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 the problems of reducing purification costs, reducing purification steps, difficult to remove alanine, etc., and achieve the effect of increasing yield, simplifying separation, purification and refining process

Active Publication Date: 2010-09-08
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] Examples 1-3 , remove L-alanine in L-2-aminobutyric acid solution

[0024] To 500ml of a solution containing L-2-aminobutyric acid 30g / L and L-alanine 5g / L, alanine racemase and D- Amino acid oxidase, then carry out the conversion reaction with the conditions shown in Table 1, and detect the content of L-2-aminobutyric acid and alanine in the reaction solution respectively, and the detection results are shown in Table 2. Embodiment 1-3 The alanine racemase used in the method is respectively alanine racemase derived from Bacillus subtilis; the D-amino acid oxidase used is the D-amino acid oxidase derived from Rhodotorula, Trichoderma and Trichoderma, respectively, Wherein, the D-amino acid oxidase in Example 3 was the immobilized Triceratops D-amino acid oxidase purchased from Hunan Flegre Biotechnology Co., Ltd.; the catalase added in Example 2-3 was purchased from Zaozhuang City Jie NovoBio Enzyme Co., Ltd.

[0025] Table 1. Transformation conditions

[0026] ...

Embodiment 4-5

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

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

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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, non-natural 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, in addition to having most of the functions of natural amino acids, non-natural chiral α-amino acids also have excellent properties that non-natural amino acids do not have, and are widely used in drug synthesis (medicine and pesticides), 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 L...

Claims

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Application Information

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Patent Type & Authority Applications(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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