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Production method of feed gamma-aminobutyric acid

A technology of aminobutyric acid and its production method, which is applied in the production field of γ-aminobutyric acid for feed, can solve the problems of difficult control of glutamic acid decarboxylase activity, level and high yield, etc., so as to improve production efficiency and stabilize production Effect

Inactive Publication Date: 2019-03-22
上海邦成生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, in the production process of γ-aminobutyric acid (GABA), the GABA yield is mainly related to the activity of glutamic acid decarboxylase produced by the bacteria. The activity of decarboxylase is difficult to control, and it is difficult to control the level and high yield of subsequent production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] (1) Insert the Escherichia coli sclerotin tube seeds into the seed medium, the seed medium: yeast powder 4g / L, peptone 8g / L, sodium chloride 10g / L, pH7.0; seed culture conditions: temperature 36°C, Air volume 0.6vvm, stirring 350rpm, cultivate to OD1.5; inoculate in fermentation medium according to 4% seed inoculum, fermentation medium: (NH4)2SO49g / L, glucose 6g / L, K2HPO417g / L, KH2PO43.0g / L, MgSO4 18g / L, glutamic acid 0.04g / L; air volume 1: 0.5vvm, temperature 37°C, stirring 300rpm to grow to OD3.0 and cool down to 29°C;

[0032] (2) Add 8g / L lactose to induce the expression of glutamic acid decarboxylase. When OD600 reaches 15, start to reduce the culture temperature to 25°C, reduce the rotation speed by 20% to 240, and reduce the air volume by 50%. After that, reduce the temperature every 1 hour 0.5°C, the stirring speed was reduced by 10% from the last time, and the air flow rate was reduced to 50% of the previous time. At the same time, the enzyme activity rapid as...

Embodiment 2

[0036](1) Insert tube seeds of Escherichia coli sclerotin into seed medium, seed medium: yeast powder 7g / L, peptone 5g / L, sodium chloride 0.8g / L, pH 7.0. Seed culture conditions: temperature 35°C, air volume 0.8VVM, agitation 250rpm, cultivate to OD1.5; inoculate 2% seed in the fermentation medium, fermentation medium: peptone 12g / L, sucrose 4g / L, K2HPO414g / L, KH2PO42g / L, MgSO428g / L, glutamic acid 0.11g / L; air volume 0.7vvm, temperature 35°C, stirring at 400rpm and culturing until OD2.5 and cooling down to 28°C;

[0037] (2) Add 4.5g / L lactose to induce the expression of glutamic acid decarboxylase. When the OD600 reaches 18, start to reduce the culture temperature to 25°C, reduce the rotation speed by 20% to 240, and reduce the air volume by 50% at the same time, and then reduce it every 1 hour The temperature is 0.5°C, the stirring speed is reduced by 10% from the previous time, and the air flow rate is reduced to 50% of the previous time. At the same time, the enzyme activ...

Embodiment 3

[0041] (1) Insert Escherichia coli sclerotin tube seeds into the seed medium, seed medium: yeast powder 3g / L, peptone 11g / L, sodium chloride 0.9g / L, pH7.0; seed culture conditions: temperature 34°C , air volume 1.0VVM, stirring 400rpm, cultivated to OD1.1; according to the seed 1.5% inoculum size inoculated in the fermentation medium, fermentation medium: (NH4)2SO411g / L, sucrose 4g / L, K2HPO413g / L, KH2PO41.7g / L, MgSO4 25g / L, glutamic acid 0.08g / L; air volume 1: 1.0vvm, temperature 35°C, stirring at 350rpm, culture until OD1.5~2.5 and cool down to 25°C;

[0042] (2) Add 6g / L lactose to induce the expression of glutamic acid decarboxylase. When OD600 reaches 12, start to reduce the culture temperature to 25°C, reduce the rotation speed by 20% to 240, and reduce the air volume by 50%. After that, reduce the temperature every 1 hour 0.5°C, the stirring speed was reduced by 10% from the last time, and the air flow rate was reduced to 50% of the previous time. At the same time, the ...

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Abstract

The invention discloses a production method of feed gamma-aminobutyric acid. The production method comprises the following steps: (1) inoculating genetic engineering escherichia coli into a fermentation medium with a seed solution cultured by a seed medium at an inoculation amount of 1%-4%; (2) adding 4-8g / L of an inducer for inducing the expression of a glutamic acid decarboxylase, performing gradient adjustment on culture conditions of fermentation liquor when OD600 reaches 10-18, detecting the enzyme activity of thalli constantly at the same time and ending fermentation when the enzyme activity is 55U or above; (3) heating the completed fermentation liquor to 30-38 DEG C with a heating system, stirring the fermentation liquor at a rotation speed of 100-200rpm and adding a conversion rawmaterial; and (4) directly performing spray drying on the converted fermentation liquor to obtain the feed gamma-aminobutyric acid. According to the production method of the feed gamma-aminobutyric acid, by optimizing production conditions of producing strains of glutamic acid, the enzyme activity of the glutamic acid decarboxylase can be quickly detected and controlled, a method for controllingthe enzyme activity of the glutamic acid decarboxylase is improved, and the capacities of improving production efficiency and stabilizing production can be achieved.

Description

technical field [0001] The invention relates to the technical field of feed processing, in particular to a production method of gamma-aminobutyric acid for feed. Background technique [0002] As a non-protein amino acid, γ-aminobutyric acid is a good raw material for medical drugs and health products. It has the functions of lowering blood pressure, maintaining nerve stability, improving brain function, protecting liver and kidney, and can be used as a drug for treating brain damage. It has anti-heat stress and growth-promoting effects on livestock and poultry. It has been developed into a new type of functional factor and is widely used in medicine, food health care, chemical industry and agriculture. In recent years, the demand for γ-aminobutyric acid in domestic and foreign markets has increased year by year. The production methods of γ-aminobutyric acid mainly include chemical synthesis, solid-state fermentation and biological enzyme conversion. [0003] The chemical s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/00C12N9/88C12R1/19
CPCC12N9/88C12P13/005C12Y401/01015
Inventor 张继学陆克文
Owner 上海邦成生物工程有限公司
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