A kind of isopropylmalate synthase and its application

A malate synthase, isopropyl technology, applied in the field of metabolic engineering, can solve the problems of unstable fermentation performance, slow growth, unstable genetic traits, etc.

Active Publication Date: 2020-07-24
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the existing industrial production strains of L-leucine are mainly obtained by mutagenesis, and have deficiencies such as nutritional deficiencies, slow growth, unstable genetic traits, etc., resulting in long fermentation cycles, unstable fermentation performance, low yield and conversion rate And other issues

Method used

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  • A kind of isopropylmalate synthase and its application
  • A kind of isopropylmalate synthase and its application
  • A kind of isopropylmalate synthase and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: The gene leuA encoding 2-isopropylmalate synthase that relieves L-leucine feedback inhibition M the acquisition

[0021] (1) Screening of mutants resistant to L-leucine structural analogues

[0022] ① Preparation of Corynebacterium glutamicum (Corynebacterium glutamicum) ATCC13032 bacterial suspension

[0023] 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.

[0024] ② Plasma mutagenesis at atmospheric pressure and room temperature

[0025] 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 20s.

[0026] ③ Screening of mutant strains resistant to L-leucine structural analog leu...

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Abstract

The invention relates to a 2-isopropylmalate synthetase for removing feedback inhibition of L-leucine and an application of the 2-isopropylmalate synthetase, and belongs to the field of metabolic engineering. The leuA<M> gene coded 2-isopropylmalate synthetase has the following characteristics: the synthetase removes the feedback inhibition effect of the L-leucine on the synthetase; under the condition that the concentration of the L-leucine is 0-15 mmol/L, the LEUA<M> activity is not obviously changed; the activity of the 2-isopropylmalate synthetase is not obviously reduced compared with that of a wild LeuA<M> coded 2-isopropylmalate synthetase; and the 2-isopropylmalate synthetase can be applied to the production of the L-leucine.

Description

Technical field: [0001] The invention relates to an isopropylmalate synthetase for releasing L-leucine feedback inhibition and application thereof, which belongs to the field of metabolic engineering. Background technique: [0002] L-leucine belongs to branched-chain amino acids and is one of the eight essential amino acids for humans and vertebrates. In living organisms, L-leucine is used for proteases, hormones, etc. In addition, L-leucine also has functions such as promoting muscle synthesis. Therefore, L-leucine is widely used in medicine, food, animal husbandry and other fields, and has broad market and application prospects. [0003] The existing synthesis methods of L-leucine include hair hydrolysis extraction method and fermentation method, wherein fermentation method is the main synthesis method. However, isopropylmalate synthase, a key enzyme in the L-leucine synthesis pathway, is feedback-inhibited by L-leucine, making it difficult for L-leucine to accumulate e...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/10C12N15/54C12P13/06
CPCC12N9/1025C12P13/06C12Y203/03013
Inventor 张成林徐庆阳李燕军张宇李英滋朱福周卢楠韩世宝董解荣王子申徐昊李子翼
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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