Title:
ALKALINE PROTEASE MUTANT HAVING HIGH ACTIVITY AT LOW TEMPERATURE AND USE THEREOF
Document Type and Number:
WIPO Patent Application WO/2024/077702
Kind Code:
A1
Abstract:
An alkaline protease mutant having an improved enzyme activity obtained by means of iterative saturation mutagenesis using overlapping PCR technology, and a preparation therefor and the use thereof. The alkaline protease gene apr from Bacillus clausii is subjected to iterative saturation mutagenesis by using overlapping PCR technology, and the mutant, which has high levels of activity at a low temperature, is screened out and is efficiently expressed and prepared in Bacillus amyloliquefaciens, so that the problem whereby alkaline protease has a low level of activity in a low-temperature washing environment is solved.
Inventors:
LU FUPING (CN)
LIU YIHAN (CN)
LI YU (CN)
MA XIANGYANG (CN)
LIU FUFENG (CN)
ZHANG HUITU (CN)
WANG HONGBIN (CN)
LIU YIHAN (CN)
LI YU (CN)
MA XIANGYANG (CN)
LIU FUFENG (CN)
ZHANG HUITU (CN)
WANG HONGBIN (CN)
Application Number:
PCT/CN2022/131434
Publication Date:
April 18, 2024
Filing Date:
November 11, 2022
Export Citation:
Assignee:
TIANJIN UNIV OF SCIENCE AND TECHNOLOGY (CN)
International Classes:
C12N1/21; C12N9/54; C12N15/10; C12N15/57; C12N15/75; C12R1/07; C12R1/10
Foreign References:
| CN112501149A | 2021-03-16 | |||
| CN112458072A | 2021-03-09 | |||
| CN111373039A | 2020-07-03 | |||
| CN112662652A | 2021-04-16 |
Other References:
LI XINYUE, REN SHAODONG, SONG GUANGCHAO, LIU YIHAN, LI YU, LU FUPING: "Novel Detection Method for Evaluating the Activity of an Alkaline Serine Protease from Bacillus clausii", JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, AMERICAN CHEMICAL SOCIETY, US, vol. 70, no. 12, 30 March 2022 (2022-03-30), US , pages 3765 - 3774, XP093158376, ISSN: 0021-8561, DOI: 10.1021/acs.jafc.2c00358
TEPLYAKOV, A.V. ET AL.: "Protein Engineering of The High-alkaline Serine Protease PB92 from Bacillus Alcalophilus: Functional and Structural Consequences of Mutation at The S4 Substrate Binding Pocket", PROTEIN ENGINEERING, vol. 5, no. 5, 31 July 1992 (1992-07-31), XP008048592
LI JIALIN, JIANG LUYING, CAO XUE, WU YIFAN, LU FUPING, LIU FUFENG, LI YU, LIU YIHAN: "Improving the activity and stability of Bacillus clausii alkaline protease using directed evolution and molecular dynamics simulation", ENZYME AND MICROBIAL TECHNOLOGY, STONEHAM, MA, US, vol. 147, 1 June 2021 (2021-06-01), US , pages 109787, XP093158378, ISSN: 0141-0229, DOI: 10.1016/j.enzmictec.2021.109787
TEPLYAKOV, A.V. ET AL.: "Protein Engineering of The High-alkaline Serine Protease PB92 from Bacillus Alcalophilus: Functional and Structural Consequences of Mutation at The S4 Substrate Binding Pocket", PROTEIN ENGINEERING, vol. 5, no. 5, 31 July 1992 (1992-07-31), XP008048592
LI JIALIN, JIANG LUYING, CAO XUE, WU YIFAN, LU FUPING, LIU FUFENG, LI YU, LIU YIHAN: "Improving the activity and stability of Bacillus clausii alkaline protease using directed evolution and molecular dynamics simulation", ENZYME AND MICROBIAL TECHNOLOGY, STONEHAM, MA, US, vol. 147, 1 June 2021 (2021-06-01), US , pages 109787, XP093158378, ISSN: 0141-0229, DOI: 10.1016/j.enzmictec.2021.109787
Attorney, Agent or Firm:
BEIJING RISINGMARK INTELLECTUAL PROPERTY AGENCY (CN)
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