Method for promoting microbial enzyme process to synthesizing glutathione

A glutathione and microbial enzyme technology, applied in the direction of fermentation, can solve the problems of reducing the biocatalytic efficiency of microbial cells, cumbersome processing process, cell lysis, etc., and achieve the effect of reducing the permeability barrier

Inactive Publication Date: 2006-08-09
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These methods are mainly permeabilization pretreatment, and the treatment procedures are mainly permeabilization treatment-centrifugation-washing and other operations. The treatment process is relatively cumbersome. Phenomena such as cracking, resulting in the loss of microbial intracellular substances during centrifugation, reducing the efficiency of microbial cell biocatalysis

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Recombinant Escherichia coli WSH-KE1 is not permeabilized, that is, in the basic reaction solution of enzymatically synthesizing glutathione, low concentration of permeabilizing agent is not added, and glutathione is synthesized enzymatically by its bacteria Peptides, the other conditions were as described in the instructions, and the final glutathione production was 1.4g / L.

Embodiment 2

[0024] Traditional pretreatment-centrifugation-washing procedures were performed on recombinant Escherichia coli WSH-KE1 with different concentrations of toluene, xylene or polyoxyethylene stearic acid amide, and glutathione was synthesized by its bacterial cell enzyme method, and the rest of the conditions were As stated in the instructions, its optimal glutathione production is only 2.5g / L.

Embodiment 3

[0026] In the reaction system of recombinant Escherichia coli WSH-KE1 enzymatically synthesizing glutathione, 0.25% to 2% (v / v) different final concentrations of xylene were directly added, and the rest of the conditions were as described in the instructions. When the final concentration reaches 1% (v / v), the optimal glutathione production is 3.5g / L.

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Abstract

This invention relates to a method for synthesizing glutathions by a microbe enzyme acceleration method, which takes an example of synthesizing glutathions by reorganiziong colibacillus enzymes to take the reorganized cplibacillus E. coli WSH-KE1 cells as the enzyme source and adds a low concentrated organic solvent or a surface active agent into a reactor directly to reduce the permeability barrier of ectoblasts of cells then to catalyze L-Glu, L-Cys and Gly in the existence of ATP to synthesize glutathions, the synthesized volume of which can reach to 4.8g/L in two hours of reaction.

Description

technical field [0001] A method for promoting the synthesis of glutathione by microbial enzymatic method, by directly adding a penetrating agent to the reaction system, reducing the permeability barrier of the outer membrane of cells, promoting the synthesis of glutathione by microbial cell enzymatic method, and increasing the concentration of glutathione. The peptide yield relates to the technical field of synthesizing high-value useful compounds by using microbial cell enzymes. Background technique [0002] The use of microbial whole-cell biocatalysis to synthesize high-value useful compounds has attracted more and more attention because of its potential for large-scale industrial applications. Such as the enzymatic synthesis of glutathione by genetically engineered bacteria. By cloning the genes gsh I and gsh II encoding γ-glutamylcysteine ​​synthetase and glutathione synthase, and constructing a recombinant Escherichia coli with high glutathione synthesis activity, and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/02
Inventor 堵国成廖鲜艳陈坚朱至
Owner JIANGNAN UNIV
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