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Biotransformation method of L-cysteine

A technology of biotransformation and cysteine, applied in the field of biotransformation of L-cysteine, can solve the problems of limited source of raw materials, complex process, environmental pollution, etc. good effect

Inactive Publication Date: 2018-10-23
武汉茵茂特生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the main process in China is the reduction method after hydrolysis of hair. However, the source of raw materials is limited, the energy consumption is high, and a large amount of irritating gas and waste acid are produced during the production process, which seriously pollutes the environment.
The DL-ATC metabolic pathway method is complicated in process and high in cost. Therefore, it is very meaningful to develop a conversion method of L-cysteine ​​with low cost, less pollution and suitable for industrial production.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The preparation process of L-cysteine ​​comprises the following steps:

[0040] The biotransformation method of L-cysteine, reaction is carried out in 1L shaking flask, and reaction system is controlled as 300mL, with 3-chloro-pyruvate of 30g as substrate, with MOPS buffer solution (with 3-morpholine propanesulfonate Acid and Na 2 PO 4 The normal saline solution of the buffer pair) is a solvent, with 23g / L derived from the 3-chloro-L-alanine dehydrogenase of Saccharomyces cerevisiae, the genetically engineered bacterium that derives from the formate dehydrogenase co-expression of Candida boidinii Cells as catalysts, exogenous 3-chloro-L-alanine dehydrogenation in genetically engineered bacteria co-expressed with 3-chloro-L-alanine dehydrogenase from Saccharomyces cerevisiae and formate dehydrogenase from Candida boidinii The coding gene of the enzyme is the nucleotide sequence shown in SEQ ID NO: 1 in the sequence listing; the 3-chloro-L-alanine dehydrogenase derived ...

Embodiment 2

[0043] The preparation process of L-cysteine ​​comprises the following steps:

[0044] The biotransformation method of L-cysteine, reaction is carried out in 1L shaking flask, and reaction system is controlled as 300mL, with 3-chloro-pyruvate of 30g as substrate, with carbonate buffer solution as solvent, with 25g / The 3-chloro-L-alanine dehydrogenase derived from Saccharomyces cerevisiae and the whole cell of genetically engineered bacteria co-expressed with formate dehydrogenase derived from Candida boidinii were used as catalysts, and the 3-chloro-L-alanine derived from Saccharomyces cerevisiae The coding gene of exogenous 3-chloro-L-alanine dehydrogenase in the genetically engineered bacteria co-expressed with alanine dehydrogenase and formate dehydrogenase derived from Candida boidinii is shown in SEQ ID NO: 1 in the sequence listing The nucleotide sequence; The 3-chloro-L-alanine dehydrogenase derived from Saccharomycescerevisiae, the coding gene of exogenous formate deh...

Embodiment 3

[0047] The preparation process of L-cysteine ​​comprises the following steps:

[0048]The biotransformation method of L-cysteine, the reaction is carried out in a 1L shake flask, the reaction system is controlled to 300mL, with 30g of 3-chloro-pyruvate as the substrate, phosphate buffer solution as the solvent, and 20g / L 3-Chloro-L-alanine dehydrogenase derived from Saccharomyces cerevisiae and formate dehydrogenase derived from Candida boidinii were co-expressed as whole cells of genetically engineered bacteria as catalysts, and 3-chloro-L-alanine derived from Saccharomyces cerevisiae The coding gene of exogenous 3-chloro-L-alanine dehydrogenase in the genetically engineered bacteria co-expressed with amino acid dehydrogenase and formate dehydrogenase derived from Candida boidinii is shown in SEQ ID NO: 1 in the sequence listing Nucleotide sequence; the coding gene of exogenous formate dehydrogenase in the genetically engineered bacteria derived from the 3-chloro-L-alanine de...

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Abstract

The invention discloses a biotransformation method of L-cysteine. The biotransformation method comprises: forming a reaction system by 3-chloro-pyruvic acid as a substrate and a solvent, adding a biocatalyst, coenzyme, an additive and an ammonium salt to the reaction system, carrying out a biotransformation reaction to obtain a transformed liquid containing 3-chloro-L-alanine, purifying to obtaina 3-chloro-L-alanine crystal, and carrying out a beta-substitution reaction on the 3-chloro-L-alanine as a substrate by using L-cysteine desulfhydrase as a catalyst to generate the L-cysteine. According to the present invention, the process is simple, has no special requirements on equipment, and is suitable for industrial production.

Description

technical field [0001] The invention relates to the field of amino acid preparation, in particular to a biotransformation method of L-cysteine. Background technique [0002] The efficacy of L-cysteine ​​and its complexes in medical treatment has attracted increasing attention. In recent years, studies have suggested that L-cysteine ​​mainly has the following physiological functions: 1. L-cysteine ​​in vivo Acid has effective detoxification effect on many harmful substances and toxins such as formaldehyde, acetaldehyde, chloroform, carbon tetrachloride, lead, cadmium, mercury chloride, peroxide ester and tetrodotoxin; 2. Use L-cysteine It can effectively prevent and improve the damage of radioactive substances to the body; 3. When inflammation or allergy occurs, the activity of thiol enzymes such as cholinesterase in the body decreases, and the supply of L-cysteine ​​can maintain the activity of thiol enzymes and improve Inflammation and allergic skin symptoms; 4. Because L-...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/12
CPCC12P13/12
Inventor 胡磊徐灿陈建华胡鹏高
Owner 武汉茵茂特生物技术有限公司
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