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Alcohol dehydrogenase LC3 and gene and application thereof

A technology of alcohol dehydrogenase and glucose dehydrogenase, which is applied to alcohol dehydrogenase LC3 and its gene and application fields, and can solve problems such as rare conversion rate of enzymes

Active Publication Date: 2016-05-04
百开盛(上海)生物科技有限公司
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Problems solved by technology

[0007] Most of the oxidoreductases that can catalyze 4-chloro-3-hydroxybutyrate ethyl (CHBE) from 4-chloroacetoacetate (COBE) to optical alcohols are enzymes that generate (S)-CHBE. The enzymes that generate (R)-CHBE are relatively rare and the conversion rate is relatively low

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  • Alcohol dehydrogenase LC3 and gene and application thereof
  • Alcohol dehydrogenase LC3 and gene and application thereof
  • Alcohol dehydrogenase LC3 and gene and application thereof

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Embodiment Construction

[0037] The present invention will be further described below in conjunction with specific embodiments. It should be understood that the following examples are only used to illustrate the present invention but not to limit the scope of the present invention.

[0038] 1. Amplification of the alcohol dehydrogenase gene lc3 derived from Lactobacillus reetii

[0039] Alcohol dehydrogenase gene lc3 of the present invention is derived from CCTCCM2011381 Lactobacillus curieae sp.Nov., based on the homology comparison of NCBI (http: / / www.ncbi.nlm.nih.gov / Blast.cgi), The inventors of the present application found a diacetyl reductase derived from Lactobacillus parafarraginis with the highest similarity of 81%.

[0040] The alcohol dehydrogenase LC3 of the present invention belongs to the family of short-chain dehydrogenases, and the nucleotide sequence of the alcohol dehydrogenase gene lc3 encoding the alcohol dehydrogenase LC3 is shown in SEQ ID NO: 1, with a size of 777bp.

[0041] ...

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Abstract

The invention provides an alcohol dehydrogenase LC3 and a gene and application thereof. An amino acid sequence of the alcohol dehydrogenase LC3 is shown in SEQ ID NO:2. An alcohol dehydrogenase gene lc 3 is extracted from lactobacillus curieae, a gene engineering strain is established, the heterologous expression of the alcohol dehydrogenase gene lc 3 is realized, and the enzymatic characteristic of the produced alcohol dehydrogenase LC3 is excellent. A co-expression gene engineering strain containing a glucose dehydrogenase gdh gene and the alcohol dehydrogenase gene lc3 is also successfully established, so as to realize the co-expression of glucose dehydrogenase and alcohol dehydrogenase. The alcohol dehydrogenase LC3 has the characteristics that the alcohol dehydrogenase LC3 and the gene engineering strain are applied to the production of chiral alcohol(R)-4-chloride-3hydroxy ethyl butyrate and other chiral alcohol products by a biological enzyme synthesizing method; the substrate concentration is high, the reaction rate is high, and the convenience is realized.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to an alcohol dehydrogenase LC3 and its gene and application. Background technique [0002] Ketoreductase widely exists in higher animals, plants and lower microorganisms. Ketoreductase exhibits high chemical, regio, and enantioselectivity in catalyzing ketones and alcohols. It has been favored by many scholars and has become an indispensable catalyst in the field of biocatalysis. [0003] Alcohol dehydrogenase (ADH) family-derived enzymes play an important role in the synthesis of chiral compounds. Alcohol dehydrogenase has redox ability to aldehydes, ketones, monosaccharides and ketone steroids. Alcohol dehydrogenases can be divided into two superfamilies, zinc ion-dependent ADH and ion-independent ADH (short-chain alcohol dehydrogenase). The most studied is the short-chain alcohol dehydrogenase family. Ketoreductase, as a branch of the oxidoreductase system, uses reduc...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/04C12N15/53C12N15/70C12N1/21C12P41/00C12P7/62C12R1/19
CPCC12N9/0006C12N9/0008C12N15/70C12N2800/101C12P7/62C12P41/002C12Y101/01118C12Y101/01119
Inventor 王华磊张一平魏东芝
Owner 百开盛(上海)生物科技有限公司
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