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Method for synthesizing L-carnosine by using microorganism enzyme method

A microbial enzyme and carnosine technology, applied in the field of carnosine preparation, can solve the problems of reducing the biocatalytic efficiency of microbial cells, excessive damage to the biological enzyme cell membrane, loss of microbial intracellular substances, etc., to improve the reaction conversion rate, low cytotoxicity, and enzymatic The effect of improving the reaction efficiency

Inactive Publication Date: 2019-09-27
SUZHOU FUSHILAI PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, both physical treatment and chemical treatment are permeabilization pretreatments, and the treatment procedures are mainly permeabilization treatment → centrifugation → washing and other operations, and the treatment process is relatively cumbersome
In addition, during the permeability pretreatment, excessive damage to the cell membrane of the biological enzyme and lysis of the cell often occur, resulting in the loss of the microbial intracellular substance during centrifugation, which reduces the efficiency of microbial cell biocatalysis

Method used

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  • Method for synthesizing L-carnosine by using microorganism enzyme method

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Experimental program
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Effect test

Embodiment 1

[0016] In the L-carnosine reaction system that is equipped with a stirring device such as a reaction kettle and adding microbial enzymes, directly add an organic solvent for activating microbial enzyme activity and as a penetrating agent and directly add the same for activating microbial enzymes Active divalent metal ions are reacted for 480min at a stirring speed of 140rpm and a reaction temperature of 40°C in the stirring device. After the reaction is completed, the reaction conversion rate is determined by high-performance liquid chromatography analysis, and the reaction solution is collected and centrifuged. The whole cells obtained by centrifugation can continue to be catalyzed to realize recycling, and 15.1 g / L of L-carnosine can be obtained. In the present embodiment, the L-carnosine reaction system is the L-carnosine basic reaction solution, and the volume percent concentration of the organic solvent in the L-carnosine reaction solution is 0.1% (V / V) , the mol concentr...

Embodiment 2

[0019] In the L-carnosine reaction system that is equipped with a stirring device such as a reaction kettle and adding microbial enzymes, directly add an organic solvent for activating microbial enzyme activity and as a penetrating agent and directly add the same for activating microbial enzymes Active divalent metal ions are reacted for 520min at a stirring speed of 160rpm and a reaction temperature of 35°C in the stirring device. After the reaction is completed, the reaction conversion rate is determined by HPLC analysis, and the reaction solution is collected and centrifuged. The whole cells obtained by centrifugation can continue to be catalyzed to realize recycling and obtain 15.2g / L L-carnosine. In the present embodiment, the L-carnosine reaction system is the L-carnosine basic reaction solution, and the volume percent concentration of the organic solvent in the L-carnosine reaction solution is 1.3% (V / V) , the mol concentration (molar concentration) of described divalen...

Embodiment 3

[0021] In the L-carnosine reaction system that is equipped with a stirring device such as a reaction kettle and adding microbial enzymes, directly add an organic solvent for activating microbial enzyme activity and as a penetrating agent and directly add the same for activating microbial enzymes Active divalent metal ions are reacted for 450min at a stirring speed of 150rpm and a reaction temperature of 38°C in the stirring device. After the reaction is completed, the reaction conversion rate is determined by HPLC analysis, and the reaction solution is collected and centrifuged. The whole cells obtained by centrifugation can continue to be catalyzed to realize recycling and obtain 15.15g / L L-carnosine. In the present embodiment, the L-carnosine reaction system is the L-carnosine basic reaction solution, and the volume percent concentration of the organic solvent in the L-carnosine reaction solution is 2.5% (V / V) , the mol concentration (molar concentration) of described divale...

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Abstract

The invention discloses a method for synthesizing L-carnosine by using a microorganism enzyme method and belongs to the technical field of carnosine preparation. The method comprises the following steps: directly putting an organic solvent which is used for activating activity of a microorganism enzyme and is used as a permeation agent into an L-carnosine reaction system with the microorganism enzyme in a reaction container provided with a stirring device, directly adding divalent metal ions for activating the activity of the microorganism enzyme, carrying out a reaction under stirring of the stirring device, and carrying out centrifugal separation after the reaction is completed, thereby obtaining L-carnosine. The microorganism enzyme can be activated, the permeation barrier of extraneous coats of cells can be reduced, later crystal separation can be facilitated, the synthesis cost can be lowered, the reaction speed can be remarkably increased, and the productivity can be improved.

Description

technical field [0001] The invention belongs to the technical field of carnosine preparation, and in particular relates to a method for synthesizing L-carnosine by microbial enzymatic method. Background technique [0002] L-carnosine is a natural β,α-dipeptide composed of L-histidine and β-alanine, which has strong antioxidant capacity. The currently reported L-carnosine synthesis processes are mainly divided into two categories: one is chemical synthesis, and the other is biocatalytic synthesis. Due to the severe reaction conditions and complicated steps of the chemical synthesis method, it is necessary to activate the carboxyl group of β-alanine or its derivatives, and at the same time, it is necessary to protect some other sensitive groups. After the synthesis reaction is completed, it is necessary to protect the protective group. The target product is obtained by deprotecting the protective group. The by-products in the reaction process are relatively complicated. The r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/10
CPCC12P17/10
Inventor 丁建飞浦佳春钱庆陆建刚邢健
Owner SUZHOU FUSHILAI PHARMA CO LTD
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