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Escherichia coli strain for high yield of Gamma-aminobutyric acid and method for producing Gamma-aminobutyric acid therefrom

A technology of aminobutyric acid and Escherichia coli, which is applied in the field of biochemistry, can solve the problems of low product concentration, low enzyme activity, and long reaction cycle, and achieve the effects of simple reaction system, simplified extraction process, and reduced production cost

Inactive Publication Date: 2011-02-16
ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The results of published patents and non-patent literatures using glutamate decarboxylase of Escherichia coli to prepare GABA have problems such as low enzyme activity, long reaction cycle, and low product concentration in the reaction solution.

Method used

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  • Escherichia coli strain for high yield of Gamma-aminobutyric acid and method for producing Gamma-aminobutyric acid therefrom
  • Escherichia coli strain for high yield of Gamma-aminobutyric acid and method for producing Gamma-aminobutyric acid therefrom
  • Escherichia coli strain for high yield of Gamma-aminobutyric acid and method for producing Gamma-aminobutyric acid therefrom

Examples

Experimental program
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Effect test

Embodiment 1

[0021] After the CGMCC No.3562E.coli ZS-07 strain was activated on the slant medium, it was transferred to the seed medium with a pH of 7.0-7.8. After shaking and culturing at 37°C for 6-9 hours, it was inoculated with an inoculum of 3-12%. Fermentation medium, pH 7.0-7.8, 37°C, 220r / min, shake culture for 8-14h, then reduce the pH to 3.5-3.8 0.5-1h before the end to obtain the fermentation broth containing bacteria, centrifuge and collect at 4000r / min bacteria. Resuspend the bacteria in water equivalent to 1 / 2 times the volume of the original fermentation broth, add 15g of L-glutamic acid to each 100mL of the bacterial suspension, add 0.1mmol / L of pyridoxal phosphate, and control the pH with hydrochloric acid or sulfuric acid. 3.5-3.8, shaking reaction at 41°C, when there is no insoluble substrate, add substrate L-glutamic acid 15g / 100mL. When reacting for about 20 hours, add pyridoxal phosphate 0.1mmol / L, then add substrate L-glutamic acid 15g / 100mL, and add bacteria (prepa...

Embodiment 2

[0023] After the CGMCC No.3562E.coli ZS-07 bacterial strain is activated on the slant medium, it is transferred to the seed medium, and after shaking and culturing at 37°C for 6-9h, it is inoculated with a 3-12% inoculum amount to contain pyridoxal phosphate ( 0.02mmol / L) fermentation medium, pH 7.0-7.8, 37°C shaking culture for 8-14h, reduce the pH to 3.5-3.8 0.5-1h before the end of the culture, add 28g L-glutamic acid per 100mL fermentation broth , use hydrochloric acid or sulfuric acid to control the pH to 3.5-3.8, continue to vibrate or stir until the substrate is completely converted, and centrifuge to obtain a reaction solution containing γ-aminobutyric acid, and the conversion rate can reach 100%.

Embodiment 3

[0025] After the CGMCC No.3562E.coli ZS-07 strain was activated on the slant medium, it was transferred to the seed medium with a pH of 7.0-7.8. After shaking and culturing at 37°C for 6-9 hours, it was inoculated with an inoculum of 3-12%. Fermentation medium, pH 7.0-7.8, 37°C, 220r / min, shake culture for 8-14h, then reduce the pH to 3.5-3.8 0.5-1h before the end of culture, obtain the fermentation broth containing bacteria, centrifuge at 4000r / min Separate and collect bacteria. Resuspend the bacteria in an acetic acid-sodium acetate buffer solution with a pH of 3.5 equivalent to 1 / 2 the volume of the original fermentation broth, add 5 g of L-sodium glutamate per 100 mL of the bacterial suspension, and control the pH to 3.5 with hydrochloric acid or sulfuric acid -3.8, shake the reaction at 41°C until the substrate is completely converted, centrifuge to obtain a reaction solution containing γ-aminobutyric acid, and the conversion rate reaches 100%. The purity of GABA reaches...

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Abstract

The invention discloses a safe escherichia coli ZS-07 strain for high yield of Gamma-aminobutyric acid as well as a method for synthesizing Gamma-aminobutyric acid by utilizing the same, belonging to the field of biochemistry. The preservation number of the escherichia coli strain is CGMCC No.3562, and glutamate decarboxylase contained in the escherichia coli is acted on L-glutamic acid, L-glutamate or matters containing the L-glutamic acid or the L-glutamate to synthesize the Gamma-aminobutyric acid. The strain has the advantages of high glutamate decarboxylase activity and strong acid resistance, and the method has the advantages of mild reaction condition, fewer byproducts, simplicity, low cost and high production efficiency and is easy to operate. In addition, the Gamma-aminobutyric acid has high purity and can be used in the fields of chemical engineering, foods, feeds, medicines, and the like.

Description

technical field [0001] The invention relates to a high-yielding gamma-aminobutyric acid Escherichia coli ZS-07 strain and a microorganism and enzymatic preparation method of gamma-aminobutyric acid, belonging to the field of biochemistry. Background technique [0002] γ-aminobutyric acid (GABA for short) is a non-protein amino acid that widely exists in nature and is an important inhibitory neurotransmitter in the central nervous system. Long-term lack of GABA in the brain will lead to epilepsy, Parkinson and other diseases. At the same time, GABA is also related to brain aging, and its deficiency will lead to "deafness and blindness" in the elderly. Therefore, GABA not only has calming and antidepressant effects, but also enhances brain function and long-term memory. In recent years, with the in-depth research on the physiological activity of GABA, it has been found that its physiological functions are extremely extensive. For example, it can increase the secretion of gr...

Claims

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Application Information

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IPC IPC(8): C12P13/00C12R1/19C12N1/20
Inventor 戴桂馥吴健周新钦王卫富马文艳王廷王春阳谭印成蒋盛耀李季伦
Owner ZHENGZHOU UNIV
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