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Flora for producing L-theanine and application of flora

A technology of theanine and flora, applied in the field of microorganisms to achieve the effect of good practicability and efficient synthesis and secretion

Inactive Publication Date: 2019-05-24
ANHUI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The currently reported microorganisms that can synthesize theanine include Escherichia coli, Bacillus subtilis, and Pseudomonas, but these are free microorganisms in nature and are not isolated from tea trees.

Method used

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  • Flora for producing L-theanine and application of flora
  • Flora for producing L-theanine and application of flora
  • Flora for producing L-theanine and application of flora

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1) Isolation and cultivation of the endophytic flora of the four-year-old Anhui tea tree variety "Shuchazao"

[0026] Aseptic operation of ultra-clean workbench: Take a few leaves of tea tree aseptic tissue culture seedlings, cut them into 0.5cm×1.0cm pieces with scissors, insert them into solid LB medium at random obliquely, seal with parafilm, and culture at 28°C Cultivate the box upside down for 2 to 3 days, and after a rich flora grows around the leaves (such as figure 1 ), which were inoculated into liquid LB medium for further cultivation.

[0027] 2) Sample preparation and detection process of theanine synthesized by tea tree endophytic flora

[0028] Shake culture at 28°C to OD 600 When it is 1.0, add the final concentration of 20mM glutamine and 20mM ethylamine to the LB culture solution simultaneously, and use the same concentration bacterial solution without adding any substrate and the blank medium adding the same concentration substrate as control experim...

Embodiment 2

[0032] 1) Isolation and cultivation of the endophytic flora of the ten-year-old tea tree variety "Longjing 43" in Zhejiang Province

[0033] Aseptic operation of ultra-clean workbench: Take a few pieces of aseptic tissue-cultured seedling leaves of the ten-year-old Zhejiang tea tree variety "Longjing 43", cut them into 0.5cm×1.0cm pieces with scissors, and randomly insert them into solid LB culture medium obliquely , sealed with a parafilm, and cultured upside down in a constant temperature incubator at 28°C for 2 to 3 days. After a rich flora grew around the leaves, they were inoculated into liquid LB medium for further cultivation.

[0034] 2) Sample preparation and detection process of theanine synthesized by tea tree endophytic flora

[0035] Shake culture at 28°C to OD 600 When it is 1.0, add the final concentration of 20mM glutamine and 20mM ethylamine to the LB culture solution simultaneously, and use the same concentration bacterial solution without adding any substra...

Embodiment 3

[0039] 1) Isolation and cultivation of the endophytic flora of the four-year-old Zhejiang tea tree variety "Wuniuzao"

[0040] Aseptic operation of ultra-clean workbench: Take a few leaves of aseptic tissue-cultured seedlings of the four-year-old Zhejiang tea tree variety "Wu Niuzao", cut them into 0.5cm×1.0cm pieces with scissors, and randomly insert them into solid LB for culture In the culture medium, seal it with a parafilm, and incubate it upside down in a constant temperature incubator at 28°C for 2 to 3 days. After a rich flora grows around the leaves, inoculate it into the liquid LB medium for further cultivation.

[0041] 2) Sample preparation and detection process of theanine synthesized by tea tree endophytic flora

[0042] Shake culture at 28°C to OD 600 When it is 1.0, add the final concentration of 20mM glutamine and 20mM ethylamine to the LB culture solution simultaneously, and use the same concentration bacterial solution without adding any substrate and the b...

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Abstract

The invention discloses a flora for producing L-theanine and application of the flora. The flora is a tea tree endophyte separated from tea tree aseptic tissue culture saplings, and belongs to Rhodanobacteraceae Luteibacter sp.. The flora and the application have the advantages that the flora for producing the L-theanine is the endophyte separated from tea trees for the first time, and can excellently grow on LB culture media, floras are cultured at the temperatures of 28 DEG C until the floras are in logarithmic growth phases, glutamine and ethylamine are added into the culture media, then the floras continue to be cultured for 7 days, synthesis of the L-theanine can be detected in experiment groups as compared with control groups, synthesized theanine can be quickly secreted out of cells, the secretion efficiency can reach 90% at least, and secretion theanine can be efficiently synthesized; the glutamine and the ethylamine are used as substrates by L-theanine production methods, operation methods are simple, and the flora is high in theanine production efficiency and has an excellent industrial application prospect.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a kind of bacterial group producing L-theanine and its application. Background technique [0002] Theanine (L-Theanine), which is N-ethyl-γ-glutamine, accounts for more than 70% of the total amino acids in tea, is a unique non-protein free amino acid in tea trees, and is also the main substance that gives tea special flavor , its content directly determines the quality of tea. Theanine has medicinal effects such as relaxing nerves, anti-tumor, anti-depression, protecting blood vessels, lowering blood pressure, suppressing obesity and improving immune function of the body. In addition, it can also be used as a functional food additive in the food field. Therefore, theanine is more and more favored by people, and there is a great demand for it in the international market. [0003] Currently, there are three main methods for producing theanine. One is extracted from ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P13/04C12R1/01
CPCC12N1/20C12P13/04C12N1/205C12R2001/01
Inventor 孙俊常慢慢宛晓春张照亮张正竹
Owner ANHUI AGRICULTURAL UNIVERSITY
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