Method for screening lactic acid bacteria producing gamma-amino butyric acid

A technology of aminobutyric acid lactic acid bacteria and aminobutyric acid, applied in the biological field, can solve the problems of severe chemical synthesis reaction conditions, toxicity and corrosiveness, poor safety, etc., and achieve the effects of easy operation, simple method and mild reaction conditions

Inactive Publication Date: 2011-01-19
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In contrast, the chemical synthesis reaction conditions are severe, the chemical solvents used are toxic and corrosive, there are many by-products, and the safety is poor. It is mainly used in the chemical industry

Method used

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  • Method for screening lactic acid bacteria producing gamma-amino butyric acid
  • Method for screening lactic acid bacteria producing gamma-amino butyric acid
  • Method for screening lactic acid bacteria producing gamma-amino butyric acid

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Embodiment Construction

[0036] (1) Enrichment culture of lactic acid bacteria

[0037] Take 10-20mL of fermented dairy products (Shanghai Guangming Dairy Co., Ltd., Nanjing Weigang Dairy Co., Ltd.), fresh milk (from the Animal Husbandry Science and Technology Park of Anhui University of Science and Technology), fermented fruit and vegetable products (Yangzhou Sanhe pickles), etc. as raw materials. After diluting with sterilized distilled water, take 10-20 mL and add it to 100-20 mL skim milk or MRS liquid medium for enrichment culture, and culture at 37°C for 48 hours to obtain an enriched culture solution.

[0038] The skimmed milk medium: add 10 g of skim milk powder to 100 mL of distilled water, and sterilize at 115° C. for 10 min.

[0039] The MRS liquid medium: peptone 10.0g, beef extract 10.0g, yeast extract 5.0g, glucose 20.0g, sodium acetate 5.0g, triammonium citrate 2.0g, dipotassium hydrogen phosphate 2.0g, magnesium sulfate heptahydrate 0.58 g, manganese sulfate monohydrate 0.20g, TWEEN-8...

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Abstract

The invention discloses a method for screening lactic acid bacteria producing gamma-amino butyric acid, belonging to the biotechnical field. The method comprises the following steps: taking the separated and purified lactic acid bacteria; inoculating in an MRS culture medium; centrifuging and collecting thallus when the thallus growth reaches the stationary phase; carrying out whole-cell catalysis; taking L-glutamic acid or L-sodium glutamate as a unique substrate; taking acetic acid-sodium acetate as a buffer system; stirring and reacting for 1-24 hours at the temperature of 20-80 DEG C and with the pH of 3.2-8.0; utilizing glutamate decarboxylase producing GAD lactic acid bacteria to lead the L-glutamic acid or the L-sodium glutamate to play decarboxylization, so as to generate the gamma-aminobutyric acid; analyzing whether the reaction system contains gamma-amino butyric acid or not through paper chromatography to screen the lactic acid bacteria producing the gamma-amino butyric acid; and identifying the strains producing the gamma-amino butyric acid by the morphological characteristics, physiological biochemical characteristics and a 16SrDNA sequence. The method of the invention can greatly improve the directional screening efficiency of the functional probiotic bacteria.

Description

1. Technical field [0001] The invention relates to a screening method for microorganisms, in particular to a screening method for biosynthesizing γ-aminobutyric acid-producing lactic acid bacteria, and belongs to the field of biotechnology. 2. Background technology [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, 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, inducing human sperm acrosome, and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12Q1/68C12Q1/04C12R1/46
Inventor 陆兆新李远宏吕凤霞别小妹邹晓葵
Owner NANJING AGRICULTURAL UNIVERSITY
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