Recombinant plasmid for producing pipecolinic acid, genetic engineering strain and method

A technology of genetically engineered bacteria and piperic acid, applied in the field of bioengineering, can solve the problem of no patent application, etc., and achieve the effect of high growth rate and high yield

Active Publication Date: 2017-09-01
CHONGQING UNIV
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  • Abstract
  • Description
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Problems solved by technology

There is no relevant literature report on the production of L-piperidinecarboxylic acid by high-density fermentation, and there is no related patent application

Method used

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  • Recombinant plasmid for producing pipecolinic acid, genetic engineering strain and method
  • Recombinant plasmid for producing pipecolinic acid, genetic engineering strain and method
  • Recombinant plasmid for producing pipecolinic acid, genetic engineering strain and method

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Embodiment Construction

[0044] For the purpose of the invention, the present invention provides the following technical solutions:

[0045] On the one hand, the present invention provides a kind of recombinant plasmid, and this recombinant plasmid is to carry L-lysine α oxidase gene (LysOX), and L-lysine is catalyzed by L-lysine α oxidase, is converted into Δ 1 - piperidine-2-carboxylic acid (Pip2C).

[0046] Preferably, the recombinant plasmid is a pLMAIP-01 vector carrying a complete coding sequence including the L-lysine alpha oxidase gene (LysOX).

[0047] On the other hand, the present invention provides a kind of recombinant plasmid, this recombinant plasmid is after the aforementioned recombinant plasmid pLMAIP-01 and PCR product glucose dehydrogenase gene (gcd) are cut with KpnI and SacI double-digestion, and gene is cleaved by T4DNA ligase The fragment was ligated with the vector fragment (pLMAIP-02).

[0048] Preferably, the recombinant plasmid is a pLMAIP-02 vector carrying complete codi...

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Abstract

The invention discloses a recombinant plasmid for producing L-pipecolinic acid, a high-yield genetic engineering strain capable of enhancing heterogonous synthesis metabolic pathways of L-pipecolinic acid, and a production method of pipecolinic acid. The invention further discloses a preparation method of the recombinant plasmid and the genetic engineering strain. The strategy of constructing the high-yield genetic engineering strain comprises the following steps: adopting pTrc99a single-plasmid multi-gene tandem expression, linking L-lysine-alpha-oxidase genes (LysoX), glucose dehydrogenase genes (gcd), Pip2C reductase genes (DpkA) and lysine transportase genes (lysQ), optimizing a regulatory element for controlling transcription and translation in front of each gene, transforming the constructed plasmid pLMAIP-05 into competent cells of BL21 (DE3) of essential genes (cadA) in a knockout lysine degradation pathway, thereby obtaining the high-yield genetic engineering strain (MLPR08) capable of enhancing heterogonous synthesis metabolic pathways of L-pipecolinic acid. Due to expression of the lysine transportase, the rate of transporting a substrate L-lysine into the cells is improved, the fermentation time is shortened, and synthesis of the L-pipecolinic acid is further promoted.

Description

technical field [0001] The invention belongs to the field of biological engineering. The invention relates to a recombinant plasmid and a genetically engineered bacterium enhanced with L-piperidinecarboxylic acid heterologous synthesis and metabolism pathway. The present application also relates to the use of the above-mentioned recombinant plasmid and genetic engineering bacteria in the preparation of biological products for the production of L-piperidine carboxylic acid by genetic engineering fermentation. Background technique [0002] Energy and environmental protection are important issues facing the world today. The use of microbial fermentation to produce L-piperidine carboxylic acid can reduce environmental pollution and is a new manufacturing method that is in line with the sustainable development of the country and the nation. [0003] L-Pipecolic acid (L-PA) is an important rigid cyclic non-protein amino acid, which can not only limit the conformation of polypept...

Claims

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Application Information

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IPC IPC(8): C12N15/70C12N1/21C12P17/16
CPCC12N9/0004C12N9/0006C12N9/0022C12N9/50C12N15/70C12P17/16C12Y101/9901C12Y104/03014
Inventor 王丹米乐陈五九
Owner CHONGQING UNIV
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