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New linoleic acid isomerase gene, and vector and strain containing same

A technology of linoleic acid isomerase and conjugated linoleic acid, which is applied in the field of bioengineering, can solve the problems of low CLA production, achieve the effects of increasing production, improving catalytic efficiency, and reliable biological safety

Inactive Publication Date: 2013-05-29
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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Problems solved by technology

[0004] Although the expression conditions of PAI have been explored in various systems, the yield of CLA is relatively low, mainly because most organisms tend to use some of these codons in the process of expressing heterologous genes, almost every All species exhibit some degree of preference for codon usage

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  • New linoleic acid isomerase gene, and vector and strain containing same
  • New linoleic acid isomerase gene, and vector and strain containing same
  • New linoleic acid isomerase gene, and vector and strain containing same

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Embodiment Construction

[0025] Below in conjunction with embodiment, following embodiment is illustrative, not limiting, can not limit protection scope of the present invention with following embodiment.

[0026] The content of the present invention includes: an expression plasmid with the copai gene after codon optimization, a Saccharomyces cerevisiae strain capable of producing t10,c12-CLA containing the copai expression plasmid, a construction method of the Saccharomyces cerevisiae strain and the Saccharomyces cerevisiae strain Method for producing t10,c12-CLA.

[0027] The natural linoleic acid isomerase gene pai of the present invention is derived from Propionibacterium acnes, the strain preservation number is ATCC 6919 (purchased from Guangdong Microbial Culture Collection Center), and the GenBank number of the gene is AX062088 .1, analyzing the codon bias of Saccharomyces cerevisiae, performing codon optimization, and obtaining the new gene sequence copai, the nucleotide sequence of which is s...

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Abstract

The invention relates to a method for expressing a linoleic acid isomerase by using a Saccharomyces cerevisiae surface display system and synthesizing t10, c12-conjugated linoleic acid through the catalytic action of the enzyme. The invention performs codon optimization on a linoleic acid isomerase gene (pai) derived from Propionibacterium acnes to obtain a brand new linoleic acid isomerase gene (copai), construct an expression vector containing the copai and obtain a Saccharomyces cerevisiae strain containing the expression vector. According to the invention, a microbial surface display system is used to express the linoleic acid isomerase (coPAI) on the surface of a Saccharomyces cerevisiae cell, so that an extracellular substrate can be in full contact with the enzyme, thereby improving the catalytic efficiency; and the single isomer having physiological activity can be obtained, the separation and purification step can be omitted, and the production cost can be lowered.

Description

technical field [0001] The invention belongs to the technical field of bioengineering and relates to the optimization and construction of genes, in particular to a new linoleic acid isomerase gene, a carrier and a bacterial strain containing the gene. Background technique [0002] Conjugated linoleic acid (CLA) is octadecadienoic acid with a conjugated double bond, and is a series of positions and conformations derived from the essential fatty acid linoleic acid (LA) The general term for isomers. CLA has various physiological activities such as anti-cancer, anti-atherosclerosis, lowering cholesterol, regulating blood sugar, enhancing the body's immunity, and promoting growth. Among them, t10,c12-CLA is one of the most physiologically active isomers. In nature, CLA is widely found in meat and dairy products of ruminants, but the content is extremely low. [0003] At present, the industrial production of CLA mainly adopts the chemical isomerization method, that is, the base...

Claims

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Application Information

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IPC IPC(8): C12N15/61C12N15/63C12N1/19C12N9/90C12P7/64C12R1/865
Inventor 刘浩何希宏李凡
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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