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Process for preparing gamma-amino butyric acid through enzymatic conversion

A technology of aminobutyric acid and enzymatic conversion, which is applied in the field of enzymatic conversion and preparation of γ-aminobutyric acid, can solve the problems of low yield, complicated process, incomplete separation and the like, and achieves high reaction rate, simple technological process, specific effect

Inactive Publication Date: 2007-02-07
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 1993, people such as Ji Qingfang (Journal of Amino Acids, 1993, (1) 4-7) adopted the method that isoelectric point precipitation and ion-exchange resin combine to separate L-glutamic acid and L-aspartic acid, and the technique is complicated, obtains Low rate, incomplete separation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The enzymatic conversion preparation method of gamma-aminobutyric acid, the preparation steps are as follows:

[0030] 1. Cultivate the E.coli AS 1.505 strain in the following 1000mL medium: 1% peptone, 0.5% NaCl, 0.3% beef extract, 0.1% L-glutamic acid, pH7.2. Shake the flask at 35°C for 12 hours , Centrifuge at 4000rpm for 15min to obtain wet cells.

[0031] 2. Add the cells to 600mL of transformation solution, the transformation solution contains 10% L-glutamic acid and L-aspartic acid mixture (ratio is 50:50), 6mL 0.5% Tween-80, 600mL pH4. 8 acetic acid buffer, react at 35°C for 36h. The concentration of L-aspartic acid in the reaction solution is 5%, the concentration of γ-aminobutyric acid is 3.5%, and the molar conversion rate of L-glutamic acid is 100%.

[0032] 3. Adjust the pH of the reaction solution to 5.5 with 6N NaOH, stir and heat up to 60°C, add activated carbon to decolorize and remove bacteria, adjust the pH of the filtrate to 2.8 with 5N HCl, concen...

Embodiment 2

[0034] The enzymatic conversion preparation method of gamma-aminobutyric acid, the preparation steps are as follows:

[0035] 1. Cultivate the E.coli AS 1.505 strain in the following 1000mL medium: peptone 1%, NaCl 0.5%, beef extract 0.3%, lactose 1.5%, L-glutamic acid 0.1%, pH 7.0. Shake at 35°C Shake the bottle for 12 hours and centrifuge at 4000 rpm for 15 minutes to obtain wet cells.

[0036] 2. Add the cells to 600mL of transformation solution, the transformation solution contains 10% L-glutamic acid and L-aspartic acid mixture (ratio is 60:40), 6mL 0.5% CTAB, 600mL Macllvaine pH4.7 Buffer, react at 35°C for 36h. The concentration of L-aspartic acid in the reaction solution is 4%, the concentration of γ-aminobutyric acid is 4.2%, and the molar conversion rate of L-glutamic acid is 100%.

[0037] 3. Adjust the pH of the reaction solution to 5.5 with 6N NaOH, stir and heat up to 60°C, add activated carbon to decolorize and remove bacteria, adjust the pH of the filtrate to...

Embodiment 3

[0039] The enzymatic conversion preparation method of gamma-aminobutyric acid, the preparation steps are as follows:

[0040]1. Cultivate the E.coli AS 1.505 strain in the following 1000mL medium: 0.5% corn steep liquor, 0.5% NaCl, 2% beef extract, 0.5% maltose, 0.1% L-glutamic acid, pH7.2. 36°C Shake the flask for 12 hours and centrifuge at 4000 rpm for 15 minutes to obtain wet cells.

[0041] 2. Add the cells to 600mL of transformation solution containing 10% L-glutamic acid and L-aspartic acid mixture (ratio of 40:60), 6mL 0.5% OP, 600mL pH4.5 citric acid -Sodium citrate buffer, react at 35°C for 36h. The concentration of L-aspartic acid in the reaction solution is 6%, the concentration of γ-aminobutyric acid is 2.8%, and the molar conversion rate of L-glutamic acid is 100%.

[0042] 3. Adjust the pH of the reaction solution to 5.5 with 6N NaOH, stir and raise the temperature to 60°C, add activated carbon to decolorize and remove bacteria, adjust the pH of the filtrate to...

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Abstract

The invention discloses an enzyme conversion method for preparing gamma-butyric acid. The preparation method comprises employing two mixed acidic amino acids of L-glutamic acid and L-aspartic acid as raw material, mixing the cells of Escherichia.coli AS1.505 with highly active L-glutamic acid decarboxylases and the conversion liquid containing the mixture of L-glutamic acid and L-aspartic acid, implementing enzymatic reaction under the temperature of 28-45 DEG C, then separating the conversion products by isoelectric point crystallization process or isoelectric point crystallization and ion exchange resin combined method to obtain high purity gamma-butyric acid and L-aspartic acid. The invention solves the problem of the highly effective separation of two acidic mixed amino acids, and obtains gamma-butyric acid with higher additional value, and has advantages of low price, simple operation, short conversion time, and low production cost.

Description

1. Technical field [0001] The invention relates to the technical field of biochemical pharmacy, in particular to an enzymatic conversion preparation method of gamma-aminobutyric acid. 2. Technical background [0002] γ-aminobutyric acid is a commonly used drug in clinical practice. It can reduce blood ammonia in the human body and treat various types of hepatic coma. Supplements and boosters, nutrition for the elderly. In addition, L-aspartic acid is the synthetic precursor of L-lysine and purine and pyrimidine bases, as K + , Mg 2+ Ionophores transport electrolytes to the myocardium and improve myocardial function. It is also the main raw material of the new sweetener Aspartame. [0003] According to current literature reports, the preparation method of gamma-aminobutyric acid mainly contains following two kinds: [0004] 1. Enzyme method [0005] In 1961, Chen Zhimin et al. (Acta Biochemistry and Biophysics, Vol.1, No.2, 1961) used the decarboxylase produced by Escher...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P13/00C12N9/10C12R1/185
Inventor 焦庆才吴晓燕李加友钱绍松陈然
Owner NANJING UNIV
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