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Method for preparing theanine by biological method

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: 2017-01-11
NANTONG LICHENG BIOLOGICAL ENG
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  • Summary
  • Abstract
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  • Claims
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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 method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1 produces the separation of glutamylase bacterium

[0027] Take a little soil to be separated and add it to the separation medium (in g / L): glucose 15, yeast extract 12, corn steep liquor 10, MgSO 4 ·7H 2 O 0.5, K 2 HPO 4 1, pH7.0. Cultivate at 37°C and 200rpm for 24 hours, after dilution and coating, obtain a pure culture, cultivate at 37°C for 30 hours, and obtain 23 strains with high enzyme activity, and finally select a strain whose enzyme activity can reach 0.2U / mL strain was used as the experimental strain.

Embodiment 2

[0028] Embodiment 2 obtains thalline

[0029] Using the nitroreductor Pseudomonas SK16.004 obtained in Example 1, in the fermentation medium (in g / L): glucose 12, yeast extract 0.9, sodium glutamate 12, MgSO 4 ·7H 2 O0.5, K 2 HPO 4 0.6, KH 2 PO 4 0.6, EDTA-Fe 0.1, pH 6.8-7.0, 35-37°C, 200rpm, cultivated for 16h; refrigerated and centrifuged to obtain bacteria, which is the enzyme source.

Embodiment 3

[0030] Embodiment 3 enzyme reaction

[0031] Use the enzyme source of embodiment 2, in the 50mM sodium borate buffer (Na 2 B 4 o 7 -NaOH, pH 10), theanine was synthesized by performing an enzyme reaction at 37°C for 2 to 8 hours.

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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 nitroreductor pseudomonas strain which can be used for efficient production of theanine. Background technique [0002] Theanine (L-theanine) is a unique amino acid in tea, and its content directly determines the quality of tea. In addition, theanine also has the effects of lowering blood pressure, anti-tumor, protecting nerves, relaxing, anti-depression, anti-fatigue, improving immunity, and reducing body fat. The various physiological functions, good stability and safety of L-theanine make it widely demanded and applied in food and health care products. [0003] Therefore, people have tried various methods of manufacturing theanine, and currently there are mainly three manufacturing methods. The first is the method of extraction from tea leaves. But because the content of theanine in dry tea leaves is only about 1.5%, and photosynthesis will cause th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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