal; and (iii) oxidizing the prop-2- enal in the presence of a catalyst to produce the propenoic acid.

[0043] In some of any embodiments, acidifying the production medium containing 3-hydroxypropanal includes acidifying the production medium containing 3-hydroxypropanal with a mineral acid. In some of any embodiment, acidifying the production medium containing 3-hydroxypropanal or the separated production medium containing 3-hydroxypropanal includes acidifying the production medium containing 3- hydroxypropanal or the separated production medium containing 3-hydroxypropanal with a mineral acid. In some of any embodiments, the mineral acid is selected from the group consisting of phosphoric acid, hydrochloric acid, sulfuric acid, nitric acid, boric acid, hydrofluoric acid, hydroiodic acid, hydrobromic acid, and perchloric acid.

[0044] In some of any embodiments, converting the 3-hydroxypropanal in the acidified production medium into gaseous prop-2-enal includes heating the acidified production medium. In some embodiments, the heating is performed under reduced pressure. In some embodiments, the heating is performed under reduced pressure as compared to atmospheric pressure, e.g., approximately 1 bar. In some embodiments, the heating is performed under reduced pressure of less than 1 bar, such as between or between about 40 and 400 mbar.

[0045] In some of any embodiments, oxidizing the prop-2-enal in the presence of a catalyst to produce the propenoic acid includes the use of vapor-phase oxidation. In some of any embodiments, the catalyst includes a two-metal catalyst. In some of any embodiments, oxidizing the prop-2-enal in the presence of a catalyst to produce the propenoic acid includes reacting the prop-2-enal with an oxygen-containing gas at a temperature that is between or between about 125 and 320 degrees C in the presence of a two-metal catalyst. In some of any embodiments, the two-metal catalyst contains palladium and copper or silver. [0046] Provided herein is 3-hydroxypropanal (reuterin) produced by some of any embodiments. Provided herein is propenoic acid produced by some of any embodiments.

[0047] Provided herein is an isolated strain of Lactobacillus reuteri, wherein the isolated strain was isolated from feces of an animal. In some embodiments, the animal is selected from the group consisting of a human, a mouse, a rat, a pig, a hog, a boar, a chicken, a turkey, a duck, a goose, a fowl, a pheasant, a cow, a steer, a bull, a buffalo, and a bison. In some of any embodiments, the isolated strain of Lactobacillus reuteri was isolated by a method including: a) suspending the feces in buffer, thereby resulting in a fecal suspension, and b) culturing the fecal suspension in an isolation medium that promotes the growth of Lactobacillus reuteri. In some embodiments, the isolation medium contains glucose, dipotassium hydrogen phosphate, magnesium sulfate heptahydrate, manganous sulfate tetrahydrate, peptone, yeast extract, meat extract, triammonium citrate, and sodium acetate trihydrate. In some embodiments, the isolation medium further contains glycerol and vancomycin.

[0048] Provided herein is a kit containing an isolated strain of Lactobacillus reuteri, and instructions for use thereof. Provided herein is a kit containing the isolated strain of Lactobacillus reuteri of some of any embodiments, and instructions for use thereof. In some embodiments, the kit further contains a growth medium and/or a production medium. In some embodiments, the kit further contains one or more components for producing a growth medium and/or production medium.

[0049] Provided herein is a composition containing an isolated strain of Lactobacillus reuteri. Provided herein is a composition containing the isolated strain of Lactobacillus reuteri of some of any embodiments. [0050] Provided herein is a composition containing the 3-hydroxypropanal (reuterin) of some of any embodiments. Provided herein is a composition containing the propenoic acid of some of any embodiments. Provided herein is a kit containing the 3-hydroxypropanal (reuterin) of some of any embodiments. Provided herein is a kit containing the propenoic acid of some of any embodiments, Provided herein is a kit containing the composition of some of any embodiments.

Brief Description of the Drawings

[0051] FIG. 1 depicts the chemical structures of propenoic acid (a), prop-2-enal (b), and 3- hydroxypropanal (c).

[0052] FIG. 2 illustrates the conventional pathways to propenoic acid, from propene, glucose, or glycerol. For propene, the vast majority of propenoic acid is conventionally produced from petroleum by industrial companies by the route highlighted in grey, i.e., from propene to prop-2-enal via vapor-phase oxidation catalysis and then to propenoic acid via vapor-phase oxidation catalysis. For glucose, some companies have tried feeding genetically modified bacteria to produce lactic acid, and other companies have tried using genetically modified yeast. For glycerol, some companies have tried to go directly from glycerol to propenoic acid, and other companies have explored the glycerol to allyl alcohol route to propenoic acid. These methods have limitations.

[0053] FIG. 3 illustrates an exemplary method for the production of propenoic acid using the methods described herein. The isolated strain of Lactobacillus reuteri as described herein can grow on several carbon sources, including lactose (a disaccharide). The cells of the isolated strain of Lactobacillus reuteri are allowed to age and are then fed glycerol. The cells then convert the glycerol to 3-hydroxypropanal, which is secreted into the medium, e.g., production medium. The 3-hydroxypropanal is then converted into prop-2- enal, which can then enter established industrial production processes for its conversion into propenoic acid. The 3-hydroxypropanal can also be converted to 1,3 -propanediol or 3-hydroxypropanoic acid.

[0054] FIG. 4 illustrates an exemplary method for the production of propenoic acid by fermentation, distillation, and catalysis, as described herein. The isolated strain of Lactobacillus reuteri is inoculated into a growth medium and is grown to exhaustion. The depleted growth medium is removed and a production medium is introduced. The cells of the population of the isolated strain of Lactobacillus reuteri begin producing 3-hydroxypropanal from a glycerol source in the production medium. The cells may deplete the production medium and may be separated from the production medium, or the production medium may be continuously drawn off, i.e., removed and saved, while fresh production medium is introduced, i.e., a draw and fill or continuous fermentation. The production medium that includes the 3-hydroxypropanal is then heated and placed under reduced pressure as compared to atmospheric pressure to distill prop-2 -enal. The prop-2-enal is then passed over a catalyst to produce propenoic acid.

Detailed Description

[0055] Provided herein are methods of producing reuterin and/or propenoic acid, along with isolated strains of Lactobacillus reuteri, e.g., for use in such methods, and kits and compositions comprising an isolated strain of Lactobacillus reuteri.

[0056] Propenoic acid (also known as acrylic acid) is a simple carboxylic acid that is used to make thousands of specialized chemicals, and is a source of vinyls, polishes, paints, clear plastics, coatings, adhesives, and elastomers. Conventionally, propenoic acid is obtained by two successive oxidations of propene gas, which itself is created during gasoline refinement. See Intratec Solutions, Acrylic acid production via propene oxidation. February 2016, accessed at: https://www.chemengonline.com/acrylic-acid- production-via-propylene-oxidation-intratec- solutions/). Propenoic acid can be made from prop-2-enal, 3- hydroxypropanoic acid, glycerol, allyl alcohol, or lactic acid. Prop-2 -enal is toxic and dangerous. It is a barely glimpsed intermediate between propene and propenoic acid during industrial synthesis. 3- hydroxypropanal dehydrates to prop-2-enal, and prop-2 -enal can be rehydrated to 3-hydroxypropanal.

[0057] Josef Redtenbacher first synthesized propenoic acid via biomass glycerol from yeast in the 1850s. Since then, there have been efforts to produce propenoic acid from various sources and methods, such as shown in FIG. 2.

[0058] Direct biomass conversion of glycerol into propenoic acid has been attempted. Arkema prototyped a catalyst to convert glycerol (also called glycerin) directly into propenoic acid, but this effort was abandoned because the major source of glycerol was biodiesel waste, which had to be highly refined, thereby rendering their product at least 15% more expensive. See Chemical & Engineering News, Hunting for biobased acrylic acid. November 2013, accessed at: https://cen.acs.org/articles/91/i46/Hunting-Biobased-Acrylic -Acid.html). Myriant explored an alternative path from glycerol through allyl alcohol, and another path from 3- hydroxypropionic acid (see, e.g., WO 2015034948), but faced the same feedstock problem.

[0059] Direct fermentation has also been attempted. However, propenoic acid is not a candidate for direct biomanufacturing. Polyacrylic acid is beneficial for use in diapers because it is toxic to microbes, but it is very difficult to make. See Chu et al., Metabolic Engineering, Direct fermentation route for the production of acrylic acid, Metabolic Engineering, 2015, Vol. 32, pages 23-29. Some pseudomonads have been fed acrylonitrile and made 390 g/L propenoic acid, but, unfortunately, acrylonitrile is produced by ammoxidation of propene, so this would not be a feasible replacement for propene.

[0060] Precursor fermentation has also been attempted. Myriant fed E. coli refined glucose to make lactic acid, which could then be catalytically converted to propenoic acid. See BusinessWire, Myriant Develops Proprietary Process to Produce Bio-Acrylic Acid. March 2012, accessed at: https://www.businesswire.eom/news/home/20120328005035/en/Myr iant-Develops-Proprietary- Process-to- Produce-Bio-Acrylic-Acid. However, this process suffered from the reluctance of E. coli to make high yields of lactic acid. Cargill partnered with Novozymes and BASF to ferment 3 -hydroxypropionic acid from refined sugar, but this effort was stopped after three years. See Plastics Today, BASF exits bio-acrylic acid partnership with Novozymes and Cargill. January 2015, accessed at: https://www.plasticstoday.com/basf- exits-bio-acrylic-acid-partnership-novozymes-and- cargill.

[0061] As such, the demand for the production of propenoic acid is high, but the economics of previous methods have fallen short. Two failure points relate to refinement and novel catalysts. [0062] With regards to refinement, this requires that the microbes have very pure and very consistent sources of carbon, typically glucose, which is often derived from corn. Moreover, fermentation output must be refined, with anything that goes over a chemical catalyst having to be as pure as possible. Further, if the microbes do not secrete the target chemical, then it must be carefully separated from the microbes themselves and from the fermentation medium. Finally, if the catalytic input is not pure, then neither is the propenoic acid, and the catalyst gets wasted.

[0063] With regards to novel catalysts, the precursor molecules being generated all require different catalyst chemistries. The additional capital cost of catalyst development and scale faces the fully depreciated infrastructure deployed for petroleum conversion.

[0064] As of today, there are no industrial scale biomass challengers to petroleum-derived propenoic acid. [0065] Accordingly, there is a need for a more economically efficient and more environmentally friendly method for producing propenoic acid, which the present disclosure provides a solution for. The present disclosure describes, inter alia, methods of producing 3-hydroxypropanal (reuterin) from glycerol using isolated strains of Lactobacillus reuteri, including those isolated as described herein, along with engineered, e.g., genetically engineered, forms of such isolated strains. Reuterin producing using these methods can then be converted to propenoic acid using standard industrial methods. The methods provided herein allow for a more economically efficient and environmentally friendly method of producing propenoic acid in multiple ways. First, strains isolated using the methods described herein can be grown on a variety of societal byproducts allowing usage of waste streams. Further, the isolated strains of Lactobacillus reuteri described herein can advantageously produce reuterin using a production medium having a concentration of between or between about 0.1 mM and 5 mM, 10 mM, 15 mM, 20 mM, or 25mM. For instance, it was surprisingly discovered that decreasing the concentration of an exemplary carbohydrate source, lactose, from 25 mM to 1.5 mM in the production medium actually increased reuterin yield by approximately 5-15%. The isolated microbe is also capable of using biodiesel glycerol as it is resistant to the levels of methanol endogenous to the stream. Finally, distillation allows for impurity removal which will allow for direct incorporation into current catalyst streams for the production of propenoic acid. Additional advantages are apparent to those skilled in the art in view of the disclosure provided herein.

[0066] All publications, including patent documents, scientific articles and databases, referred to in this application are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication were individually incorporated by reference. If a definition set forth herein is contrary to or otherwise inconsistent with a definition set forth in the patents, applications, published applications and other publications that are herein incorporated by reference, the definition set forth herein prevails over the definition that is incorporated herein by reference.

[0067] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

I. METHODS OF PRODUCING REUTERIN AND PROPENOIC ACID

[0068] Provided herein are methods of producing 3-hydroxypropanal (reuterin), comprising: (a) culturing an isolated strain of Lactobacillus reuteri in a growth medium, thereby resulting in a population of the isolated strain of Lactobacillus reuteri; and (b) culturing the population in a production medium comprising a glycerol source to produce 3-hydroxypropanal. This production of 3-hydroxypropanal (reuterin) is depicted, for instance, in FIG. 4. In some embodiments, the method further comprises separating the production medium comprising 3-hydroxypropanal from the population, thereby resulting in a separated production medium comprising 3-hydroxypropanal. In some embodiments, the 3-hydroxypropanal (reuterin) that is produced is further converted into propenoic acid, such as by any method known in the art or as described herein. In some embodiments, the isolated strain is an isolated strain that is further engineered, e.g., genetically engineered.

[0069] Also provided herein are methods of producing propenoic acid, comprising: (a) culturing an isolated strain of Lactobacillus reuteri in a growth medium, thereby resulting in a population of the isolated strain of Lactobacillus reuteri; (b) culturing the population in a production medium comprising a glycerol source to produce 3-hydroxypropanal, thereby resulting in a production medium comprising 3-hydroxypropanal; (c) converting the 3-hydroxypropanal into propenoic acid. This production of propenoic acid is depicted, for instance, in FIG. 4. In some embodiments, the isolated strain is an isolated strain that is further engineered, e.g., genetically engineered.

A. _ Culturing an isolated strain of Lactobacillus reuteri in a growth medium

[0070] In some embodiments, the methods provided herein comprise culturing an isolated strain of Lactobacillus reuteri in a growth medium, thereby resulting in a population of the isolated strain of Lactobacillus reuteri. In some embodiments, the isolated strain is an isolated strain that is further engineered, e.g., genetically engineered. 1. Isolated strains of Lactobacillus reuteri

[0071] Isolated strains of Lactobacillus reuteri can be isolated using any available method, and can be isolated from any available source that contains Lactobacillus reuteri, such as feces.

[0072] In some embodiments, the isolated strain of Lactobacillus reuteri is isolated from feces of an animal. The animal is not particularly limited and may be any animal, e.g., any animal whose feces contains Lactobacillus reuteri. In some embodiments, the animal is a mammal.

[0073] In some embodiments, the animal is selected from the group consisting of humans, porcine, poultry, rodents, and cattle. In some embodiments, the animal is a porcine animal. In some embodiments, the porcine animal is a pig, hog, or boar. In some embodiments, the animal is a poultry animal. In some embodiments, the poultry animal is a chicken, turkey, duck, goose, fowl, or pheasant. In some embodiments, the animal is a cattle animal. In some embodiments, the cattle animal is a cow, steer, or bull.

[0074] In some embodiments, the animal is selected from the group consisting of a human, a mouse, a rat, a pig, a hog, a boar, a chicken, a turkey, a duck, a goose, a fowl, a horse, a donkey, a sheep, a pheasant, a cow, a steer, a bull, a buffalo, and a bison.

[0075] In some embodiments, the animal is a livestock animal. In some embodiments, the livestock animal is selected from the group consisting of a pig, a hog, a boar, a chicken, a turkey, a horse, a donkey, a sheep, a pheasant, a cow, a steer, a bull, a buffalo, and a bison.

[0076] In some embodiments, the animal is a pig. In some embodiments, the animal is a hog. In some embodiments, the animal is a duck. In some embodiments, the animal is a goose. In some embodiments, the animal is a mouse. In some embodiments, the animal is a rat. In some embodiments, the animal is a chicken. In some embodiments, the animal is a turkey. In some embodiments, the animal is a horse. In some embodiments, the animal is a sheep. In some embodiments, the animal is a pheasant. In some embodiments, the animal is a cow. In some embodiments, the animal is a steer. In some embodiments, the animal is a bull. In some embodiments, the animal is a buffalo. In some embodiments, the animal is a bison.

[0077] In some embodiments, the isolated strain of Lactobacillus reuteri was isolated by a method comprising culturing the feces or a fecal suspension in a medium that promotes the growth of Lactobacillus reuteri.

[0078] In some embodiments, the isolated strain of Lactobacillus reuteri was isolated by a method comprising: a) suspending the feces in buffer, thereby resulting in a fecal suspension, and b) culturing the fecal suspension in an isolation medium that promotes the growth of Lactobacillus reuteri. [0079] In some embodiments, the buffer can be any buffer suitable for suspending microbes. In some embodiments, the buffer is MOPS, such as MOPS pH 7 buffer. In some embodiments, the buffer comprises a salt, such as sodium chloride (NaCl). In some embodiments, the buffer comprises NaCl.

[0080] The amount of feces that is used for isolating a strain of Lactobacillus reuteri is not particularly limited. In some embodiments, between or between about 0.1 g and 10 g of feces is used for the isolation. In some embodiments, between or between about 0.1 g and 10 g, 0.1 g and 5 g, 0.5 g and 10 g, 0.5 g and 5 g, 1 g and 10 g, 1 g and 5 g, 1 g and 4 g, 1 g and 3 g, or 1 g and 2 g of feces is used for the isolation. In some embodiments, between or between about 0.01 g and 2 g of feces per mL of buffer is used for the isolation. In some embodiments, between or between about 0.01 g and 2 g, 0.01 g and 1.5 g, 0.01 g and 1 g, 0.01 g and 0.5 g, 0.01 g and 0.1 g, 0.05 g and 2 g, 0.05 g and 1.5 g, 0.05 g and 1 g, 0.05 g and 0.5 g, 0.05 g and 0.1 g, 0.1 g and 2 g, 0.1 g and 1.5 g, 0.1 g and 1 g, 0.1 g and 0.5 g, 0.25 g and 2 g, 0.25 g and 1.5 g, 0.25 g and 1 g, or 0.25 g and 0.5 g of feces per mL of buffer is used for the isolation. In some embodiments, at or about 0.01 g, 0.025 g, 0.05 g, 0.075 g, 0.1 g, 0.125 g, 0.15 g, 0.175 g, 0.2 g, 0.225 g, 0.25 g, 0.275 g, 0.3 g, 0.325 g, 0.35 g, 0.375 g, 0.4 g, 0.425 g, 0.45 g, 0.475 g, 0.5 g, 0.525 g, 0.55 g, 0.575 g, 0.6 g, 0.625 g, 0.65 g, 0.675 g, 0.7 g, 0.725 g, 0.75 g, 0.775 g, 0.8 g, 0.825 g, 0.85 g, 0.875 g, 0.9 g, 0.925 g, 0.95 g, 0.975 g, or 1.0 g per mL of buffer is used for the isolation. In some embodiments, at or about 0.1 g, 0.125 g, 0.15 g, 0.175 g, 0.2 g, 0.225 g, 0.25 g, 0.275 g, 0.3 g, 0.325 g, 0.35 g, 0.375 g, 0.4 g, 0.425 g, 0.45 g, 0.475 g, or 0.5 g of feces per mL of buffer is used for the isolation.

[0081] In some embodiments, the isolation medium can be any medium suitable for promoting the selective growth of Lactobacillus reuteri. In some embodiments, the isolation medium comprises MRS broth (Sigma, Product No. 69966). In some embodiments, the isolation medium comprises glucose, dipotassium hydrogen phosphate, magnesium sulfate heptahydrate, manganous sulfate tetrahydrate, peptone, yeast extract, meat extract, triammonium citrate, and sodium acetate trihydrate.

[0082] In some embodiments, the isolation medium further comprises components that promote the selective growth of Lactobacillus reuteri over other microbial strains. In some embodiments, the components that promote the selective growth of Lactobacillus reuteri over other microbial strains comprise glycerol and vancomycin. Glycerol is known to promote the growth of Lactobacillus reuteri, and Lactobacillus reuteri is known to be resistant to the antibiotic vancomycin, thereby promoting the selective growth of Lactobacillus reuteri.

[0083] In some embodiments, following the selective growth of Lactobacillus reuteri in the isolation medium, dilutions are plated onto agar plates comprising vancomycin, and reuterin production is detected to identify colonies of Lactobacillus reuteri. In some embodiments, reuterin-producing colonies are identified using an agar overlay assay with a colorimetric aldehyde-detecting reagent under acidic conditions, followed by neutralizing with sodium hydroxide to develop color. In some embodiments, the colorimetric aldehyde- detecting reagent is 2,4-dinitrophenylhydrazine (DNPH).

[0084] In some embodiments, the isolated strain of Lactobacillus reuteri grows on lactose.

[0085] In some embodiments, lactose, e.g., unpurified lactose from a dairy source, is sufficient to promote the growth of the isolated strain of Lactobacillus reuteri. In some embodiments, the isolated strain of Lactobacillus reuteri does not require refined sugar to grow.

[0086] In some embodiments, the isolated strain is an isolated strain that is further engineered, e.g., genetically engineered, as described herein. In some embodiments, the isolated strain of Lactobacillus reuteri has been further genetically engineered after being isolated. In some embodiments, the isolated strain is further genetically modified after being isolated from the feces of the animal.

[0087] In some embodiments, the isolated strain may be genetically engineered to alter the DNA sequence of the isolated organism for any of many reasons, including but not limited to: a) decreasing the cost of the media used for growth of the isolated strain or production of 3-hydroxypropanal, b) increasing the production of 3-hydroxypropanal by the isolated strain, and/or c) enabling growth of the isolated strain in a media containing a carbon or nitrogen source the isolated strain is natively unable to grow on.

[0088] For instance, in some embodiments, the isolated strain, following isolation, is further genetically engineered. In some embodiments, the isolated strain is genetically engineered to increase the production of 3-hydroxypropanal. In some embodiments, the isolated strain is genetically engineered to enable or increase growth of the isolated strain in a media containing a carbon or nitrogen source that the isolated strain is natively unable to grow in or inefficiently grows in. In some embodiments, the isolated strain is genetically engineered to decrease the cost of the media used for growth of the isolated strain (growth medium) or for production of 3-hydroxypropanal (production medium), such as by allowing for a reduced amount of medium components and/or for less costly medium components.

[0089] In some embodiments, the engineering, e.g., genetic engineering, comprises introducing a gene disruption and/or introducing an exogenous nucleic acid into the isolated strain. In some embodiments, the engineering, e.g., genetic engineering, comprises introducing a gene disruption and introducing an exogenous nucleic acid into the isolated strain. In some embodiments, the engineering, e.g., genetic engineering, comprises introducing a gene disruption into the isolated strain. In some embodiments, the gene disruption comprises a genetic alteration that renders a gene product encoded by the disrupted gene inactive or attenuated. In some embodiments, the gene disruption comprises deletion of an entire gene, deletion of a regulatory sequence required for transcription or translation, deletion of a portion of the gene that results in a truncated gene product, or by a mutation that inactivates or attenuates a gene product encoded by the gene. In some embodiments, the gene disruption comprises a complete gene deletion. In some embodiments, the gene disruption comprises a partial gene deletion. In some embodiments, the gene disruption comprises a mutation, e.g., a null mutation.

[0090] In some embodiments, the engineering, e.g., genetic engineering, comprises introducing an exogenous nucleic acid into the isolated strain. The exogenous nucleic acid is not particularly limited and can, in some embodiments, be any exogenous nucleic acid that encodes a polypeptide of interest. In some embodiments, the polypeptide of interest enables or increases growth of the isolated strain. In some embodiments, the polypeptide of interest enables or increases growth of the isolated strain in a media containing a carbon or nitrogen source that the isolated strain is natively unable to grow in or inefficiently grows in. In some embodiments, the polypeptide of interest promotes increased production of 3- hydroxypropanal. In some embodiments, the polypeptide of interest promotes increased growth and/or survival of the isolated strain in a media, wherein the growth and/or survival is increased as compared to the isolated strain without the engineering, e.g., genetic engineering, in the same media. In some embodiments, the exogenous nucleic acid encodes a non-coding RNA, such as small interfering RNA (siRNA) or microRNA (miRNA) that repress or suppress gene expression, such as a gene of interest.

2. Growth medium

[0091] The growth medium comprises components that promote the growth of Lactobacillus reuteri, e.g., the isolated strain of Lactobacillus reuteri.

[0092] In some embodiments, the growth medium comprises vitamins, amino acids, nucleotides, metal salts, unpurified carbon source, glycerol, sodium acetate, and polysorbate 80. In some embodiments, the growth medium does not comprise glucose. In some embodiments, the growth medium comprises vitamins, amino acids, nucleotides, metal salts, acid whey, glycerol, sodium acetate, and polysorbate 80, and does not comprise glucose. In some embodiments, the unpurified carbon source is an organic waste product or byproduct.

[0093] In some embodiments, the vitamins comprise one or more vitamins. In some embodiments, the vitamins comprise one or more vitamins selected from the group consisting of lipoate, pyridoxine, ascorbic acid, pyridoxal, pyridoxamine, biotin, folic acid, cobalamin, pantothenate, niacin, and riboflavin. In some embodiments, the vitamins comprise lipoate, pyridoxine, pantothenate, niacin, and riboflavin.

[0094] In some embodiments, the growth medium comprises an organic waste product or byproduct, such as acid whey or delactosed permeate. In some embodiments, the growth medium comprises acid whey. In some embodiments, the growth medium comprises delactosed permeate. In some embodiments, the organic waste product or byproduct is from a source that is selected from the group consisting of biodiesel production, bioethanol fermentation, and soap manufacturing.

3. Culturing conditions for growth medium

[0095] In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature that allows for growth of the isolated strain of Lactobacillus reuteri in the growth medium. In some embodiments, the temperature is selected based on the duration of incubation time that is desired. For instance, in some embodiments, the incubating the isolated strain in the growth medium occurs at a temperature that is or is about 37 degrees C for a duration of time that is or is about 10, 11, or 12 hours; or the incubating the isolated strain in the growth medium occurs at a temperature that is or is about 45° C for a duration of time that is or is about 5, 6, or 7 hours.

[0096] In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 28° C and 50° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 30° C and 48° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 32° C and 48° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 34° C and 48° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 34° C and 46° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 36° C and 46° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 34° C and 40° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 40° C and 50° C. [0097] In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of: (i) between or between about 28° C, 29° C, 31° C, 32° C, 33° C, or 34° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C; or (ii) between or between about 35° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45 ⁰ C„ 46 ⁰ C„ 47° C, 48° C, 49° C, or 50° C; or (iii) between or between about 36° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C; or (iv) between or between about 37° C and 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C; or (v) between or between about 37° C and 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, or 45° C; or (vi) at or about 28° C, 29° C, 30° C, 31° C, 32° C, 33° C, 34° C, 35° C, 36° C, 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C; or (vii) at or about 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, or 45° C; or (viii) at or about 37° C; or (ix) at or about 45° C.

[0098] In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 28° C, 29° C, 30° C, 31° C, 32° C, 33° C, or 34° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 28° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 29° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 30° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 31° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 32° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C. In some embodiments, culturing the isolated strain of

Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 33° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 34° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C.

[0099] In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 35° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 36° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 37° C and 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 37° C and 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, or 45° C.

[0100] In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of at or about 28° C, 29° C, 30° C, 31° C, 32° C, 33° C, 34° C, 35° C, 36° C, 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of at or about 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, or 45° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of at or about 35° C, 36° C, 37° C, 38° C, or 39° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of at or about 37° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of at or about 43° C, 44° C, 45° C, 46° C, or 47° C. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of at or about 45° C.

[0101] In some embodiments, the culturing the isolated strain of Lactobacillus reuteri in the growth medium occurs for a duration of time sufficient to result in a suitable optical density (OD), e.g., a suitable OD for producing 3-hydroxypropanal. In some embodiments, the suitable OD is between or between about 10 and 50. In some embodiments, the suitable OD is between or between about 10 and 12. In some embodiments, the suitable OD is or is about 10, 15, 20, 25, 30, 35, 40, 45, or 50. In some embodiments, the suitable OD is or is about 10, 11, or 12. Accordingly, in some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium occurs for a duration of time sufficient to result in an optical density (OD) within the range of or of about 10-50. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium occurs for a duration of time sufficient to result in an optical density (OD) within the range of or of aboutlO-12. In some embodiments, the duration of time depends on the incubation temperature, e.g., a shorter duration may be sufficient with a temperature above 37 degrees C, such as 40° C, 41° C, 42° C, 43° C, 44° C, or 45° C, as compared to 37° C.

[0102] In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium occurs for a duration of time that is: (i) at least or at least about 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 22 hours, 24 hours, 30 hours, 36 hours, 42 hours, or 48 hours; or (ii) is or is about 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, or 20 hours; or (iii) is between or between about 4 and 10 hours, 4 and 9 hours, 4 and 8 hours, 4 and 7 hours, 4 and 6 hours, 5 and 10 hours, 5 and 9 hours, 5 and 8 hours, or 5 and 7 hours; or (iv) is or is about 5 hours, 6 hours, 7 hours, or 8 hours; or (v) is between or between about 8 and 14 hours, 8 and 13 hours, 8 and 12 hours, 9 and 14 hours, 9 and 13 hours, 9 and 12 hours, 10 and 14 hours, 10 and 13 hours, or 10 and 12 hours; or (vi) is or is about 9 hours, 10 hours, 11 hours, 12 hours, or 13 hours.

[0103] In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium occurs for a duration of time that is at least or at least about 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 22 hours, 24 hours, 30 hours, 36 hours, 42 hours, or 48 hours. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium occurs for a duration of time that is or is about 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, or 20 hours. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium occurs for a duration of time that is between or between about 4 and 10 hours, 4 and 9 hours, 4 and 8 hours, 4 and 7 hours, 4 and 6 hours, 5 and 10 hours, 5 and 9 hours, 5 and 8 hours, or 5 and 7 hours. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium occurs for a duration of time that is or is about 5 hours, 6 hours, 7 hours, or 8 hours. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium occurs for a duration of time that is between or between about 8 and 14 hours, 8 and 13 hours, 8 and 12 hours, 9 and 14 hours, 9 and 13 hours, 9 and 12 hours, 10 and 14 hours, 10 and 13 hours, or 10 and 12 hours. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium occurs for a duration of time that is or is about 9 hours, 10 hours, 11 hours, 12 hours, or 13 hours. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium occurs for a duration of time that is or is about 5 hours, 6 hours, or 7 hours. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium occurs for a duration of time that is or is about 10 hours, 11 hours, or 12 hours.

[0104] In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature at or about 37° C for a duration of time that is or is about 10 hours, 11 hours, or 12 hours. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature at or about 45° C for a duration of time that is or is about 5 hours, 6 hours, 7 hours, or 8 hours.

[0105] In some embodiments, the isolated strain of Lactobacillus reuteri is cultured in the growth medium until CO2 generation by the isolated strain has reduced below a threshold level. In some embodiments, the threshold level is based on a level as compared to the level at peak CO2 generation. For instance, in some embodiments, the threshold level is 20%, 25%, 30%, 33%, 35%, 40%, 45%, 55%, 60%, 65%, 66%, or 70% of peak CO2 generation. In some embodiments, the threshold level is at or about 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, or 55% of peak CO2 generation. In some embodiments, the threshold level is a percentage that is between or between about 25% and 60%, such as between or between about 25% and 55%, 25% and 50%, 30% and 60%, 30% and 55%, 30% and 50%, 35% and 60%, 35% and 55%, 35% and 50%, 40% and 60%, 40% and 55%, 45% and 50%, 50% and 60%, or 55% and 60%. In some embodiments, the threshold level is a percentage that is between or between about 33% and 50%. In some embodiments, the threshold level is 1/5, 1/4, 1/3, or 1/2 of peak CO2 generation. In some embodiments, the threshold level is 1/3 of peak CO2 generation. In some embodiments, the threshold level is 1/2 of peak CO2 generation. In some embodiments, the threshold level is 1/3 to 1/2 of peak CO2 generation. In some embodiments, the threshold is at or about 50% of peak CO2 generation.

[0106] In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium until CO2 generation has reduced to between or between about 1/3 to 1/2 of peak CO2 generation. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium until CO2 generation has reduced to between or between about 33% to 50% of peak CO2 generation. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium until CO2 generation has reduced to at or below 1/2 of peak CO2 generation. In some embodiments, culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium until CO2 generation has reduced to about Vi of peak CO2 generation.

[0107] In some embodiments, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an optical density (OD) that is suitable for producing 3-hydroxypropanal. In some embodiments, an OD that is suitable for producing 3- hydroxypropanal is at least or at least about 8, 9, 10, 11, or 12. In some embodiments, an OD that is suitable for producing 3-hydroxypropanal is at least or at least about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50. In some embodiments, an OD that is suitable for producing 3-hydroxypropanal is at least or at least about 25, 30, 35, 40, 45, or 50. In some embodiments, an OD that is suitable for producing 3-hydroxypropanal is at least or at least about 10. In some embodiments, an OD that is suitable for producing 3-hydroxypropanal is at least or at least about 15. In some embodiments, an OD that is suitable for producing 3-hydroxypropanal is at least or at least about 20. In some embodiments, an OD that is suitable for producing 3-hydroxypropanal is at least or at least about 25. In some embodiments, an OD that is suitable for producing 3-hydroxypropanal is at least or at least about 30. In some embodiments, an OD that is suitable for producing 3-hydroxypropanal is at least or at least about 35. In some embodiments, an OD that is suitable for producing 3-hydroxypropanal is at least or at least about 40. In some embodiments, an OD that is suitable for producing 3-hydroxypropanal is at least or at least about 45. In some embodiments, an OD that is suitable for producing 3-hydroxypropanal is at least or at least about 50. In some embodiments, an OD that is suitable for producing 3-hydroxypropanal is at least or at least about is at least or at least about 25-50.

[0108] In some embodiments, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an optical density (OD) between or between about 9 and 15, 10 and 14, 12 and 20, 18 and 25, 20 and 28, or 25 and 50. In some embodiments, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an OD that is between or between about 8 and 15, 8 and 14, 8 and 13, 8 and 12, 9 and 15, 9 and 14, 9 and 13, 9 and 12, 10 and 15, 10 and 14, 10 and 13, or 10 and 12. In some embodiments, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an OD that is between or between about 9 and 50, 10 and 50, 11 and 50, 12 and 50, 13 and 50, 14 and 50, 9 and 45, 10 and 45, 11 and 45, 12 and 45, 13 and 45, 14 and 45, 9 and 40, 10 and 40, 11 and 40, 12 and 40, 13 and 40, 14 and 40, 9 and 35, 10 and 35, 11 and 35, 12 and 35, 13 and 35, 14 and 35, 9 and 30, 10 and 30, 11 and 30, 12 and 30, 13 and 30, 14 and 30, 9 and 25, 10 and 25, 11 and 25, 12 and 25, 13 and 25, 14 and 25, 9 and 20, 10 and 20, 11 and 20, 12 and 20, 13 and 20, 14 and 20, 15 and 50, 20 and 50, 25 and 50, 30 and 50, 35 and 50, 40 and 50, 15 and 45, 20 and 45, 25 and 45, 30 and 45, 35 and 45, 40 and 45, 15 and 40, 20 and 40, 25 and 40, 30 and 40, 35 and 40, 15 and 35, 20 and 35, 25 and 35, 30 and 35, 15 and 30, 20 and 30, or 25 and 30.

[0109] In some embodiments, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an OD that is between or between about 10 and 14. In some embodiments, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an OD that is between or between about 10 and 25. In some embodiments, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an OD that is between or between about 15 and 25. In some embodiments, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an OD that is between or between about 18 and 25. In some embodiments, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an OD that is between or between about 20 and 40. In some embodiments, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an OD that is between or between about 25 and 50. In some embodiments, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an OD that is between or between about 18 and 26. In some embodiments, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an OD that is between or between about 22 and 50. In some embodiments, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an OD that is between or between about 10 and 50.

[0110] In some embodiments, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an OD that is or is about 10, 11, 12, 13, or 14. In some embodiments, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an OD that is or is about 10, 11, or 12. In some embodiments, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an OD that is or is about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50. In some embodiments, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an OD that is or is about 18, 20, 25, 30, 35, 40, 45, or 50. In some embodiments, following the culturing of the isolated or engineered from isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated or engineered from isolated strain of Lactobacillus reuteri has an OD that is or is about 18, 22, or 25. In some embodiments, following the culturing of the isolated or engineered from isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated or engineered from isolated strain of Lactobacillus reuteri has an OD that is or is about 35, 40, 45 or 50.

4. Separating the population from the growth medium

[0111] In some embodiments, the methods provided herein further comprise, following culturing the isolated strain of Lactobacillus reuteri in the growth medium, separating the population of the isolated strain of Lactobacillus reuteri from the growth media.

[0112] In some embodiments, the separating the population of the isolated strain of Lactobacillus reuteri from the growth media can be performed using any suitable separation technique. In some embodiments, separating the population of the isolated strain of Lactobacillus reuteri from the growth media comprises the use of tangential flow filtration and/or centrifugation. In some embodiments, separating comprises the use of tangential flow filtration, followed by centrifuging the population. In some embodiments, separating comprises the use of tangential flow filtration, followed by centrifuging the population, and followed by removing the growth media. In some embodiments, separating the population of the isolated strain of Lactobacillus reuteri from the growth media comprises removal of the growth media from the population of the isolated strain of Lactobacillus reuteri. B. _ Culturing a population of Lactobacillus reuteri in a production medium

[0113] In some embodiments, the methods provided herein comprise culturing the population, i.e., the population of the isolated strain of Lactobacillus reuteri, in a production medium comprising a glycerol source to produce 3-hydroxypropanal, thereby resulting in a production medium comprising 3- hydroxypropanal. In some embodiments, the production medium comprises a carbohydrate source, such as lactose, galactose, sucrose, glucose, maltose, arabinose, or fructose, or any other suitable carbohydrate source.

[0114] In some embodiments, the culturing the population, i.e., the population of the isolated strain of Lactobacillus reuteri, in a production medium comprising a glycerol source to produce 3-hydroxypropanal, occurs following the culturing an isolated strain of Lactobacillus reuteri in a growth medium. In some embodiments, the culturing the population, i.e., the population of the isolated strain of Lactobacillus reuteri, in a production medium comprising a glycerol source to produce 3-hydroxypropanal, occurs following the separating the population of the isolated strain of Lactobacillus reuteri from the growth media. Accordingly, in some embodiments, the methods provided herein comprise, following the culturing an isolated strain of Lactobacillus reuteri in a growth medium, culturing the population, i.e., the population of the isolated strain of Lactobacillus reuteri, in a production medium comprising a glycerol source to produce 3- hydroxypropanal, thereby resulting in a production medium comprising 3-hydroxypropanal. In some embodiments, the methods provided herein comprise, following the separating the population of the isolated strain of Lactobacillus reuteri from the growth media, culturing the population, i.e., the population of the isolated strain of Lactobacillus reuteri, in a production medium comprising a glycerol source to produce 3- hydroxypropanal, thereby resulting in a production medium comprising 3-hydroxypropanal. In some embodiments, the isolated strain is cultured in a production medium that comprises a carbohydrate source. In some embodiments, the carbohydrate source comprises a carbohydrate selected from the group consisting of lactose, galactose, sucrose, glucose, maltose, arabinose, and fructose, or any combination thereof.

[0115] In some embodiments, the isolated strain can be cultured in a production medium that comprises a low concentration of a carbohydrate source. For instance, it was surprisingly discovered that decreasing the concentration of lactose from 25 mM to 1.5 mM in the production medium actually increased reuterin yield by 5-15%, as described in Example 2. Accordingly, in some embodiments, the isolated strain is cultured in a production medium comprising a concentration of a carbohydrate source, e.g., lactose, that is between or between about 0.1 mM and 25 mM, 0.1 mM and 20 mM, 0.1 mM and 15 mM, 0.1 mM and 10 mM, 0.1 mM and 5 mM, 0.1 mM and 3 mM, 0.1 mM and 2 mM, 0.5 mM and 25 mM, 0.5 mM and 20 mM, 0.5 mM and 15 mM, 0.5 mM and 10 mM, 0.5 mM and 5 mM, 0.5 0.5 mM and 2 mM, 1 mM and 25 mM, 1 mM and 20 mM, 1 mM and 15 mM, 1 mM and 10 mM, 1 mM and 5 mM, 1 mM and 3 mM, or 1 mM and 2 mM.

1. Production medium

[0116] In some embodiments, the methods provided herein comprise the use of a production medium comprising a glycerol source, e.g., by culturing the population of the isolated strain of Lactobacillus reuteri in a production medium comprising a glycerol source. The glycerol source acts as a source of the glycerol that is converted into 3-hydroxypropanal (reuterin) by the isolated strain of Lactobacillus reuteri.

[0117] In some embodiments, the glycerol source can be any source of glycerol that is suitable for conversion into 3-hydroxypropanal (reuterin) by Lactobacillus reuteri. In some embodiments, the glycerol source is glycerol. In some embodiments, the glycerol source is from an organic waste source. In some embodiments, the organic waste source has been pasteurized.

[0118] In some embodiments, the production medium is any medium that allows for the conversion of glycerol to 3-hydroxypropanal (reuterin) by the isolated strain of Lactobacillus reuteri.

[0119] In some embodiments, the production medium comprises a carbohydrate source. In some embodiments, the carbohydrate source comprises a carbohydrate selected from the group consisting of lactose, galactose, sucrose, glucose, maltose, arabinose, and fructose, or any combination thereof.

[0120] In some embodiments, the production medium comprises lactose. In some embodiments, the production medium comprises galactose. In some embodiments, the production medium comprises sucrose. In some embodiments, the production medium comprises glucose. In some embodiments, the production medium comprises maltose. In some embodiments, the production medium comprises arabinose. In some embodiments, the production medium comprises fructose.

[0121] In some embodiments, the production medium comprises glycerol, a carbohydrate source, and a buffering agent. In some embodiments, the production medium comprises glycerol, a carbohydrate source, and a buffering agent, and one or more of sodium chloride (NaCl), potassium chloride, metals, and iron(II) sulfate, and any combination thereof.

[0122] In some embodiments, the production medium comprises glycerol, a carbohydrate source, a buffering agent, sodium chloride (NaCl), potassium chloride, metals, and iron(II) sulfate.

[0123] In some embodiments, the production medium comprises glycerol, a carbohydrate source, MOPS pH 7, sodium chloride (NaCl), potassium chloride, metals, and iron(II) sulfate. [0124] In some embodiments, the production medium comprises glycerol, a carbohydrate source, a buffering agent, polysorbate 80, sodium chloride (NaCl), potassium chloride, metals, and iron(II) sulfate. In some embodiments, the production medium comprises glycerol, a carbohydrate source, a buffering agent, sodium chloride (NaCl), potassium chloride, metals, and iron(II) sulfate. In some embodiments, the production medium optionally comprises polysorbate 80.

[0125] In some embodiments, the production medium comprises glycerol, a carbohydrate source, MOPS pH 7, polysorbate 80, sodium chloride (NaCl), potassium chloride, metals, and iron(II) sulfate. In some embodiments, the production medium comprises glycerol, a carbohydrate source, MOPS pH 7, sodium chloride (NaCl), potassium chloride, metals, and iron(II) sulfate. In some embodiments, the production medium optionally comprises polysorbate 80.

[0126] In some embodiments, the buffering agent is MOPS, e.g., MOPS pH 7, or phosphate buffer, e.g., phosphate buffer pH 7. In some embodiments, the buffering agent is MOPS, e.g., MOPS pH 7. In some embodiments, the buffering agent is phosphate buffer, e.g., phosphate buffer pH 7.

[0127] In some embodiments, the carbohydrate source is selected from the group consisting of lactose, galactose, sucrose, glucose, maltose, arabinose, and fructose, or any combination thereof.

[0128] In some embodiments, the production medium comprises glycerol, lactose, a buffering agent, sodium chloride (NaCl), potassium chloride, metals, and iron(II) sulfate.

[0129] In some embodiments, the production medium comprises glycerol, lactose, MOPS pH 7, sodium chloride (NaCl), potassium chloride, metals, and iron(II) sulfate.

[0130] In some embodiments, the production medium comprises glycerol, lactose, a buffering agent, polysorbate 80, sodium chloride (NaCl), potassium chloride, metals, and iron(II) sulfate. In some embodiments, the production medium comprises glycerol, lactose, a buffering agent, sodium chloride (NaCl), potassium chloride, metals, and iron(II) sulfate. In some embodiments, the production medium optionally comprises polysorbate 80. In some embodiments, the buffering agent is MOPS, e.g., MOPS pH 7, or phosphate buffer, e.g., phosphate buffer pH 7. In some embodiments, the buffering agent is MOPS, e.g., MOPS pH 7. In some embodiments, the buffering agent is phosphate buffer, e.g., phosphate buffer pH 7.

[0131] In some embodiments, the production medium comprises glycerol, lactose, MOPS pH 7, polysorbate 80, sodium chloride (NaCl), potassium chloride, metals, and iron(II) sulfate. In some embodiments, the production medium comprises glycerol, lactose, MOPS pH 7, sodium chloride (NaCl), potassium chloride, metals, and iron(II) sulfate. In some embodiments, the production medium optionally comprises polysorbate 80. [0132] In some embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol. In some embodiments, the production medium comprises between or between about 50 mM and 750 mM, 700 mM, 650 mM, 600 mM, 550 mM, 500 mM, 450 mM, 400 mM, 350 mM, 300 mM, 350 mM, or 200 mM glycerol; or between or between about 100 mM and 750 mM, 700 mM, 650 mM, 600 mM, 550 mM, 500 mM, 450 mM, 400 mM, 350 mM, 300 mM, 350 mM, or 200 mM glycerol; or between or between about 150 mM and 750 mM, 700 mM, 650 mM, 600 mM, 550 mM, 500 mM, 450 mM, 400 mM, 350 mM, 300 mM, 350 mM, or 200 mM glycerol. In some embodiments, the production medium comprises between or between about 100 mM and 300 mM glycerol. In some embodiments, the production medium comprises between or between about 150 mM and 250 mM glycerol. In some embodiments, the production medium comprises at or about 100 mM, 105 mM, 110 mM, 115 mM, 120 mM, 125 mM, 130 mM, 135 mM, 140 mM, 145 mM, 150 mM, 155 mM, 160 mM, 165 mM, 170 mM, 175 mM, 180 mM, 185 mM, 190 mM,

195 mM, 200 mM, 205 mM, 210 mM, 215 mM, 220 mM, 225 mM, 230 mM, 235 mM, 240 mM, 245 mM,

250 mM, 255 mM, 260 mM, 265 mM, 270 mM, 275 mM, 280 mM, 285 mM, 290 mM, 295 mM, 300 mM,

305 mM, 310 mM, 315 mM, 320 mM, 325 mM, 330 mM, 335 mM, 340 mM, 345 mM, 350 mM, 355 mM,

360 mM, 365 mM, 370 mM, 375 mM, 380 mM, 385 mM, 390 mM, 395 mM, or 400 mM glycerol. In some embodiments, the production medium comprises at or about 200 mM glycerol.

[0133] In some embodiments, the production medium comprises between or between about 0.1 mM and 60 mM of a carbohydrate source. In some embodiments, the production medium comprises between or between about 0.5 mM and 60 mM of a carbohydrate source. In some embodiments, the production medium comprises between or between about 1 mM and 60 mM of a carbohydrate source. In some embodiments, the production medium comprises between or between about 0.1 mM and 60 mM, 0.1 mM and 50 mM, 0.1 mM and 40 mM, 0.1 mM and 30 mM, 0.1 mM and 25 mM, 0.1 mM and 20 mM, 0.1 mM and 15 mM, 0.1 mM and 10 mM, 0.1 mM and 5 mM, 0.1 mM and 3 mM, 0.1 mM and 2.5 mM, 0.1 mM and 2 mM, O.lmM and 1.5 mM, 0.5 mM and 60 mM, 0.5 mM and 50 mM, 0.5 mM and 40 mM, 0.5 mM and 30 mM, 0.5 mM and 25 mM, 0.5 mM and 20 mM, 0.5 mM and 15 mM, 0.5 mM and 10 mM, 0.5 mM and 5 mM, 0.5 mM and 3 mM, 0.5 mM and 2.5 mM, 0.5 mM and 2 mM, 0.5 mM and 1.5 mM, 1 mM and 60 mM, 1 mM and 50 mM, 1 mM and 40 mM, 1 mM and 30 mM, 1 mM and 25 mM, 1 mM and 20 mM, 1 mM and 15 mM, 1 mM and 10 mM, 1 mM and 5 mM, 1 mM and 3 mM, 1 mM and 2.5 mM, 1 mM and 2 mM, ImM and 1.5 mM, 3 mM and 60 mM, 3 mM and 50 mM, 3 mM and 40 mM, 3 mM and 30 mM, 3 mM and 25 mM, 3 mM and 20 mM, 3 mM and 15 mM, 3 mM and 10 mM, 3 mM and 5 mM, 5 mM and 60 mM, 5 mM and 50 mM, 5 mM and 40 mM, 5 mM and 30 mM, 5 mM and 25 mM, 5 mM and 20 mM, 5 mM and 15 mM, or 5 mM and 10 mM of a carbohydrate source. [0134] In some embodiments, the production medium comprises less than 25 mM of the carbohydrate source. In some embodiments, the production medium comprises less than 25 mM of the carbohydrate source, but at least 0.1 mM, 0.2 mM, 0.3 mM, 0.4 mM, 0.5 mM, 0.6 mM, 0.7 mM, 0.8 mM, 0.9 mM, or 1 mM of the carbohydrate source. In some embodiments, the production medium comprises between or between about 0.5 mM and 20 mM of the carbohydrate source. In some embodiments, the production medium comprises between or between about 0.5 mM and 15 mM of the carbohydrate source. In some embodiments, the production medium comprises between or between about 0.5 mM and 10 mM of the carbohydrate source. In some embodiments, the production medium comprises between or between about 0.5 mM and 5 mM of the carbohydrate source. In some embodiments, the production medium comprises between or between about 0.5 mM and 3 mM of the carbohydrate source. In some embodiments, the production medium comprises between or between about 0.5 mM and 2 mM of the carbohydrate source. In some embodiments, the production medium comprises between or between about 1 mM and 3 mM of the carbohydrate source. In some embodiments, the production medium comprises between or between about 1 mM and 2 mM of the carbohydrate source.

[0135] In some embodiments, the carbohydrate source is lactose. In some embodiments, the carbohydrate source is galactose. In some embodiments, the carbohydrate source is sucrose. In some embodiments, the carbohydrate source is glucose. In some embodiments, the carbohydrate source is maltose. In some embodiments, the carbohydrate source is arabinose. In some embodiments, the carbohydrate source is fructose.

[0136] In some embodiments, the production medium comprises between or between about 0.1 mM and 60 mM lactose. In some embodiments, the production medium comprises between or between about 0.5 mM and 60 mM lactose. In some embodiments, the production medium comprises between or between about 1 mM and 60 mM lactose. In some embodiments, the production medium comprises between or between about 0.1 mM and 60 mM, 0.1 mM and 50 mM, 0.1 mM and 40 mM, 0.1 mM and 30 mM, 0.1 mM and 25 mM, 0.1 mM and 20 mM, 0.1 mM and 15 mM, 0.1 mM and 10 mM, 0.1 mM and 5 mM, 0.1 mM and 3 mM, 0.1 mM and 2.5 mM, 0.1 mM and 2 mM, O.lmM and 1.5 mM, 0.5 mM and 60 mM, 0.5 mM and 50 mM, 0.5 mM and 40 mM, 0.5 mM and 30 mM, 0.5 mM and 25 mM, 0.5 mM and 20 mM, 0.5 mM and 15 mM, 0.5 mM and 10 mM, 0.5 mM and 5 mM, 0.5 mM and 3 mM, 0.5 mM and 2.5 mM, 0.5 mM and 2 mM, 0.5 mM and 1.5 mM, 1 mM and 60 mM, 1 mM and 50 mM, 1 mM and 40 mM, 1 mM and 30 mM, 1 mM and 25 mM, 1 mM and 20 mM, 1 mM and 15 mM, 1 mM and 10 mM, 1 mM and 5 mM, 1 mM and 3 mM, 1 mM and 2.5 mM, 1 mM and 2 mM, ImM and 1.5 mM, 3 mM and 60 mM, 3 mM and 50 mM, 3 mM and 40 mM, mM, 5 mM and 60 mM, 5 mM and 50 mM, 5 mM and 40 mM, 5 mM and 30 mM, 5 mM and 25 mM, 5 mM and 20 mM, 5 mM and 15 mM, or 5 mM and 10 mM lactose.

[0137] In some embodiments, the production medium comprises less than 25 mM lactose. In some embodiments, the production medium comprises between or between about 0.5 mM and 20 mM lactose. In some embodiments, the production medium comprises between or between about 0.5 mM and 15 mM lactose. In some embodiments, the production medium comprises between or between about 0.5 mM and 10 mM lactose. In some embodiments, the production medium comprises between or between about 0.5 mM and 5 mM lactose. In some embodiments, the production medium comprises between or between about 0.5 mM and 3 mM lactose. In some embodiments, the production medium comprises between or between about 0.5 mM and 2 mM lactose. In some embodiments, the production medium comprises between or between about 1 mM and 3 mM lactose. In some embodiments, the production medium comprises between or between about 1 mM and 2 mM lactose.

[0138] In some embodiments, the production medium comprises between or between about 1 mM and 20 mM lactose. In some embodiments, the production medium comprises between or between about 1 mM and 15 mM lactose. In some embodiments, the production medium comprises between or between about 1 mM and 10 mM lactose. In some embodiments, the production medium comprises between or between about 1 mM and 5 mM lactose. In some embodiments, the production medium comprises between or between about 1 mM and 3 mM lactose. In some embodiments, the production medium comprises between or between about 1 mM and 2 mM lactose. In some embodiments, the production medium comprises between or between about 1.5 mM and 20 mM lactose. In some embodiments, the production medium comprises between or between about 1.5 mM and 15 mM lactose. In some embodiments, the production medium comprises between or between about 1.5 mM and 10 mM lactose. In some embodiments, the production medium comprises between or between about 1.5 mM and 5 mM lactose. In some embodiments, the production medium comprises between or between about 1.5 mM and 3 mM lactose. In some embodiments, the production medium comprises between or between about 1.5 mM and 2 mM lactose.

[0139] In some embodiments, the production medium comprises at or about 0.1 mM lactose. In some embodiments, the production medium comprises at or about 0.5 mM lactose. In some embodiments, the production medium comprises at or about 1 mM lactose. In some embodiments, the production medium comprises at or about 1.5 mM lactose. In some embodiments, the production medium comprises at or about 2 mM lactose. In some embodiments, the production medium comprises at or about 3 mM lactose. In some embodiments, the production medium comprises at or about 4 mM lactose. In some embodiments, the production medium comprises at or about 5 mM lactose. In some embodiments, the production medium comprises at or about 6 mM lactose. In some embodiments, the production medium comprises at or about 7 mM lactose. In some embodiments, the production medium comprises at or about 8 mM lactose. In some embodiments, the production medium comprises at or about 9 mM lactose. In some embodiments, the production medium comprises at or about 10 mM lactose.

[0140] In some embodiments, the production medium comprises between or between about 10 mM and 60 mM lactose. In some embodiments, the production medium comprises between or between about 10 mM and 55 mM, 10 mM and 50 mM, 10 mM and 45 mM, 10 mM and 40 mM, 10 mM and 35 mM, 10 mM and 30 mM, 10 mM and 25 mM, 15 mM and 55 mM, 15 mM and 50 mM, 15 mM and 45 mM, 15 mM and 40 mM, 15 mM and 35 mM, 15 mM and 30 mM, 15 mM and 25 mM, 20 mM and 55 mM, 20 mM and 50 mM, 20 mM and 45 mM, 20 mM and 40 mM, 20 mM and 35 mM, 20 mM and 30 mM, 20 mM and 25 mM lactose. In some embodiments, the production medium comprises between or between about 20 mM and 30 mM lactose. In some embodiments, the production medium comprises at or about 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29 mM, 30 mM, 31 mM, 32 mM, 33 mM, 34 mM, 35 mM, 36 mM, 37 mM, 38 mM, 39 mM, 40 mM, 41 mM, 42 mM, 43 mM, 44 mM, 45 mM, 46 mM, 47 mM, 48 mM, 49 mM, 50 mM, 51 mM, 52 mM, 53 mM, 54 mM, 55 mM, 56 mM, 57 mM, 58 mM, 59 mM, or 60 mM lactose. In some embodiments, the production medium comprises at or about 25 mM lactose. In some embodiments, the production medium comprises at or about 10 mM lactose. In some embodiments, the production medium comprises at or about 11 mM lactose. In some embodiments, the production medium comprises at or about 12 mM lactose. In some embodiments, the production medium comprises at or about 13 mM lactose. In some embodiments, the production medium comprises at or about 14 mM lactose. In some embodiments, the production medium comprises at or about 15 mM lactose. In some embodiments, the production medium comprises at or about 16 mM lactose. In some embodiments, the production medium comprises at or about 17 mM lactose. In some embodiments, the production medium comprises at or about 18 mM lactose. In some embodiments, the production medium comprises at or about 19 mM lactose. In some embodiments, the production medium comprises at or about 20 mM lactose. In some embodiments, the production medium comprises at or about 21 mM lactose. In some embodiments, the production medium comprises at or about 22 mM lactose. In some embodiments, the production medium comprises at or about 23 mM lactose. In some embodiments, the production medium comprises at or about 24 mM lactose. In some embodiments, the production medium comprises at or about 25 mM lactose.

[0141] In some embodiments, the production medium comprises a buffering agent. [0142] In some embodiments, the production medium comprises between or between about 20 mM and 110 mM, 20 mM and 100 mM, 20 mM and 90 mM, 20 mM and 80 mM, 20 mM and 70 mM, 20 mM and 60 mM, 20 mM and 50 mM, 30 mM and 110 mM, 30 mM and 100 mM, 30 mM and 90 mM, 30 mM and 80 mM, 30 mM and 70 mM, 30 mM and 60 mM, 30 mM and 50 mM, 40 mM and 110 mM, 40 mM and 100 mM, 40 mM and 90 mM, 40 mM and 80 mM, 40 mM and 70 mM, 40 mM and 60 mM, or 40 mM and 50 mM of the buffering agent. In some embodiments, the production medium comprises between or between about 30 mM and 70 mM of the buffering agent. In some embodiments, the production medium comprises between or between about 40 mM and 60 mM of the buffering agent. In some embodiments, the production medium comprises at or about 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29 mM, 30 mM, 31 mM, 32 mM, 33 mM, 34 mM, 35 mM, 36 mM, 37 mM, 38 mM, 39 mM, 40 mM, 41 mM, 42 mM, 43 mM, 44 mM, 45 mM, 46 mM, 47 mM, 48 mM, 49 mM, 50 mM, 51 mM, 52 mM, 53 mM, 54 mM, 55 mM, 56 mM, 57 mM, 58 mM, 59 mM, 60 mM, 61 mM, 62 mM, 63 mM, 64 mM, 65 mM, 66 mM, 67 mM, 68 mM, 69 mM, 70 mM, 71 mM, 72 mM, 73 mM, 74 mM, 75 mM, 76 mM, 77 mM, 78 mM, 79 mM, 80 mM, 81 mM, 82 mM, 83 mM, 84 mM, 85 mM, 86 mM, 87 mM, 88 mM, 89 mM, 90 mM, 91 mM, 92 mM, 93 mM, 94 mM, 95 mM, 96 mM, 97 mM, 98 mM, 99 mM, 100 mM, 101 mM, 102 mM, 103 mM, 104 mM, 105 mM, 106 mM, 107 mM, 108 mM, 109 mM, 110 mM, 111 mM, 112 mM, 113 mM, 114 mM, 115 mM, 116 mM, 117 mM, 118 mM, 119 mM, 120 mM, 121 mM, 122 mM, 123 mM, 124 mM, or 125 mM of the buffering agent. In some embodiments, the production medium comprises at or about 50 mM of the buffering agent.

[0143] In some embodiments, the buffering agent is MOPS or a phosphate buffer. In some embodiments, the buffering agent is MOPS pH 7, or a phosphate buffer pH 7.

[0144] In some embodiments, the buffering agent is MOPS, e.g., MOPS pH 7. In some embodiments, the production medium comprises between or between about 20 mM and 125 mM MOPS pH 7. In some embodiments, the production medium comprises between or between about 20 mM and 110 mM, 20 mM and 100 mM, 20 mM and 90 mM, 20 mM and 80 mM, 20 mM and 70 mM, 20 mM and 60 mM, 20 mM and 50 mM, 30 mM and 110 mM, 30 mM and 100 mM, 30 mM and 90 mM, 30 mM and 80 mM, 30 mM and 70 mM, 30 mM and 60 mM, 30 mM and 50 mM, 40 mM and 110 mM, 40 mM and 100 mM, 40 mM and 90 mM, 40 mM and 80 mM, 40 mM and 70 mM, 40 mM and 60 mM, or 40 mM and 50 mM MOPS pH 7. In some embodiments, the production medium comprises between or between about 30 mM and 70 mM MOPS pH 7. In some embodiments, the production medium comprises between or between about 40 mM and 60 mM MOPS pH 7. In some embodiments, the production medium comprises at or about 20 mM, 21 mM, 22 35 mM, 36 mM, 37 mM, 38 mM, 39 mM, 40 mM, 41 mM, 42 mM, 43 mM, 44 mM, 45 mM, 46 mM, 47 mM, 48 mM, 49 mM, 50 mM, 51 mM, 52 mM, 53 mM, 54 mM, 55 mM, 56 mM, 57 mM, 58 mM, 59 mM, 60 mM, 61 mM, 62 mM, 63 mM, 64 mM, 65 mM, 66 mM, 67 mM, 68 mM, 69 mM, 70 mM, 71 mM, 72 mM, 73 mM, 74 mM, 75 mM, 76 mM, 77 mM, 78 mM, 79 mM, 80 mM, 81 mM, 82 mM, 83 mM, 84 mM, 85 mM, 86 mM, 87 mM, 88 mM, 89 mM, 90 mM, 91 mM, 92 mM, 93 mM, 94 mM, 95 mM, 96 mM, 97 mM, 98 mM, 99 mM, 100 mM, 101 mM, 102 mM, 103 mM, 104 mM, 105 mM, 106 mM, 107 mM, 108 mM, 109 mM, 110 mM, 111 mM, 112 mM, 113 mM, 114 mM, 115 mM, 116 mM, 117 mM, 118 mM, 119 mM, 120 mM, 121 mM, 122 mM, 123 mM, 124 mM, or 125 mM MOPS pH 7. In some embodiments, the production medium comprises at or about 50 mM MOPS pH 7.

[0145] In some embodiments, the buffering agent is a phosphate buffer, e.g., a phosphate buffer pH 7. In some embodiments, the production medium comprises between or between about 20 mM and 125 mM phosphate buffer pH 7. In some embodiments, the production medium comprises between or between about 20 mM and 110 mM, 20 mM and 100 mM, 20 mM and 90 mM, 20 mM and 80 mM, 20 mM and 70 mM, 20 mM and 60 mM, 20 mM and 50 mM, 30 mM and 110 mM, 30 mM and 100 mM, 30 mM and 90 mM, 30 mM and 80 mM, 30 mM and 70 mM, 30 mM and 60 mM, 30 mM and 50 mM, 40 mM and 110 mM, 40 mM and 100 mM, 40 mM and 90 mM, 40 mM and 80 mM, 40 mM and 70 mM, 40 mM and 60 mM, or 40 mM and 50 mM phosphate buffer pH 7. In some embodiments, the production medium comprises between or between about 30 mM and 70 mM phosphate buffer pH 7. In some embodiments, the production medium comprises between or between about 40 mM and 60 mM phosphate buffer pH 7. In some embodiments, the production medium comprises at or about 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29 mM, 30 mM, 31 mM, 32 mM, 33 mM, 34 mM, 35 mM, 36 mM, 37 mM, 38 mM, 39 mM, 40 mM, 41 mM, 42 mM, 43 mM, 44 mM, 45 mM, 46 mM, 47 mM, 48 mM, 49 mM, 50 mM, 51 mM, 52 mM, 53 mM, 54 mM, 55 mM, 56 mM, 57 mM, 58 mM, 59 mM, 60 mM, 61 mM, 62 mM, 63 mM, 64 mM, 65 mM, 66 mM, 67 mM, 68 mM, 69 mM, 70 mM, 71 mM, 72 mM, 73 mM, 74 mM, 75 mM, 76 mM, 77 mM, 78 mM, 79 mM, 80 mM, 81 mM, 82 mM, 83 mM, 84 mM, 85 mM, 86 mM, 87 mM, 88 mM, 89 mM, 90 mM, 91 mM, 92 mM, 93 mM, 94 mM, 95 mM, 96 mM, 97 mM, 98 mM, 99 mM, 100 mM, 101 mM, 102 mM, 103 mM, 104 mM, 105 mM, 106 mM, 107 mM, 108 mM, 109 mM, 110 mM, 111 mM, 112 mM, 113 mM, 114 mM, 115 mM, 116 mM, 117 mM, 118 mM, 119 mM, 120 mM, 121 mM, 122 mM, 123 mM, 124 mM, or 125 mM phosphate buffer pH 7. In some embodiments, the production medium comprises at or about 50 mM phosphate buffer pH 7.

[0146] In some embodiments, the production medium comprises polysorbate 80 (also known as Tween 80). In some embodiments, the production medium does not comprise polysorbate 80. In some embodiments, the production medium optionally comprises polysorbate 80. In some embodiments, the production medium comprises between or between about 0.1 g/L and 10 g/L polysorbate 80. In some embodiments, the production medium comprises between or between about 0.1 g/L and 10 g/L, 0.1 g/L and 9 g/L, 0.1 g/L and 8 g/L, 0.1 g/L and 7 g/L, 0.1 g/L and 6 g/L, 0.1 g/L and 5 g/L, 0.1 g/L and 4 g/L, 0.1 g/L and

3 g/L, 0.1 g/L and 2 g/L, 0.1 g/L and 1.5 g/L, 0.1 g/L and 1.4 g/L, 0.1 g/L and 1.3 g/L, 0.1 g/L and 1.2 g/L, 0.1 g/L and 1.1 g/L, 0.1 g/L and 1 g/L, 0.2 g/L and 10 g/L, 0.2 g/L and 9 g/L, 0.2 g/L and 8 g/L, 0.2 g/L and 7 g/L, 0.2 g/L and 6 g/L, 0.2 g/L and 5 g/L, 0.2 g/L and 4 g/L, 0.2 g/L and 3 g/L, 0.2 g/L and 2 g/L, 0.2 g/L and 1.5 g/L, 0.2 g/L and 1.4 g/L, 0.2 g/L and 1.3 g/L, 0.2 g/L and 1.2 g/L, 0.2 g/L and 1.1 g/L, 0.3 g/L and 10 g/L, 0.3 g/L and 9 g/L, 0.3 g/L and 8 g/L, 0.3 g/L and 7 g/L, 0.3 g/L and 6 g/L, 0.3 g/L and 5 g/L, 0.3 g/L and

4 g/L, 0.3 g/L and 3 g/L, 0.3 g/L and 2 g/L, 0.3 g/L and 1.5 g/L, 0.3 g/L and 1.4 g/L, 0.3 g/L and 1.3 g/L, 0.3 g/L and 1.2 g/L, 0.3 g/L and 1.1 g/L, 0.4 g/L and 10 g/L, 0.4 g/L and 9 g/L, 0.4 g/L and 8 g/L, 0.4 g/L and 7 g/L, 0.4 g/L and 6 g/L, 0.4 g/L and 5 g/L, 0.4 g/L and 4 g/L, 0.4 g/L and 3 g/L, 0.4 g/L and 2 g/L, 0.4 g/L and

1.5 g/L, 0.4 g/L and 1.4 g/L, 0.4 g/L and 1.3 g/L, 0.4 g/L and 1.2 g/L, 0.4 g/L and 1.1 g/L, 0.5 g/L and 10 g/L, 0.5 g/L and 9 g/L, 0.5 g/L and 8 g/L, 0.5 g/L and 7 g/L, 0.5 g/L and 6 g/L, 0.5 g/L and 5 g/L, 0.5 g/L and 4 g/L, 0.5 g/L and 3 g/L, 0.5 g/L and 2 g/L, 0.5 g/L and 1.5 g/L, 0.5 g/L and 1.4 g/L, 0.5 g/L and 1.3 g/L, 0.5 g/L and 1.2 g/L, 0.5 g/L and 1.1 g/L, 0.6 g/L and 10 g/L, 0.6 g/L and 9 g/L, 0.6 g/L and 8 g/L, 0.6 g/L and 7 g/L, 0.6 g/L and 6 g/L, 0.6 g/L and 5 g/L, 0.6 g/L and 4 g/L, 0.6 g/L and 3 g/L, 0.6 g/L and 2 g/L, 0.6 g/L and

1.5 g/L, 0.6 g/L and 1.4 g/L, 0.6 g/L and 1.3 g/L, 0.6 g/L and 1.2 g/L, 0.6 g/L and 1.1 g/L, 0.7 g/L and 10 g/L, 0.7 g/L and 9 g/L, 0.7 g/L and 8 g/L, 0.7 g/L and 7 g/L, 0.7 g/L and 6 g/L, 0.7 g/L and 5 g/L, 0.7 g/L and 4 g/L, 0.7 g/L and 3 g/L, 0.7 g/L and 2 g/L, 0.7 g/L and 1.5 g/L, 0.7 g/L and 1.4 g/L, 0.7 g/L and 1.3 g/L, 0.7 g/L and 1.2 g/L, 0.7 g/L and 1.1 g/L, 0.8 g/L and 10 g/L, 0.8 g/L and 9 g/L, 0.8 g/L and 8 g/L, 0.8 g/L and 7 g/L, 0.8 g/L and 6 g/L, 0.8 g/L and 5 g/L, 0.8 g/L and 4 g/L, 0.8 g/L and 3 g/L, 0.8 g/L and 2 g/L, 0.8 g/L and

1.5 g/L, 0.8 g/L and 1.4 g/L, 0.8 g/L and 1.3 g/L, 0.8 g/L and 1.2 g/L, 0.8 g/L and 1.1 g/L, 0.9 g/L and 10 g/L, 0.8 g/L and 9 g/L, 0.9 g/L and 8 g/L, 0.9 g/L and 7 g/L, 0.9 g/L and 6 g/L, 0.9 g/L and 5 g/L, 0.9 g/L and 4 g/L, 0.9 g/L and 3 g/L, 0.9 g/L and 2 g/L, 0.9 g/L and 1.5 g/L, 0.9 g/L and 1.4 g/L, 0.9 g/L and 1.3 g/L, 0.9 g/L and 1.2 g/L, or 0.9 g/L and 1.1 g/L polysorbate 80. In some embodiments, the production medium comprises between or between about 0.5 g/L and 2 g/L polysorbate 80. In some embodiments, the production medium comprises at or about 0.1 g/L, 0.2 g/L, 0.3 g/L, 0.4 g/L, 0.5 g/L, 0.6 g/L, 0.7 g/L, 0.8 g/L, 0.9 g/L, 1 g/L, 1.1 g/L, 1.2 g/L, 1.3 g/L, 1.4 g/L, 1.5 g/L, 1.6 g/L, 1.7 g/L, 1.8 g/L, 1.9 g/L, 2 g/L, 2.1 g/L, 2.2 g/L, 2.3 g/L, 2.4 g/L, 2.5 g/L, 2.6 g/L, 2.7 g/L, 2.8 g/L, 2.9 g/L, 3 g/L, 3.5 g/L, 4 g/L, 4.5 g/L, 5 g/L, 5.5 g/L, 6 g/L,

6.5 g/L, 7 g/L, 7.5 g/L, 8 g/L, 8.5 g/L, 9 g/L, 9.5 g/L, or 10 g/L polysorbate 80. In some embodiments, the production medium comprises at or about 1 g/L polysorbate 80. [0147] In some embodiments, the production medium comprises between or between about 10 and 60 mM sodium chloride (NaCl). In some embodiments, the production medium comprises between or between about 10 mM and 55 mM, 10 mM and 50 mM, 10 mM and 45 mM, 10 mM and 40 mM, 10 mM and 35 mM, 10 mM and 30 mM, 10 mM and 25 mM, 15 mM and 55 mM, 15 mM and 50 mM, 15 mM and 45 mM, 15 mM and 40 mM, 15 mM and 35 mM, 15 mM and 30 mM, 15 mM and 25 mM, 20 mM and 55 mM, 20 mM and 50 mM, 20 mM and 45 mM, 20 mM and 40 mM, 20 mM and 35 mM, 20 mM and 30 mM, 20 mM and 25 mM NaCl. In some embodiments, the production medium comprises between or between about 15 mM and 40 mM NaCl. In some embodiments, the production medium comprises at or about 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29 mM, 30 mM, 31 mM, 32 mM, 33 mM, 34 mM, 35 mM, 36 mM, 37 mM, 38 mM, 39 mM, 40 mM, 41 mM, 42 mM, 43 mM, 44 mM, 45 mM, 46 mM, 47 mM, 48 mM, 49 mM, 50 mM, 51 mM, 52 mM, 53 mM, 54 mM, 55 mM, 56 mM, 57 mM, 58 mM, 59 mM, or 60 mM NaCl. In some embodiments, the production medium comprises at or about 25 mM NaCl.

[0148] In some embodiments, the production medium comprises between or between about 1 mM and 8 mM potassium chloride (KC1). In some embodiments, the production medium comprises between or between about 1 mM and 7 mM, 1 mM and 6 mM, 1 mM and 5 mM, 1 mM and 4 mM, 1 mM and 3.5 mM, 1.5 mM and 7 mM, 1.5 mM and 6 mM, 1.5 mM and 5 mM, 1.5 mM and 4 mM, 1.5 mM and 3.5 mM, 2 mM and 7 mM, 2 mM and 6 mM, 2 mM and 5 mM, 2 mM and 4 mM, 2 mM and 3.5 mM, 2.5 mM and 7 mM, 2.5 mM and 6 mM, 2.5 mM and 5 mM, 2.5 mM and 4 mM, 2.5 mM and 3.5 mM, 2.75 mM and 3.25 mM, or 3 mM and 3.25 mM KC1. In some embodiments, the production medium comprises between or between about 2.5 and 4 mM KC1. In some embodiments, the production medium comprises at or about 1 mM, 1.1 mM, 1.2 mM, 1.3 mM, 1.4 mM, 1.5 mM, 1.6 mM, 1.7 mM, 1.8 mM, 1.9 mM, 2 mM, 2.1 mM, 2.2 mM, 2.3 mM, 2.4 mM, 2.5 mM, 2.6 mM, 2.7 mM, 2.8 mM, 2.9 mM, 3 mM, 3.1 mM, 3.125 mM, 3.2 mM, 3.3 mM, 3.4 mM, 3.5 mM, 3.6 mM, 3.7 mM, 3.8 mM, 3.9 mM, 4 mM, 4.1 mM, 4.2 mM, 4.3 mM, 4.4 mM, 4.5 mM, 4.6 mM,

4.7 mM, 4.8 mM, 4.9 mM, 5 mM, 5.1 mM, 5.2 mM, 5.3 mM, 5.4 mM, 5.5 mM, 5.6 mM, 5.7 mM, 5.8 mM,

5.9 mM, 6 mM, 6.1 mM, 6.2 mM, 6.3 mM, 6.4 mM, 6.5 mM, 6.6 mM, 6.7 mM, 6.8 mM, 6.9 mM, or 7 mM

KC1. In some embodiments, the production medium comprises at or about 3.125 mM KC1.

[0149] In some embodiments, the production medium comprises between or between about 2 pM and 40 pM iron(II) sulfate. In some embodiments, the production medium comprises between or between about 2 pM and 40 pM, 2 pM and 35 pM, 2 pM and 30 pM, 2 pM and 25 pM, 2 pM and 20 pM, 2 pM and 15 pM, 2 pM and 12 pM, 2 pM and 10 pM, 4 pM and 40 pM, 4 pM and 35 pM, 4 pM and 30 pM, 4 pM and 25 35 pM, 5 pM and 30 pM, 5 pM and 25 pM, 5 pM and 20 pM, 5 pM and 15 pM, 5 pM and 12 pM, 5 pM and 10 pM, 6 pM and 40 pM, 6 pM and 35 pM, 6 pM and 30 pM, 6 pM and 25 pM, 6 pM and 20 pM, 6 pM and 15 pM, 6 pM and 12 pM, 6 pM and 10 pM, 8 pM and 40 pM, 8 pM and 35 pM, 8 pM and 30 pM, 8 pM and 25 pM, 8 pM and 20 pM, 8 pM and 15 pM, 8 pM and 12 pM, or 8 pM and 10 pM iron(II) sulfate. In some embodiments, the production medium comprises between or between about 5 and 15 pM iron(II) sulfate. In some embodiments, the production medium comprises at or about 2 pM, 3 pM, 4 pM, 5 pM, 6 pM, 7 pM, 8 pM, 9 pM, 10 pM, 11 pM, 12 pM, 13 pM, 14 pM, 15 pM, 16 pM, 17 pM, 18 pM, 19 pM, 20 pM, 21 pM, 22 pM, 23 pM, 24 pM, 25 pM, 26 pM, 27 pM, 28 pM, 29 pM, 30 pM, 31 pM, 32 pM, 33 pM, 34 pM, 35 pM, 36 pM, 37 pM, 38 pM, 39 pM, or 40 pM iron(II) sulfate. In some embodiments, the production medium comprises at or about 10 pM iron(II) sulfate.

[0150] In some embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 mM and 60 mM lactose, between or between about 20 mM and 125 mM of a buffering agent, e.g., MOPS pH 7, between or between about 10 and 60 mM sodium chloride, between or between about 1 mM and 8 mM potassium chloride, one or more metals, and between or between about 2 pM and 40 pM iron(II) sulfate. In some embodiments, the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 mM and 60 mM lactose, between or between about 20 mM and 125 mM of a buffering agent, e.g., MOPS pH 7, between or between about 10 and 60 mM sodium chloride, between or between about 1 mM and 8 mM potassium chloride, metals comprising Al, B, Ca, Co, Cu, Fe, Mg, Mn, Mo, Ni, Se, V, W, and Zn, and between or between about 2 pM and 40 pM iron(II) sulfate.

[0151] In some embodiments, the production medium comprises between or between about 100 mM and 300 mM glycerol, between or between about 0.5 mM and 30 mM lactose, between or between about 30 mM and 70 mM of a buffering agent, e.g., MOPS pH 7, between or between about 15 and 40 mM sodium chloride, between or between about 2.5 mM and 4 mM potassium chloride, one or more metals, and between or between about 5 pM and 15 pM iron(II) sulfate. In some embodiments, the production medium comprises between or between about 100 mM and 300 mM glycerol, between or between about 0.5 mM and 30 mM lactose, between or between about 30 mM and 70 mM of a buffering agent, e.g., MOPS pH 7, between or between about 15 and 40 mM sodium chloride, between or between about 2.5 mM and 4 mM potassium chloride, metals comprising Al, B, Ca, Co, Cu, Fe, Mg, Mn, Mo, Ni, Se, V, W, and Zn, and between or between about 5 pM and 15 pM iron(II) sulfate.

[0152] In some embodiments, the production medium comprises between or between about 100 mM and 300 mM glycerol, between or between about 1 mM and 2 mM lactose, between or between about 30 mM and 70 mM of a buffering agent, e.g., MOPS pH 7, between or between about 15 and 40 mM sodium chloride, between or between about 2.5 mM and 4 mM potassium chloride, one or more metals, and between or between about 5 pM and 15 pM iron(II) sulfate. In some embodiments, the production medium comprises between or between about 100 mM and 300 mM glycerol, between or between about 1 mM and 2 mM lactose, between or between about 30 mM and 70 mM of a buffering agent, e.g., MOPS pH 7, between or between about 15 and 40 mM sodium chloride, between or between about 2.5 mM and 4 mM potassium chloride, metals comprising Al, B, Ca, Co, Cu, Fe, Mg, Mn, Mo, Ni, Se, V, W, and Zn, and between or between about 5 pM and 15 pM iron(II) sulfate.

[0153] In some embodiments, the production medium comprises 200 mM glycerol, 1.5 mM lactose, 50 mM MOPS pH 7, 25 mM sodium chloride, 3.125 mM potassium chloride, metals, and 10 pM iron(II) sulfate. In some embodiments, the production medium comprises 200 mM glycerol, 1.5 mM lactose, 50 mM MOPS pH 7, 25 mM sodium chloride, 3.125 mM potassium chloride, metals comprising Al, B, Ca, Co, Cu, Fe, Mg, Mn, Mo, Ni, Se, V, W, and Zn, and 10 pM iron(II) sulfate.

[0154] In some embodiments, the production medium comprises 200 mM glycerol, 25 mM lactose, 50 mM MOPS pH 7, 25 mM sodium chloride, 3.125 mM potassium chloride, metals, and 10 pM iron(II) sulfate. In some embodiments, the production medium comprises 200 mM glycerol, 25 mM lactose, 50 mM MOPS pH 7, 1 g/L polysorbate 80, 25 mM sodium chloride, 3.125 mM potassium chloride, metals, and 10 pM iron(II) sulfate.

[0155] In some embodiments, the production medium comprises one or more metals. In some embodiments, the one or more metals comprises one or more metals selected from the group consisting of aluminum (Al), boron (B), calcium (Ca), cobalt (Co), copper (Cu), iron (Fe), magnesium (Mg), manganese (Mn), molybdenum (Mo), nickel (Ni), selenium (Se), vanadium (V), tungsten (W), and zinc (Zn). In some embodiments, the one or more metals comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 metals selected from the group consisting of Al, B, Ca, Co, Cu, Fe, Mg, Mn, Mo, Ni, Se, V, W, and Zn. In some embodiments, the one or more metals comprises Al, B, Ca, Co, Cu, Fe, Mg, Mn, Mo, Ni, Se, V, W, and Zn. [0156] In some embodiments, the one or more metals comprise aluminum (Al), e.g., aluminum potassium sulfate, at a concentration of between or between about 1 pM and 20 pM. In some embodiments, the one or more metals comprise aluminum (Al), e.g., aluminum potassium sulfate, at a concentration of between or between about 1 pM and 20 pM, 1 pM and 18 pM, 1 pM and 16 pM, 1 pM and 14 pM, 1 pM and 12 pM, 1 pM and 10 pM, 1 pM and 8 pM, 1 pM and 6 pM, 1 pM and 4 pM, 1 pM and 2 pM, 2 pM and 20 pM, 2 pM and 18 pM, 2 pM and 16 pM, 2 pM and 14 pM, 2 pM and 12 pM, 2 pM and 10 pM, 2 pM and 8 pM, 2 pM and 12 pM, 4 pM and 10 pM, 4 pM and 8 pM, 4 pM and 6 pM, 6 pM and 20 pM, 6 pM and 18 pM, 6 pM and 16 pM, 6 pM and 14 pM, 6 pM and 12 pM, 6 pM and 10 pM, 6 pM and 8 pM, 8 pM and 20 pM, 8 pM and 18 pM, 8 pM and 16 pM, 8 pM and 14 pM, 8 pM and 12 pM, 8 pM and 10 pM, 10 pM and 20 pM, 10 pM and 18 pM, 10 pM and 16 pM, 10 pM and 14 pM, 10 pM and 12 pM, 12 pM and 20 pM, 12 pM and 18 M, 12 uM and 16 uM, or 12 uM and 14 uM. In some embodiments, the one or more metals comprise aluminum (Al), e.g., aluminum potassium sulfate, at a concentration at or about 1 pM, 2 pM, 3 pM, 4 pM, 5 pM, 6 pM, 7 pM, 8 pM, 9 pM, 10 pM, 11 pM, 12 pM, 13 pM, 14 pM, 15 pM, 16 pM, 17 pM, 18 pM, 19 pM, or 20 pM. In some embodiments, the aluminum metal is aluminum potassium sulfate.

[0157] In some embodiments, the one or more metals comprise boron (B), e.g., boric acid, at a concentration of between or between about 1 and 20 pM. In some embodiments, the one or more metals comprise boron (B), e.g., boric acid, at a concentration of between or between about 1 pM and 20 pM, 1 pM and 18 pM, 1 pM and 16 pM, 1 pM and 14 pM, 1 pM and 12 pM, 1 pM and 10 pM, 1 pM and 8 pM, 1 pM and 6 pM, 1 pM and 4 pM, 1 pM and 2 pM, 2 pM and 20 pM, 2 pM and 18 pM, 2 pM and 16 pM, 2 pM and 14 pM, 2 pM and 12 pM, 2 pM and 10 pM, 2 pM and 8 pM, 2 pM and 6 pM, 2 pM and 4 pM, 4 pM and 20 pM, 4 pM and 18 pM, 4 pM and 16 pM, 4 pM and 14 pM, 4 pM and 12 pM, 4 pM and 10 pM, 4 pM and 8 pM, 4 pM and 6 pM, 6 pM and 20 pM, 6 pM and 18 pM, 6 pM and 16 pM, 6 pM and 14 pM, 6 pM and 12 pM, 6 pM and 10 pM, 6 pM and 8 pM, 8 pM and 20 pM, 8 pM and 18 pM, 8 pM and 16 pM, 8 pM and 14 pM, 8 pM and 12 pM, 8 pM and 10 pM, 10 pM and 20 pM, 10 pM and 18 pM, 10 pM and 16 pM, 10 pM and 14 pM, 10 pM and 12 pM, 12 pM and 20 pM, 12 pM and 18 pM, 12 pM and 16 pM, or 12 pM and 14 pM. In some embodiments, the one or more metals comprise boron (B), e.g., boric acid, at a concentration at or about 1 pM, 2 pM, 3 pM, 4 pM, 5 pM, 6 pM, 7 pM, 8 pM, 9 pM, 10 pM, 11 pM, 12 pM, 13 pM, 14 pM, 15 pM, 16 pM, 17 pM, 18 pM, 19 pM, or 20 pM. In some embodiments, the boron metal is boric acid.

[0158] In some embodiments, the one or more metals comprise calcium (Ca), e.g., calcium chloride, at a concentration of between or between about 50 pM and 600 pM. In some embodiments, the one or more metals comprise calcium (Ca), e.g., calcium chloride, at a concentration of between or between about 50 pM and 600 pM, 50 pM and 550 pM, 50 pM and 500 pM, 50 pM and 450 pM, 50 pM and 400 pM, 50 pM and 350 pM, 50 pM and 300 pM, 50 pM and 250 pM, 50 pM and 200 pM, 50 pM and 150 pM, 50 pM and 100 pM, 100 pM and 600 pM, 100 pM and 550 pM, 100 pM and 500 pM, 100 pM and 450 pM, 100 pM and 400 pM, 100 pM and 350 pM, 100 pM and 300 pM, 100 pM and 250 pM, 100 pM and 200 pM, 100 pM and 150 pM, 150 pM and 600 pM, 150 pM and 550 pM, 150 pM and 500 pM, 150 pM and 450 pM, 150 pM and 400 pM, 150 pM and 350 pM, 150 pM and 300 pM, 150 pM and 250 pM, 150 pM and 200 pM, 200 pM and 600 pM, 200 pM and 550 pM, 200 pM and 500 pM, 200 pM and 450 pM, 200 pM and 400 pM, 200 pM and 350 pM, 200 pM and 300 pM, 200 pM and 250 pM, 250 pM and 600 pM, 250 pM and 550 pM, 250 pM and 500 pM, 250 pM and 450 pM, 250 pM and 400 pM, 250 pM and 350 pM, 250 pM and 300 pM, 300 pM and 600 pM, 300 pM and 550 pM, 300 pM and 500 pM, 300 pM and 450 pM, 300 pM and 400 pM, 300 pM and 350 pM, 400 pM and 600 pM, 400 pM and 550 pM, 400 pM and 500 pM, or 400 uM and 450 uM. In some embodiments, the one or more metals comprise calcium (Ca), e.g., calcium chloride, at a concentration of at or about 50 pM, 75 pM, 100 pM, 125 pM, 150 pM, 175 pM, 200 pM, 225 pM, 250 pM, 275 pM, 300 pM, 325 pM, 350 pM, 375 pM, 400 pM, 425 pM, 450 pM, 475 pM, 500 pM, 525 pM, 550 pM, 575 pM, or 600 pM. In some embodiments, the calcium metal is calcium chloride.

[0159] In some embodiments, the one or more metals comprise cobalt (Co), e.g., cobalt chloride, at a concentration of between or between about 0.02 pM to 50 pM. In some embodiments, the one or more metals comprise cobalt (Co), e.g., cobalt chloride, at a concentration of between or between about 0.02 pM and 50 pM, 0.1 pM and 50 pM, 0.25 pM and 50 pM, 0.5 pM and 50 pM, 1 pM and 50 pM, 2 pM and 50 pM, 5 pM and 50 pM, 10 pM and 50 pM, 15 pM and 50 pM, 20 pM and 50 pM, 25 pM and 50 pM, 0.02 pM and 40 pM, 0.1 pM and 40 pM, 0.25 pM and 40 pM, 0.5 pM and 40 pM, 1 pM and 40 pM, 2 pM and 40 pM, 5 pM and 40 pM, 10 pM and 40 pM, 15 pM and 40 pM, 20 pM and 40 pM, 25 pM and 40 pM, 0.02 pM and 30 pM, 0.1 pM and 30 pM, 0.25 pM and 30 pM, 0.5 pM and 30 pM, 1 pM and 30 pM, 2 pM and 30 pM, 5 pM and 30 pM, 10 pM and 30 pM, 15 pM and 30 pM, 20 pM and 30 pM, 25 pM and 30 pM, 0.02 pM and 25 pM, 0.1 pM and 25 pM, 0.25 pM and 25 pM, 0.5 pM and 25 pM, 1 pM and 25 pM, 2 pM and 25 pM, 5 pM and 25 pM, 10 pM and 25 pM, 15 pM and 25 pM, 20 pM and 25 pM, 0.02 pM and 20 pM, 0.1 pM and 20 pM, 0.25 pM and 20 pM, 0.5 pM and 20 pM, 1 pM and 20 pM, 2 pM and 20 pM, 5 pM and 20 pM, 10 pM and 20 pM, 15 pM and 20 pM, 0.02 pM and 15 pM, 0.1 pM and 15 pM, 0.25 pM and 15 pM, 0.5 pM and 15 pM, 1 pM and 15 pM, 2 pM and 15 pM, 5 pM and 15 pM, 10 pM and 15 pM, 0.02 pM and 10 pM, 0.1 pM and 10 pM, 0.25 pM and 10 pM, 0.5 pM and 10 pM, 1 pM and 10 pM, 2 pM and 10 pM, 5 pM and 10 pM, 0.02 pM and 5 pM, 0.1 pM and 5 pM, 0.25 pM and 5 pM, 0.5 pM and 5 pM, 1 pM and 5 pM, or 2 pM and 5 pM.

[0160] In some embodiments, the one or more metals comprise cobalt (Co), e.g., cobalt chloride, at a concentration of between or between about 0.02 pM to 1 pM. In some embodiments, the one or more metals comprise cobalt (Co), e.g., cobalt chloride, at a concentration of between or between about 0.02 pM and 1 pM, 0.02 pM and 0.9 pM, 0.02 pM and 0.8 pM, 0.02 pM and 0.7 pM, 0.02 pM and 0.6 pM, 0.02 pM and 0.5 pM, 0.02 pM and 0.4 pM, 0.02 pM and 0.3 pM, 0.02 pM and 0.2 pM, 0.02 pM and 0.1 pM, 0.04 pM and 1 pM, 0.04 pM and 0.9 pM, 0.04 pM and 0.8 pM, 0.04 pM and 0.7 pM, 0.04 pM and 0.6 pM, 0.04 pM and 0.5 pM, 0.04 pM and 0.4 pM, 0.04 pM and 0.3 pM, 0.04 pM and 0.2 pM, 0.04 pM and 0.1 pM, 0.06 pM and 1 pM, 0.06 pM and 0.9 pM, 0.06 pM and 0.8 pM, 0.06 pM and 0.7 pM, 0.06 pM and 0.6 pM, 0.06 pM and 0.5 pM, 0.06 pM and 0.4 pM, 0.06 pM and 0.3 pM, 0.06 pM and 0.2 pM, 0.06 pM and 0.1 pM, 0.08 pM and 1 pM, 0.08 pM and 0.9 pM, 0.08 pM and 0.8 pM, 0.08 pM and 0.7 pM, 0.08 pM and 0.6 pM, 0.08 pM and 0.5 pM, 0.08 pM and 0.4 pM, 0.08 pM and 0.3 pM, 0.08 pM and 0.2 pM, 0.08 pM and 0.1 pM, 0.1 pM and 1 pM, 0.1 pM and 0.9 pM, 0.1 pM and 0.8 pM, 0.1 pM and 0.7 pM, 0.1 pM and 0.6 pM, 0.1 pM and 0.5 pM, 0.1 pM and 0.4 pM, 0.1 pM and 0.3 pM, 0.1 pM and 0.2 pM, 0.2 pM and 1 pM, 0.2 pM and 0.9 pM, 0.2 pM and 0.8 pM, 0.2 pM and 0.7 pM, 0.2 pM and 0.6 pM, 0.2 pM and 0.5 pM, 0.2 pM and 0.4 pM, 0.2 pM and 0.3 pM, 0.4 pM and 1 pM, 0.4 pM and 0.9 pM, 0.4 pM and 0.8 pM, 0.4 pM and 0.7 pM, 0.4 pM and 0.6 pM, 0.4 pM and 0.5 pM, 2.5 pM and 5 pM, 5 pM and 10 pM, 10 pM and 30 pM, or 30 uM and 50 uM. In some embodiments, the one or more metals comprise cobalt (Co), e.g., cobalt chloride, at a concentration at or about 0.02 pM, 0.04 pM, 0.06 pM, 0.08 pM, 0.1 pM, 0.15 pM, 0.2 pM, 0.25 pM, 0.3 pM, 0.35 pM, 0.4 pM, 0.45 pM, 0.5 pM, 0.55 pM, 0.6 pM, 0.65 pM, 0.7 pM, 0.75 pM, 0.8 pM, 0.85 pM, 0.9 pM, 0.95 pM, 1 pM, 1.5 pM, 2 pM, 2.5 pM, 3 pM, 3.5 pM, 3.6 pM, 4 pM, 4.5 pM, 5 pM, 7.5 pM, 10 pM, 15 pM, 20 pM, 25 pM, 30 pM, 40 pM, or 50 pM. In some embodiments, the cobalt metal is cobalt chloride.

[0161] In some embodiments, the one or more metals comprise copper (Cu), e.g., copper sulfate, at a concentration of between or between about 0.05 pM and 2 pM. In some embodiments, the one or more metals comprise copper (Cu), e.g., copper sulfate, at a concentration of between or between about 0.05 pM and 2 pM, 0.05 pM and 1.5 pM, 0.05 pM and 1 pM, 0.05 pM and 0.5 pM, 0.05 pM and 0.1 pM, 0.1 pM and 2 pM, 0.1 pM and 1.5 pM, 0.1 pM and 1 pM, 0.1 pM and 0.5 pM, 0.2 pM and 2 pM, 0.2 pM and 1.5 pM, 0.2 pM and 1 pM, 0.2 pM and 0.5 pM, 0.5 pM and 2 pM, 0.5 pM and 1.5 pM, 0.5 pM and 1 pM, 1 pM and 2 pM, or 1 pM and 1.5 pM. In some embodiments, the one or more metals comprise copper (Cu), e.g., copper sulfate, at a concentration at or about 0.05 pM, 0.1 pM, 0.15 pM, 0.2 pM, 0.25 pM, 0.3 pM, 0.35 pM, 0.4 pM, 0.45 pM, 0.5 pM, 0.55 pM, 0.6 pM, 0.65 pM, 0.7 pM, 0.75 pM, 0.8 pM, 0.85 pM, 0.9 pM, 0.95 pM, 1 pM, 1.05 pM, 1.1 pM, 1.15 pM, 1.2 pM, 1.25 pM, 1.3 pM, 1.35 pM, 1.4 pM, 1.45 pM, 1.5 pM, 1.55 pM, 1.6 pM, 1.65 pM, 1.7 pM, 1.75 pM, 1.8 pM, 1.85 pM, 1.9 pM, 1.95 pM, or 2 pM. In some embodiments, the copper metal is copper sulfate.

[0162] In some embodiments, the one or more metals comprise iron (Fe), e.g., iron sulfate, at a concentration of between or between about 1 pM and 30 pM. In some embodiments, the one or more metals comprise iron (Fe), e.g., iron sulfate, at a concentration of between or between about 1 pM and 30 pM, 1 pM 7.5 pM, 1 pM and 5 pM, 1 pM and 2.5 pM, 2 pM and 30 pM, 2 pM and 25 pM, 2 pM and 20 pM, 2 pM and 15 pM, 2 pM and 10 pM, 2 pM and 7.5 pM, 2 pM and 5 pM, 2 pM and 2.5 pM, 4 pM and 30 pM, 4 pM and 25 pM, 4 pM and 20 pM, 4 pM and 15 pM, 4 pM and 10 pM, 4 pM and 7.5 pM, 4 pM and 5 pM, 6 pM and 30 pM, 6 pM and 25 pM, 6 pM and 20 pM, 6 pM and 15 pM, 6 pM and 10 pM, 6 pM and 7.5 pM, 8 pM and 30 pM, 8 pM and 25 pM, 8 pM and 20 pM, 8 pM and 15 pM, 8 pM and 10 pM, 10 pM and 30 pM, 10 pM and 25 pM, 10 pM and 20 pM, 10 pM and 15 pM, 12 pM and 30 pM, 12 pM and 25 pM, 12 pM and 20 pM, 12 pM and 15 pM, 15 pM and 30 pM, 15 pM and 25 pM, 15 pM and 20 pM, 20 pM and 30 pM, or 20 pM and 25 pM. In some embodiments, the one or more metals comprises iron (Fe), e.g., iron sulfate, at a concentration of at or about 1 pM, 2 pM, 3 pM, 4 pM, 5 pM, 6 pM, 7 pM, 8 pM, 9 pM, 10 pM, 11 pM, 12 pM, 13 pM, 14 pM, 15 pM, 16 pM, 17 pM, 18 pM, 19 pM, 20 pM, 21 pM, 22 pM, 23 pM, 24 pM, 25 pM, 26 pM, 27 pM, 28 uM, 29 uM, or 30 uM. In some embodiments, the iron metal is iron sulfate.

[0163] In some embodiments, the one or more metals comprises magnesium (Mg), e.g., magnesium sulfate, at a concentration of between or between about 250 pM and 2000 pM. In some embodiments, the one or more metals comprises magnesium (Mg), e.g., magnesium sulfate, at a concentration of between or between about 250 pM and 2000 pM, 250 pM and 1800 pM, 250 pM and 1600 pM, 250 pM and 1400 pM, 250 pM and 1200 pM, 250 pM and 1000 pM, 250 pM and 800 pM, 250 pM and 700 pM, 250 pM and 600 pM, 500 pM and 2000 pM, 500 pM and 1800 pM, 500 pM and 1600 pM, 500 pM and 1400 pM, 500 pM and 1200 pM, 500 pM and 1000 pM, 500 pM and 800 pM, 500 pM and 700 pM, 750 pM and 2000 pM, 750 pM and 1800 pM, 750 pM and 1600 pM, 750 pM and 1400 pM, 750 pM and 1200 pM, 750 pM and 1000 pM, 1000 pM and 2000 pM, 1000 pM and 1800 pM, 1000 pM and 1600 pM, 1000 pM and 1400 pM, or 1000 pM and 1200 pM. In some embodiments, the one or more metals comprises magnesium (Mg), e.g., magnesium sulfate, at a concentration of at or about 250 pM, 275 pM, 300 pM, 325 pM, 350 pM, 375 pM, 400 pM, 425 pM, 450 pM, 475 pM, 500 pM, 525 pM, 550 pM, 575 pM, 600 pM, 625 pM, 650 pM, 675 pM, 700 pM, 725 pM, 750 pM, 775 pM, 800 pM, 825 pM, 850 pM, 875 pM, 900 pM, 925 pM, 950 pM, 975 pM, 1000 pM, 1025 pM, 1050 pM, 1075 pM, 1100 pM, 1125 pM, 1150 pM, 1175 pM, 1200 pM, 1225 pM, 1250 pM, 1275 pM, 1300 pM, 1325 pM, 1350 pM, 1375 pM, 1400 pM, 1425 pM, 1450 pM, 1475 pM, 1500 pM, 1525 pM, 1550 pM, 1575 pM, 1600 pM, 1625 pM, 1650 pM, 1675 pM, 1700 pM, 1725 pM, 1750 pM, 1775 pM, 1800 pM, 1825 pM, 1850 pM, 1875 pM, 1900 pM, 1925 pM, 1950 pM, 1975 pM, or 2000 pM. In some embodiments, the magnesium metal is magnesium sulfate.

[0164] In some embodiments, the one or more metals comprise manganese (Mn), e.g., manganese sulfate, at a concentration of between or between about 0.02 pM and 5 pM. In some embodiments, the one or more metals comprise manganese (Mn), e.g., manganese sulfate, at a concentration of between or between about 0.02 pM and 5 pM, 0.02 pM and 4 pM, 0.02 pM and 3 pM, 0.02 pM and 2 pM, 0.02 pM and 1 pM, 0.02 pM and 0.75 pM, 0.02 pM and 0.5 pM, 0.02 pM and 0.1 pM, 0.02 pM and 0.05 pM, 0.05 pM and 5 pM, 0.05 pM and 4 pM, 0.05 pM and 3 pM, 0.05 pM and 2 pM, 0.05 pM and 1 pM, 0.05 pM and 0.75 pM, 0.05 pM and 0.5 pM, 0.05 pM and 0.1 pM, 0.1 pM and 5 pM, 0.1 pM and 4 pM, 0.1 pM and 3 pM, 0.1 pM and 2 pM, 0.1 pM and 1 pM, 0.1 pM and 0.75 pM, 0.1 pM and 0.5 pM, 0.3 pM and 5 pM, 0.3 pM and 4 pM, 0.3 pM and 3 pM, 0.3 pM and 2 pM, 0.3 pM and 1 pM, 0.3 pM and 0.75 pM, 0.3 pM and 0.5 pM, 0.5 pM and 5 pM, 0.5 pM and 4 pM, 0.5 pM and 3 pM, 0.5 pM and 2 pM, 0.5 pM and 1 pM, 0.5 pM and 0.75 pM, 1 pM and 5 pM, 1 pM and 4 pM, 1 pM and 3 pM, or 1 pM and 2 pM. In some embodiments, the one or more metals comprises manganese (Mn), e.g., manganese sulfate, at a concentration of at or about 0.02 uM, 0.04 pM, 0.06 pM, 0.08 pM, 0.1 pM, 0.15 pM, 0.2 pM, 0.25 pM, 0.3 pM, 0.35 pM, 0.4 pM, 0.45 pM, 0.5 pM, 0.55 pM, 0.6 pM, 0.65 pM, 0.7 pM, 0.75 pM, 0.8 pM, 0.85 pM, 0.9 pM, 0.95 pM, 1 pM, 1.25 pM, 1.5 pM, 1.75 pM, 2 pM, 2.25 pM, 2.5 pM, 2.75 pM, 3 pM, 3.25 pM, 3.5 pM, 3.75 pM, 4 pM, 4.25 pM, 4.5 uM, 4.75 uM, or 5 uM. In some embodiments, the manganese metal is manganese sulfate.

[0165] In some embodiments, the one or more metals comprise molybdenum (Mo), e.g., sodium molybdate, at a concentration of between or between about 0.02 pM and 2 pM. In some embodiments, the one or more metals comprise molybdenum (Mo), e.g., sodium molybdate, at a concentration of between or between about 0.02 pM and 2 pM, 0.02 pM and 1.5 pM, 0.02 pM and 1 pM, 0.02 pM and 0.5 pM, 0.02 pM and 0.1 pM, 0.02 pM and 0.05 pM, 0.04 pM and 2 pM, 0.04 pM and 1.5 pM, 0.04 pM and 1 pM, 0.04 pM and 0.5 pM, 0.04 pM and 0.1 pM, 0.04 pM and 0.05 pM, 0.1 pM and 2 pM, 0.1 pM and 1.5 pM, 0.1 pM and 1 pM, 0.1 pM and 0.5 pM, 0.2 pM and 2 pM, 0.2 pM and 1.5 pM, 0.2 pM and 1 pM, 0.2 pM and 0.5 pM, 0.5 pM and 2 pM, 0.5 pM and 1.5 pM, 0.5 pM and 1 pM, 1 pM and 2 pM, or 1 pM and 1.5 pM. In some embodiments, the one or more metals comprise molybdenum (Mo), e.g., sodium molybdate, at a concentration at or about 0.02 pM, 0.04 pM, 0.06 pM, 0.08 pM, 0.1 pM, 0.12 pM, 0.14 pM, 0.15 pM, 0.175 pM, 0.2 pM, 0.25 pM, 0.3 pM, 0.35 pM, 0.4 pM, 0.45 pM, 0.5 pM, 0.55 pM, 0.6 pM, 0.65 pM, 0.7 pM, 0.75 pM, 0.8 pM, 0.85 pM, 0.9 pM, 0.95 pM, 1 pM, 1.05 pM, 1.1 pM, 1.15 pM, 1.2 pM, 1.25 pM, 1.3 pM, 1.35 pM, 1.4 pM, 1.45 pM, 1.5 pM, 1.55 pM, 1.6 pM, 1.65 pM, 1.7 pM, 1.75 pM, 1.8 pM, 1.85 pM, 1.9 pM, 1.95 pM, or 2 pM. In some embodiments, the molybdenum metal is sodium molybdate.

[0166] In some embodiments, the one or more metals comprise nickel (Ni), e.g., nickel chloride, at a concentration of between or between about 0.02 pM and 2 pM. In some embodiments, the one or more metals comprise nickel (Ni), e.g., nickel chloride, at a concentration of between or between about 0.02 pM and 2 pM, 0.02 pM and 1.5 pM, 0.02 pM and 1 pM, 0.02 pM and 0.5 pM, 0.02 pM and 0.1 pM, 0.02 pM and 0.05 pM, 0.04 pM and 2 pM, 0.04 pM and 1.5 pM, 0.04 pM and 1 pM, 0.04 pM and 0.5 pM, 0.04 pM and 0.1 pM, 0.04 pM and 0.05 pM, 0.1 pM and 2 pM, 0.1 pM and 1.5 pM, 0.1 pM and 1 pM, 0.1 pM and 0.5 pM, 0.2 pM and 2 pM, 0.2 pM and 1.5 pM, 0.2 pM and 1 pM, 0.2 pM and 0.5 pM, 0.5 pM and 2 pM, 0.5 pM and 1.5 pM, 0.5 pM and 1 pM, 1 pM and 2 pM, or 1 pM and 1.5 pM. In some embodiments, the one or more metals comprise nickel (Ni), e.g., nickel chloride, at a concentration at or about 0.02 uM, 0.04 uM, 0.06 pM, 0.08 pM, 0.1 pM, 0.12 pM, 0.14 pM, 0.15 pM, 0.175 pM, 0.2 pM, 0.25 pM, 0.3 pM, 0.35 pM, 0.4 pM, 0.45 pM, 0.5 pM, 0.55 pM, 0.6 pM, 0.65 pM, 0.7 pM, 0.75 pM, 0.8 pM, 0.85 pM, 0.9 pM, 0.95 pM, 1 pM, 1.05 pM, 1.1 pM, 1.15 pM, 1.2 pM, 1.25 pM, 1.3 pM, 1.35 pM, 1.4 pM, 1.45 pM, 1.5 pM, 1.55 pM, 1.6 pM, 1.65 pM, 1.7 pM, 1.75 pM, 1.8 pM, 1.85 pM, 1.9 pM, 1.95 pM, or 2 pM. In some embodiments, the nickel metal is nickel chloride.

[0167] In some embodiments, the one or more metals comprise selenium (Se), e.g., sodium selenite, at a concentration of between or between about 0.02 pM and 2 pM. In some embodiments, the one or more metals comprise selenium (Se), e.g., sodium selenite, at a concentration of between or between about 0.02 pM and 2 pM, 0.02 pM and 1.5 pM, 0.02 pM and 1 pM, 0.02 pM and 0.5 pM, 0.02 pM and 0.1 pM, 0.02 pM and 0.05 pM, 0.04 pM and 2 pM, 0.04 pM and 1.5 pM, 0.04 pM and 1 pM, 0.04 pM and 0.5 pM, 0.04 pM and 0.1 pM, 0.04 pM and 0.05 pM, 0.1 pM and 2 pM, 0.1 pM and 1.5 pM, 0.1 pM and 1 pM, 0.1 pM and 0.5 pM, 0.2 pM and 2 pM, 0.2 pM and 1.5 pM, 0.2 pM and 1 pM, 0.2 pM and 0.5 pM, 0.5 pM and 2 pM, 0.5 pM and 1.5 pM, 0.5 pM and 1 pM, 1 pM and 2 pM, or 1 pM and 1.5 pM. In some embodiments, the one or more metals comprise selenium (Se), e.g., sodium selenite, at a concentration at or about 0.02 pM, 0.04 pM, 0.06 pM, 0.08 pM, 0.1 pM, 0.12 pM, 0.14 pM, 0.15 pM, 0.175 pM, 0.2 pM, 0.25 pM, 0.3 pM, 0.35 pM, 0.4 pM, 0.45 pM, 0.5 pM, 0.55 pM, 0.6 pM, 0.65 pM, 0.7 pM, 0.75 pM, 0.8 pM, 0.85 pM, 0.9 pM, 0.95 pM, 1 pM, 1.05 pM, 1.1 pM, 1.15 pM, 1.2 pM, 1.25 pM, 1.3 pM, 1.35 pM, 1.4 pM, 1.45 pM, 1.5 pM, 1.55 pM, 1.6 pM, 1.65 pM, 1.7 pM, 1.75 pM, 1.8 pM, 1.85 pM, 1.9 pM, 1.95 pM, or 2 pM. In some embodiments, the selenium metal is sodium selenite.

[0168] In some embodiments, the one or more metals comprise vanadium (V), e.g., vanadium chloride, at a concentration of between or between about 0.5 pM and 10 pM. In some embodiments, the one or more metals comprise vanadium (V), e.g., vanadium chloride, at a concentration of between or between about 0.5 pM and 10 pM, 0.5 pM and 9 pM, 0.5 pM and 8 pM, 0.5 pM and 7 pM, 0.5 pM and 6 pM, 0.5 pM and 5 pM, 0.5 pM and 4 pM, 0.5 pM and 3 pM, 0.5 pM and 2 pM, 0.5 pM and 1 pM, 0.75 pM and 10 pM, 0.75 pM and 9 pM, 0.75 pM and 8 pM, 0.75 pM and 7 pM, 0.75 pM and 6 pM, 0.75 pM and 5 pM, 0.75 pM and 4 pM, 0.75 pM and 3 pM, 0.75 pM and 2 pM, 0.75 pM and 1 pM, 1 pM and 10 pM, 1 pM and 9 pM, 1 pM and 8 pM, 1 pM and 7 pM, 1 pM and 6 pM, 1 pM and 5 pM, 1 pM and 4 pM, 1 pM and 3 pM, 1 pM and 2 2 pM and 4 pM, 2 pM and 3 pM, 4 pM and 10 pM, 4 pM and 9 pM, 4 pM and 8 pM, 4 pM and 7 pM, 4 pM and 6 pM, 4 pM and 5 pM, 6 pM and 10 pM, 6 pM and 9 pM, 6 pM and 8 pM, or 6 pM and 7 pM. In some embodiments, the one or more metals comprise vanadium (V), e.g., vanadium chloride, at a concentration at or about 0.5 pM, 0.6 pM, 0.7 pM, 0.8 pM, 0.9 pM, 1 pM, 1.1 pM, 1.2 pM, 1.3 pM, 1.4 pM, 1.5 pM, 1.6 pM, 1.7 pM, 1.8 pM, 1.9 pM, 2 pM, 2.1 pM, 2.2 pM, 2.3 pM, 2.4 pM, 2.5 pM, 2.6 pM, 2.7 pM, 2.8 pM, 2.9 pM, 3 pM, 3.1 pM, 3.2 pM, 3.3 pM, 3.4 pM, 3.5 pM, 3.6 pM, 3.8 pM, 4 pM, 4.2 pM, 4.4 pM, 4.6 pM, 4.8 pM, 5 pM, 5.2 pM, 5.4 pM, 5.6 pM, 5.8 pM, 6 pM, 6.2 pM, 6.4 pM, 6.6 pM, 6.8 pM, 7 pM, 7.2 pM, 7.4 pM, 7.6 pM, 7.8 pM, 8 pM, 8.2 pM, 8.4 pM, 8.6 pM, 8.8 pM, 9 pM, 9.2 pM, 9.4 pM, 9.6 pM, 9.8 pM, or 10 pM. In some embodiments, the vanadium metal is vanadium chloride.

[0169] In some embodiments, the one or more metals comprise tungsten (W), e.g., sodium tungstate, at a concentration of between or between about 0.001 pM and 0.5 pM. In some embodiments, the one or more metals comprise tungsten (W), e.g., sodium tungstate, at a concentration of between or between about 0.001 pM and 0.5 pM, 0.001 pM and 0.4 pM, 0.001 pM and 0.3 pM, 0.001 pM and 0.2 pM, 0.001 pM and 0.1 pM, 0.001 pM and 0.05 pM, 0.001 pM and 0.01 pM, 0.001 pM and 0.005 pM, 0.002 pM and 0.5 pM, 0.002 pM and 0.4 pM, 0.002 pM and 0.3 pM, 0.002 pM and 0.2 pM, 0.002 pM and 0.1 pM, 0.002 pM and 0.05 pM, 0.002 pM and 0.01 pM, 0.002 pM and 0.005 pM, 0.005 pM and 0.5 pM, 0.005 pM and 0.4 pM, 0.005 pM and 0.3 pM, 0.005 pM and 0.2 pM, 0.005 pM and 0.1 pM, 0.005 pM and 0.05 pM, 0.005 pM and 0.01 pM, 0.01 pM and 0.5 pM, 0.01 pM and 0.4 pM, 0.01 pM and 0.3 pM, 0.01 pM and 0.2 pM, 0.01 pM and 0.1 pM, 0.01 pM and 0.05 pM, 0.02 pM and 0.5 pM, 0.02 pM and 0.4 pM, 0.02 pM and 0.3 pM, 0.02 pM and 0.2 pM, 0.02 pM and 0.1 pM, 0.02 pM and 0.05 pM, 0.05 pM and 0.5 pM, 0.05 pM and 0.4 pM, 0.05 pM and 0.3 pM, 0.05 and 0.2 pM, 0.05 pM and 0.1 pM, 0.1 pM and 0.5 pM, 0.1 pM and 0.4 pM, 0.1 pM and 0.3 pM, 0.1 pM and 0.2 pM, 0.25 pM and 0.5 pM, 0.25 pM and 0.4 pM, or 0.25 pM and 0.3 pM. In some embodiments, the one or more metals comprise tungsten (W), e.g., sodium tungstate, at a concentration at or about 0.001 pM, 0.002 pM, 0.003 pM, 0.004 pM, 0.005 pM, 0.006 pM, 0.007 pM, 0.008 pM, 0.009 pM, 0.01 pM, 0.015 pM, 0.02 pM, 0.025 pM, 0.03 pM, 0.035 pM, 0.04 pM, 0.045 pM, 0.05 pM, 0.055 pM, 0.06 pM, 0.065 pM, 0.07 pM, 0.075 pM, 0.08 pM, 0.085 pM, 0.09 pM, 0.1 pM, 0.11 pM, 0.12 pM, 0.13 pM, 0.14 pM, 0.15 pM, 0.16 pM, 0.17 pM, 0.18 pM, 0.19 pM, 0.2 pM, 0.21 pM, 0.22 pM, 0.23 pM,

0.24 pM, 0.25 pM, 0.26 pM, 0.27 pM, 0.28 pM, 0.29 pM, 0.3 pM, 0.31 pM, 0.32 pM, 0.33 pM, 0.34 pM,

0.35 pM, 0.36 pM, 0.37 pM, 0.38 pM, 0.39 pM, 0.4 pM, 0.41 pM, 0.42 pM, 0.43 pM, 0.44 pM, 0.45 pM,

0.46 pM, 0.47 pM, 0.48 pM, 0.49 pM, or 0.5 pM. In some embodiments, the tungsten metal is sodium tungstate. [0170] In some embodiments, the one or more metals comprise zinc (Zn), e.g., zinc chloride, at a concentration of between or between about 5 uM and 45 uM. In some embodiments, the one or more metals comprise zinc (Zn), e.g., zinc chloride, at a concentration of between or between about 5 pM and 45 pM, 5 pM and 40 pM, 5 pM and 35 pM, 5 pM and 30 pM, 5 pM and 25 pM, 5 pM and 20 pM, 5 pM and 15 pM, 5 pM and 10 pM, 7.5 pM and 45 pM, 7.5 pM and 40 pM, 7.5 pM and 35 pM, 7.5 pM and 30 pM, 7.5 pM and 25 pM, 7.5 pM and 20 pM, 7.5 pM and 15 pM, 7.5 pM and 10 pM, 10 pM and 45 pM, 10 pM and 40 pM, 10 pM and 35 pM, 10 pM and 30 pM, 10 pM and 25 pM, 10 pM and 20 pM, 10 pM and 15 pM, 12.5 pM and 45 pM, 12.5 pM and 40 pM, 12.5 pM and 35 pM, 12.5 pM and 30 pM, 12.5 pM and 25 pM, 12.5 pM and 20 pM, 12.5 pM and 15 pM, 15 pM and 45 pM, 15 pM and 40 pM, 15 pM and 35 pM, 15 pM and 30 pM, 15 pM and 25 pM, 15 pM and 20 pM, 17.5 pM and 45 pM, 17.5 pM and 40 pM, 17.5 pM and 35 pM, 17.5 pM and 30 pM, 17.5 pM and 25 pM, 17.5 pM and 20 pM, 20 pM and 45 pM, 20 pM and 40 pM, 20 pM and 35 pM, 20 pM and 30 pM, 20 pM and 25 pM, 25 pM and 45 pM, 25 pM and 40 pM, 25 pM and 35 pM, 25 pM and 30 pM, 30 pM and 45 pM, 30 pM and 40 pM, or 30 pM and 35 pM. In some embodiments, the one or more metals comprise zinc (Zn), e.g., zinc chloride, at a concentration at or about 5 pM, 5.5 pM, 6 pM, 6.5 pM, 7 pM, 7.5 pM, 8 pM, 8.5 pM, 9 pM, 9.5 pM, 10 pM, 10.5 pM, 11 pM, 11.5 pM, 12 pM, 12.5 pM, 13 pM, 13.5 pM, 14 pM, 14.5 pM, 15 pM, 15.5 pM, 16 pM, 16.5 pM, 17 pM, 17.5 pM, 18 pM, 18.5 pM, 19 pM, 19.5 pM, 20 pM, 21 pM, 22 pM, 23 pM, 24 pM, 25 pM, 26 pM, 27 pM, 28 pM, 29 pM, 30 pM, 31 pM, 32 pM, 33 pM, 34 pM, 35 pM, 36 pM, 37 pM, 38 pM, 39 pM, 40 pM, 41 pM, 42 pM, 43 pM, 44 pM, or 45 pM.

[0171] In some embodiments, the metals comprise Mg, Mn, Ca, Fe, Zn, Co, Cu, Al, B, Mo, Ni, W, V, and Se. In some embodiments, the metals comprise between or between about 250 pM and 2000 pM Mg, between or between about 0.02 pM and 5 pM Mn, between or between about 50 pM and 600 pM Ca, between or between about 1 pM and 30 pM Fe, between or between about 5 pM and 45 pM Zn, between or between about 0.02 pM and 1 pM Co, between or between about 0.05 pM and 2 pM Cu, between or between about 1 pM and 20 pM Al, between or between about 1 pM and 20 pM B, between or between about 0.02 pM and 2 pM Mo, between or between about 0.02 pM and 2 pM Ni, between or between about 0.001 pM and 0.5 pM W, between or between about 0.5 pM and 10 pM V, and between or between about 0.02 pM and 2 pM Se.

[0172] In some embodiments, the metals comprise magnesium sulfate, manganese sulfate, calcium chloride, iron sulfate, zinc chloride, cobalt chloride, copper sulfate, aluminum potassium sulfate, boric acid, sodium molybdate, nickel chloride, sodium tungstate, vanadium chloride, and sodium selenite. In some embodiments, the metals comprise between or between about 250 pM and 2000 pM magnesium sulfate, between or between about 0.02 uM and 5 uM manganese sulfate, between or between about 50 uM and 600 uM calcium chloride, between or between about 1 uM and 30 uM iron sulfate, between or between about 5 uM and 45 uM zinc chloride, between or between about 0.02 uM and 1 uM cobalt chloride, between or between about 0.05 uM and 2 uM copper sulfate, between or between about 1 pM and 20 pM aluminum potassium sulfate, between or between about 1 pM and 20 pM boric acid, between or between about 0.02 pM and 2 pM sodium molybdate, between or between about 0.02 pM and 2 pM nickel chloride, between or between about 0.001 pM and 0.5 pM sodium tungstate, between or between about 0.5 pM and 10 pM vanadium chloride, and between or between about 0.02 pM and 2 pM sodium selenite.

2. Culturing conditions for production medium

[0173] In some embodiments, the methods provided herein comprise culturing the population, e.g., the population of the isolated strain of Lactobacillus reuteri, in a production medium comprising a glycerol source to produce 3-hydroxypropanal, thereby resulting in a production medium comprising 3- hydroxypropanal. 3-hydroxypropanal is also known as reuterin, which can be used for the production of propenoic acid through conversion of 3-hydroxypropanal to prop-2-enal (also known as acrolein) followed by conversion of prop-2-enal to propenoic acid.

[0174] In some embodiments, culturing the population in the production medium occurs for a duration of time that results in converting the glycerol source to 3-hydroxypropanal. In some embodiments, culturing the population in the production medium occurs for a duration of time that results in converting at least or at least about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the glycerol source to 3-hydroxypropanal. In some embodiments, culturing the population in the production medium occurs for a duration of time that results in converting at least or at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the glycerol source to 3-hydroxypropanal. In some embodiments, culturing the population in the production medium occurs for a duration of time that results in converting at least or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the glycerol source to 3- hydroxypropanal. In some embodiments, culturing the population in the production medium occurs for a duration of time that results in converting at or about 100% of the glycerol source to 3-hydroxypropanal.

[0175] In some embodiments, the duration of time is not particularly limited and can be adjusted to accommodate different conditions, so long as the duration of time is sufficient to allow for conversion of the glycerol source to 3-hydroxypropanal. In some embodiments, the duration of time may depend on the temperature that the culturing is performed in. [0176] In some embodiments, culturing the population in the production medium occurs for a duration of time that is or is at least 30 minutes, 45 minutes, 60 minutes, 75 minutes, 90 minutes, 2 hours, 2.5 hours, 3 hours, or 4 hours. In some embodiments, culturing the population in the production medium occurs for a duration of time that is or is at least 30 minutes. In some embodiments, culturing the population in the production medium occurs for a duration of time that is between or between about 30 minutes and 5 hours. In some embodiments, culturing the population in the production medium occurs for a duration of time that is between or between about 1 hour and 4 hours. In some embodiments, culturing the population in the production medium occurs for a duration of time that is between or between about 1 hour and 4 hours, 1 hour and 3.75 hours, 1 hour and 3.5 hours, 1 hour and 3.25 hours, 1 hour and 3 hours, 1 hour and 2.75 hours, 1 hour and 2.5 hours, 1 hour and 2.25 hours, 1 hour and 2 hours, 1.25 hours and 4 hours, 1.25 hours and 3.75 hours, 1.25 hours and 3.5 hours, 1.25 hours and 3.25 hours, 1.25 hours and 3 hours, 1.25 hours and 2.75 hours, 1.25 hours and 2.5 hours, 1.25 hours and 2.25 hours, 1.25 hours and 2 hours, 1.5 hours and 4 hours,

1.5 hours and 3.75 hours, 1.5 hours and 3.5 hours, 1.5 hours and 3.25 hours, 1.5 hours and 3 hours, 1.5 hours and 2.75 hours, 1.5 hours and 2.5 hours, 1.5 hours and 2.25 hours, 1.5 hours and 2 hours, 1.75 hours and 4 hours, 1,75 hours and 3.75 hours, 1.75 hours and 3.5 hours, 1.75 hours and 3.25 hours, 1.75 hours and 3 hours, 1.75 hours and 2.75 hours, 1.75 hours and 2.5 hours, 1.75 hours and 2.25 hours, 1.75 hours and 2 hours, 2 hours and 4 hours, 2 hours and 3.75 hours, 2 hours and 3.5 hours, 2 hours and 3.25 hours, 2 hours and 3 hours, 2 hours and 2.75 hours, 2 hours and 2.5 hours, 2 hours and 2.25 hours, 2.25 hours and 4 hours, 2.25 hours and 3.75 hours, 2.25 hours and 3.5 hours, 2.25 hours and 3.25 hours, 2.25 hours and 3 hours, 2.25 hours and 2.75 hours, 2.25 hours and 2.5 hours, 2.5 hours and 4 hours, 2.5 hours and 3.75 hours, 2.5 hours and 3.5 hours, 2.5 hours and 3.25 hours, 2.5 hours and 3 hours, or 2.5 hours and 2.75 hours. In some embodiments, culturing the population in the production medium occurs for a duration of time that is between or between about 2 hours and 3 hours. In some embodiments, culturing the population in the production medium occurs for a duration of time that is or is at least 30 minutes, 45 minutes, 1 hour, 1.25 hours, 1.5 hours, 1.75 hours, 2 hours, 2.25 hours, 2.5 hours, 2.75 hours, 3 hours, 3.25 hours, 3.5 hours, or 4 hours. In some embodiments, culturing the population in the production medium occurs for a duration of time that is or is about 30 minutes, 45 minutes, 1 hour, 1.25 hours, 1.5 hours, 1.75 hours, 2 hours, 2.25 hours,

2.5 hours, 2.75 hours, 3 hours, 3.25 hours, 3.5 hours, 4 hours, 4.25 hours, 4.5 hours, 4.75 hours, or 5 hours. [0177] In some embodiments, culturing the population in the production medium occurs for a duration of time that is or is at least 30 minutes, 45 minutes, 60 minutes, 75 minutes, 90 minutes, 2 hours, 2.5 hours, 3 hours, 4 hours, 6 hours, 12 hours, 18 hours, 24 hours, 30 hours, 36 hours, 42 hours, or 48 hours. In some embodiments, culturing the population in the production medium occurs for a duration of time that is between or between about 30 minutes and 48 hours, 30 minutes and 42 hours, 30 minutes and 36 hours, 30 minutes and 30 hours, 30 minutes and 24 hours, 30 minutes and 18 hours, 30 minutes and 12 hours, 1 hour and 48 hours, 1 hour and 42 hours, 1 hour and 36 hours, 1 hour and 30 hours, 1 hour and 24 hours, 1 hour and 18 hours, 1 hour and 12 hours, 2 hours and 48 hours, 2 hours and 42 hours, 2 hours and 36 hours, 2 hours and 30 hours, 2 hours and 24 hours, 2 hours and 18 hours, or 2 hours and 12 hours. In some embodiments, culturing the population in the production medium occurs for a duration of time that is or is about 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 8 hours, 10 hours, 12 hours, 16 hours, 20 hours, 25 hours, 30 hours, 36 hours, 42 hours, or 48 hours. In some embodiments, culturing the population in the production medium occurs for a duration of time that is or is about 30 minutes, 1 hour, 2 hours, 5 hours, 10 hours, 20 hours, 30 hours, or 48 hours.

[0178] In some embodiments, culturing the population in the production medium occurs at a temperature that is suitable for the production of 3-hydroxypropanal by the population of the isolated strain of Lactobacillus reuteri, e.g., at or about 37 degrees C. In some embodiments, culturing the population in the production medium occurs at a temperature that is between or between about 28° C and 45° C. In some embodiments, culturing the population in the production medium occurs at a temperature that is or is about 28° C, 29° C, 30° C, 31° C, 32° C, 33° C, 34° C, 35° C, 36° C, 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, or 45° C. In some embodiments, culturing the population in the production medium occurs at a temperature that is at or about 37° C.

[0179] In some embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or at least 1 g/L/hour, 2 g/L/hour, 3 g/L/hour, 4 g/L/hour, 5 g/L/hour, 6 g/L/hour, 7 g/L/hour, 8 g/L/hour, 9 g/L/hour, 10 g/L/hour, 11 g/L/hour, 12 g/L/hour, 13 g/L/hour, 14 g/L/hour, 15 g/L/hour, 20 g/L/hr, 25 g/L/hr, or 30 g/L/hr. In some embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or at least 1 g/L/hour, 2 g/L/hour, 3 g/L/hour, 4 g/L/hour, 5 g/L/hour, 6 g/L/hour, 7 g/L/hour, or 8 g/L/hour. In some embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or about 3 g/L/hour, 4 g/L/hour, 5 g/L/hour, 6 g/L/hour, 7 g/L/hour, 8 g/L/hour, 9 g/L/hour, or 10 g/L/hour.

[0180] In some embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or at least 6 g/L/hour. In some embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or at least 10 g/L/hour. In some embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or at least 15-22 g/L/hour. In some embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or at least 15 g/L/hour. In some embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or at least 18 g/L/hour. In some embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or at least 20 g/L/hour. In some embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or at least 22 g/L/hour.

[0181] In some embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of between or between about 3 g/L/hour and 10 g/L/hour, 3 g/L/hour and 9 g/L/hour, 3 g/L/hour and 8 g/L/hour, 3 g/L/hour and 7 g/L/hour, 4 g/L/hour and 10 g/L/hour, 4 g/L/hour and 9 g/L/hour, 4 g/L/hour and 8 g/L/hour, 4 g/L/hour and 7 g/L/hour, 5 g/L/hour and 10 g/L/hour, 5 g/L/hour and 9 g/L/hour, 5 g/L/hour and 8 g/L/hour, 5 g/L/hour and 7 g/L/hour, 6 g/L/hour and 10 g/L/hour, 6 g/L/hour and

9 g/L/hour, 6 g/L/hour and 8 g/L/hour, or 6 g/L/hour and 7 g/L/hour. In some embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or about 1 g/L/hour, 1.25 g/L/hour, 1.5 g/L/hour, 1.75 g/L/hour, 2 g/L/hour, 2.25 g/L/hour, 2.5 g/L/hour, 2.75 g/L/hour, 3 g/L/hour, 3.25 g/L/hour, 3.5 g/L/hour, 3.75 g/L/hour, 4 g/L/hour, 4.25 g/L/hour, 4.5 g/L/hour, 4.75 g/L/hour, 5 g/L/hour, 5.25 g/L/hour, 5.5 g/L/hour, 5.75 g/L/hour, 6 g/L/hour, 6.25 g/L/hour, 6.5 g/L/hour, 6.75 g/L/hour, 7 g/L/hour, 7.25 g/L/hour, 7.5 g/L/hour, 7.75 g/L/hour, 8 g/L/hour, 8.25 g/L/hour, 8.5 g/L/hour, 8.75 g/L/hour, 9 g/L/hour, 9.25 g/L/hour, 9.5 g/L/hour, 9.75 g/L/hour, or 10 g/L/hour. In some embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of between or between about 1 g/L/hour and 50 g/L/hour, 2 g/L/hour and 50 g/L/hour, 5 g/L/hour and 50 g/L/hour, 10 g/L/hour and 50 g/L/hour, 15 g/L/hour and 50 g/L/hour, 20 g/L/hour and 50 g/L/hour, 1 g/L/hour and 40 g/L/hour, 2 g/L/hour and 40 g/L/hour, 5 g/L/hour and 40 g/L/hour, 10 g/L/hour and 40 g/L/hour, 15 g/L/hour and 40 g/L/hour, 20 g/L/hour and 40 g/L/hour, 1 g/L/hour and 30 g/L/hour, 2 g/L/hour and 30 g/L/hour, 5 g/L/hour and 30 g/L/hour, 10 g/L/hour and 30 g/L/hour, 15 g/L/hour and 30 g/L/hour, 20 g/L/hour and 30 g/L/hour, 1 g/L/hour and 25 g/L/hour, 2 g/L/hour and 25 g/L/hour, 5 g/L/hour and 25 g/L/hour, 10 g/L/hour and 25 g/L/hour, 15 g/L/hour and 25 g/L/hour, 20 g/L/hour and 25 g/L/hour, 1 g/L/hour and 20 g/L/hour, 2 g/L/hour and 20 g/L/hour, 5 g/L/hour and 20 g/L/hour, 10 g/L/hour and 20 g/L/hour, or 15 g/L/hour and 20 g/L/hour.

[0182] In some embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of between or between about 3 and 30 g/L/hour, 3 and 25 g/L/hour, 3 and 20 g/L/hour, 3 and 15 g/L/hour, 5 and 30 g/L/hour, 5 and 25 g/L/hour, 5 and 20 g/L/hour, 5 and 15 g/L/hour, 7 and 30 g/L/hour, 7 and 25 g/L/hour, 7 and 20 g/L/hour, 7 and 15 g/L/hour, 10 and 30 g/L/hour, 10 and 25 g/L/hour,

10 and 20 g/L/hour, 10 and 15 g/L/hour, 15 and 30 g/L/hour, 15 and 25 g/L/hour, 15 and 20 g/L/hour, 20 and 30 g/L/hour, or 20 and 25 g/L/hour. [0183] In some embodiments, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or about 6 g/L/hour, 6.25 g/L/hour, 6.5 g/L/hour, 6.75 g/L/hour, 7 g/L/hour, 7.25 g/L/hour, 7.5 g/L/hour, 7.75 g/L/hour, 8 g/L/hour, 8.25 g/L/hour, 8.5 g/L/hour, 8.75 g/L/hour, 9 g/L/hour, 9.25 g/L/hour, 9.5 g/L/hour, 9.75 g/L/hour, 10 g/L/hour, 11 g/L/hr, 12 g/L/hr, 13 g/L/hr, 14 g/L/hr, 15 g/L/hr, 16 g/L/hr, 17 g/L/hr, 18 g/L/hr, 19 g/L/hr, 20 g/L/hr, 21 g/L/hr, 22 g/L/hr, 23 g/L/hr, 24 g/L/hr, 25 g/L/hr, 26 g/L/hr, 27 g/L/hr, 28 g/L/hr, 29 g/L/hr, or 30 g/L/hr,.

C. _ Separating reuterin-containing production medium from the population of Lactobacillus reuteri

[0184] In some embodiments, the methods provided herein further comprise separating the production medium comprising 3-hydroxypropanal from the population, thereby resulting in a separated production medium comprising 3-hydroxypropanal. For instance, in some embodiments of methods of producing reuterin, the method further comprises, after the culturing the population in the production medium to produce 3-hydroxypropanal, separating the production medium comprising 3-hydroxypropanal from the population, thereby resulting in a separated production medium comprising 3-hydroxypropanal.

[0185] In some embodiments, the methods provided herein further comprise separating the production medium comprising 3-hydroxypropanal from the population prior to the converting, thereby resulting in a separated production medium comprising the 3-hydroxypropanal. For instance, in some embodiments of methods of producing reuterin, the method further comprises, after the culturing the population in the production medium to produce 3-hydroxypropanal, separating the production medium comprising 3- hydroxypropanal from the population prior to the converting, thereby resulting in a separated production medium comprising the 3-hydroxypropanal.

[0186] In some embodiments, separating the production medium comprising 3-hydroxypropanal from the population comprises centrifuging the population and removing the production medium comprising 3- hydroxypropanal. The removed production medium comprising 3-hydroxypropanal can then be used in subsequent steps for the conversion of 3-hydroxypropanal (reuterin) into propenoic acid.

[0187] In some embodiments, the separated production medium comprising 3-hydroxypropanal comprises 3-hydroxypropanal. In some embodiments, the separated production medium comprising 3-hydroxypropanal comprises at or at least 6 g/L, 7 g/L, 8 g/L, 9 g/L, 10 g/L, 11 g/L, 12 g/L, 13 g/L, 14 g/L, 15 g/L, 16 g/L, 17 g/L, 18 g/L, 19 g/L, 20 g/L, 21 g/L, 22 g/L, 23 g/L, 24 g/L, 25 g/L, 30 g/L, 40 g/L, 50 g/L, 75 g/L, 100 g/L, 125 g/L, 150 g/L, 175 g/L, or 200 g/L of 3-hydroxypropanal. In some embodiments, the separated production medium comprising 3-hydroxypropanal comprises between or between about 1 g/L and 50 g/L of 3- hydroxypropanal. In some embodiments, the separated production medium comprising 3-hydroxypropanal comprises between or between about 1 g/L and 50 g/L, 1 g/L and 45 g/L, 1 g/L and 40 g/L, 1 g/L and 35 g/L, 1 g/L and 30 g/L, 1 g/L and 25 g/L, 1 g/L and 20 g/L, 1 g/L and 15 g/L, 1 g/L and 10 g/L, 5 g/L and 50 g/L, 5 g/L and 45 g/L, 5 g/L and 40 g/L, 5 g/L and 35 g/L, 5 g/L and 30 g/L, 5 g/L and 25 g/L, 5 g/L and 20 g/L, 5 g/L and 15 g/L, 5 g/L and 10 g/L, 10 g/L and 50 g/L, 10 g/L and 45 g/L, 10 g/L and 40 g/L, 10 g/L and 35 g/L, 10 g/L and 30 g/L, 10 g/L and 25 g/L, 10 g/L and 20 g/L, 10 g/L and 15 g/L, 15 g/L and 50 g/L, 15 g/L and 45 g/L, 15 g/L and 40 g/L, 15 g/L and 35 g/L, 15 g/L and 30 g/L, 15 g/L and 25 g/L, 15 g/L and 20 g/L, 20 g/L and 50 g/L, 20 g/L and 45 g/L, 20 g/L and 40 g/L, 20 g/L and 35 g/L, 20 g/L and 30 g/L, or 20 g/L and 25 g/L of 3-hydroxypropanal. In some embodiments, the separated production medium comprising 3- hydroxypropanal comprises between or between about 10 g/L and 20 g/L of 3-hydroxypropanal. In some embodiments, the separated production medium comprising 3-hydroxypropanal comprises between or between about 12 g/L and 18 g/L of 3-hydroxypropanal.

[0188] In some embodiments, the separated production medium comprising 3-hydroxypropanal comprises between or between about 6 and 200 g/L, 6 and 150 g/L, 6 and 100 g/L, 6 and 80 g/L, 10 and 200 g/L, 10 and 150 g/L, 10 and 100 g/L, 10 and 80 g/L, 15 and 200 g/L, 15 and 150 g/L, 15 and 100 g/L, 15 and 80 g/L, 20 and 200 g/L, 20 and 150 g/L, 20 and 100 g/L, 20 and 80 g/L, 30 and 200 g/L, 30 and 150 g/L, 30 and 100 g/L, 30 and 80 g/L, 40 and 200 g/L, 40 and 150 g/L, 40 and 100 g/L, 40 and 80 g/L, 50 and 200 g/L, 50 and 150 g/L, 50 and 100 g/L, 50 and 80 g/L, 60 and 200 g/L, 60 and 150 g/L, 60 and 100 g/L, or 60 and 80 g/L. [0189] In some embodiments, the separated production medium comprising 3-hydroxypropanal comprises between or between about 6 g/L and 50 g/L, 6 g/L and 45 g/L, 6 g/L and 40 g/L, 6 g/L and 36 g/L, 6 g/L and 32 g/L, 6 g/L and 28 g/L, 6 g/L and 24 g/L, 6 g/L and 20 g/L, 9 g/L and 50 g/L, 9 g/L and 45 g/L, 9 g/L and 40 g/L, 9 g/L and 36 g/L, 9 g/L and 32 g/L, 9 g/L and 28 g/L, 9 g/L and 24 g/L, 9 g/L and 20 g/L, 12 g/L and 50 g/L, 12 g/L and 45 g/L, 12 g/L and 40 g/L, 12 g/L and 36 g/L, 12 g/L and 32 g/L, 12 g/L and 28 g/L, 12 g/L and 24 g/L, 12 g/L and 20 g/L, 15 g/L and 50 g/L, 15 g/L and 45 g/L, 15 g/L and 40 g/L, 15 g/L and 36 g/L, 15 g/L and 32 g/L, 15 g/L and 28 g/L, 15 g/L and 24 g/L, 15 g/L and 20 g/L, 18 g/L and 50 g/L, 18 g/L and 45 g/L, 18 g/L and 40 g/L, 18 g/L and 36 g/L, 18 g/L and 32 g/L, 18 g/L and 28 g/L, 18 g/L and 24 g/L, or 18 g/L and 20 g/L of 3-hydroxypropanal.

1. Converting reuterin into propenoic acid

[0190] In some embodiments, the methods provided herein comprise converting the 3-hydroxypropanal into propenoic acid. In some embodiments, converting the 3-hydroxypropanal into propenoic acid comprises converting the 3-hydroxypropanal into prop-2-enal (also known as acrolein) and converting the prop-2-enal into propenoic acid. In some embodiments, these conversion methods can include any known conversion methods available in the art. In some embodiments, acrolein is converted into propenoic acid (also known as acrylic acid) using methods as described in US 4,107,204.

[0191] In some embodiments, converting the 3-hydroxypropanal into prop-2 -enal comprises acidifying the production medium comprising 3-hydroxypropanal, thereby resulting in an acidified production medium; and converting the 3-hydroxypropanal in the acidified production medium into gaseous prop-2 -enal.

[0192] In some embodiments, converting the 3-hydroxypropanal into prop-2 -enal comprises acidifying the production medium comprising 3-hydroxypropanal or the separated production medium comprising 3- hydroxypropanal, thereby resulting in an acidified production medium; and converting the 3- hydroxypropanal in the acidified production medium into gaseous prop-2-enal.

[0193] In some embodiments, converting prop-2-enal into propenoic acid comprises oxidizing the prop-2 - enal in the presence of a catalyst to produce the propenoic acid. In some embodiments, the catalyst can be any suitable catalyst.

[0194] In some embodiments, the converting the 3-hydroxypropanal into propenoic acid comprises: (i) acidifying the production medium comprising 3-hydroxypropanal, thereby resulting in an acidified production medium; (ii) converting the 3-hydroxypropanal in the acidified production medium into gaseous prop-2-enal; and (iii) oxidizing the prop-2 -enal in the presence of a catalyst to produce the propenoic acid. In some embodiments, the converting the 3-hydroxypropanal into propenoic acid comprises: (i) acidifying the production medium comprising 3-hydroxypropanal or the separated production medium comprising 3- hydroxypropanal, thereby resulting in an acidified production medium; (ii) converting the 3-hydroxypropanal in the acidified production medium into gaseous prop-2-enal; and (iii) oxidizing the prop-2-enal in the presence of a catalyst to produce the propenoic acid.

[0195] In some embodiments, acidifying the production medium comprising 3-hydroxypropanal comprises acidifying the production medium comprising 3-hydroxypropanal with a mineral acid. In some embodiments, acidifying the production medium comprising 3-hydroxypropanal or the separated production medium comprising 3-hydroxypropanal comprises acidifying the production medium comprising 3-hydroxypropanal or the separated production medium comprising 3-hydroxypropanal with a mineral acid.

[0196] In some embodiments, the mineral acid is selected from the group consisting of phosphoric acid, hydrochloric acid, sulfuric acid, nitric acid, boric acid, hydrofluoric acid, hydroiodic acid, hydrobromic acid, and perchloric acid. In some embodiments, the mineral acid is phosphoric acid.

[0197] In some embodiments, converting the 3-hydroxypropanal in the acidified production medium into gaseous prop-2-enal comprises heating the acidified production medium. In some embodiments, the heating is performed under reduced pressure. In some embodiments, the heating is performed under reduced pressure as compared to atmospheric pressure, which is approximately 1 bar. In some embodiments, the heating is performed under reduced pressure of less than about 1 bar, such as between or between about 40 and 400 millibar (mbar). In some embodiments, the heating is performed at a pressure of between or between about 10 and 900 mbar, 10 and 800 mbar, 10 and 700 mbar, 10 and 600 mbar, 10 and 500 mbar, 10 and 400 mbar,

20 and 900 mbar, 20 and 800 mbar, 20 and 700 mbar, 20 and 600 mbar, 20 and 500 mbar, 20 and 400 mbar,

40 and 900 mbar, 40 and 800 mbar, 40 and 700 mbar, 40 and 600 mbar, 40 and 500 mbar, 40 and 400 mbar,

80 and 900 mbar, 80 and 800 mbar, 80 and 700 mbar, 80 and 600 mbar, 80 and 500 mbar, 80 and 400 mbar,

150 and 900 mbar, 150 and 800 mbar, 150 and 700 mbar, 150 and 600 mbar, 150 and 500 mbar, 150 and 400 mbar, 250 and 900 mbar, 250 and 800 mbar, 250 and 700 mbar, 250 and 600 mbar, 250 and 500 mbar, or 250 and 400 mbar. In some embodiments, the heating is performed at a pressure of between or between about 40 and 400 mbar.

[0198] In some embodiments, heating is performed at a temperature of between about 30° C and about 95° C, such as at or about 35° C, 40° C, 45° C, 50° C, 55° C, 60° C, 65° C, 70° C, 75° C, 80° C, 85° C, or 90° C, or any value between any of the foregoing.

[0199] In some embodiments, prop-2 -enal (acrolein) is converted into propenoic acid (also known as acrylic acid) using methods as described in US 4,107,204, the contents of which are hereby incorporated by reference in their entirety.

[0200] In some embodiments, oxidizing the prop-2 -enal in the presence of a catalyst to produce the propenoic acid comprises the use of vapor-phase oxidation. In some embodiments, the catalyst comprises a two-metal catalyst. In some embodiments, oxidizing the prop-2-enal in the presence of a catalyst to produce the propenoic acid comprises reacting the prop-2 -enal with an oxygen-containing gas at a temperature that is between or between about 125 and 320 degrees C in the presence of a two-metal catalyst. In some embodiments, the two-metal catalyst comprises palladium and copper or silver.

II. REUTERIN, PROPENOIC ACID, ISOLATED STRAINS, KITS, AND COMPOSITIONS

[0201] Also provided herein are reuterin and propenoic acid produced by the methods described herein, and isolated strains of Lactobacillus reuteri, kits, and compositions.

[0202] Provided herein is 3-hydroxypropanal (reuterin) produced by any of the methods described herein. [0203] Provided herein is propenoic acid produced by any of the methods described herein.

[0204] Provided herein are isolated strains of Lactobacillus reuteri, such as any of the isolated strains of

Lactobacillus reuteri described herein. [0205] In some embodiments, the isolated strain of Lactobacillus reuteri was isolated from feces of an animal. The animal is not particularly limited and may be any animal, e.g., any animal whose feces contains Lactobacillus reuteri. In some embodiments, the animal is a mammal.

[0206] In some embodiments, the animal is selected from the group consisting of humans, porcine, poultry, rodents, and cattle. In some embodiments, the animal is a porcine animal. In some embodiments, the porcine animal is a pig, hog, or boar. In some embodiments, the animal is a poultry animal. In some embodiments, the poultry animal is a chicken, turkey, duck, goose, fowl, or pheasant. In some embodiments, the animal is a cattle animal. In some embodiments, the cattle animal is a cow, steer, or bull.

[0207] In some embodiments, the animal is selected from the group consisting of a human, a mouse, a rat, a pig, a hog, a boar, a chicken, a turkey, a duck, a goose, a fowl, a horse, a donkey, a sheep, a pheasant, a cow, a steer, a bull, a buffalo, and a bison.

[0208] In some embodiments, the animal is a livestock animal. In some embodiments, the livestock animal is selected from the group consisting of a pig, a hog, a boar, a chicken, a turkey, a horse, a donkey, a sheep, a pheasant, a cow, a steer, a bull, a buffalo, and a bison.

[0209] In some embodiments, the animal is a pig. In some embodiments, the animal is a hog. In some embodiments, the animal is a duck. In some embodiments, the animal is a goose. In some embodiments, the animal is a mouse. In some embodiments, the animal is a rat. In some embodiments, the animal is a chicken. In some embodiments, the animal is a turkey. In some embodiments, the animal is a horse. In some embodiments, the animal is a sheep. In some embodiments, the animal is a pheasant. In some embodiments, the animal is a cow. In some embodiments, the animal is a steer. In some embodiments, the animal is a bull. In some embodiments, the animal is a buffalo. In some embodiments, the animal is a bison.

[0210] In some embodiments, the isolated strain of Lactobacillus reuteri was isolated by a method comprising: a) suspending the feces in buffer, thereby resulting in a fecal suspension, and b) culturing the fecal suspension in an isolation medium that promotes the growth of Lactobacillus reuteri. In some embodiments, the isolation medium comprises glucose, dipotassium hydrogen phosphate, magnesium sulfate heptahydrate, manganous sulfate tetrahydrate, peptone, yeast extract, meat extract, triammonium citrate, and sodium acetate trihydrate. In some embodiments, the isolation medium further comprises glycerol and vancomycin.

[0211] Also provided herein is a kit comprising an isolated strain of Lactobacillus reuteri, such as any of the isolated strains of Lactobacillus reuteri described herein, and instructions for use thereof. In some embodiments, the instructions for use thereof comprise instructions for producing reuterin and/or propenoic acid, such as by any of the methods described herein. In some embodiments, the instructions for use thereof comprise instructions for culturing the isolated strain of Lactobacillus reuteri in a growth medium, thereby resulting in a population of the isolated strain of Lactobacillus reuteri; and/or (b) culturing the population in a production medium comprising a glycerol source to produce 3-hydroxypropanal, such as by any of the methods described herein.

[0212] In some embodiments, the kit further comprises a growth medium and/or a production medium, such as any of the growth mediums and/or production mediums described herein. In some embodiments, the kit further comprises one or more components for producing a growth medium and/or production medium, such as any of the growth mediums and/or production mediums described herein.

[0213] Also provided herein is a kit comprising the 3-hydroxypropanal (reuterin) produced by any of the methods described herein.

[0214] Also provided herein is a kit comprising the propenoic acid produced by any of the methods described herein.

[0215] Also provided herein is a composition comprising an isolated strain of Lactobacillus reuteri, such as any of the isolated strains of Lactobacillus reuteri described herein.

[0216] Also provided herein is a composition comprising the 3-hydroxypropanal (reuterin) produced by any of the methods described herein.

[0217] Also provided herein is a composition comprising the propenoic acid produced by any of the methods described herein.

[0218] Also provided herein is a kit comprising a composition comprising the 3-hydroxypropanal (reuterin) produced by any of the methods described herein.

[0219] Also provided herein is a kit comprising a composition comprising the propenoic acid produced by any of the methods described herein.

III. DEFINITIONS

[0220] Unless defined otherwise, all terms of art, notations and other technical and scientific terms or terminology used herein are intended to have the same meaning as is commonly understood by one of ordinary skill in the art to which the claimed subject matter pertains. In some cases, terms with commonly understood meanings are defined herein for clarity and/or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a substantial difference over what is generally understood in the art [0221] Terms and phrases used in this disclosure, and variations thereof, unless otherwise expressly stated, should be construed as open ended as opposed to limiting. As examples of the foregoing: the term “including” should be read as meaning “ including, without limitation” or the like; the term “example” is used to provide exemplary instances of the item in discussion, not an exhaustive or limiting list thereof; and adjectives such as “conventional,” “traditional,” “normal,” “standard,” “known,” and terms of similar meaning, should not be construed as limiting the item described to a given time period, or to an item available as of a given time. Instead, these terms should be read to encompass conventional, traditional, normal, or standard technologies that may be available, known now, or at any time in the future. Likewise, a group of items linked with the conjunction “and” should not be read as requiring that each and every one of those items be present in the grouping, but, rather, should be read as “ and/or” unless expressly stated otherwise. Similarly, a group of items linked with the conjunction “or” should not be read as requiring mutual exclusivity among that group, but rather should also be read as “and/or” unless expressly stated otherwise. Furthermore, although items, elements, or components of the invention may be described or claimed in the singular, the plural is contemplated to be within the scope thereof unless limitation to the singular is explicitly stated. The presence of broadening words and phrases, such as “one or more,” “at least,” “but not limited to,” or other like phrases in some instances, shall not be read to mean that the narrower case is intended or required in instances where such broadening phrases may be absent.

[0222] As used herein, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. For example, “a” or “an” means “at least one” or “one or more.”

[0223] The term “about” as used herein refers to the usual error range for the respective value readily known to the skilled person in this technical field. Reference to “about” a value or parameter herein includes (and describes) embodiments that are directed to that value or parameter per se.

[0224] As used herein, a composition refers to any mixture of two or more products, substances, or compounds, including cells. It may be a solution, a suspension, liquid, powder, a paste, aqueous, nonaqueous or any combination thereof.

[0225] As used herein, “growth medium” refers to a medium that promotes the growth of the isolated strain of Lactobacillus reuteri, e.g., to achieve sufficient biomass for use in converting a glycerol source to 3- hydroxypropanal during a subsequent production stage.

[0226] As used herein, “production medium” refers to a medium that allows for the conversion of a glycerol source to 3-hydroxypropanal by the isolated strain of Lactobacillus reuteri, e.g., promoting conversion of a glycerol source to 3-hydroxypropanal by the isolated strain of Lactobacillus reuteri without promoting its growth. [0227] As used herein, “growth” refers to growth in number by proliferation of individual cells of the isolated strain of Lactobacillus reuteri into a larger population of cells of the isolated strain, such as growth that occurs when the isolated strain of Lactobacillus reuteri is cultured in growth medium.

[0228] As used herein, the term “isolated” when used in reference to a microbial organism, e.g., Lactobacillus reuteri, such as an isolated strain of Lactobacillus reuteri, is intended to mean an organism that is substantially free of at least one component as the referenced microbial organism is found in nature, and can, in some embodiments, include an isolated microbial organism, e.g., an isolated strain of Lactobacillus reuteri, that is further engineered, e.g., genetically engineered. The term includes a microbial organism, e.g., Lactobacillus reuteri, that is removed from some or all components as it is found in its natural environment. The term also includes a microbial organism, e.g., Lactobacillus reuteri, that is removed from some or all components as the microbial organism is found in non-naturally occurring environments. As such, an isolated microbial organism is partly or completely separated from other substances as it is found in nature or as it is grown, stored, or subsisted in non-naturally occurring environments. Specific examples of isolated microbial organisms include partially pure microbes, substantially pure microbes, and microbes cultured in a medium that is non-naturally occurring. Isolated strains that are further engineered, e.g., genetically engineered, are still referred to herein as being “isolated” or as being an “isolated strain.” Accordingly, reference to an “isolated strain” also encompasses an isolated strain that has been further engineered, e.g., genetically engineered, such as by any engineering described herein or known or practiced in the art.

[0229] As used herein, the term “exogenous” means that that the referenced molecule, e.g., nucleic acid, such as an exogenous nucleic acid, is introduced into a microbial organism, e.g., Lactobacillus reuteri, such as an isolated strain of Lactobacillus reuteri, by genetic engineering. The exogenous nucleic acid can be introduced, for example, by introduction of a nucleic acid into the host microbial organism, such as by integration into a chromosome of the host microbial organism or by introduction of a nucleic acid as non- chromosomal genetic material, such as via a plasmid. As such, the term “exogenous” as it is used in reference to expression of an encoding nucleic acid refers to introduction of the encoding nucleic acid in an expressible form into the host microbial organism, e.g., an isolated strain of Lactobacillus reuteri.

[0230] As used herein, the term “gene disruption,” or grammatical equivalents thereof, is intended to mean a genetic alteration that renders the encoded gene product inactive or attenuated. The genetic alteration can be, for example, deletion of the entire gene, deletion of a regulatory sequence required for transcription or translation, deletion of a portion of the gene that results in a truncated gene product, or by any of various mutation strategies that inactive or attenuate the encoded gene product. For instance, in some embodiments, the method of gene disruption can be complete gene deletion because it reduces or eliminates the occurrence of genetic reversions in the isolated microbial organism being engineered. A gene disruption can, in some embodiments, include a null mutation, which is a mutation within a gene or region of a gene that results in the gene not being transcribed into RNA and/or translated into a functional gene product. Null mutations can arise from various types of mutations including, for example, an inactivating point mutation, deletion of a portion of a gene, deletion of an entire gene, or deletion of a chromosomal segment.

IV. EXEMPLARY EMBODIMENTS

[0231] Among the provided embodiments are:

1. A method of producing 3-hydroxypropanal (reuterin), comprising: (a) culturing an isolated strain of Lactobacillus reuteri in a growth medium, thereby resulting in a population of the isolated strain of Lactobacillus reuteri; and (b) culturing the population in a production medium comprising a glycerol source to produce 3-hydroxypropanal.

2. The method of embodiment 1, further comprising separating the production medium comprising 3-hydroxypropanal from the population, thereby resulting in a separated production medium comprising 3- hy droxypropanal .

3. A method of producing propenoic acid, comprising: (a) culturing an isolated strain of Lactobacillus reuteri in a growth medium, thereby resulting in a population of the isolated strain of Lactobacillus reuteri; (b) culturing the population in a production medium comprising a glycerol source to produce 3-hydroxypropanal, thereby resulting in a production medium comprising 3-hydroxypropanal; (c) converting the 3-hydroxypropanal into propenoic acid.

4. The method of embodiment 3, further comprising separating the production medium comprising 3-hydroxypropanal from the population prior to the converting, thereby resulting in a separated production medium comprising the 3-hydroxypropanal.

5. The method of any one of embodiments 1-4, wherein the isolated strain of Lactobacillus reuteri was isolated from feces of an animal.

6. The method of embodiment 5, wherein the animal is selected from the group consisting of humans, porcine, poultry, rodents, and cattle. 7. The method of embodiment 5, wherein the animal is a livestock animal.

8. The method of embodiment 7, wherein the livestock animal is selected from the group consisting of porcine, poultry, and cattle.

9. The method of any one of embodiments 5-8, wherein the animal is a porcine animal.

10. The method of embodiment 9, wherein the porcine animal is a pig, hog, or boar.

11. The method of any one of embodiments 5-8, wherein the animal is a poultry animal.

12. The method of embodiment 11, wherein the poultry animal is a chicken, turkey, duck, goose, fowl, or pheasant.

13. The method of any one of embodiments 5-8, wherein the animal is a cattle animal.

14. The method of embodiment 13, wherein the cattle animal is a cow, steer, or bull.

15. The method of any one of embodiments 1-5, wherein the animal is selected from the group consisting of a human, a mouse, a rat, a pig, a hog, a boar, a chicken, a turkey, a duck, a goose, a fowl, a pheasant, a cow, a steer, a bull, a buffalo, and a bison.

16. The method of any one of embodiments 1-15, wherein the isolated strain of Lactobacillus reuteri was isolated by a method comprising: a) suspending the feces in buffer, thereby resulting in a fecal suspension, and b) culturing the fecal suspension in an isolation medium that promotes the growth of Lactobacillus reuteri.

17. The method of embodiment 16, wherein the isolation medium comprises glucose, dipotassium hydrogen phosphate, magnesium sulfate heptahydrate, manganese sulfate tetrahydrate, peptone, yeast extract, meat extract, triammonium citrate, and sodium acetate trihydrate.

18. The method of embodiment 17, wherein the isolation medium further comprises glycerol and vancomycin.

19. The method of any one of embodiments 1-18, wherein the growth medium comprises components that promote the growth of Lactobacillus reuteri. 20. The method of any one of embodiments 1-19, wherein the growth medium comprises vitamins, amino acids, nucleotides, metal salts, acid whey, glycerol, sodium acetate, and polysorbate 80.

21. The method of any one of embodiments 1-20, wherein the growth medium does not comprise glucose.

22. The method of any one of embodiments 1-21, wherein culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 28 degrees C and 50 degrees C.

23. The method of any one of embodiments 1-21, wherein culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of: (i) between or between about 28° C, 29° C, 31° C, 32° C, 33° C, or 34° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C ; or (ii) between or between about 35° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C; or (iii) between or between about 36° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C; or (iv) between or between about 37° C and 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C; or (v) between or between about 37° C and 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, or 45° C; or (vi) at or about 28° C, 29° C, 30° C, 31° C, 32° C, 33° C, 34° C, 35° C, 36° C, 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C; or (vii) at or about 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, or 45° C; or (viii) at or about 37° C; or (ix) at or about 45° C.

24. The method of any one of embodiments 1-23, wherein culturing the isolated strain of Lactobacillus reuteri in the growth medium occurs for a duration of time sufficient to result in an optical density (OD) within the range of or of about 10-12.

25. The method of any one of embodiments 1-24, wherein culturing the isolated strain of Lactobacillus reuteri in the growth medium occurs for a duration of time that is: (i) at least or at least about 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 22 hours, 24 hours, 30 hours, 36 hours, 42 hours, or 48 hours; or (ii) is or is about 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, or 20 hours; or (iii) is between or between about 4 hours and 10 hours, 4 hours and 9 hours, 4 hours and 8 hours, 4 hours and 7 hours, 4 hours and 6 hours, 5 hours and 10 hours, 5 hours and 9 hours, 5 hours and 8 hours, or 5 hours and 7 hours; or (iv) is or is about 5 hours, 6 hours, 7 hours, or 8 hours; or (v) is between or between about 8 hours and 14 hours, 8 hours and 13 hours, 8 hours and 12 hours, 9 hours and 14 hours, 9 hours and 13 hours, 9 hours and 12 hours, 10 hours and 14 hours, 10 hours and 13 hours, or 10 hours and 12 hours; or (vi) is or is about 9 hours, 10 hours, 11 hours, 12 hours, or 13 hours.

26. The method of any one of embodiments 1-25, wherein culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature at or about 37 degrees C for a duration of time that is or is about 10 hours, 11, or 12 hours.

27. The method of any one of embodiments 1-25, wherein culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature at or about 45 degrees C for a duration of time that is or is about 5 hours, 6 hours, 7 hours, or 8 hours.

28. The method of any one of embodiments 1-27, wherein culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium until CO2 generation has reduced to between or between about 1/3 to 1/2 of peak CO2 generation.

29. The method of any one of embodiments 1-27, wherein culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium until CO2 generation has reduced to about 1/2 of peak CO2 generation.

30. The method of any one of embodiments 1-29, wherein, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an optical density (OD) between or between about 9 and 15 or 10 and 14.

31. The method of embodiment 30, wherein the OD is between or between about 10 and 14.

32. The method of embodiment 30 or embodiment 31, wherein the OD is or is about 10, 11, 12, 13, or 14.

33. The method of any one of embodiments 1-32, further comprising separating the population of the isolated strain of Lactobacillus reuteri from the growth media. 34. The method of embodiment 33, wherein the separating comprises the use of tangential flow filtration, followed by centrifuging the population, and followed by removing the growth media.

35. The method of any one of embodiments 1-34, wherein the glycerol source is glycerol.

36. The method of any one of embodiments 1-35, wherein the glycerol source is from an organic waste source.

37. The method of embodiment 36, wherein the organic waste source has been pasteurized.

38. The method of any one of embodiments 1-37, wherein the production medium comprises glycerol, lactose, MOPS pH 7, polysorbate 80, sodium chloride, potassium chloride, metals, and iron(II) sulfate.

39. The method of embodiment 38, wherein the production medium comprises 200 rnM glycerol, 25 mM lactose, 50 mM MOPS pH 7, 1 g/L polysorbate 80, 25 mM sodium chloride, 3.125 mM potassium chloride, metals, and 10 pM iron(II) sulfate.

40. The method of embodiment 38 or embodiment 39, wherein the metals comprise one or more metals selected from the group consisting of Mg, Mn, Ca, Fe, Zn, Co, Cu, Al, B, Mo, Ni, W, V, and Se.

41. The method of any one of embodiments 38-40, wherein the metals comprise Mg, Mn, Ca, Fe, Zn, Co, Cu, Al, B, Mo, Ni, W, V, and Se.

42. The method of any one of embodiments 38-41, wherein the metals comprise between or between about 250 pM and 2000 pM Mg, between or between about 0.02 pM and 5 pM Mn, between or between about 50 pM and 600 pM Ca, between or between about 1 pM and 30 pM Fe, between or between about 5 pM and 45 pM Zn, between or between about 0.02 pM and 1 pM Co, between or between about 0.05 pM and 2 pM Cu, between or between about 1 pM and 20 pM Al, between or between about 1 pM and 20 pM B, between or between about 0.02 pM and 2 pM Mo, between or between about 0.02 pM and 2 pM Ni, between or between about 0.001 pM and 0.5 pM W, between or between about 0.5 pM and 10 pM V, and between or between about 0.02 pM and 2 pM Se.

43. The method of any one of embodiments 1-42, wherein culturing the population in the production medium occurs for a duration of time that results in converting at least or at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the glycerol source to 3-hydroxypropanal.

44. The method of embodiment 43, wherein culturing the population in the production medium occurs for a duration of time that results in converting at least or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the glycerol source to 3-hydroxypropanal.

45. The method of embodiment 43 or embodiment 44, wherein culturing the population in the production medium occurs for a duration of time that results in converting at or about 100% of the glycerol source to 3-hydroxypropanal.

46. The method of any one of embodiments 1-45, wherein culturing the population in the production medium occurs for a duration of time that is or is at least 30 minutes.

47. The method of embodiment 46, wherein culturing the population in the production medium occurs for a duration of time that is between or between about 30 minutes and 48 hours.

48. The method of embodiment 46 or embodiment 47, wherein culturing the population in the production medium occurs for a duration of time that is between or between about 1 hour and 4 hours.

49. The method of embodiment 46 or embodiment 47, wherein culturing the population in the production medium occurs for a duration of time that is between or between about 2 and 3 hours.

50. The method of embodiment 46, wherein culturing the population in the production medium occurs for a duration of time that is or is at least 30 minutes, 45 minutes, 1 hour, 1.25 hours, 1.5 hours, 1.75 hours, 2 hours, 2.25 hours, 2.5 hours, 2.75 hours, 3 hours, 3.25 hours, 3.5 hours, or 4 hours.

51. The method of any one of embodiments 1-50, wherein culturing the population in the production medium comprises incubating the population in the production medium at a temperature at or about 37 degrees C.

52. The method of any one of embodiments 1-50, wherein, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or at least 2, 3, 4, 5, 6, 7, or 8 g/L/hour.

53. The method of any one of embodiments 1-50, wherein, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or about 3, 4, 5, 6, 7, 8, 9, or 10 g/L/hour. 54. The method of any one of embodiments 1-50, wherein, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or at least 6 g/L/hour.

55. The method of any one of embodiments 1-50, wherein, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of between or between about 3 and 10 g/L/hour, 3 and 9 g/L/hour, 3 and 8 g/L/hour, 3 and 7 g/L/hour, 4 and 10 g/L/hour, 4 and 9 g/L/hour, 4 and 8 g/L/hour, 4 and 7 g/L/hour, 5 and 10 g/L/hour, 5 and 9 g/L/hour, 5 and 8 g/L/hour, 5 and 7 g/L/hour, 6 and 10 g/L/hour, 6 and 9 g/L/hour, 6 and 8 g/L/hour, or 6 and 7 g/L/hour.

56. The method of any one of embodiments 4-55, wherein separating the production medium comprising 3-hydroxypropanal from the population comprises centrifuging the population and removing the production medium comprising 3-hydroxypropanal.

57. The method of any one of embodiments 4-56, wherein the separated production medium comprising 3-hydroxypropanal comprises at or at least 6 g/L, 7 g/L, 8 g/L, 9 g/L, 10 g/L, 11 g/L, 12 g/L, 13 g/L, 14 g/L, 15 g/L, 16 g/L, 17 g/L, 18 g/L, 19 g/L, 20 g/L, 21 g/L, 22 g/L, 23 g/L, or 24 g/L of 3- hy droxypropanal .

58. The method of any one of embodiments 4-57, wherein the separated production medium comprising 3-hydroxypropanal comprises between or between about 6 g/L and 40 g/L, 6 g/L and 36 g/L, 6 g/L and 32 g/L, 6 g/L and 28 g/L, 6 g/L and 24 g/L, 6 g/L and 20 g/L, 9 g/L and 40 g/L, 9 g/L and 36 g/L, 9 g/L and 32 g/L, 9 g/L and 28 g/L, 9 g/L and 24 g/L, 9 g/L and 20 g/L, 12 g/L and 40 g/L, 12 g/L and 36 g/L, 12 g/L and 32 g/L, 12 g/L and 28 g/L, 12 g/L and 24 g/L, 12 g/L and 20 g/L, 15 g/L and 40 g/L, 15 g/L and 36 g/L, 15 g/L and 32 g/L, 15 g/L and 28 g/L, 15 g/L and 24 g/L, 15 g/L and 20 g/L, 18 g/L and 40 g/L, 18 g/L and 36 g/L, 18 g/L and 32 g/L, 18 g/L and 28 g/L, 18 g/L and 24 g/L, or 18 g/L and 20 g/L of 3- hy droxypropanal .

59. The method of any one of embodiments 4-58, wherein the separated production medium comprising 3-hydroxypropanal comprises between or between about 12 g/L and 18 g/L of 3- hy droxypropanal .

60. The method of any one of embodiments 3-59, wherein converting the 3-hydroxypropanal into propenoic acid comprises converting the 3-hydroxypropanal into prop-2-enal and converting the prop-2 -enal into propenoic acid. 61. The method of embodiment 60, wherein converting the 3 -hydroxy prop anal into prop-2 -enal comprises acidifying the production medium comprising 3-hydroxypropanal, thereby resulting in an acidified production medium; and converting the 3-hydroxypropanal in the acidified production medium into gaseous prop-2-enal.

62. The method of embodiment 60, wherein converting the 3-hydroxypropanal into prop-2 -enal comprises acidifying the production medium comprising 3-hydroxypropanal or the separated production medium comprising 3-hydroxypropanal, thereby resulting in an acidified production medium; and converting the 3-hydroxypropanal in the acidified production medium into gaseous prop-2-enal.

63. The method of any one of embodiments 60-62, wherein converting prop-2-enal into propenoic acid comprises oxidizing the prop-2-enal in the presence of a catalyst to produce the propenoic acid.

64. The method of any one of embodiments 3-63, wherein the converting the 3-hydroxypropanal into propenoic acid comprises: (i) acidifying the production medium comprising 3-hydroxypropanal, thereby resulting in an acidified production medium; (ii) converting the 3-hydroxypropanal in the acidified production medium into gaseous prop-2-enal; and (iii) oxidizing the prop-2-enal in the presence of a catalyst to produce the propenoic acid.

65. The method of any one of embodiments 3-63, wherein the converting the 3-hydroxypropanal into propenoic acid comprises: (i) acidifying the production medium comprising 3-hydroxypropanal or the separated production medium comprising 3-hydroxypropanal, thereby resulting in an acidified production medium; (ii) converting the 3-hydroxypropanal in the acidified production medium into gaseous prop-2 -enal; and (iii) oxidizing the prop-2 -enal in the presence of a catalyst to produce the propenoic acid.

66. The method of embodiment 61 or embodiment 64, wherein acidifying the production medium comprising 3-hydroxypropanal comprises acidifying the production medium comprising 3-hydroxypropanal with a mineral acid.

67. The method of embodiment 62 or embodiment 65, wherein acidifying the production medium comprising 3-hydroxypropanal or the separated production medium comprising 3-hydroxypropanal comprises acidifying the production medium comprising 3-hydroxypropanal or the separated production medium comprising 3-hydroxypropanal with a mineral acid. 68. The method of embodiment 66 or embodiment 67, wherein the mineral acid is selected from the group consisting of phosphoric acid, hydrochloric acid, sulfuric acid, nitric acid, boric acid, hydrofluoric acid, hydroiodic acid, hydrobromic acid, and perchloric acid.

69. The method of any one of embodiments 64-68, wherein converting the 3-hydroxypropanal in the acidified production medium into gaseous prop-2-enal comprises heating the acidified production medium.

70. The method of embodiment 69, wherein the heating is performed under pressure.

71. The method of any one of embodiments 63-70, wherein oxidizing the prop-2-enal in the presence of a catalyst to produce the propenoic acid comprises the use of vapor -phase oxidation.

72. The method of any one of embodiments 63-70, wherein the catalyst comprise a two-metal catalyst.

73. The method of any one of embodiments 63-70, wherein oxidizing the prop-2-enal in the presence of a catalyst to produce the propenoic acid comprises reacting the prop-2 -enal with an oxygencontaining gas at a temperature that is between or between about 125 and 320 degrees C in the presence of a two-metal catalyst.

74. The method of embodiment 72 or embodiment 73, wherein the two-metal catalyst comprises palladium and copper or silver.

75. 3-hydroxypropanal (reuterin) produced by the method of any one of embodiments 1-74.

76. Propenoic acid produced by the method of any one of embodiments 3-74.

77. An isolated strain of Lactobacillus reuteri, wherein the isolated strain was isolated from feces of an animal.

78. The isolated strain of embodiment 77, wherein the animal is selected from the group consisting of a human, a mouse, a rat, a pig, a hog, a boar, a chicken, a turkey, a duck, a goose, a fowl, a pheasant, a cow, a steer, a bull, a buffalo, and a bison.

79. The isolated strain of embodiment 77 or embodiment 78, wherein the isolated strain of Lactobacillus reuteri was isolated by a method comprising: a) suspending the feces in buffer, thereby resulting in a fecal suspension, and b) culturing the fecal suspension in an isolation medium that promotes the growth of Lactobacillus reuteri.

80. The isolated strain of embodiment 79, wherein the isolation medium comprises glucose, dipotassium hydrogen phosphate, magnesium sulfate heptahydrate, manganous sulfate tetrahydrate, peptone, yeast extract, meat extract, triammonium citrate, and sodium acetate trihydrate.

81. The isolated strain of embodiment 79 or embodiment 80, wherein the isolation medium further comprises glycerol and vancomycin.

82. A kit comprising an isolated strain of Lactobacillus reuteri, and instructions for use thereof.

83. A kit comprising the isolated strain of Lactobacillus reuteri of any one of embodiments 77-81, and instructions for use thereof.

84. The kit of embodiment 82 or embodiment 83, further comprising a growth medium and/or a production medium.

85. The kit of embodiment 82 or embodiment 83, further comprising one or more components for producing a growth medium and/or production medium.

86. A composition comprising an isolated strain of Lactobacillus reuteri.

87. A composition comprising the isolated strain of Lactobacillus reuteri of any one of embodiments 77-81.

88. A composition comprising the 3-hydroxypropanal (reuterin) of embodiment 75.

89. A composition comprising the propenoic acid of embodiment 76.

90. A kit comprising the 3-hydroxypropanal (reuterin) of embodiment 75.

91. A kit comprising the propenoic acid of embodiment 76.

92. A kit comprising the composition of any one of embodiments 86-89.

[0232] Also among the provided embodiments are: 1. A method of producing 3-hydroxypropanal (reuterin), comprising: (a) culturing an isolated strain of Lactobacillus reuteri in a growth medium, thereby resulting in a population of the isolated strain of Lactobacillus reuteri; and (b) culturing the population in a production medium comprising a glycerol source to produce 3-hydroxypropanal.

2. The method of embodiment 1, further comprising separating the production medium comprising 3- hydroxypropanal from the population, thereby resulting in a separated production medium comprising 3- hydroxypropanal .

3. A method of producing propenoic acid, comprising: (a) culturing an isolated strain of Lactobacillus reuteri in a growth medium, thereby resulting in a population of the isolated strain of Lactobacillus reuteri;

(b) culturing the population in a production medium comprising a glycerol source to produce 3- hydroxypropanal, thereby resulting in a production medium comprising 3-hydroxypropanal; (c) converting the 3-hydroxypropanal into propenoic acid.

4. The method of embodiment 3, further comprising separating the production medium comprising 3- hydroxypropanal from the population prior to the converting, thereby resulting in a separated production medium comprising the 3-hydroxypropanal.

5. The method of any one of embodiments 1-4, wherein the isolated strain of Lactobacillus reuteri was isolated from feces of an animal.

6. The method of embodiment 5, wherein the animal is selected from the group consisting of humans, porcine, poultry, rodents, and cattle.

7. The method of embodiment 5, wherein the animal is a livestock animal.

8. The method of embodiment 7, wherein the livestock animal is selected from the group consisting of porcine, poultry, and cattle.

9. The method of any one of embodiments 5-8, wherein the animal is a porcine animal.

10. The method of embodiment 9, wherein the porcine animal is a pig, hog, or boar.

11. The method of any one of embodiments 5-8, wherein the animal is a poultry animal.

12. The method of embodiment 11, wherein the poultry animal is a chicken, turkey, duck, goose, fowl, or pheasant.

13. The method of any one of embodiments 5-8, wherein the animal is a cattle animal.

14. The method of embodiment 13, wherein the cattle animal is a cow, steer, or bull.

15. The method of any one of embodiments 1-5, wherein the animal is selected from the group consisting of a human, a mouse, a rat, a pig, a hog, a boar, a chicken, a turkey, a duck, a goose, a fowl, a pheasant, a cow, a steer, a bull, a buffalo, and a bison. 16. The method of any one of embodiments 1-15, wherein the isolated strain of Lactobacillus reuteri was isolated by a method comprising: a) suspending the feces in buffer, thereby resulting in a fecal suspension, and b) culturing the fecal suspension in an isolation medium that promotes the growth of Lactobacillus reuteri.

17. The method of embodiment 16, wherein the isolation medium comprises glucose, dipotassium hydrogen phosphate, magnesium sulfate heptahydrate, manganese sulfate tetrahydrate, peptone, yeast extract, meat extract, triammonium citrate, and sodium acetate trihydrate.

18. The method of embodiment 17, wherein the isolation medium further comprises glycerol and vancomycin.

19. The method of any one of embodiments 1-18, wherein the growth medium comprises components that promote the growth of Lactobacillus reuteri.

20. The method of any one of embodiments 1-19, wherein the growth medium comprises vitamins, amino acids, nucleotides, metal salts, acid whey, glycerol, sodium acetate, and polysorbate 80.

21. The method of any one of embodiments 1-20, wherein the growth medium does not comprise glucose.

22. The method of any one of embodiments 1-21, wherein culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of between or between about 28 degrees C and 50 degrees C.

23. The method of any one of embodiments 1-21, wherein culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature of: (i) between or between about 28° C, 29° C, 31° C, 32° C, 33° C, or 34° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C; or (ii) between or between about 35° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C; or (iii) between or between about 36° C and 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C; or

(iv) between or between about 37° C and 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C; or (v) between or between about 37° C and 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, or 45° C; or (vi) at or about 28° C, 29° C, 30° C, 31° C, 32° C, 33° C, 34° C, 35° C, 36° C, 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, or 50° C; or (vii) at or about 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, or 45° C; or (viii) at or about 37° C; or (ix) at or about 45° C. 24. The method of any one of embodiments 1-23, wherein culturing the isolated strain of Lactobacillus reuteri in the growth medium occurs for a duration of time sufficient to result in an optical density (OD) within the range of or of about 10-50.

25. The method of any one of embodiments 1-24, wherein culturing the isolated strain of Lactobacillus reuteri in the growth medium occurs for a duration of time sufficient to result in an optical density (OD) within the range of or of about 10-12.

26. The method of any one of embodiments 1-25, wherein culturing the isolated strain of Lactobacillus reuteri in the growth medium occurs for a duration of time that is: (i) at least or at least about 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 22 hours, 24 hours, 30 hours, 36 hours, 42 hours, or 48 hours; or (ii) is or is about 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, or 20 hours; or (iii) is between or between about 4 hours and 10 hours, 4 hours and 9 hours, 4 hours and 8 hours, 4 hours and 7 hours, 4 hours and 6 hours, 5 hours and 10 hours, 5 hours and 9 hours, 5 hours and 8 hours, or 5 hours and 7 hours; or (iv) is or is about 5 hours, 6 hours, 7 hours, or 8 hours; or (v) is between or between about 8 hours and 14 hours, 8 hours and 13 hours, 8 hours and 12 hours, 9 hours and 14 hours, 9 hours and 13 hours, 9 hours and 12 hours, 10 hours and 14 hours, 10 hours and 13 hours, or 10 hours and 12 hours; or (vi) is or is about 9 hours, 10 hours, 11 hours, 12 hours, or 13 hours.

27. The method of any one of embodiments 1-26, wherein culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature at or about 37 degrees C for a duration of time that is or is about 10 hours, 11, or 12 hours.

28. The method of any one of embodiments 1-26, wherein culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium at a temperature at or about 45 degrees C for a duration of time that is or is about 5 hours, 6 hours, 7 hours, or 8 hours.

29. The method of any one of embodiments 1-28, wherein culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium until CO2 generation has reduced to between or between about 1/3 to 1/2 of peak CO2 generation.

30. The method of any one of embodiments 1-28, wherein culturing the isolated strain of Lactobacillus reuteri in the growth medium comprises incubating the isolated strain in the growth medium until CO2 generation has reduced to about 1/2 of peak CO2 generation. 31. The method of any one of embodiments 1-30, wherein, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an optical density (OD) between or between about 9 and 50.

32. The method of embodiment 31, wherein the OD is between or between about 9 and 45, 10 and 45, 11 and 45, 12 and 45, 13 and 45, 14 and 45, 9 and 40, 10 and 40, 11 and 40, 12 and 40, 13 and 40, 14 and 40, 9 and 35, 10 and 35, 11 and 35, 12 and 35, 13 and 35, 14 and 35, 9 and 30, 10 and 30, 11 and 30, 12 and 30, 13 and 30, 14 and 30, 9 and 25, 10 and 25, 11 and 25, 12 and 25, 13 and 25, 14 and 25, 9 and 20, 10 and 20, 11 and 20, 12 and 20, 13 and 20, 14 and 20, 15 and 50, 20 and 50, 25 and 50, 30 and 50, 35 and 50, 40 and 50,

15 and 45, 20 and 45, 25 and 45, 30 and 45, 35 and 45, 40 and 45, 15 and 40, 20 and 40, 25 and 40, 30 and

40, 35 and 40, 15 and 35, 20 and 35, 25 and 35, 30 and 35, 15 and 30, 20 and 30, or 25 and 30.

33. The method of embodiment 31, wherein the OD is between or between about 9 and 15, 10 and 14, 12 and 20, 18 and 26, 20 and 30, or 22 and 50.

34. The method of any one of embodiments 1-33, wherein, following the culturing of the isolated strain of Lactobacillus reuteri in the growth medium, the population of the isolated strain of Lactobacillus reuteri has an optical density (OD) between or between about 9 and 15 or 10 and 14.

35. The method of embodiment 34, wherein the OD is between or between about 9 and 15.

36. The method of embodiment 34, wherein the OD is between or between about 10 and 14.

37. The method of embodiment 31 or embodiment 32, wherein the OD is between or between about 25 and 50.

38. The method of any one of embodiments 31-34, wherein the OD is or is about 10, 11, 12, 13, or 14.

39. The method of any one of embodiments 1-38, further comprising separating the population of the isolated strain of Lactobacillus reuteri from the growth media.

40. The method of embodiment 39, wherein the separating comprises the use of tangential flow filtration, followed by centrifuging the population, and followed by removing the growth media.

41. The method of any one of embodiments 1-40, wherein the glycerol source is glycerol.

42. The method of any one of embodiments 1-41, wherein the glycerol source is from an organic waste source.

43. The method of embodiment 42, wherein the organic waste source has been pasteurized.

44. The method of any one of embodiments 1-43, wherein the production medium comprises glycerol, a carbohydrate source, and a buffering agent.

45. The method of any one of embodiments 1-44, wherein: the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 60 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 25 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 20 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 15 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 10 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 5 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.5 and 10 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.5 and 5 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.5 and 3 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.5 and 2 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 1 and 10 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 1 and 5 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 1 and 3 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 1 and 2 mM of a carbohydrate source, and between or between about 20 mM and 125 mM of a buffering agent.

46. The method of any one of embodiments 1-45, wherein the production medium comprises glycerol, a carbohydrate source, a buffering agent, sodium chloride, potassium chloride, metals, and iron(II) sulfate. 47. The method of any one of embodiments 1-46, wherein the production medium comprises between or between about 50 rnM and 800 mM glycerol, between or between about 0.1 and 60 mM of a carbohydrate source, between or between about 20 mM and 125 rnM of a buffering agent, between or between about 10 and 60 mM sodium chloride, between or between about 1 mM and 8 mM potassium chloride, one or more metals, and between or between about 2 pM and 40 pM iron(II) sulfate.

48. The method of any one of embodiments 1-47, wherein the production medium comprises between or between about 50 rnM and 800 mM glycerol, between or between about 0.1 and 60 mM lactose, between or between about 20 rnM and 125 mM of a buffering agent, optionally wherein the buffering agent is MOPS or phosphate buffer, between or between about 10 and 60 mM sodium chloride, between or between about 1 mM and 8 mM potassium chloride, metals comprising Al, B, Ca, Co, Cu, Fe, Mg, Mn, Mo, Ni, Se, V, W, and Zn, and between or between about 2 pM and 40 pM iron(II) sulfate.

49. The method of any one of embodiments 44-48, wherein the carbohydrate source comprises a carbohydrate selected from the group consisting of lactose, galactose, sucrose, glucose, maltose, arabinose, and fructose, or any combination thereof.

50. The method of any one of embodiments 44-49, wherein the carbohydrate source is lactose.

51. The method of any one of embodiments 1-44, wherein: the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 60 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 25 rnM of lactose, and between or between about 20 rnM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 rnM and 800 mM glycerol, between or between about 0.1 and 20 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 15 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 rnM glycerol, between or between about 0.1 and 10 mM of lactose, and between or between about 20 rnM and 125 rnM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 rnM glycerol, between or between about 0.1 and 5 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.5 and 10 rnM of lactose, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 rnM and 800 mM glycerol, between or between about 0.5 and 5 rnM of lactose, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.5 and 3 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.5 and 2 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 1 and 10 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 1 and 5 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 1 and 3 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent; or the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 1 and 2 mM of lactose, and between or between about 20 mM and 125 mM of a buffering agent.

52. The method of any one of embodiments 44-51, wherein the buffering agent is MOPS or phosphate buffer.

53. The method of any one of embodiments 44-52, wherein the buffering agent is MOPS pH 7.

54. The method of any one of embodiments 1-53, wherein the production medium comprises glycerol, lactose, MOPS pH 7, polysorbate 80, sodium chloride, potassium chloride, metals, and iron(II) sulfate.

55. The method of any one of embodiments 1-54, wherein the production medium comprises 200 mM glycerol, 1.5 mM lactose, 50 mM MOPS pH 7, 25 mM sodium chloride, 3.125 mM potassium chloride, metals, and 10 pM iron(II) sulfate.

56. The method of any one of embodiments 1-54, wherein the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.5 and 10 mM lactose, between or between about 20 mM and 125 mM of a buffering agent, optionally wherein the buffering agent is MOPS or phosphate buffer, between or between about 10 and 60 mM sodium chloride, between or between about 1 mM and 8 mM potassium chloride, one or more metals, and between or between about 2 pM and 40 pM iron(II) sulfate.

57. The method of any one of embodiments 1-54, wherein the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 60 mM lactose, between or between about 20 mM and 125 mM of a buffering agent, optionally wherein the buffering agent is MOPS or phosphate buffer, between or between about 10 and 60 mM sodium chloride, between or between about 1 mM and 8 mM potassium chloride, one or more metals, and between or between about 2 uM and 40 uM iron(II) sulfate.

58. The method of any one of embodiments 1-54, wherein the production medium comprises between or between about 50 mM and 800 mM glycerol, between or between about 0.1 and 60 mM lactose, between or between about 20 mM and 125 mM of a buffering agent, optionally wherein the buffering agent is MOPS or phosphate buffer, between or between about 10 and 60 mM sodium chloride, between or between about 1 mM and 8 mM potassium chloride, metals comprising Al, B, Ca, Co, Cu, Fe, Mg, Mn, Mo, Ni, Se, V, W, and Zn, and between or between about 2 pM and 40 pM iron(II) sulfate.

59. The method of any one of embodiments 46-57, wherein the metals comprise one or more metals selected from the group consisting of Mg, Mn, Ca, Fe, Zn, Co, Cu, Al, B, Mo, Ni, W, V, and Se.

60. The method of any one of embodiments 46-57, wherein the metals comprise Mg, Mn, Ca, Fe, Zn, Co, Cu, Al, B, Mo, Ni, W, V, and Se.

61. The method of any one of embodiments 46-57, 59, and 60, wherein the metals comprise between or between about 250 pM and 2000 pM Mg, between or between about 0.02 pM and 5 pM Mn, between or between about 50 pM and 600 pM Ca, between or between about 1 pM and 30 pM Fe, between or between about 5 pM and 45 pM Zn, between or between about 0.02 pM and 50 pM Co, between or between about 0.05 pM and 2 pM Cu, between or between about 1 pM and 20 pM Al, between or between about 1 pM and 20 pM B, between or between about 0.02 pM and 2 pM Mo, between or between about 0.02 pM and 2 pM Ni, between or between about 0.001 pM and 0.5 pM W, between or between about 0.5 pM and 10 pM V, and between or between about 0.02 pM and 2 pM Se.

62. The method of any one of embodiments 46-57 and 59-61, wherein the metals comprise between or between about 250 pM and 2000 pM Mg, between or between about 0.02 pM and 5 pM Mn, between or between about 50 pM and 600 pM Ca, between or between about 1 pM and 30 pM Fe, between or between about 5 pM and 45 pM Zn, between or between about 0.02 pM and 1 pM Co, between or between about 0.05 pM and 2 pM Cu, between or between about 1 pM and 20 pM Al, between or between about 1 pM and 20 pM B, between or between about 0.02 pM and 2 pM Mo, between or between about 0.02 pM and 2 pM Ni, between or between about 0.001 pM and 0.5 pM W, between or between about 0.5 pM and 10 pM V, and between or between about 0.02 pM and 2 pM Se.

63. The method of any one of embodiments 1-62, wherein culturing the population in the production medium occurs for a duration of time that results in converting at least or at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the glycerol source to 3-hydroxypropanal. 64. The method of embodiment 63, wherein culturing the population in the production medium occurs for a duration of time that results in converting at least or at least about 90%, 91%, 92%, 93%, 94%, 95%,

96%, 97%, 98%, or 99% of the glycerol source to 3-hydroxypropanal.

65. The method of embodiment 63 or embodiment 64, wherein culturing the population in the production medium occurs for a duration of time that results in converting at or about 100% of the glycerol source to 3-hydroxypropanal.

66. The method of any one of embodiments 1-65, wherein culturing the population in the production medium occurs for a duration of time that is or is at least 30 minutes.

67. The method of embodiment 66, wherein culturing the population in the production medium occurs for a duration of time that is between or between about 30 minutes and 48 hours.

68. The method of embodiment 66 or embodiment 67, wherein culturing the population in the production medium occurs for a duration of time that is between or between about 30 minutes and 5 hours.

69. The method of any one of embodiments 66-68, wherein culturing the population in the production medium occurs for a duration of time that is between or between about 1 hour and 4 hours.

70. The method of any one of embodiments 66-69, wherein culturing the population in the production medium occurs for a duration of time that is between or between about 2 and 3 hours.

71. The method of embodiment 66 or embodiment 67, wherein culturing the population in the production medium occurs for a duration of time that is between or between about 1 and 48 hours, 1 and 42 hours, 1 and 36 hours, 1 and 30 hours, 1 and 24 hours, 1 and 18 hours, 1 and 12 hours, 1 and 6 hours, 2 and 48 hours, 2 and 42 hours, 2 and 36 hours, 2 and 30 hours, 2 and 24 hours, 2 and 18 hours, 2 and 12 hours, 2 and 6 hours, 4 and 48 hours, 4 and 42 hours, 4 and 36 hours, 4 and 30 hours, 4 and 24 hours, 4 and 18 hours, 4 and 12 hours, 8 and 48 hours, 8 and 42 hours, 8 and 36 hours, 8 and 30 hours, 8 and 24 hours, 8 and 18 hours, 8 and 12 hours, 12 and 48 hours, 12 and 42 hours, 12 and 36 hours, 12 and 30 hours, 12 and 24 hours, 12 and 18 hours, 16 and 48 hours, 16 and 42 hours, 16 and 36 hours, 16 and 30 hours, 16 and 24 hours, 20 and 48 hours, 20 and 42 hours, 20 and 36 hours, 20 and 30 hours, 20 and 24 hours, 24 and 48 hours, 24 and 42 hours, 24 and 36 hours, or 24 and 30 hours.

72. The method of embodiment 66, wherein culturing the population in the production medium occurs for a duration of time that is or is at least 30 minutes, 45 minutes, 1 hour, 1.25 hours, 1.5 hours, 1.75 hours, 2 hours, 2.25 hours, 2.5 hours, 2.75 hours, 3 hours, 3.25 hours, 3.5 hours, or 4 hours.

73. The method of embodiment 66, wherein culturing the population in the production medium occurs for a duration of time that is or is at least 4 hours, 5 hours, 6 hours, 8 hours, 10 hours, 12 hours, 16 hours, 20 hours, 25 hours, 30 hours, 36 hours, 42 hours, or 48 hours. 74. The method of embodiment 66, wherein culturing the population in the production medium occurs for a duration of time that is or is about 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 8 hours, 10 hours, 12 hours, 16 hours, 20 hours, 25 hours, 30 hours, 36 hours, 42 hours, or 48 hours.

75. The method of any one of embodiments 1-74, wherein culturing the population in the production medium comprises incubating the population in the production medium at a temperature at or about 37 degrees C.

76. The method of any one of embodiments 1-75, wherein, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or at least 2, 3, 4, 5, 6, 7, or 8 g/L/hour.

77. The method of any one of embodiments 1-76, wherein, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or about 3, 4, 5, 6, 7, 8, 9, or 10 g/L/hour.

78. The method of any one of embodiments 1-76, wherein, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or at least 6 g/L/hour.

79. The method of any one of embodiments 1-76, wherein, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of between or between about 3 and 10 g/L/hour, 3 and 9 g/L/hour, 3 and 8 g/L/hour, 3 and 7 g/L/hour, 4 and 10 g/L/hour, 4 and 9 g/L/hour, 4 and 8 g/L/hour, 4 and 7 g/L/hour, 5 and 10 g/L/hour, 5 and 9 g/L/hour, 5 and 8 g/L/hour, 5 and 7 g/L/hour, 6 and 10 g/L/hour, 6 and 9 g/L/hour, 6 and 8 g/L/hour, or 6 and 7 g/L/hour.

80. The method of any one of embodiments 1-76 and 78, wherein, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of at or at least 10 g/L/hour.

81. The method of any one of embodiments 1-76 and 78, wherein, during the culturing in the production medium, 3-hydroxypropanal is produced at a rate of between or between about 3 and 30 g/L/hour, 3 and 25 g/L/hour, 3 and 20 g/L/hour, 3 and 15 g/L/hour, 5 and 30 g/L/hour, 5 and 25 g/L/hour, 5 and 20 g/L/hour, 5 and 15 g/L/hour, 7 and 30 g/L/hour, 7 and 25 g/L/hour, 7 and 20 g/L/hour, 7 and 15 g/L/hour, 10 and 30 g/L/hour, 10 and 25 g/L/hour, 10 and 20 g/L/hour, 10 and 15 g/L/hour, 15 and 30 g/L/hour, 15 and 25 g/L/hour, 15 and 20 g/L/hour, 20 and 30 g/L/hour, or 20 and 25 g/L/hour.

82. The method of any one of embodiments 4-81, wherein separating the production medium comprising 3-hydroxypropanal from the population comprises centrifuging the population and removing the production medium comprising 3-hydroxypropanal.

83. The method of any one of embodiments 4-82, wherein the separated production medium comprising 3-hydroxypropanal comprises at or at least 6 g/L, 7 g/L, 8 g/L, 9 g/L, 10 g/L, 11 g/L, 12 g/L, 13 g/L, 14 g/L, 15 g/L, 16 g/L, 17 g/L, 18 g/L, 19 g/L, 20 g/L, 21 g/L, 22 g/L, 23 g/L, 24 g/L, 25 g/L, 30 g/L, 40 g/L, 50 g/L, 75 g/L, 100 g/L, 125 g/L, 150 g/L, 175 g/L, or 200 84. The method of any one of embodiments 4-83, wherein the separated production medium comprising 3-hydroxypropanal comprises at or at least 6 g/L, 7 g/L, 8 g/L, 9 g/L, 10 g/L, 11 g/L, 12 g/L, 13 g/L, 14 g/L,

15 g/L, 16 g/L, 17 g/L, 18 g/L, 19 g/L, 20 g/L, 21 g/L, 22 g/L, 23 g/L, or 24 g/L of 3-hydroxypropanal.

85. The method of any one of embodiments 4-84, wherein the separated production medium comprising

3-hydroxypropanal comprises between or between about 6 and 200 g/L, 6 and 150 g/L, 6 and 100 g/L, 6 and 80 g/L, 6 and 60 g/L, 6 and 50 g/L, 6 and 30 g/L, 6 and 20 g/L, 6 and 15 g/L, 10 and 200 g/L, 10 and 150 g/L, 10 and 100 g/L, 10 and 80 g/L, 10 and 60 g/L, 10 and 50 g/L, 10 and 30 g/L, 10 and 20 g/L, 10 and 15 g/L, 15 and 200 g/L, 15 and 150 g/L, 15 and 100 g/L, 15 and 80 g/L, 15 and 60 g/L, 15 and 50 g/L, 15 and 30 g/L, 15 and 20 g/L, 20 and 200 g/L, 20 and 150 g/L, 20 and 100 g/L, 20 and 80 g/L, 20 and 60 g/L, 20 and 50 g/L, 20 and 30 g/L, 30 and 200 g/L, 30 and 150 g/L, 30 and 100 g/L, 30 and 80 g/L, 30 and 60 g/L, 30 and 50 g/L, 40 and 200 g/L, 40 and 150 g/L, 40 and 100 g/L, 40 and 80 g/L, 40 and 60 g/L, 40 and 50 g/L, 50 and

200 g/L, 50 and 150 g/L, 50 and 100 g/L, 50 and 80 g/L, 50 and 60 g/L, 60 and 200 g/L, 60 and 150 g/L, 60 and 100 g/L, or 60 and 80 g/L.

86. The method of any one of embodiments 4-85, wherein the separated production medium comprising 3-hydroxypropanal comprises between or between about 6 g/L and 40 g/L, 6 g/L and 36 g/L, 6 g/L and 32 g/L, 6 g/L and 28 g/L, 6 g/L and 24 g/L, 6 g/L and 20 g/L, 9 g/L and 40 g/L, 9 g/L and 36 g/L, 9 g/L and 32 g/L, 9 g/L and 28 g/L, 9 g/L and 24 g/L, 9 g/L and 20 g/L, 12 g/L and 40 g/L, 12 g/L and 36 g/L, 12 g/L and 32 g/L, 12 g/L and 28 g/L, 12 g/L and 24 g/L, 12 g/L and 20 g/L, 15 g/L and 40 g/L, 15 g/L and 36 g/L, 15 g/L and 32 g/L, 15 g/L and 28 g/L, 15 g/L and 24 g/L, 15 g/L and 20 g/L, 18 g/L and 40 g/L, 18 g/L and 36 g/L, 18 g/L and 32 g/L, 18 g/L and 28 g/L, 18 g/L and 24 g/L, or 18 g/L and 20 g/L of 3-hydroxypropanal.

87. The method of any one of embodiments 4-86, wherein the separated production medium comprising 3-hydroxypropanal comprises between or between about 12 g/L and 18 g/L of 3-hydroxypropanal.

88. The method of any one of embodiments 3-87, wherein converting the 3-hydroxypropanal into propenoic acid comprises converting the 3-hydroxypropanal into prop-2-enal and converting the prop-2 -enal into propenoic acid.

89. The method of embodiment 88, wherein converting the 3-hydroxypropanal into prop-2 -enal comprises acidifying the production medium comprising 3-hydroxypropanal, thereby resulting in an acidified production medium; and converting the 3-hydroxypropanal in the acidified production medium into gaseous prop-2-enal.

90. The method of embodiment 88, wherein converting the 3-hydroxypropanal into prop-2 -enal comprises acidifying the production medium comprising 3-hydroxypropanal or the separated production medium comprising 3-hydroxypropanal, thereby resulting in an acidified production medium; and converting the 3-hydroxypropanal in the acidified production medium into gaseous prop-2-enal.

91. The method of any one of embodiments 88-90, wherein converting prop-2-enal into propenoic acid comprises oxidizing the prop-2-enal in the presence of a catalyst to produce the propenoic acid.

92. The method of any one of embodiments 3-91, wherein the converting the 3-hydroxypropanal into propenoic acid comprises: (i) acidifying the production medium comprising 3-hydroxypropanal, thereby resulting in an acidified production medium; (ii) converting the 3-hydroxypropanal in the acidified production medium into gaseous prop-2-enal; and (iii) oxidizing the prop-2-enal in the presence of a catalyst to produce the propenoic acid.

93. The method of any one of embodiments 3-91, wherein the converting the 3-hydroxypropanal into propenoic acid comprises: (i) acidifying the production medium comprising 3-hydroxypropanal or the separated production medium comprising 3-hydroxypropanal, thereby resulting in an acidified production medium; (ii) converting the 3-hydroxypropanal in the acidified production medium into gaseous prop-2 -enal; and (iii) oxidizing the prop-2 -enal in the presence of a catalyst to produce the propenoic acid.

94. The method of any one of embodiments 89-92, wherein acidifying the production medium comprising 3-hydroxypropanal comprises acidifying the production medium comprising 3-hydroxypropanal with a mineral acid.

95. The method of any one of embodiments 90-93, wherein acidifying the production medium comprising 3-hydroxypropanal or the separated production medium comprising 3-hydroxypropanal comprises acidifying the production medium comprising 3-hydroxypropanal or the separated production medium comprising 3-hydroxypropanal with a mineral acid.

96. The method of embodiment 94 or embodiment 95, wherein the mineral acid is selected from the group consisting of phosphoric acid, hydrochloric acid, sulfuric acid, nitric acid, boric acid, hydrofluoric acid, hydroiodic acid, hydrobromic acid, and perchloric acid.

97. The method of any one of embodiments 92-96, wherein converting the 3-hydroxypropanal in the acidified production medium into gaseous prop-2-enal comprises heating the acidified production medium.

98. The method of embodiment 97, wherein the heating is performed under pressure.

99. The method of embodiment 97 or embodiment 98, wherein the heating is performed under reduced pressure as compared to atmospheric pressure.

100. The method of any one of embodiments 97-99, wherein the heating is performed under a pressure between or between about 40 and 400 millibar (mbar). 101. The method of any one of embodiments 91-100, wherein oxidizing the prop-2 -enal in the presence of a catalyst to produce the propenoic acid comprises the use of vapor-phase oxidation.

102. The method of any one of embodiments 91-101, wherein the catalyst comprise a two-metal catalyst.

103. The method of any one of embodiments 91-100, wherein oxidizing the prop-2 -enal in the presence of a catalyst to produce the propenoic acid comprises reacting the prop-2-enal with an oxygen-containing gas at a temperature that is between or between about 125 and 320 degrees C in the presence of a two-metal catalyst.

104. The method of embodiment 102 or embodiment 103, wherein the two-metal catalyst comprises palladium and copper or silver.

105. The method of any one of embodiments 1-104, wherein the isolated strain of Lactobacillus reuteri has been further genetically engineered after being isolated.

106. The method of embodiment 105, wherein the genetic engineering comprises introducing a gene disruption and/or introducing an exogenous nucleic acid into the isolated strain.

107. 3-hydroxypropanal (reuterin) produced by the method of any one of embodiments 1-106.

108. Propenoic acid produced by the method of any one of embodiments 3-106.

109. An isolated strain of Lactobacillus reuteri, wherein the isolated strain was isolated from feces of an animal.

110. The isolated strain of embodiment 109, wherein the animal is selected from the group consisting of a human, a mouse, a rat, a pig, a hog, a boar, a chicken, a turkey, a duck, a goose, a fowl, a pheasant, a cow, a steer, a bull, a buffalo, and a bison.

111. The isolated strain of embodiment 109 or embodiment 110, wherein the isolated strain of Lactobacillus reuteri was isolated by a method comprising: a) suspending the feces in buffer, thereby resulting in a fecal suspension, and b) culturing the fecal suspension in an isolation medium that promotes the growth of Lactobacillus reuteri.

112. The isolated strain of embodiment 111, wherein the isolation medium comprises glucose, dipotassium hydrogen phosphate, magnesium sulfate heptahydrate, manganous sulfate tetrahydrate, peptone, yeast extract, meat extract, triammonium citrate, and sodium acetate trihydrate.

113. The isolated strain of embodiment 111 or embodiment 112, wherein the isolation medium further comprises glycerol and vancomycin.

114. The isolated strain of any one of embodiments 109-113, wherein the isolated strain is further genetically modified after being isolated from the feces of the animal.

115. A kit comprising an isolated strain of Lactobacillus reuteri, and instructions for use thereof. 116. A kit comprising the isolated strain of Lactobacillus reuteri of any one of embodiments 109-114, and instructions for use thereof.

117. The kit of embodiment 115 or embodiment 116, further comprising a growth medium and/or a production medium.

118. The kit of embodiment 115 or embodiment 116, further comprising one or more components for producing a growth medium and/or production medium.

119. A composition comprising an isolated strain of Lactobacillus reuteri.

120. A composition comprising the isolated strain of Lactobacillus reuteri of any one of embodiments 109-114.

121. A composition comprising the 3-hydroxypropanal (reuterin) of embodiment 107.

122. A composition comprising the propenoic acid of embodiment 108.

123. A kit comprising the 3-hydroxypropanal (reuterin) of embodiment 107.

124. A kit comprising the propenoic acid of embodiment 108.

125. A kit comprising the composition of any one of embodiments 119-122.

V. EXAMPLES

[0233] The following examples are included for illustrative purposes only and are not intended to limit the scope of the invention.

Example 1 : Production of 3-hvdroxyDropanal using an isolated bacterial strain of Lactobacillus reuteri [0234] This example describes an exemplary workflow for the production of 3-hydroxypropanal (reuterin) using an isolated strain of Lactobacillus reuteri, which can then be converted into prop-2-enal under acidic conditions, which can then be converted into propenoic acid using established industry production processes. [0235] A wildtype strain of Lactobacillus reuteri was isolated and selected for use in producing 3- hydroxypropanal (reuterin), which is an intermediate compound in the synthesis of propenoic acid. This strain was isolated from livestock feces by the following process: Feces (1 -2g) from livestock were suspended in buffer (4-10 mL of 50 mM MOPS pH 7 and 50 mM NaCl) and then inoculated into liquid growth in MRS broth (Sigma, Product No. 69966) supplemented with high concentrations of vancomycin and 50 mM glycerol to reduce non-lactobacilli growth and encourage L. reuteri growth. Using densely grown cultures, dilutions were plated to single colonies and those were resuspended and spot plated in duplicate onto new MRS-vancomycin plates and incubated at 37C under N2 atmosphere overnight. Using the grown plates, reuterin was detected using an agar overlay assay with the colorimetric aldehyde-detecting reagent, 2,4-dinitrophenylhydrazine (DNPH), under acidic conditions. Finally, color was developed by neutralizing with sodium hydroxide and reuterin-producing colonies isolated for growth.

[0236] For fermentation using a 14L fermentation tank, two seedstocks (approximately 1.1 mL each) of the isolated strain of Lactobacillus reuteri were thawed on ice for 5 minutes and then 2 mL of the total seedstock was inoculated into 280 mL of an MRS-like broth comprising 10 g/L peptone, 8 g/L meat extract, 4 g/L yeast extract, 100 rnM carbohydrate (glucose, sucrose, lactose, galactose, maltose, arabinose, or fructose), 5 g/L sodium acetate trihydrate, 1 g/L polysorbate 80, 2 g/L dipotassium hydrogen phosphate, 2 g/L triammonium citrate, 0.2 g/L magnesium sulfate heptahydrate, and 0.05 g/L manganese sulfate tetrahydrate. The inoculated seedstock was mixed to homogeneity and then the seedstock was incubated at 37 degrees C overnight for approximately 12-15 hours. For fermentation using a 1.5 L fermentation tank, 100 pL of seedstock was inoculated into 30 mL of a modified form of MRS broth where lactose is supplemented in place of glucose, and incubated at 37 degrees C overnight for approximately 14-15 hours. Following incubation, the seed cultures are confirmed to have an optical density (OD) of approximately 10-12.

[0237] During the seed growth, the 14L tank was inoculated with 8.5 L of acid whey (AW) and pasteurized at 75 degrees C for 40 minutes, followed by cooling to approximately room temperature at 30 degrees C. Pasteurization was carried out either in an autoclave (for 1.5 L fermentations) or inside the fermentation tank (for 15 L fermentations). The AW was used at approximately 0.6x of the total growth media. The AW supplies all of the lactose (sugar) and many of the amino acids that are needed for microbial growth. The AW was then stirred rapidly and slowly fed 12.5 M NaOH and was allowed to equilibrate in order to reach a pH of approximately 5.5, resulting in pH-adjusted AW (pH-AW).

[0238] To provide the remaining components for robust growth, growth supplements were then added to the pH-AW, thereby resulting in a growth media.

[0239] The temperature of the growth media was then acclimated to 37 degrees C while sparging with N2 to remove O2.

[0240] The grown cultures were then mixed to homogeneity. The 14L tank was then inoculated with the 280 mL overnight culture grown from the seedstock, which had an OD of approximately 10-12, or, for the 1.5 L tank, it was then inoculated with the 30 mL overnight culture grown from the seedstock, which had an OD of approximately 10-12.

[0241] For the growth phase, the culture in the growth media was then incubated at 37 degrees C while stirring at 300 RPM under N2 with a controlled pH of 5.5. In some productions, the culture was incubated at 45 degrees C, which increases microbial growth rate and reduces the time of the growth phase from approximately 10-12 hours down to less than 7 hours. In some productions, N2 is only supplied to the culture for part of the growth phase since the microbe does not require N2 and, instead, the N2 is being used to remove the solvated O2 from the media.

[0242] Peak CO2 generation is expected to occur at > 6.5 hours. If performing fed-batch fermentation, at the peak of CO2 generation, neutralized AW media was added at a feed rate of approximately 83 mL/hr for the 14 L tank. For batch and fed-batch, once the CO2 generation has reduced to approximately 1/2 of the max CO2 generation, which occurred after approximately 10-12 hours, the cells were harvested for the production phase. A normalized optical density (OD) was also determined, which is typically 10-15 (2.5-3.75 g/L cell dry weight (CDW)), and the volume of production media that was needed to reach an OD value of 10 was calculated.

[0243] For the 14 L tank, 1.5 mL of FeSO4 (100 mM) was freshly added to the production media as calculated.

[0244] Cells were separated from the growth media using (1) tangential flow filtration (TFF) to reduce the volume of cells to 1/10 of the tank size, and (2) one step of centrifugation to remove the growth media, by centrifuging the sample at approximately 8,000 RPM for 10 min. The cells were then resuspended in room temperature production media at an OD of approximately 10 and were returned to the fermenter at 37 degrees C under N2 while stirring at 100 RPM for 2-3 hours. This allows for the conversion of glycerol to reuterin (3-hydroxypropanal), with a production rate of over 6 g/L/hr of reuterin. The production media is a nutrient-limited media that prevents the microbes from growing, and includes glycerol which is converted into reuterin (3-hydroxypropanal). The composition of the production media used is provided in Table E2, below. The production media can optionally include Tween, such as included in the production medium of Table E2 in this Example.

Table E2. [0245] The cells were then separated from the production media using centrifugation, and the reuterin (3- hydroxypropanal) was distilled under acidic conditions from broth via conversion to acrolein (prop-2-enal). Cell-free production media is acidified with 1 M H2SO4 and distilled under reduced pressure at 50C as compared to atmospheric pressure.

[0246] The acrolein (prop-2 -enal) can then be converted to propenoic acid for industrial applications by oxidizing the prop-2-enal in the presence of catalyst using established industry processes.

Example 2 : Production of 3-hydroxypropanal using an isolated bacterial strain of Lactobacillus reuteri and a reduced concentration of lactose in the production medium

[0247] The isolated strain of Lactobacillus reuteri from Example 1 was cultured under similar conditions with a similar workflow, including similar growth medium and similar production medium, as described in Example 1, except that the concentration of lactose in the production medium was reduced from 25 mM to 1.5 mM, as shown in Table E3. The production media of Table E3 includes a lower concentration of lactose (1.5 mM). The production media can optionally include Tween, such as included in the production medium of Table E2 in this Example.

Table E3. [0248] It was found that reducing the concentration of lactose from 25 mM (as used in Example 1) to 1.5 mM unexpectedly resulted in an approximately 5-15% increase in reu terin yield.

[0249] The present invention is not intended to be limited in scope to the particular disclosed embodiments, which are provided, for example, to illustrate various aspects of the invention. Various modifications to the compositions and methods described will become apparent from the description and teachings herein. Such variations may be practiced without departing from the true scope and spirit of the disclosure and are intended to fall within the scope of the present disclosure.