Method for high-efficiently producing L-theanine through production of coli [gamma]-glutamylmethylamine synthetase with escherichia coli

A technology of glutamine, Escherichia coli, applied in the field of bioengineering

Inactive Publication Date: 2014-12-17
SHANGHAI KING SHINE BIOLOGY TECH
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Problems solved by technology

However, there is no literature report on the high-efficiency enzymatic preparation of theanine using γ-glutamine synthetase derived from genetic engineering as a catalyst and lower-cost glutamic acid and ethylamine as substrates

Method used

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  • Method for high-efficiently producing L-theanine through production of coli [gamma]-glutamylmethylamine synthetase with escherichia coli
  • Method for high-efficiently producing L-theanine through production of coli [gamma]-glutamylmethylamine synthetase with escherichia coli
  • Method for high-efficiently producing L-theanine through production of coli [gamma]-glutamylmethylamine synthetase with escherichia coli

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Embodiment Construction

[0018] 1 Materials and Methods

[0019] 1.1 Strains and plasmids

[0020] Escherichia coli ( Escherichia coli ) DH5α, BL21 (DE3) strains and plasmid vectors pMD18-T simple vector, pET28a are all preserved in our laboratory.

[0021] 1.2 Reagents and materials

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Abstract

The invention relates to a genetic engineered bacterium which contains a recombinant plasmid pET28a-GMAS, is 6585 bp in a plasmid size, is 1308 bp in a fragment size of a GMAS, is named as Escherichia coli and has a kanapenecilin resistance. Genes of the [gamma]-glutamylmethylamine synthetase are inducibly expressed by isopropylsulpho-[beta]-D-galactoside. The genetic engineered bacterium has a capability of biologically synthesizing theanine. By means of glutamic acid and ethylamine as substrates, a method in the invention is simple in operation method, is convenient to control in production, is high in conversion and production efficiency of the theanine and has an excellent industrial application prospect.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a method for catalyzing and synthesizing L-theanine by using gamma-glutamine synthetase produced by Escherichia coli genetic engineering bacteria. Background technique [0002] Theanine (N-ethyl-γ-glutamine) is a unique amino acid in tea and one of the taste substances of tea. Its content directly determines the quality of tea. At the same time, theanine has medicinal effects such as relaxing nerves, anti-depression, anti-tumor, lowering blood pressure, and improving immunity. It can also be used in the food field, such as an additive for functional foods. Therefore, more and more people pay attention to it, and the demand in domestic and foreign markets continues to increase. [0003] At present, there are mainly three methods for the production of theanine. One is extracted from tea leaves. The content of theanine in tea is low, and the sources of raw mate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12P13/04C12R1/19
Inventor 马承国程占冰王开成
Owner SHANGHAI KING SHINE BIOLOGY TECH
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