Lactic acid bacterium for high-yield gamma-aminobutyric acid and application of lactic acid bacterium

A technology of aminobutyric acid and Lactobacillus brevis, applied in the direction of microorganism-based methods, bacteria, microorganisms, etc., can solve the problems of low fermentation yield and difficult separation and purification, and achieve high fermentation yield of strains, high safety, and reduced cost effect

Active Publication Date: 2017-12-15
BLOOMAGE BIOTECHNOLOGY CORP LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the problems of low fermentation yield and difficult separation and purification, the present invention provides a lactic acid bacterium screened from nature and a method for producing high-purity GABA by fermentation of the strain

Method used

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  • Lactic acid bacterium for high-yield gamma-aminobutyric acid and application of lactic acid bacterium
  • Lactic acid bacterium for high-yield gamma-aminobutyric acid and application of lactic acid bacterium

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Example 1 Isolation, purification and identification of bacterial strains

[0030] Enrichment culture: Take a small amount of fresh vegetables, homemade sauerkraut, vegetable soil samples, purchased sauerkraut, fresh milk and other samples and add lactic acid bacteria medium (MRS medium, component g / L: peptone 10.0, beef extract powder 4.0, yeast extract powder 4.0, glucose 20.0, dipotassium hydrogen phosphate 2.0, triammonium citrate 2.0, sodium acetate 5.0, magnesium sulfate 0.2, manganese sulfate 0.05, Tween-801.0), 30 ℃ static culture for 2 days.

[0031] Qualitative detection: paper chromatography (or thin-layer chromatography), spotting the diluted fermentation broth on qualitative filter paper (or silica gel prefabricated plate), and developing with chromatographic fluid. The chromatographic solution uses n-butanol: glacial acetic acid: water (60:15:25), and adds 0.4% ninhydrin. After unfolding, let it dry naturally, develop color at 85 ºC for 10 minutes, dilu...

Embodiment 2

[0033] Example 2 Preparation of high-purity γ-aminobutyric acid

[0034] (1) Seed solution preparation: Inoculate the HX12-19 strain into the culture medium, shake the flask for fermentation, and obtain the seed solution;

[0035] (2) Fermentation: Culture for 9 hours, OD 600 The seed solution >0.8 was added to the liquid medium (10L) in a 10L fermenter, and the medium components were: peptone 10.0 g / L, yeast powder 10.0 g / L, glucose 10.0 g / L, Tween 80 1.0 mL / L , sodium acetate 5.0g / L, dipotassium hydrogen phosphate 2.0g / L, magnesium sulfate 0.5g / L, manganese sulfate 0.2g / L, monosodium glutamate (sodium glutamate ≥ 95%) 10.0g / L. Ventilation and agitation culture When cultured for 18 hours, OD600=2.9, the fermentation was completed, and the fermentation liquid was obtained;

[0036] (3) Transformation: collect the bacteria by centrifugation, add purified water to about 5L, add sodium glutamate, control the temperature at 30±2°C, start stirring, and carry out the reaction. Du...

Embodiment 3

[0040] Example 3 Preparation of high-purity γ-aminobutyric acid

[0041] (1) Seed liquid preparation: Inoculate the HX12-19 strain into the culture medium, shake the flask for fermentation, and obtain the seed liquid;

[0042] (2) Fermentation: Culture for 8 hours, OD 600The seed solution >0.8 was added to the liquid medium (10L) in a 10L fermenter, and the medium components were: peptone 10.0 g / L, yeast powder 10.0 g / L, glucose 10.0 g / L, Tween 80 1.0 mL / L , sodium acetate 5.0g / L, dipotassium hydrogen phosphate 2.0g / L, magnesium sulfate 0.5g / L, manganese sulfate 0.3g / L, monosodium glutamate (sodium glutamate ≥ 95%) 10.0g / L. Ventilation and agitation culture When cultured for 20 hours, OD600=3.1, the fermentation was completed, and the fermentation liquid was obtained;

[0043] (3) Transformation: collect the bacteria by centrifugation, add purified water to about 5L, add sodium glutamate, control the temperature at 30±2°C, start stirring, and carry out the reaction. During ...

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Abstract

The invention screens out lactobacillus parabrevis HX12-19 for high-yield gamma-aminobutyric acid. The lactobacillus parabrevis HX12-19 is preserved in China Center for Type Culture Collection (CCTCC), the preservation number is CCTCCM2017307. The lactobacillus parabrevis HX12-19 is obtained through isolating and screening out foodstuff materials, has high safety and can be used for fields such as food; traditional fermentation and bioconversion subdivision control are adopted, bio-enzyme conversion is fully utilized to obtain a product gamma-aminobutyric acid, the production efficiency is greatly improved, the cost is reduced, the fermentation yield can be up to 235g/L, so that the gamma-aminobutyric acid has productive competitiveness; a crystalline method is adopted to purify products, the purity can be up to 98%, and the lactobacillus parabrevis HX12-19 is suitable for industrial production.

Description

technical field [0001] The invention relates to a strain of lactic acid bacteria and the application of the strain in producing gamma-aminobutyric acid, belonging to the field of microbial fermentation engineering. Background technique [0002] Gamma-aminobutyric acid (GABA for short) is widely distributed in prokaryotes and eukaryotes. In September 2009, the Ministry of Health of my country approved γ-aminobutyric acid as a new resource food. [0003] GABA has the functions of anti-hypertension, promoting sleep, improving brain function and liver function, etc. In cosmetics, it can also play anti-wrinkle, whitening, moisturizing and other functions. GABA is widely used in cosmetics, medicines, health care products and food abroad. In recent years, as developed countries such as Europe and the United States have put forward the development trend of green and healthy raw materials, foods and cosmetics containing the highly safe functional factor GABA will be more popular wi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P13/00C12R1/225
CPCC12P13/005C12N1/205C12R2001/225
Inventor 张天萌陆震孙劭靖郭文逸石艳丽郭学平栾贻宏
Owner BLOOMAGE BIOTECHNOLOGY CORP LTD
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