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Semicontinuous fermentation method for producing gamma-aminobutyric acid by lactic acid bacteria

A technology of aminobutyric acid and lactic acid bacteria, applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc., can solve problems that have not been seen, and achieve the effects of reduced dosage, safe methods, and mild reaction conditions

Inactive Publication Date: 2011-06-29
山东水晶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

Based on these factors, in order to maximize the yield of GABA, it is necessary to combine segmental oxygen control and segmental temperature control during the fermentation process, but there is no report on this aspect so far.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] 1. Preparation of seed solution

[0030] Put the freeze-dried powder of Yihuan lactic acid bacteria NBRC 12005 into 100mL sterilized MRS medium, and culture it in a shaker flask at 30°C and 160r / min for 24h. Then inoculate 100 mL of sterilized MRS medium with 5% inoculum amount, and culture in shake flask at 33° C. and 160 r / min for 24 hours to obtain seed solution.

[0031] Among them: the components of the MRS medium are: casein peptone 10g / L, beef extract 10g / L, yeast extract 5g / L, glucose 5g / L, sodium acetate 5g / L, diamine citrate 0.2g / L, Tween 0.1g / L, dipotassium hydrogen phosphate 0.2g / L, magnesium sulfate 0.2g / L, manganese sulfate 0.05g / L, pH 6.5.

[0032] 2. Production of γ-aminobutyric acid by semi-continuous fermentation

[0033] Inoculate the seed liquid into 1L MRSG medium with an inoculum amount of 5%, and ferment by adding substrates in batches, temperature control in stages, and oxygen control in stages: add 5g at 12h, 30h, 50h, and 70h of fermentation ...

Embodiment 2

[0037] 1. Preparation of seed solution

[0038] Step is identical with embodiment 1.

[0039] 2. Production of γ-aminobutyric acid by semi-continuous fermentation

[0040] The seed solution was inoculated into 1L MRSG medium with an inoculum amount of 5%, and the combined fermentation was carried out by adding substrates in batches, temperature control in stages, and oxygen control in stages: add 10g at 16h, 36h, 56h, and 76h of fermentation of sodium glutamate into the fermentation broth. From the start of fermentation to 16 hours, the temperature is controlled at 37°C for facultative anaerobic fermentation; from the 16th to 96th hour of fermentation, the temperature is controlled at 30°C for anaerobic fermentation.

[0041] Among them, the MRSG medium is: glucose 10.92g / L, peptone 29.67g / L, beef extract 39.66g / L, manganese sulfate 0.06g / L, vitamin B 6 0.02g / L, sodium glutamate 2g / L, pH4.8.

[0042]The total fermentation time was 96h, the fermentation was terminated, and...

Embodiment 3

[0044] 1. Preparation of seed solution

[0045] Step is identical with embodiment 1.

[0046] 2. Production of γ-aminobutyric acid by semi-continuous fermentation

[0047] The seed solution was inoculated into 1L of MRSG medium with an inoculation amount of 5%, and the combined fermentation was carried out by adding substrates in batches, temperature control in stages, and oxygen control in stages: 20g were added at 24h, 42h, 62h, and 82h of fermentation of sodium glutamate into the fermentation broth. From the start of fermentation to 24 hours, the temperature is controlled at 37°C for facultative anaerobic fermentation; from the 24th to 106h of fermentation, the temperature is controlled at 30°C for anaerobic fermentation.

[0048] Among them, the MRSG medium is: glucose 12g / L, peptone 30g / L, beef extract 40g / L, manganese sulfate 1g / L, vitamin B 6 0.05g / L, sodium glutamate 2g / L, pH4.8.

[0049] The total fermentation time was 106h, the fermentation was terminated, and th...

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Abstract

The invention discloses a semicontinuous fermentation method for producing gamma-aminobutyric acid by lactic acid bacteria, which comprises the following steps: inoculating lactic acid bacteria onto an Man, Rogosa and Sharpe gelatin (MRSG) culture medium; and fermenting in a mode of segmental substrate addition, segmental temperature control and segmental oxygen control to obtain gamma-aminobutyric acid, wherein the solvent of the MRSG culture medium is water, and the solute comprises the following materials at concentration: 7 to 12g / L glucose, 20 to 30g / L peptone, 30 to 40g / L beef extract, 0.05 to 1g / L manganese sulfate, 0.01 to 0.05g / L vitamin B, and 2 to 5g / L sodium glutamate. When the method provided by the invention is adopted, the yield of the gamma-aminobutyric acid can reach 13.1to 38.7g / L.

Description

technical field [0001] The invention relates to a semi-continuous fermentation method for producing gamma-aminobutyric acid by using lactic acid bacteria. Background technique [0002] γ-aminobutyric acid is widely distributed in mammals to single-celled organisms, and can be converted into L-glutamic acid or glutamate by glutamic acid decarboxylase. Studies have proved that γ-aminobutyric acid is an inhibitory neurotransmitter in the central nervous system, and has important physiological functions, such as lowering blood pressure, promoting mental stability, promoting blood flow in the brain, increasing brain vitality, nourishing nerve cells, and increasing growth hormone secrete, invigorate the liver and kidney, improve menopausal syndrome, etc. In 2007, Japan's Coca-Cola Company launched the γ-aminobutyric acid functional drink "AQUARIUS SHARPCHARGE" for the public, which has relaxation and anti-stress effects. PFI GABA RelaxS (gamma-aminobutyric acid tablets) imported...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/00C12R1/225
Inventor 孙君社刘萍张秀清范杰张京声
Owner 山东水晶生物科技股份有限公司
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