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Preparation method of enzymatic conversion of L-2-propalanine

A technology of aminobutyric acid and aminobutyric acid, applied in the field of L-2-aminobutyric acid enzymatic transformation and preparation, can solve problems such as L-2-aminobutyric acid not reported in literature, and achieve enzymatic separation efficiency High, easily controllable scale and yield, and low cost of production

Active Publication Date: 2009-09-23
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0024] So far, the method for preparing L-2-aminobutyric acid by aminoacylase derived from microbial sources or genetic engineering sources has not been reported in the literature

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] L-2-aminobutyric acid enzymatic transformation preparation method, its preparation steps are as follows:

[0041] 1. Weigh 50g of DL-2-aminobutyric acid and dissolve it in 150mL of water, adjust the pH to 9.0 with 30% NaOH aqueous solution, then add acetic anhydride and 30% NaOH aqueous solution dropwise at the same time to control the pH to about 10.0, and the acylation is completed After vacuum concentration, the concentrated solution was adjusted to pH 2.0, cooled and crystallized, and dried to obtain 69 g of N-acetyl-DL-2-aminobutyric acid with a content of 95% and a molar yield of 93%.

[0042] 2. Add 15 g of Pseudomonas maltophilia obtained by centrifuging 1000 mL of fermented liquid into 1000 mL of transformation liquid, which contains 50 g of N-acetyl-DL-2-aminobutyric acid, 10 mL of mass concentration 0.5% Tween-80, 1 mL. .1mol / L cobalt ion, pH7.0, react at 40℃ for 35h. The concentration of L-2-aminobutyric acid in the reaction solution was 16.2 g / L, and the m...

Embodiment 2

[0046] L-2-aminobutyric acid enzymatic transformation preparation method, its preparation steps are as follows:

[0047] 1. Weigh 100g of DL-2-aminobutyric acid and dissolve it in 300mL of water, adjust the pH to 9.0 with 30% NaOH aqueous solution, then add acetic anhydride and 30% NaOH aqueous solution dropwise at the same time to control the pH to about 10.0, and the acylation is completed After vacuum concentration, the concentrated solution was adjusted to pH 2.0, cooled and crystallized, and dried to obtain 136 g of N-acetyl-DL-2-aminobutyric acid with a content of 95% and a molar yield of 92%.

[0048] 2. Add 20 g of Alcaligenes obtained by centrifuging 1000 mL of fermentation broth into 1000 mL of transformation liquid, which contains 100 g of N-acetyl-DL-2-aminobutyric acid and 10 mL of 1% polyethylene glycol octylphenyl ether , 1mL 0.1mol / L magnesium ion, pH7.5, react at 40℃ for 60h. The concentration of L-2-aminobutyric acid in the reaction solution was 33g / L, and t...

Embodiment 3

[0052] L-2-aminobutyric acid enzymatic transformation preparation method, its preparation steps are as follows:

[0053] 1. Weigh 120g of DL-2-aminobutyric acid and dissolve it in 400mL of water, adjust the pH to 9.0 with 30% NaOH aqueous solution, then add acetic anhydride and 30% NaOH aqueous solution dropwise at the same time to control the pH to about 10.0, and the acylation is completed After vacuum concentration, the concentrated solution was adjusted to pH 2.0, cooled and crystallized, and dried to obtain 165 g of N-acetyl-DL-2-aminobutyric acid with a content of 94% and a molar yield of 92%.

[0054] 2. Add 15 g of mycobacteria obtained by centrifuging 1000 mL of fermentation broth into 1000 mL of transformation liquid, which contains 150 g of N-acetyl-DL-2-aminobutyric acid and 10 mL of 1% hexadecyltrimethyl bromide Ammonium, 1mL 0.1mol / L calcium ion, pH7.2, react at 40℃ for 90h. The concentration of L-2-aminobutyric acid in the reaction solution was 47.6 g / L, and th...

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Abstract

The invention belongs to the field of biotechnology, in particular relates to a preparation method of enzymatic conversion of L-2-propalanine. The preparation method takes DL-2-propalanine as a starting material; N-acetyl-DL-2-propalanine is synthesized by acetic anhydride; somatic cells containing aminoacylase or extract of aminoacylase is mixed with N-acetyl-DL-2- propalanine water solution to carry out enzymatic reaction at temperature of 30 to 60 DEG C; the converted product is separated by isoelectric point crystallization or the combination of isoelectric point crystallization and ion exchange resin to obtain high-purity L-2-propalanine; and the N-acetyl-DL-2-propalanine remained in the reaction is used for enzymatic resolution again after chemical racemization. The method has the advantages of low material price, easy operation, short conversion time, low production cost, and the like.

Description

1. Technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an enzymatic transformation and preparation method of L-2-aminobutyric acid. 2. Background technology [0002] 2-Aminobutyric acid is an aliphatic amino acid that inhibits the transmission of human nerve information. It can be used clinically for sequelae of cerebrovascular diseases and has a hypotensive effect. It is also used for painless caries removal in dentistry. In addition, 2-aminobutyric acid is also an important chemical raw material and pharmaceutical intermediate, which has been widely used in drug synthesis, such as ethambutol hydrochloride, levetiracetam, etc. Ethambutol hydrochloride is an anti-tuberculosis drug discovered by R.G.Wilkinson in 1962. The drug has a strong inhibitory effect on Mycobacterium tuberculosis, but has no obvious effect on other bacteria, and has no cross-resistance with other anti-tuberculosis drugs. Applicable to various types ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/04C12R1/69C12R1/645C12R1/05C12R1/32C12R1/38
Inventor 焦庆才刘均忠陈争依董凯赵根海刘茜
Owner NANJING UNIV
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