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Enzymatic conversion preparation method of gamma-L-glutamoyl n-propylamine

A technology for glutamyl-n-propylamine and enzymatic conversion, which is applied in the directions of biochemical equipment and methods, acyltransferases, enzymes, etc., can solve the problems of preparing γ-L-glutamyl-n-propylamine and the like, and achieves low cost, high enzyme Strong stereoselectivity and mild reaction conditions

Inactive Publication Date: 2018-09-28
宿州学院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there are reports on the synthesis of γ-D-glutamyl-L-tryptophan, glutathione, theanine, γ-glutamyl-taurine and other compounds catalyzed by γ-glutamyl transpeptidase There are many, but there is no report of using this enzyme to prepare γ-L-glutamine n-propylamine

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] A kind of enzymatic conversion preparation method of γ-L-glutamyl n-propylamine of the present embodiment comprises the following steps:

[0027] (1) Put 1000mL Escherichia coli ATCC15489 in culture medium (10g / L glucose, 5g / L beef extract, 1.5g / L citric acid, 2.0g / L ammonium sulfate, 4.0g / L K 2 HPO 4 , 2.0g / L MgSO 4 , 0.05g / L CaCl 2 , 0.001g / L CoCl 2 and 0.0002g / L MnC 4 h 6 o 4 4H 2 Cultivate and ferment in (0), the fermented broth is centrifuged to obtain 12g wet thallus, joins in the 500mL transformation liquid (contains 40g L-glutamic acid-γ-methyl ester, 60g n-propylamine and 0.005g / L ethyl acetate in the transformation liquid) , pH 6.0, enzymatic reaction at 35°C for 12 hours, after the reaction, the molar conversion rate of L-glutamic acid-γ-methyl ester was 91%;

[0028] (2) Centrifuge the transformation solution at 3000r / min for 10min to remove the bacterial cells; heat, decolorize with activated carbon, filter with suction, absorb the decolorization so...

Embodiment 2

[0030] A kind of enzymatic conversion preparation method of γ-L-glutamyl n-propylamine of the present embodiment comprises the following steps:

[0031] (1) Put 1000mL Pseudomonas aeruginosa CGMCC NO: 1.1129 in the medium (45g / L maltose, 40g / L yeast extract, 1.5g / L citric acid, 2.0g / L ammonium sulfate, 4.0g / L K 2 HPO 4 , 2.0g / L MgSO 4 , 0.05g / L CaCl 2 , 0.001g / LCoCl 2 and 0.0002g / L MnC 4 h 6 o 4 4H 2 Cultivate and ferment in (0), fermented liquid centrifugation obtains 15g wet thallus, joins in the 500mL conversion liquid, contains the acetic acid of the L-glutamic acid-γ-methyl ester of 20g, the n-propylamine of 30g and 0.01g / L in the conversion liquid Butyl ester, pH 11, enzymatic reaction at 50°C for 12 hours, after the reaction, the molar conversion rate of L-glutamic acid-γ-methyl ester is 90%;

[0032] (2) Centrifuge the transformation solution at 5000r / min for 20min, remove the bacterial cells, heat, decolorize with activated carbon, filter with suction, absorb ...

Embodiment 3

[0034] A kind of enzymatic conversion preparation method of γ-L-glutamyl n-propylamine of the present embodiment comprises the following steps:

[0035] (1) Put 1000mL Pseudomonas stutzeri CGMCC NO:1.202 in the medium (20g / L sucrose, 20g / L corn steep liquor, 1.5g / L citric acid, 2.0g / L ammonium sulfate, 4.0g / L K 2 HPO 4 , 2.0g / L MgSO 4 , 0.05g / L CaCl 2 , 0.001g / LCoCl 2 and 0.0002g / L MnC 4 h 6 o 4 4H 2 (0) to cultivate and ferment, the fermented liquid is centrifuged to obtain wet thallus 20g, joins in the 500mL transformation liquid, contains the n-propylamine of 40g, the n-propylamine of 60g and the octane of 0.02g / L in the transformation liquid Alcohol, pH 7.0, enzymatic reaction at 40°C for 12 hours, after the reaction, the molar conversion rate of L-glutamic acid-γ-methyl ester was 89%;

[0036] (2) Centrifuge the transformation solution at 4000r / min for 15min, remove the bacterial cells, heat, decolorize with activated carbon, and suction filter, absorb the decolor...

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PUM

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Abstract

The invention provides an enzymatic conversion preparation method of gamma-L-glutamoyl n-propylamine. The method comprises the following steps: carrying out culture and fermentation on a strain with the activity of gamma-glutamyl transpeptidase in a culture medium, and carrying out centrifugalizing on the fermentation liquid, thus obtaining the wet thallus containing gamma-glutamyl transpeptidase;(2) adding conversion liquid into the wet thallus, carrying out enzymatic reaction under the condition with the temperature being 35-50 DEG C and the pH being 6-11, and carrying out separation by adopting an isoelectric point crystallization method, thus obtaining gamma-L-glutamoyl n-propylamine, wherein the conversion liquid contains L-glutamic acid-gamma-methyl ester, n-propylamine, and one ofethyl acetate, butyl acetate, octanol and n-hexanol. According to the method, the specific strain of gamma-glutamyl transpeptidase is cultured in the culture medium, gamma-glutamyl transpeptidase is efficiently expressed, so that the enzymatic synthesis of gamma-L-glutamoyl n-propylamine has relatively high catalysis rate and conversion rate, in addition, the reaction conditions are mild, the stereoselectivity of enzyme is strong, the catalysis efficiency is high, the cost is low, the technical process is simple, and the method is suitable for industrial production.

Description

technical field [0001] The invention relates to the field of preparation of γ-L-glutamyl n-propylamine, in particular to an enzymatic conversion preparation method of γ-L-glutamyl-n-propylamine. Background technique [0002] γ-L-glutamyl n-propylamine is an important γ-glutamyl alkylamine, and γ-glutamyl alkylamine has important physiological activities. The γ-L-glutamyl n-propylamine can be used in the field of medicine and has a very wide application prospect. [0003] γ-Glutamyl transpeptidase (EC 2.3.2.2) is an important enzyme involved in the glutamyl cycle reaction, which can specifically catalyze the migration process of glutamyl groups, so as to obtain compounds containing glutamyl groups. So far, there are reports on the synthesis of γ-D-glutamyl-L-tryptophan, glutathione, theanine, γ-glutamyl-taurine and other compounds catalyzed by γ-glutamyl transpeptidase There are many, but there is no report of using this enzyme to prepare γ-L-glutamine n-propylamine. [00...

Claims

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Application Information

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IPC IPC(8): C12P13/02C12N9/10
CPCC12N9/104C12P13/02C12Y203/02002
Inventor 徐礼生
Owner 宿州学院
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