	US 12,215,372 B2
	Method for asymmetrically preparing L-phosphinothricin by oxidation-reduction reaction through biological multi-enzyme coupling
Yaping Xue, Hangzhou (CN); Feng Cheng, Hangzhou (CN); Liuyu Wang, Hangzhou (CN); Yuguo Zheng, Hangzhou (CN); Jianmiao Xu, Hangzhou (CN); and Shuping Zou, Hangzhou (CN)
Assigned to ZHEJIANG UNIVERSITY OF TECHNOLOGY, Hangzhou (CN)
Appl. No. 17/607,371
Filed by ZHEJIANG UNIVERSITY OF TECHNOLOGY, Hangzhou (CN)
PCT Filed Jun. 8, 2020, PCT No. PCT/CN2020/094841 § 371(c)(1), (2) Date Oct. 28, 2021, PCT Pub. No. WO2021/184557, PCT Pub. Date Sep. 23, 2021.
Claims priority of application No. 202010192774.4 (CN), filed on Mar. 18, 2020.
Prior Publication US 2022/0220516 A1, Jul. 14, 2022
Int. Cl. C12P 7/52 (2006.01); C12N 1/20 (2006.01); C12N 9/06 (2006.01); C12N 9/10 (2006.01); C12N 15/52 (2006.01); C12N 15/70 (2006.01)

CPC C12P 7/52 (2013.01) [C12N 1/20 (2013.01); C12N 9/0024 (2013.01); C12N 9/1096 (2013.01); C12N 15/52 (2013.01); C12N 15/70 (2013.01); C12Y 104/03003 (2013.01); C12Y 206/01019 (2013.01); C12N 2511/00 (2013.01); C12N 2800/101 (2013.01)]

10 Claims

1. A method for asymmetrically preparing L-phosphinothricin by oxidation-reduction reaction through biological multi-enzyme coupling, where D,L-phosphinothricin as a raw material is catalyzed by an enzyme catalysis system to obtain L-phosphinothricin, comprising the step of catalyzing D-phosphinothricin in D,L-phosphinothricin into 2-carbonyL-4-[hydroxy(methyl)phosphono] butyric acid by a D-amino acid oxidase mutant of the enzyme catalysis system and catalytically reducing the 2-carbonyl-4-[hydroxy(methyl)phosphono] butyric acid into L-phosphinothricin by a transaminase of the enzyme catalysis system, and

wherein the D-amino acid oxidase mutant is obtained by mutation of D-amino acid oxidase in wild strain Rhodotorula taiwanensis at one of the following sites:

(a) M213S-N54V-F58E-D207A;

(b) M213S-N54V-F58E-D207A-S60T.