CPC C12N 9/0016 (2013.01) [C12N 9/0006 (2013.01); C12P 13/04 (2013.01); G16B 30/10 (2019.02); C12Y 104/01002 (2013.01); C12Y 104/01003 (2013.01); C12Y 104/01004 (2013.01)] | 6 Claims |
1. A method for preparing L-phosphinothricin, characterized in that, 2-carbonyl-4-(hydroxymethylphosphono)-butyric acid as a substrate is allowed to react as catalyzed by a catalyst in the presence of an inorganic amino donor, a coenzyme regeneration and circulation material and a corresponding co-substrate to obtain L-phosphinothricin;
the catalyst is one of the following:
(1) an NADH phosphinothricin dehydrogenase mutant;
(2) a recombinant bacterium capable of expressing the NADH phosphinothricin dehydrogenase mutant or a crude enzyme liquid obtained by lysis of the recombinant bacterium;
wherein the NADH phosphinothricin dehydrogenase mutant is obtained by mutating a wild-type phosphinothricin dehydrogenase obtained from Lysinibacillus composti, wherein the NADH phosphinothricin dehydrogenase mutant has the amino acid sequence as set forth in SEQ ID No.2, excepting that the NADH-preferring phosphinothricin dehydrogenase mutant has a mutation site selected from one of the following:
(1) A144G-V375F-M91A;
(2) A144G-V345A-M91A.
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