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Method for preparing theanine by biological process

A technology of theanine and biological method, which is applied in the field of theanine preparation by biological method, can solve problems such as complex industrial production operations, and achieve the effects of facilitating industrial production, efficient manufacturing, and simple production

Active Publication Date: 2014-10-08
NANTONG LICHENG BIOLOGICAL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The enzymatic production of theanine with glutaminase from microbial sources has been reported, such as from Pseudomonas, and from Pseudomonas citronellolii GEA in 2005 and from Bacillus in 2007 In 2008, a method for producing theanine using glutamyl transpeptidase from Bacillus subtilis was disclosed, which is safe and reliable, but There are problems such as the need to prepare enzyme powder and the possible use of organic solvents, resulting in complex industrial production operations
There is no report on the production of theanine by cell transformation from Pseudomonas nitroreductors

Method used

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  • Method for preparing theanine by biological process

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Example 1 Isolation of glutamylase-producing bacteria

[0027] A little soil to be separated was added to the separation medium (calculated in g / L): glucose 15, yeast extract 12, corn steep liquor 10, MgSO4·7H2O 0.5, K2HPO41, pH7.0. Cultivated at 37°C and 200rpm for 24h, after dilution and coating, a pure culture was obtained. After culturing at 37°C for 30h, 23 strains with higher enzyme activity were obtained. Finally, a strain with enzyme activity of 0.2U / mL was selected. strains were used as experimental strains.

Embodiment 2

[0028] Example 2 Obtaining bacterial cells

[0029] Using the nitroreducing Pseudomonas SK16.004 obtained in Example 1, in the fermentation medium (in g / L): glucose 12, yeast extract 0.9, sodium glutamate 12, MgSO4 7H2O 0.5 , K2HPO4 0.6, KH2PO4 0.6, EDTA-Fe 0.1, pH 6.8-7.0, 35-37°C, 200 rpm for 16 h; freeze and centrifuge to obtain bacterial cells, which are enzyme sources.

Embodiment 3

[0030] Example 3 Enzymatic reaction

[0031] Using the enzyme source of Example 2, in 50 mM sodium borate buffer (Na2B4O7-NaOH, pH10) containing 0.3 M glutamine and 1.5 M ethylamine (or ethylamine hydrochloride) at 37 °C for 2- 8 hours of enzymatic reaction to synthesize theanine.

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Abstract

The invention discloses a method for preparing theanine by a biological process, which comprises the following steps: preparing cells by using Pseudomonas nitroreducens NTLC4.002 with the collection number of CCTCC NO:M2014254 as the initial strain, and preparing natural theanine by cell transformation. The novel theanine strain applicable to theanine production is separated and screened from the Chinese traditional fermentation product; and the glutaminase generated by the strain can be used for efficiently producing theanine by acting on glutamine and ethylamine under the conditions of pH 9-10 and 35-40 DEG C, wherein the conversion rate can reach more than 80%.

Description

technical field [0001] The invention relates to a method for preparing theanine by a biological method, in particular to a nitro-reducing Pseudomonas strain that can be used to efficiently produce theanine. Background technique [0002] L-theanine is an amino acid unique to tea, and its content directly determines the quality of tea. In addition, theanine also has the functions of lowering blood pressure, anti-tumor, protecting nerves, relaxing, anti-depression, anti-fatigue, improving immunity and reducing body fat. The various physiological functions and good stability and safety of L-theanine make it have a wide range of needs and applications in food and health care products. [0003] Therefore, various methods for producing theanine have been tried for a long time, and there are mainly three production methods at present. The first is the method of extraction from tea. However, since the content of theanine in dry tea leaves is only about 1.5%, and photosynthesis wil...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P13/04C12R1/38
Inventor 帅玉英江波张涛孙怡吴晓花沈宇峰
Owner NANTONG LICHENG BIOLOGICAL ENG
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