Method for improving conversion rate of chiral amine

Abstract

The invention discloses a method for improving conversion rate of chiral amine. The method comprises the following steps: synthesizing and amplifying an optimized ArR-omega TA nucleotide sequence containing enzyme cutting sites, constructing p2A4-ArR-omega TA, p2A4-FA and p2A4-ldh, wherein FA is a gene for expressing formic acid acetyl transferase, and correspondingly introducing escherichia colito obtain recombinant bacteria 1, FA and 6; fermenting to obtain fermentation liquor of the recombinant bacteria 1, FA and 6; centrifuging to obtain fermentation liquor precipitates of the recombinantbacteria 1, FA and 6; and correspondingly preparing crude FA enzyme liquid and crude ldh enzyme liquid; resuspending the fermentation liquor precipitate of the recombinant bacteria 1 by using PBS, adding PLP and NAD< +>, and reacting; and adding the crude FA enzyme liquid, the crude ldh enzyme liquid, formate dehydrogenase, D-alanine, a substrate ketone and CoA, adding a cosolvent, reacting, terminating, extracting and drying. According to the method disclosed by the invention, a byproduct pyruvic acid is removed, and the conversion rate of the chiral amine is improved. The formate dehydrogenase is added to recover a cofactor reduced coenzyme I.

Description

technical field

[0001] The invention belongs to the field of chemistry, in particular to a method for improving the conversion rate of chiral amines. Background technique

[0002] Enantiomerically pure amines have played a vital role in the chemical, pharmaceutical and agrochemical industries in recent years. Chiral amines are widely used in therapeutic drugs, such as neurological drugs, cardiovascular drugs, antihypertensive drugs, anti-infective drugs and vaccines. At present, chemical method, biological resolution method and biological asymmetric synthesis method are the main methods for preparing chiral amines. Asymmetric synthesis has become a research hotspot because of its low pollution to the environment and the theoretical yield can reach 100%. During the asymmetric synthesis of chiral amines using transaminase, the transamination reaction is reversible and the by-product pyruvate will hinder the reaction to produce amines. At present, the coupling system of lact...

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