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Preparation method of immobilized enzyme used for synthesizing gamma-L-glutamyl-L-cysteine

A technology of cysteine ​​and immobilized enzymes, applied in the field of preparation of immobilized enzymes for γ-L-glutamyl-L-cysteine ​​synthesis, which can solve the problem of high cost of separation and purification, unsuitability for industrial production, etc. problems, to achieve the effects of high product yield and purity, promotion of adhesion, and easy availability of raw materials

Inactive Publication Date: 2020-04-07
MUDANJIANG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this patent uses a protein purifier in the separation and purification of S-benzyl-γ-L-glutamyl-L-cysteine. A reversed-phase chromatograph is used. Although this method can improve the purity of the product, the cost of separation and purification is high and it is not suitable for industrial production.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] 1. Preparation of immobilized enzyme:

[0037] (1) Pretreatment of ZSM-5 molecular sieve: put 8g ZSM-5 molecular sieve in 5% sodium bicarbonate solution, use ultrasonic device to sonicate for 15min, ultrasonic frequency 40kHz, ultrasonic power 100W, after washing, transfer to 80°C oven for 8h , and then transferred to a horse-boiling furnace for 3 hours at 450°C;

[0038] (2) Preparation of immobilized enzyme: add 2.5g γ-glutamyl transpeptidase to 50mL water, stir and disperse for 10min, add pretreated ZSM-5 molecular sieve, heat to 45°C, keep stirring for 3h, then add 0.5 g titanate coupling agent, continue to heat and stir at 45° C. for 5 h, then filter with suction, rinse the filter residue with water, and dry in vacuum to obtain the immobilized enzyme.

[0039] 2. Synthesis of γ-L-glutamyl-L-cysteine:

[0040] (1) Preparation of S-benzyl-γ-L-glutamyl-L-cysteine: Add 2.1g S-benzyl-L-cysteine ​​to 1.6g L-glutamine (0.011mol) amino acid (0.01mol), and add 50mL of wa...

Embodiment 2

[0044] The titanate coupling agent in Example 1 was replaced with an equivalent amount of zinc dimethacrylate, and the rest of the operations were the same as in Example 1.

[0045] The yield of γ-L-glutamyl-L-cysteine ​​was 88.9%, and the purity was 98.6%.

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Abstract

The invention discloses a preparation method of an immobilized enzyme used for synthesizing gamma-L-glutamyl-L-cysteine, belonging to the technical field of biosynthesis. According to the invention, full contact between gamma-glutamyl transpeptidase and raw materials including L-glutamine and S-benzyl-L-cysteine is promoted in virtue of preparation of the immobilized enzyme, so effective exertionof catalytic activity is ensured. A synthesis method of gamma-L-glutamyl-L-cysteine has the advantages of usage of easily available raw materials, mild reaction conditions, simple and convenient reaction and post-treatment operation, high product yield and purity and low cost, and is suitable for industrial production.

Description

Technical field: [0001] The invention relates to the technical field of biosynthesis, in particular to a method for preparing an immobilized enzyme for synthesizing γ-L-glutamyl-L-cysteine. Background technique: [0002] γ-L-glutamyl-L-cysteine ​​is a biologically active compound containing γ-glutamyl and sulfhydryl groups formed by condensation of L-glutamic acid and L-cysteine ​​through peptide bonds. The dipeptide compound is the precursor compound of the non-protein sulfhydryl compound glutathione in the organism, and has a wide range of uses in the fields of medicine, food and health products. [0003] At present, the preparation methods of γ-L-glutamyl-L-cysteine ​​mainly include chemical synthesis and fermentation. The synthetic route of the chemical synthesis method is long, and it is easy to produce a racemate. At the same time, a large amount of organic solvents will pollute the environment. Although the fermentation method avoids the use of organic solvents, the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12P13/02C07C319/06C07C319/28C07C323/59
CPCC07B2200/07C07C319/06C07C319/28C12N9/1044C12N11/14C12P13/02C12Y203/02013C07C323/59
Inventor 穆寅东周鸿锁闫磊梁军刘志新董建将蔡克瑞胡江平胡静卫冬
Owner MUDANJIANG MEDICAL UNIV
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