Genetically engineered bacterium, and applications thereof in preparation of L-phosphinothricin

Abstract

The invention discloses a genetically engineered bacterium, and applications thereof in preparation of L-phosphinothricin. The genetically engineered bacterium comprises Escherichia coli and recombinant plasmids transferred into Escherichia coli; the recombinant plasmids contain transaminase gene and pyridoxal phosphate synthetic route enzyme gene; the nucleotide sequence of the transaminase geneis represented by SEQ ID NO.1; the nucleotide sequence of the pyridoxal phosphate synthetic route enzyme gene is represented by SEQ ID NO.2-5. According to the applications, genetic engineering technology is adopted to express ribose-5-phosphate pathway synthesis key gene PdxST in Escherichia coli, or enhance Escherichia coli self PLP synthesis pathway key gene epd, pdxJ, and dxs expression; preparation of the coenzyme reinforced transaminase genetically engineered bacterium through construction is capable of increasing intracellular coenzyme PLP content obviously, avoiding adding of externally-sourced PLP in production of L-phosphinothricin using the genetically engineered bacterium, increasing transaminase enzyme activity and stability obviously, prolonging transaminase half life, reducing production cost, and increasing L-phosphinothricin production efficiency.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a genetically engineered bacterium and its application in preparing L-glufosinate-ammonium. Background technique [0002] Transaminase (EC 2.6.1.X) is a class of enzymes that catalyze transamination, which can catalyze the transfer of the amino group on the amino donor (amino acid or amine) to the prochiral acceptor ketone or ketoacid to obtain a chiral amine or chiral amino acids, and by-product ketones or ketoacids ( figure 1 ), has won the favor of many researchers because of its high selectivity, high conversion rate and mild reaction conditions. [0003] Synthesis of R-sitagliptin catalyzed by transaminases (Savile CK, Janey JM, Mundorff EC, Moore JC, Tam S, Jarvis WR, Colbeck JC, Krebber A, Fleitz FJ, Brands J, Devine PN, Huisman GW, Hughes GJ. Biocatalytic asymmetric synthesis of chiral amines from ketones applied to sitagliptin manufacture. Science 20...

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Problems solved by technology

[0011] The invention provides a genetically engineered bacterium and its application in the preparation of L-glufosinate-ammonium, by constructing a genetically engineered bacterium capable of expressing pyridoxal phosphate (PLP) in a large amount in Escherichia coli cells without affecting transaminase activity. Solve the problem of exogenous addition of pyridoxal phosphate (PLP) in the preparation process of chiral amino acids in the prior art, greatly reducing production costs

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