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Mutagenesis method and screening method of lactic acid bacteria with high gamma-aminobutyric acid yield

A technology of aminobutyric acid and a screening method, applied in the field of microorganisms, can solve the problems such as no reports on the production of GABA by Weisseria viridans, and achieve the effects of low cost, stable GABA production, and simple and easy method.

Inactive Publication Date: 2019-01-11
XUZHOU UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A variety of lactic acid bacteria have been found to have GABA activity, but there is no report of Weissella viride producing GABA

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] A method for mutagenesis of lactic acid bacteria with high yield of gamma-aminobutyric acid, comprising the following steps:

[0030] (1) Preparation of fermentation broth: Pediococcus lactis ZY-6 (Weissella viridescensZY-6) preserved in the laboratory was selected as the starting strain. The preservation number is GDMCC NO: 60377. The strain is inoculated into potato or Martin's liquid medium for activation and culture for 2d-6d to obtain a fermentation broth;

[0031] The composition of described potato liquid medium is: potato 200g, glucose 20g, agar 15~20g and water 1000mL; The composition of Martin's liquid medium is: KH 2 PO41g, MgSO 4 ·7H 2 O 0.5g, peptone 5g, glucose 10g, agar 15-20g and water 1000ml;

[0032] (2) Preparation of bacterial suspension: Take 10mL of the above-mentioned fermentation broth in a conical flask, add 50 glass beads, shake for 10min-20min, and then dilute with sterile water according to the ratio of 1:10, 1:100, 1:1000 into different ...

Embodiment 2

[0040] A screening method for lactic acid bacteria with high yield of gamma-aminobutyric acid, comprising the following steps:

[0041] (1) Fermentation of lactic acid bacteria for testing: get the bacterial classification after the mutagenesis of Example 1 and inoculate it in the MRS medium, use the MRS medium to culture the lactic acid bacteria for testing at 41 DEG C for 24 hours, and use 3% inoculum Inoculate into the GYP fermentation medium containing 1% L-sodium glutamate, let it stand for anaerobic culture for 48 hours, take the fermentation broth and centrifuge at 2000r / min, centrifugation time 10min, and take the supernatant;

[0042] The composition of the MRS medium is: peptone 10g, beef extract 10g, yeast extract 5g, diammonium hydrogen citrate 2g, glucose 20g, Tween 801mL, sodium acetate 5g, dipotassium hydrogen phosphate 2g, magnesium sulfate 0.58g, sulfuric acid Manganese 0.25g, agar 18.0g and distilled water 1000mL;

[0043] (2) Preliminary screening of γ-amin...

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Abstract

The invention discloses a mutagenesis method and screening method of lactic acid bacteria with high gamma-aminobutyric acid yield, which comprises the following steps: selecting Pediococcus lactis ZY-6 (Weissellaviridescens ZY-6) as the starting strain, after activation culture, preparation of bacterial suspension, mutagenesis, culture to obtain mutagenic bacteria, and then fermentation culture ofmutagenic bacteria, gamma-Aminobutyric Acid Producing Strain Primary Screening, High Yield gamma-Screening of Aminobutyric Acid Lactic Acid Bacteria for gamma-GABA-producing strains. The method of the invention is simple and easy to operate, low in cost, high and stable in GABA production of mutant strain, and is an effective method for lactic acid bacteria to obtain high-efficient GABA production. The strain is preserved for a long time and has wide application prospect, and the reaction condition is mild. Glutamic acid or L-Sodium glutamate was used as the sole substrate, and no other medium was found in the transformation solution. Using the glutamate decarboxylase of lactic acid bacteria, L-Glutamic acid or L-Conversion of Sodium Glutamate to gamma-Aminobutyric acid, by paper chromatography analysis of the presence of GABA in the conversion solution, to improve the screening efficiency.

Description

technical field [0001] The invention belongs to the technical field of microorganisms and relates to a mutagenesis of lactic acid bacteria, in particular to a mutagenesis method and a screening method of a lactic acid bacteria with high yield of gamma-aminobutyric acid. Background technique [0002] γ-Aminobutyric acid (γ-Aminobutyric acid, GABA), also known as aminobutyric acid, is a natural amino acid composed of non-protein. It is the most important inhibitory neurotransmitter in the mammalian central nervous system and has important physiological functions. : Such as lowering blood pressure, improving brain function, prolonging memory, calming nerves, anti-anxiety effects, improving liver function, activating kidney function and lowering cholesterol. In addition to the above-mentioned physiological functions, GABA also has the functions of regulating hormone secretion, improving menopausal syndrome, promoting alcohol metabolism, deodorizing, efficient weight loss, induci...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N15/01C12P13/00C12R1/01
CPCC12N15/01C12P13/005C12N1/205C12R2001/01
Inventor 李文王陶李同祥
Owner XUZHOU UNIV OF TECH
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