Preparation method for natural theanine

A theanine and natural technology, applied in the field of natural theanine preparation, can solve the complex problems of theanine refining, and achieve the effects of safe and reliable products, short reaction time and simple production

Inactive Publication Date: 2009-01-14
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is a problem in the production of theanine by glutaminase enzymatic method, which produces glutamic acid as a by-product during the hydrolysis reaction of glutaminase, which complicates the purification of theanine.

Method used

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  • Preparation method for natural theanine

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Embodiment 1 produces the separation of glutamyl transpeptidase bacterium

[0023] Shrimp paste, a traditional Chinese fermented product, is used, and a little is added to the separation medium (in g / L): glucose 20, yeast extract 15, 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, a pure culture is obtained. After culturing at 37°C for 30 hours, 20 strains with high enzyme activity are obtained, and a strain with an enzyme activity of 2U / mL is selected. SK11.004 is the experimental strain.

Embodiment 2

[0024] The preparation of embodiment 2 crude enzyme powder

[0025] Using the Bacillus subtilis SK11.004 obtained in Example 1, in the fermentation medium (in g / L): sucrose 25, tryptone 5, corn steep liquor 15, MgSO 4 ·7H 2 O 0.5, K 2 HPO 4 1. Culture at pH 7.0, 37°C, 200 rpm for 16 hours, refrigerate and centrifuge to obtain the supernatant of the fermentation broth, which is the crude enzyme solution. After concentrating the crude enzyme liquid, add alcohol according to the ratio of enzyme liquid: alcohol = 4: 1 (V: V), and freeze-dry the precipitated protein to obtain the crude enzyme powder.

Embodiment 3

[0026] Embodiment 3 enzyme reaction

[0027] Use the crude enzyme powder of embodiment 2, join the 100mM 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 hours.

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Abstract

The invention relates to a natural theanine preparing method, which belongs to the technical field of the biologic engineering. Bacillus subtilis SK11.004 strain obtained through separating and selecting the Chinese traditional fermentation food is theanine producing strain of gamma-glutamyltransferase with the collection number CCTCC NO of M 208083; Gamma-glutamyl transpeptidase is acted on glutamine and ethylamine, and under the condition of pH 10, the theanine can be efficiently produced. The enzyme preparing method for theanine of the invention has the advantages that the reaction time is short, the preparation of the enzyme activity source is simple, the production of the theanine is ensured to be simple, and the product is safe and reliable. The preparing method is suitable for performing large-scale industrialized production.

Description

technical field [0001] The invention relates to a preparation method of natural theanine, which belongs to the technical field of bioengineering. Background technique [0002] Theanine (N-ethyl-γ-glutamine) is a unique amino acid in tea and an effective taste substance in green 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. Therefore, people pay more and more attention to theanine, and it is expected to be added to food and medicine to develop new functional food and functional medicine. [0003] So far, there are mainly three methods for the production of theanine. 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 the loss of part of theanine, resulting in very low yi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/10C12P13/02C12R1/125
Inventor 江波张涛帅玉英
Owner JIANGNAN UNIV
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