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Method for catalytically synthesizing L-carnosine through one-step method

A technology of carnosine and step method, which is applied in the field of one-step catalytic synthesis of L-carnosine, can solve the problems that the synthesis efficiency and product yield are not enough to be compared with the chemical method, and the catalytic system is complex, so as to avoid group protection and deprotection, Avoid the use of toxic and harmful solvents, eliminate the effect of high temperature, high pressure and high energy consumption

Pending Publication Date: 2021-10-08
SUZHOU FUSHILAI PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] Since the above two whole-cell one-step synthesis of carnosine is based on dipeptidase, the outstanding problem is that the hydrolysis of L-carnosine by peptidase cannot be avoided, which leads to the relative complexity of the entire catalytic system. Synthetic efficiency and product yield are not enough to compare with chemical methods

Method used

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  • Method for catalytically synthesizing L-carnosine through one-step method
  • Method for catalytically synthesizing L-carnosine through one-step method
  • Method for catalytically synthesizing L-carnosine through one-step method

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Embodiment 1

[0024] The enzyme synthesis reaction is carried out in a packed bed reactor. First, L-carnosinase is used to build a whole-cell catalyst into the packed bed, and then the reaction liquid containing β-alanine, L-histidine and catalyst is added, and the external circulation form Flow through the packed bed for conversion. After the reaction, use high-performance liquid chromatography to measure the conversion rate of the reaction, collect the reaction solution, and centrifuge. The whole cells obtained by centrifugation can continue to be catalyzed to realize recycling, and L-carnosine with a concentration of 50mM is obtained. In this embodiment, the L-carnosinase constructing the whole-cell catalyst is the intracellular expression vector of Pichia pastoris; the addition amount of the L-carnosinase constructing the whole-cell catalyst in the reaction solution is 25g / L , activity 80U / L, the described reaction liquid that contains β-alanine, L-histidine and catalyzer is aqueous solu...

Embodiment 2

[0028] The enzyme synthesis reaction is carried out in a packed bed reactor. First, L-carnosinase is used to build a whole-cell catalyst into the packed bed, and then the reaction liquid containing β-alanine, L-histidine and catalyst is added, and the external circulation form Flow through the packed bed for conversion. After the reaction, use high-performance liquid chromatography to measure the conversion rate of the reaction, collect the reaction solution, and centrifuge. The whole cells obtained by centrifugation can continue to be catalyzed to realize recycling, and L-carnosine with a concentration of 55mM is obtained. In this embodiment, the L-carnosinase constructing the whole-cell catalyst is the intracellular expression vector of Pichia pastoris; the addition amount of the L-carnosinase constructing the whole-cell catalyst in the reaction solution is 50g / L , activity 150U / L, the described reaction liquid that contains β-alanine, L-histidine and catalyzer is aqueous sol...

Embodiment 3

[0030]The enzyme synthesis reaction is carried out in a packed bed reactor. First, L-carnosinase is used to build a whole-cell catalyst into the packed bed, and then the reaction liquid containing β-alanine, L-histidine and catalyst is added, and the external circulation form Flow through the packed bed for conversion. After the reaction, use high performance liquid chromatography to measure the conversion rate of the reaction, collect the reaction solution, and centrifuge. The whole cells obtained by centrifugation can continue to be catalyzed to realize recycling, and L-carnosine with a concentration of 64mM is obtained. In this embodiment, the L-carnosinase constructing the whole-cell catalyst is the intracellular expression vector of Pichia pastoris; the addition amount of the L-carnosinase constructing the whole-cell catalyst in the reaction solution is 35g / L , activity 100U / L, the described reaction liquid that contains β-alanine, L-histidine and catalyzer is aqueous solu...

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Abstract

The invention discloses a method for catalytically synthesizing L-carnosine through a one-step method, and belongs to the technical field of organic matter synthesis. The method comprises the following steps of carrying out an enzyme synthesis reaction in a packed bed reactor, filling a packed bed with a whole-cell catalyst constructed by the L-carnosinase, adding a reaction solution containing beta-alanine, L-histidine and the catalyst, enabling the reaction solution to flow through the packed bed in an external circulation form for reaction conversion, determining the reaction conversion rate by high performance liquid chromatography analysis after the reaction is finished, collecting reaction liquid, performing centrifugal separation, and continuously catalyzing the whole cells obtained by centrifugal separation to realize cyclic utilization, thereby obtaining the high-concentration L-carnosine. The method has the advantages that group protection and deprotection dependent on a chemical synthesis method are avoided, green and environment-friendly production can be embodied without using toxic and harmful solvents, and energy conservation can be embodied by abandoning high temperature and high pressure.

Description

technical field [0001] The invention belongs to the technical field of organic compound synthesis, and in particular relates to a method for synthesizing L-carnosine by one-step catalysis. Background technique [0002] L-carnosine (β-alanyl-L-histidine) and its analogs (such as homocarnosine and anserine) are natural active dipeptides widely present in the brain, muscle and other important tissues of mammals. Strong antioxidant capacity. The reported L-carnosine synthesis process is mainly divided into two classes: one is chemical synthesis (i.e. chemical synthesis method, hereinafter the same), and the other is biocatalytic synthesis (i.e. biocatalytic synthesis method, hereinafter the same). [0003] Although there are many chemical synthesis methods, there are mainly the following two types: one is to use β-alanine to participate in the synthesis. The main route is to condense β-alanine with protected L-histidine after amino protection and carboxyl activation, and then ...

Claims

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Application Information

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