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Acetohydroxyacid synthase and application thereof

A technology of acetohydroxy acid and synthetase, applied in the field of metabolic engineering

Active Publication Date: 2019-09-13
TIANJIN UNIV OF SCI & TECH
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  • Abstract
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  • Claims
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Problems solved by technology

[0007] In order to overcome the current deficiency that the wild-type acetohydroxy acid synthase is subject to feedback inhibition by L-isoleucine, the present invention provides a mutant of acetohydroxy acid synthase capable of releasing the feedback inhibition of L-isoleucine, its coding gene and its application

Method used

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  • Acetohydroxyacid synthase and application thereof
  • Acetohydroxyacid synthase and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0027] Example 1: Gene ilvBN Encoding Acetohydroxyacid Synthase Relieving L-Isoleucine Feedback Inhibition M the acquisition of

[0028] (1) Screening of mutants resistant to L-isoleucine structural analogues

[0029] ① Preparation of Corynebacterium glutamicum (Corynebacterium glutamicum) ATCC13032 bacterial suspension

[0030] Corynebacterium glutamicum (Corynebacterium glutamicum) ATCC13032 was inoculated into LB liquid medium, cultured at 32°C, 200rpm for 12h, collected by centrifugation, washed with sterile saline for 3 times and then resuspended to make the OD 600 =0.6-0.8, take 10 μL of the bacterial suspension and spread it on the slide.

[0031] ② Plasma mutagenesis at atmospheric pressure and room temperature

[0032] The mutagenesis parameters were as follows: the slide was placed at 2mm from the airflow port, the power was 120W, the airflow was 10SLM, and the action time was 25s.

[0033] ③ Screening of mutant strains resistant to L-isoleucine structural analog...

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Abstract

The present invention relates to an acetohydroxyacid synthase capable of relieving L-isoleucine feedback inhibition, and an application thereof, and belongs to the field of metabolic engineering. Theacetohydroxyacid synthase encoded by an ilvBN<M> gene has the following characteristics: the enzyme relieves the feedback inhibition of L-isoleucine, an ilvBN<M> enzyme activity is free of obvious changes under a condition of a concentration of the L-isoleucine at 0-12 mmol / L, an activity of the acetohydroxyacid synthase is not obviously lowered compared with wild-type ilvBN-encoded acetohydroxyacid synthase, and the acetohydroxyacid synthase can be applied in production of the L-isoleucine.

Description

[0001] Technical field: [0002] The invention relates to a method for releasing L-isoleucine feedback inhibition of acetohydroxyacid synthetase and its application, belonging to the field of metabolic engineering. [0003] Background technique: [0004] L-isoleucine is a branched-chain amino acid and is one of the eight essential amino acids for the human body. L-Isoleucine is a raw material for the synthesis of human hormones and enzymes. It can promote protein synthesis and inhibit its decomposition, and plays a vital role in human life activities. Therefore, L-isoleucine has very broad market and application prospects in industries such as food and medicine. [0005] The synthesis methods of L-isoleucine include extraction method, chemical synthesis method and fermentation method. Industrial production mainly uses hair extraction method and fermentation method to synthesize L-isoleucine. Because the extraction method has the disadvantages of limited source of raw materia...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/10C12N15/54C12P13/06
CPCC12N9/1022C12P13/06C12Y202/01006
Inventor 张成林李英滋韩世宝徐庆阳李燕军张宇芦楠朱福周董解荣陈亭利
Owner TIANJIN UNIV OF SCI & TECH
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