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Method for performing fermentation production on gamma-aminobutyric acid by using high-concentration monopotassium phosphate as buffer salt

A technology of potassium dihydrogen phosphate and aminobutyric acid is applied in microorganism-based methods, biochemical equipment and methods, fermentation and other directions to achieve the effects of increased yield, simple operation and strong buffering capacity

Active Publication Date: 2014-01-15
健隆生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Potassium dihydrogen phosphate, as a commonly used buffer salt, is often added in lactic acid bacteria culture medium, but as the addition of potassium dihydrogen phosphate in the present invention reaches 20 ~ 50 g / L, the addition of this ultra-high concentration buffer salt And finally obtain the situation that the thallus volume and GABA content are more than 3 times higher than the initial conditions, so far there has been no report in the field of Lactococcus lactis producing GABA

Method used

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  • Method for performing fermentation production on gamma-aminobutyric acid by using high-concentration monopotassium phosphate as buffer salt
  • Method for performing fermentation production on gamma-aminobutyric acid by using high-concentration monopotassium phosphate as buffer salt
  • Method for performing fermentation production on gamma-aminobutyric acid by using high-concentration monopotassium phosphate as buffer salt

Examples

Experimental program
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Effect test

Embodiment 1

[0037] Example 1: Lactococcus lactis after two generations of activation by conventional methods was inoculated in a specific medium (glucose 30 g / L, yeast extract 40 g / L, potassium dihydrogen phosphate 20 g / L, Sodium glutamate 10 g / L, MgSO 4 0.6 g / L, MnSO 4 ·H 2O 0.1 g / L, Tween-80 0.5 mL / L, initial pH controlled at 6.8-7.0) in the fermenter. The culture temperature was 30 °C, the rotation speed was 50 r / min, and the culture was not ventilated. When the pH of the fermentation liquid dropped to 5.0 naturally, ammonia water or sodium hydroxide solution was added to maintain the pH at 5.0±0.5, and the fermentation liquid was added to the fermentation liquid after 24-48 h. Add sodium glutamate concentrate to maintain 5~15 g / L, add glucose during the whole fermentation process to maintain the concentration of fermentation liquid at about 5~20 g / L, and cultivate for 40~60 h to obtain high γ-amino Butyric acid lactic acid bacteria fermentation broth. After culturing for 48 hours...

Embodiment 2

[0038] Example 2: Lactococcus lactis after two generations of activation by conventional methods was inoculated in a specific medium (glucose 30 g / L, yeast extract 40 g / L, potassium dihydrogen phosphate 30 g / L, Sodium glutamate 10 g / L, MgSO 4 0.6 g / L, MnSO 4 ·H 2 O 0.1 g / L, Tween-80 0.5 mL / L, initial pH controlled at 6.8-7.0) in the fermenter. The culture temperature was 30 °C, the rotation speed was 50 r / min, and the culture was not ventilated. When the pH of the fermentation liquid dropped to 5.0 naturally, ammonia water or sodium hydroxide solution was added to maintain the pH at 5.0±0.5, and the fermentation liquid was added to the fermentation liquid after 24-48 h. Add sodium glutamate concentrate to maintain 5~15 g / L, add glucose during the whole fermentation process to maintain the concentration of fermentation liquid at about 5~20 g / L, and cultivate for 40~60 h to obtain high γ-amino Butyric acid lactic acid bacteria fermentation broth. After culturing for 48 hour...

Embodiment 3

[0039] Example 3: Lactococcus lactis after two generations of activation by conventional methods was inoculated in a specific medium (glucose 30 g / L, yeast extract 40 g / L, potassium dihydrogen phosphate 40 g / L, Sodium glutamate 10 g / L, MgSO 4 0.6 g / L, MnSO 4 ·H 2 O 0.1 g / L, Tween-80 0.5 mL / L, initial pH controlled at 6.8-7.0) in the fermenter. The culture temperature was 30 °C, the rotation speed was 50 r / min, and the culture was not ventilated. When the pH of the fermentation liquid dropped to 5.0 naturally, ammonia water or sodium hydroxide solution was added to maintain the pH at 5.0±0.5, and the fermentation liquid was added to the fermentation liquid after 24-48 h. Add sodium glutamate concentrate to maintain 5~15 g / L, add glucose during the whole fermentation process to maintain the concentration of fermentation liquid at about 5~20 g / L, and cultivate for 40~60 h to obtain high γ-amino Butyric acid lactic acid bacteria fermentation broth. After culturing for 48 hour...

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Abstract

The invention discloses a method for performing fermentation production on gamma-aminobutyric acid by using high-concentration monopotassium phosphate as a buffer salt, and belongs to the field of microbe fermentation engineering. The method comprises the following steps: using lactococcus lactis (Lactococcus lactis SYFS1.009) which can produce gamma-aminobutyric acid as a strain, culturing the lactococcus lactis in a culture medium containing high-concentration monopotassium phosphate, wherein the culture medium comprises the following components: 10-40 g / L of glucose, 20-50 g / L of yeast extract, 20-50 g / L of monopotassium phosphate, 5-20 g / L of sodium glutamate, 0-3 g / L of MgSO4, 0-3 g / L of MgSO4.H2O and 0-5mL / L of Tween-80; performing fed-batch operation on ammonia water or sodium hydroxide to regulate the pH of the fermentation liquor on the basis of the culture medium, adding a certain amount of glucose and sodium glutamate in the fermentation process, and culturing for 40-60 hours to obtain the fermentation liquor containing high-concentration gamma-aminobutyric acid. The used culture medium and method are safe, reliable and short in production cycle, the pH of the fermentation liquor is regulated by using the monopotassium phosphate as the buffer salt and performing fed-batch operation on the ammonia water or sodium hydroxide, and an inhibiting effect of lactic acid on the lactococcus lactis can be effectively relieved.

Description

technical field [0001] The invention relates to a method for fermenting and producing gamma-aminobutyric acid using high-concentration potassium dihydrogen phosphate as a buffer salt, belonging to the field of microbial fermentation engineering. Background technique [0002] γ-aminobutyric acid (γ-aminobutyric acid, GABA for short) is a naturally occurring, non-protein amino acid, mainly found in the brain and spinal cord of mammals. GABA has the functions of improving brain vitality, lowering blood pressure and cholesterol, stabilizing the spirit, preventing obesity, improving liver and kidney functions, etc. Therefore, it has broad application prospects in the field of functional foods. [0003] At present, the production methods of GABA mainly include chemical synthesis and biological methods. Although the chemical synthesis method has high purity, it has high cost and low yield, and involves dangerous solvents in the production process, which is difficult to realize in ...

Claims

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Application Information

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IPC IPC(8): C12P13/00C12R1/46
Inventor 江波沐万孟冯慧杰张涛
Owner 健隆生物科技股份有限公司
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