Title:
DEHYDROGENASE MUTANT L283V/L286V AND PREPARATION METHOD THEREFOR AND APPLICATION THEREOF
Document Type and Number:
WIPO Patent Application WO/2023/088098
Kind Code:
A1
Abstract:
Provided are a dehydrogenase mutant L283V/L286V and a preparation method therefor and an application thereof. The amino acid sequence of the mutant L283V/L286V is as shown in SEQ ID NO: 1; or leucine at position 283 and position 286 of dehydrogenase having an amino acid sequence as shown in SEQ ID NO: 3 is mutated into valine at the same time. The dehydrogenase mutant L283V/L286V shows high selectivity in the process of generating S-nornicotine by catalyzing reduction reaction of myosmine in a whole-cell system, has high activity of dehydrogenase and imine reductase, and is short in enzyme reduction time and high in conversion rate. The product S-nornicotine obtained by the reaction has high optical purity, and the difficulty of subsequent purification work can be reduced.
Inventors:
HUANG JIAJUN (CN)
LI RONGXU (CN)
JIANG RUIBING (CN)
WU ZIYING (CN)
HU HAOXUAN (CN)
BAI SHAOYU (CN)
ZHOU DAVID (CN)
LU TONY (CN)
LI RONGXU (CN)
JIANG RUIBING (CN)
WU ZIYING (CN)
HU HAOXUAN (CN)
BAI SHAOYU (CN)
ZHOU DAVID (CN)
LU TONY (CN)
Application Number:
PCT/CN2022/129268
Publication Date:
May 25, 2023
Filing Date:
November 02, 2022
Export Citation:
Assignee:
GOLDEN HEALTH GUANGDONG BIOTECHNOLOGY CO LTD (CN)
TF BIOSYN BIOTECHNOLOGY CO LTD (CN)
TF BIOSYN BIOTECHNOLOGY CO LTD (CN)
International Classes:
C12N9/02; C12N15/10; C12N15/53; C12N15/70; C12P17/16; C12P41/00
Domestic Patent References:
| WO2021180019A1 | 2021-09-16 |
Foreign References:
| CN113801858A | 2021-12-17 | |||
| CN113373188A | 2021-09-10 | |||
| JPS5881791A | 1983-05-17 | |||
| CN113046335A | 2021-06-29 |
Other References:
DATABASE Protein 4 July 2007 (2007-07-04), ANONYMOUS : "NAD(P)-dependent oxidoreductase [Mesorhizobium sp. L48C026A00] ", XP093068963, retrieved from NCBI Database accession no. WP_023799151.1
MA, LIN ET AL.: "Study on the Purification and Properties of Nicotine Dehydrogenase", INDUSTRIAL MICROBIOLOGY, vol. 37, no. 4, 27 August 2007 (2007-08-27), XP009546656
HAEFFNER-GORMLEY LORRAINE, CHEN ZENGDAO, ZALKIN HOWARD, COLMAN ROBERTA F.: "Importance of lysine-286 at the NADP site of glutamate dehydrogenase from Salmonella typhimurium", BIOCHEMISTRY, vol. 31, no. 34, 1 September 1992 (1992-09-01), pages 7807 - 7814, XP093068971, ISSN: 0006-2960, DOI: 10.1021/bi00149a010
MA, LIN ET AL.: "Study on the Purification and Properties of Nicotine Dehydrogenase", INDUSTRIAL MICROBIOLOGY, vol. 37, no. 4, 27 August 2007 (2007-08-27), XP009546656
HAEFFNER-GORMLEY LORRAINE, CHEN ZENGDAO, ZALKIN HOWARD, COLMAN ROBERTA F.: "Importance of lysine-286 at the NADP site of glutamate dehydrogenase from Salmonella typhimurium", BIOCHEMISTRY, vol. 31, no. 34, 1 September 1992 (1992-09-01), pages 7807 - 7814, XP093068971, ISSN: 0006-2960, DOI: 10.1021/bi00149a010
Attorney, Agent or Firm:
GUANGZHOU YUEXIU JILY PATENT & TRADEMARK LAW OFFICE (CN)
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