Method for removing alanine in reaction system for preparing neutral amino acid

A neutral amino acid and reaction system technology, applied in the field of bioengineering, can solve problems such as reducing purification costs, reducing purification steps, and reducing the yield of target amino acids, and achieves the effect of simplifying separation, purification and refining processes and increasing yields

Active Publication Date: 2014-02-05
SHANGHAI RES & DEV CENT OF INDAL BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The object of the present invention is to provide a method for removing and preparing neutral amino acid in response to the problem that alanine is difficult to remove in the reaction system of neutral amino acid prepared by the existing transaminase method or fermentation method, or the yield of target amino acid is reduced while removing. The method of alanine in the amino acid reaction system can specifically remove alanine in the system, and has very little impact on the yield of the target amino acid, thereby reducing subsequent purification steps and reducing purification costs

Method used

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  • Method for removing alanine in reaction system for preparing neutral amino acid
  • Method for removing alanine in reaction system for preparing neutral amino acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-3

[0023] Example 1-3 , Removal of L-alanine in L-2-aminobutyric acid solution

[0024] At 25°C, under the condition that the pH is controlled to 8.0 by acid and alkali, add alanine racemase and D- Amino acid oxidase, then carry out conversion reaction with the conditions shown in table 1, and detect the content of L-2-aminobutyric acid and alanine in the reaction liquid respectively, detection result is as shown in table 2, embodiment 1-3 The alanine racemase used in the method is respectively derived from the alanine racemase of Bacillus subtilis; the D-amino acid oxidase used is respectively derived from the D-amino acid oxidase of Rhodotorula, Trichomonas and Trichomonas, Among them, the D-amino acid oxidase in Example 3 is the immobilized Trichomonas trigonatus D-amino acid oxidase purchased from Hunan Fulaige Biotechnology Co., Ltd.; the catalase added in Example 2-3 was purchased from Zaozhuang City Jie Nuo Bioenzyme Co., Ltd.

[0025] Table 1. Transformation conditions...

Embodiment 4-5

[0030] Example 4-5 , Remove alanine in L-tert-leucine solution and L-valine solution

[0031] Add alanine to 100ml of a solution containing 30g / L of L-tert-leucine and 6g / L of L-alanine, and 500ml of a solution containing 30g / L of L-valine and 4g / L of L-alanine Acid racemase, amino acid oxidase and 5000U / L catalase, then add 20% hydrogen peroxide solution at a rate of 0.5ml / min, control the temperature at 25°C, maintain the pH at 8.0, and stir at 200rpm The conversion reaction was carried out under the condition of the speed, and then respectively in 4 hours and 6 hours after the reaction, the content of the target amino acid and alanine in the detection reaction solution, the detection results are as shown in table 3, wherein, in the embodiment 4-5 The alanine racemase used is 3000U / L from the alanine racemase of Bacillus subtilis; the amino acid oxidase used is 2000U / L from the D-amino acid oxidase from Rhodotorula yeast and 2000U / L from D-Amino Acid Oxidase in Saccharomy...

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Abstract

The invention provides a method for removing alanine in a reaction system for preparing neutral amino acid, wherein the reaction system for preparing the neutral amino acid is a reaction system adopting a transaminase method or a fermentation method to prepare the neutral amino acid. The method provided by the invention is as follows: alanine racemase and a catalyst capable of specially catalyzing single chiral amino acid for further reaction are added into the reaction system, thus specially removing the alanine in the reaction system. The method can specially remove the alanine in conversion liquid or fermentation liquid, thus simplifying the separation and purification and refining the technique of the neutral target amino acid.

Description

technical field [0001] The invention belongs to the field of bioengineering, and in particular relates to a method for removing alanine in a reaction system for preparing neutral amino acids. Background technique [0002] Neutral amino acids include unnatural chiral α-amino acids, L-valine, L-isoleucine, and L-leucine. Among them, unnatural chiral α-amino acids mainly include L-amino acids such as L-2-aminobutyric acid, L-tert-leucine, L-norvaline, and D-tryptophan, D-leucine, D- - D-amino acids such as valine, D-tert-leucine, etc. Among them, the non-natural chiral α-amino acid not only has most of the functions of natural amino acids, but also has excellent properties that non-natural amino acids do not have, and has a wide range of uses in drug synthesis (medicine and pesticide), food, cosmetics, etc. In particular, it has broad market prospects in drug synthesis and has become the fastest-growing chiral drug intermediate in recent years. And L-valine, L-isoleucine and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P13/04
Inventor 杨晟蒋宇范文超陶荣盛潘振华
Owner SHANGHAI RES & DEV CENT OF INDAL BIOTECH
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