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Process for biologically synthesizing gamma-amino butyric acid

An aminobutyric acid, biosynthesis technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as hidden dangers of food safety and hygiene, and achieve a reduction in separation and purification processes, low production costs, and simple methods. Effect

Inactive Publication Date: 2006-09-20
ZHEJIANG UNIV
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  • Application Information

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Problems solved by technology

It has been reported in the literature that Escherichia coli is usually used as the production strain for the biosynthesis of GABA, but there are hidden dangers in food safety and hygiene when using Escherichia coli as the production strain. This laboratory screened out a lactic acid bacteria strain with high-efficiency biosynthesis of GABA

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] After the strain was activated on the agar slant medium, it was transferred to the GYP seed medium for 24 hours. The seed culture liquid was inoculated in the GYP fermentation medium with 1% inoculation amount, and 1% of Sodium L-glutamate monohydrate, the liquid volume of the fermentation medium is 200mL / 500mL, and it is left to culture for 60 hours to obtain the fermentation liquid containing bacteria, and the bacteria are collected by centrifugation (8000r / min, 5min). After centrifugation, the cells were washed with sterilized deionized water, and 0.5 g of wet cells were taken, suspended in 25 mL of citric acid-disodium hydrogen phosphate buffer system, and the content of L-sodium glutamate was 10 mM. Reaction 1 hours, the reaction solution was centrifuged and analyzed by high-performance liquid chromatography, and the content of γ-aminobutyric acid was about 1g / L.

Embodiment 2

[0021] After the strain was activated on the agar slant medium, it was transferred to the GYP seed medium for 24 hours. The seed culture solution was inoculated into the GYP fermentation medium with 1% inoculation amount, and 1% of the fermented medium was added to the fermentation medium. Sodium L-glutamate monohydrate, the liquid volume of the fermentation medium is 200mL / 500mL, and it is left to culture for 60 hours to obtain the fermentation liquid containing bacteria, and the bacteria are collected by centrifugation (8000r / min, 5min). After centrifugation, the cells were washed with sterilized deionized water, and 0.5 g of wet cells were taken, suspended in 25 mL of citric acid-disodium hydrogen phosphate buffer system, and the content of L-sodium glutamate was 20 mM. Reaction 2 hours, the reaction solution was centrifuged and analyzed by high performance liquid chromatography, and the content of γ-aminobutyric acid was about 2g / L.

Embodiment 3

[0023] After the strain was activated on the agar slant medium, it was transferred to the GYP seed medium for 24 hours. The seed culture liquid was inoculated in the GYP fermentation medium with 1% inoculation amount, and 1% of Sodium L-glutamate monohydrate, the liquid volume of the fermentation medium is 200mL / 500mL, and it is left to culture for 60 hours to obtain the fermentation liquid containing bacteria, and the bacteria are collected by centrifugation (8000r / min, 5min). After centrifugation, the cells were washed with sterilized deionized water, and 0.5 g of wet cells were taken, suspended in 25 mL of citric acid-disodium hydrogen phosphate buffer system, and the content of L-sodium glutamate was 40 mM. Reaction 3 hours, the reaction solution was centrifuged and analyzed by high performance liquid chromatography, and the content of γ-aminobutyric acid was about 4g / L.

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Abstract

The process of biosynthesizing gamma-amino butyric acid includes the following steps: slant agar culture to activate Lactobacillus brevis with the preservation number of CGMCC No. 1306, inoculating to GYP or MRS seed culture medium to culture for 10-30 hr, inoculating to GYP or MRS fermenting culture medium in the amount of 0.5-5 % to stationarily culture at 25-35 deg.c for 48-120 hr to obtain fermented liquid containing thallus and centrifugally separating and collecting thallus; washing thallus with sterilized deionized water, suspending wet thallus in citric acid-disodium hydrogen phosphate buffering system with sodium L-glutamate to react for 1-10 hr, and centrifuging the reacted liquid to obtain gamma-amino butyric acid solution. The present invention has low production cost, mild reaction condition, less environmental pollution, simple process, high production efficiency, and high product purity.

Description

technical field [0001] The invention relates to a method for biosynthesizing γ-aminobutyric acid. Background technique [0002] γ-Aminobutyric acid (GABA for short) is a non-protein amino acid with important physiological activity and an important neurotransmitter in the central nervous system. A large number of literature reports that GABA can lower blood pressure and cholesterol, reduce neuron activity, and prevent nerve cell overheating. It not only has the effect of calming the nerves and anti-depression, but also enhances brain function and long-term memory, increases growth hormone secretion, prevents obesity, and strengthens the liver. Kidney benefit, improve menopausal syndrome and other functions. With the continuous in-depth research on the physiological activity of GABA, more and more physiological functions of γ-aminobutyric acid have been discovered in the latest research. GABA has also received more and more attention. Although GABA exists widely in organisms...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P13/00C12R1/24
Inventor 梅乐和黄俊武鸿
Owner ZHEJIANG UNIV
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