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Enzymatic transformation preparation method of [gamma]-L-glutamyl n-butylamine

A technology for enzymatic conversion of n-butyl glutamylamine, applied in biochemical equipment and methods, methods based on microorganisms, acyltransferase, etc., can solve the problems of preparing γ-L-n-butyl glutamylamine, etc., Achieve the effects of low cost, mild reaction conditions, high catalytic rate and conversion rate

Active Publication Date: 2017-07-04
宿州学院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The current applications and 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 on the use of this enzyme to prepare γ-L-glutamyl n-butylamine

Method used

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  • Enzymatic transformation preparation method of [gamma]-L-glutamyl n-butylamine

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Effect test

Embodiment 1

[0029] Such as figure 1 As shown, the enzymatic conversion preparation method of a kind of γ-L-glutamyl n-butylamine of the present embodiment comprises the following steps:

[0030] (1) 1000mL Escherichia coli ATCC15489 was cultivated and fermented in the culture medium, and the fermented liquid was centrifuged to obtain 12g wet thallus containing γ-glutamyl transpeptidase;

[0031] The formula of the above medium is: 20g / L carbon source substance (one or more of glucose, maltose, sucrose, fructose), 20g / L nitrogen source substance (beef extract, yeast extract, corn steep liquor, peptone, soybean cake hydrolyzed one or more in liquid), 2.0g / L citric acid, 2.5g / L ammonium sulfate, 5.0g / L K 2 HPO 4 , 2.5g / L MgSO 4 , 0.07g / L CaCl 2 , 0.002g / L CoCl 2 and 0.0001g / L MnC 4 h 6 o 4 4H 2 O.

[0032] (2) Add the above-mentioned wet bacteria into 500mL transformation solution, which contains 30g L-glutamic acid-γ-methyl ester, 50g n-butylamine and 0.0025g 3-methoxybutyl acetat...

Embodiment 2

[0039] Such as figure 1 As shown, the enzymatic conversion preparation method of a kind of γ-L-glutamyl n-butylamine of the present embodiment comprises the following steps:

[0040] (1) 1000mL of Pseudomonas aeruginosa CGMCC NO: 1.1129 was cultured and fermented in the culture medium, and the fermented liquid was centrifuged to obtain 15g of wet bacteria containing γ-glutamyl transpeptidase;

[0041] The formula of the above medium is: 15g / L carbon source substance (one or more of glucose, maltose, sucrose, fructose), 15g / L nitrogen source substance (beef extract, yeast extract, corn steep liquor, peptone, soybean cake hydrolysis one or more in liquid), 2.0g / L citric acid, 2.5g / L ammonium sulfate, 5.0g / L K 2 HPO 4 , 2.5g / L MgSO 4 , 0.07g / L CaCl 2 , 0.002g / L CoCl 2 and 0.0001g / L MnC 4 h 6 o 4 4H 2 O.

[0042] (2) Add the above-mentioned wet bacteria to 500mL transformation solution, which contains 20g L-glutamic acid-γ-methyl ester, 30g n-butylamine and 0.005g butyl ...

Embodiment 3

[0049] Such as figure 1 As shown, the enzymatic conversion preparation method of a kind of γ-L-glutamyl n-butylamine of the present embodiment comprises the following steps:

[0050] (1) 1000mL of Pseudomonas stutzeri CGMCC NO:1.202 was cultured and fermented in the culture medium, and the fermented liquid was centrifuged to obtain 20g of wet thallus containing γ-glutamyl transpeptidase;

[0051] The formula of the above medium is: 10g / L carbon source substance (one or more of glucose, maltose, sucrose, fructose), 5g / L nitrogen source substance (beef extract, yeast extract, corn steep liquor, peptone, soybean cake hydrolyzed one or more in liquid), 2.0g / L citric acid, 2.5g / L ammonium sulfate, 5.0g / L K 2 HPO 4 , 2.5g / L MgSO 4 , 0.07g / L CaCl 2 , 0.002g / L CoCl 2 and 0.0001g / L MnC 4 h 6 o 4 4H 2 O.

[0052] (2) Add the above-mentioned wet bacteria to 500mL transformation solution, which contains 2.5g L-glutamic acid-γ-methyl ester, 5g n-butylamine and 0.0005g amyl alcoho...

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Abstract

The invention discloses an enzymatic transformation preparation method of [gamma]-L-glutamyl n-butylamine. The preparation method comprises the following steps: (1) conducting culture fermentation on a strain which has a [gamma]-glutamyl transpeptidase in a medium, and centrifuging fermentation liquid so as to obtain a wet bacterium containing the [gamma]-glutamyl transpeptidase; (2) adding transformation liquid to the wet bacterium, wherein the transformation liquid consists of L-glutamic acid-[gamma]-methyl ester, n-butylamine as well as one of 3-methoxybutyl acetate, butyl acetate, pentanol and hexanol; and (3) conducting an enzymatic reaction under the following conditions: a temperature is 35-50 DEG C and pH value is at 6-11, and conducting isolation by virtue of isoelectric point crystallization so as to obtain the [gamma]-L-glutamyl n-butylamine. The enzymatic transformation preparation method of the [gamma]-L-glutamyl n-butylamine provided by the invention has the advantages of being mild in reaction conditions, strong in enzymatic stereoselectivity, high in catalysis efficiency, low in cost and simple in technological process.

Description

technical field [0001] The invention relates to a conversion method of γ-L-glutamyl n-butylamine, in particular to an enzymatic conversion preparation method of γ-L-glutamyl n-butylamine. Background technique [0002] γ-L-glutamyl n-butylamine is an important γ-glutamyl alkylamine, and γ-glutamyl alkylamine can promote the increase of T cells, thereby increasing immunity. The γ-L-glutamyl n-butylamine can be used in the field of medicine and has a very wide application prospect. [0003] γ-glutamyltranspeptidase (γ-glutamyltranspeptidase, GGT, EC 2.3.2.2) is a heterodimeric protein composed of two subunits, large and small, and the receptors that catalyze the reaction are amino acids (or peptides) ), to form γ-glutamyl amino acid or peptide; when γ-glutamyl is both the donor and the acceptor, the autotranspeptide reaction is carried out; when the nucleophile is water, the hydrolysis reaction is carried out. [0004] The current applications and reports on the synthesis of ...

Claims

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Application Information

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IPC IPC(8): C12P13/04C12N9/10C12R1/19C12R1/385C12R1/38C12R1/125
CPCC12N9/104C12P13/04C12Y203/02002
Inventor 徐礼生高贵珍卓馨赵亮曹稳根张兴桃姚沛琳
Owner 宿州学院
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