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Method for biosynthesis of glutathione by using yeast

A technology for glutathione and biosynthesis, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problem that the concentration of glutathione in the product is difficult to further increase, and achieve increased yield and reduced production cost, cost reduction effect

Inactive Publication Date: 2011-07-20
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In the past, in the process of enzymatic synthesis, the three precursor amino acids were added at the same time. The disadvantage is that a certain concentration of glutathione is generated during the reaction process, and then feedback inhibition occurs on γ-GCS, so that the product glutathione It is difficult to further increase the concentration of

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Reaction with waste yeast from brewery fermentation without permeabilization treatment

[0032] After centrifuging the fermentation liquid containing beer yeast at 1050×g for 5 minutes, take 10 g of wet yeast and add it to 100 mL of reaction liquid. The specific components of the reaction liquid are: potassium phosphate buffer 0.2 mol / L (pH value 7.0), MgCl 2 ·6H 2 O20mmol / L, CaCl 2 2H 2 O 10mmol / L, glucose 0.4mol / L, L-glutamic acid 10mmol / L, L-cysteine ​​10mmol / L, glycine 20mmol / L, reacted at 37°C for 36 hours to obtain glutathione The concentration is 750.5 mg / L, and the yield of cysteine ​​is 24.4%.

Embodiment 2

[0033] Example 2 Reaction with brewery waste yeast for permeabilization

[0034] Use the same wet yeast cells as in Example 1, but freeze in the refrigerator at -20°C for 24 hours before the reaction, and thaw at 37°C before use to improve the permeability of the cells;

[0035] The reaction steps are the same as in Example 1, the obtained glutathione concentration is 1028 mg / L, and the yield of cysteine ​​is 33.5%.

Embodiment 3

[0036] Example 3 Two-step conversion reaction with permeabilized brewery waste yeast

[0037] Yeast cells and their permeabilization treatment are the same as in Example 2;

[0038] Permeabilized cells undergo a two-step transformation reaction:

[0039] The first step of conversion reaction: the reaction solution used is the same as the reaction solution described in Example 1 except that glycine is not added, and the reaction is carried out at 37° C. for 7 hours;

[0040] The second step conversion reaction: add glycine to the reaction solution of the first step conversion reaction to make the concentration reach 20mmol / L, and continue to react for 29 hours at 37°C, the concentration of glutathione obtained is 1569mg / L, about The yield of cysteine ​​was 51.1%.

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Abstract

The present invention relates to a method for synthesizing glutathione using yeast cells. Gamma-GCS and GS in yeast cells are used, glucose is used as the energy, an intracellular glucolysis system is used to produce triphosadenine, glutamic acid, cysteine and glycin in reaction liquid are synthesized to form glutathione; saccharomyces cerevisiae, baker's yeast, candida and other yeast cells are permeabilized and then react in a reaction liquid composed of phosphate buffer, MgCl2.6H2O, CaCl2.2H2O, glucose, L-glutamic acid and L-cysteine, gamma-L-glutamoyl L- cysteine is produced, then glycin is added to produce glutathione; in the two-stage reaction, the inhabitation effect of glutathione to gamma-GCS catalytic reaction is greatly reduced; glutathione produced by the method has obviously improved product concentration and reaction yield.

Description

【Technical field】 [0001] The invention relates to the technical field of biocatalysis, in particular to a method for using yeast to biosynthesize glutathione. 【Background technique】 [0002] Glutathione (GSH) is an active substance with important physiological functions. It is composed of three amino acids: glutamic acid (L-Glu), cysteine ​​(L-Cys) and glycine (Gly). The chemical name is γ-L-glutamyl-L-cysteinyl-glycine, and the molecular formula is C 10 h 18 o 6 N 3 S. Glutathione has two forms, reduced form and oxidized form. It mainly exists in the reduced form in the body, and oxidized glutathione is reduced to reduced glutathione by glutathione reductase. Glutathione is widely distributed in animals, plants, and microorganisms. It has unique physiological functions in organisms. It participates in the synthesis of proteins and ribonucleic acids, the transport of oxygen and nutrients, the maintenance of endogenous enzymes, and the participation in tricarboxylic acid...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/02C12R1/84C12R1/72C12R1/645C12R1/865
Inventor 叶勤李志敏李娓
Owner EAST CHINA UNIV OF SCI & TECH
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