A kind of method that utilizes co-expression recombinant strain to transform and produce α-ketoglutarate

Abstract

The invention discloses a method for transforming and producing α-ketoglutarate by using a co-expression recombinant strain, and belongs to the field of biotechnology. The present invention selects a new type of L-glutamate oxidase from Streptomyces, induces expression and purifies it in Escherichia coli, and conducts enzymatic properties research. The optimum reaction temperature is 30~45℃, V max 100~150 U / mg, K m 8~10 mM. The original gene sequence of L-glutamate oxidase was codon-optimized and co-expressed with the catalase gene derived from Escherichia coli to construct a co-expression recombinant strain. The recombinant strain was used as the whole cell catalyst to convert L-glutamic acid (salt), and the yield of α-ketoglutarate reached 76.08 g / L after 9 h of reaction, and the molar conversion rate was 96.8%. The invention solves the problems of cumbersome production steps, low yield and environmental pollution of α-ketoglutaric acid, realizes high-yield and one-step production of α-ketoglutaric acid, and has high industrial application value.

Description

technical field [0001] The invention relates to a method for transforming and producing α-ketoglutarate by using a co-expression recombinant strain, and belongs to the field of biotechnology. Background technique [0002] L-glutamate oxidase (LGOX) is a flavoprotease with flavin adenine dinucleotide (FAD) as a prosthetic group, which can be used without the addition of exogenous cofactors. The specific oxidation of glutamic acid to hydrogen peroxide, ammonia and α-ketoglutarate [1] . α-Ketoglutarate (α-KG), as an important dibasic acid in the tricarboxylic acid cycle and amino acid metabolism, plays an important role in amino acid formation and nitrogen transport, and is widely used in medicine, fine chemicals, food and animals Feed and other fields [2] . [0003] At present, the production methods of α-KG include chemical synthesis, fermentation, and biocatalysis. Traditional α-KG production adopts chemical synthesis method, but the harmful reagents such as strong acid...

AI Technical Summary

Problems solved by technology

[0012] In order to solve the above-mentioned technical problems, the object of the present invention is to provide a method for transforming and producing α-ketoglutaric acid by co-expressing recombinant strains, which can efficiently convert L-glutamic acid (salt) into α-ketoglutarate Diacid, thereby solving the problems of high production cost, long cycle and low efficiency in the industrial production of α-ketoglutaric acid, which is conducive to the large-scale production of α-ketoglutaric acid.

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