A method for producing low molecular weight gamma-polyglutamic acid based on staged fermentation

A low-molecular-weight, staged fermentation technology, applied in the field of industrial microorganisms, can solve the problems of low efficiency and cumbersome production process of low-molecular-weight γ-polyglutamic acid, so as to expand the application market, enrich product categories, and achieve significant economic and social benefits. Effect

Active Publication Date: 2018-11-20
广东迪美生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims at the cumbersome and low-efficiency production process of low-molecular-weight γ-polyglutamic acid in the prior art, takes the existing industrial fermentation strain Bacillus subtilis PGA-7 as the research object, and optimizes and controls the fermentation process , allowing the strain to directly produce low molecular weight products during the fermentation stage, providing a low-cost and efficient method for producing low molecular weight γ-polyglutamic acid based on staged fermentation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Weigh each component of the medium, mix and dissolve with deionized water, so that the final concentration of each component is: citric acid 20g / L, sodium glutamate 15g / L, glycerol 50g / L, magnesium sulfate 0.05g / L, Manganese sulfate 0.05g / L, ferric chloride 0.1g / L and calcium chloride 0.1g / L, after adjusting the pH to 7.5, use 0.1μm, 0.22μm, 0.45μm microporous membranes to filter and sterilize once respectively to obtain Bacterial fermentation medium.

[0017] Add sterile fermentation medium into the fermenter, inoculate Bacillus subtilis (Bacillus subtilis) PGA-7 into the fermentation medium, the inoculum amount is 1% (volume fraction), and adopt staged fermentation for cultivation. In the first stage, the temperature is 36°C and the ventilation flow rate is 2Nm 3 / h, the stirring speed was 50rpm, and cultivated for 12h; in the second stage, the temperature was 35°C, the ventilation flow was doubled, the stirring speed was doubled, and the culture was continued for 48...

Embodiment 2

[0019] Weigh each component of the medium, mix and dissolve with deionized water, so that the final concentration of each component is: citric acid 20g / L, sodium glutamate 30g / L, glycerin 100g / L, magnesium sulfate 0.1g / L, Manganese sulfate 0.1g / L, ferric chloride 0.1g / L and calcium chloride 0.1g / L, after adjusting the pH to 7.5, use 0.1μm and 0.22μm microporous membranes to filter and sterilize once respectively to obtain sterile fermentation Use medium.

[0020] Add sterile fermentation medium into the fermenter, inoculate Bacillus subtilis (Bacillus subtilis) PGA-7 into the fermentation medium, the inoculum amount is 2% (volume fraction), and adopt staged fermentation for cultivation. In the first stage, the temperature is 34°C, and the ventilation flow rate is 2Nm 3 / h, the stirring speed was 50rpm, and cultivated for 12h; in the second stage, the temperature was 35°C, the ventilation flow rate was increased by 2 times, the stirring speed was increased by 2 times, and the ...

Embodiment 3

[0022] Weigh each component of the medium, mix and dissolve with deionized water, so that the final concentration of each component is: citric acid 20g / L, sodium glutamate 30g / L, glycerin 100g / L, magnesium sulfate 0.1g / L, Manganese sulfate 0.1g / L, ferric chloride 0.1g / L and calcium chloride 0.1g / L, after adjusting the pH to 7.5, use 0.1μm and 0.45μm microporous membranes to filter and sterilize once respectively to obtain sterile fermentation Use medium.

[0023] Add sterile fermentation medium into the fermenter, inoculate Bacillus subtilis (Bacillus subtilis) PGA-7 into the fermentation medium, the inoculum amount is 3% (volume fraction), and adopt staged fermentation for cultivation. In the first stage, the temperature is 34°C and the ventilation flow rate is 4Nm 3 / h, stirring speed 100rpm, cultured for 24h; in the second stage, the temperature was 35°C, the ventilation flow was doubled, the stirring speed was doubled, and the culture was continued for 72h to obtain a hig...

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Abstract

The invention discloses a method for producing low-molecular-weight gamma-polyglutamic acid based on segmented fermentation. According to the invention, a medium used for fermentation is prepared by a microfiltration membrane filtering degerming method, then bacillus subtilis is inoculated, and then culturing is carried out by employing segmented fermentation; in a first phase, temperature is 30-36 DEG C, an air flow rate is 2-10 Nm<3> / h, the stirring speed is 50-200rpm, culturing is carried out for 12-24 hours; and in a second phase, the temperature is 35-39 DEG C, the air flow rate is accelerated by 1-2 times, the stirring speed is accelerated by 1-2 times, culturing is carried out for 24-96 hours; and thereby a gamma-polyglutamic acid fermentation broth with high products concentration being 30-60g / L and low molecular weight being 35-260 KDa is obtained. By adjusting and optimizing the fermentation technology, the low-molecular-weight product with high density can be directly obtained at a microbial fermentation phase, and the method is the novel technology for producing the low-molecular-weight gamma-polyglutamic acid with high efficiency and low cost.

Description

technical field [0001] The invention belongs to the technical field of industrial microorganisms, and in particular relates to a method for producing low molecular weight gamma-polyglutamic acid based on staged fermentation. Background technique [0002] γ-polyglutamic acid is a glutamic acid polymer produced by microbial fermentation. It has the functions of moisturizing, maintaining skin elasticity, increasing the content of natural moisturizing factors, whitening skin color, and promoting the absorption of nutrients. It can be industrially produced through microbial fermentation . [0003] The biological function of γ-polyglutamic acid is closely related to its relative molecular weight. Low-molecular-weight products are widely used in cosmetics, but the molecular weight of direct fermentation products is relatively large, and the molecular weight must be reduced by post-acid hydrolysis (currently the main method for reducing molecular weight). Resulting in a serious red...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P13/02C12R1/125
Inventor 林万泉温灼斌陈爱美冯劲施庆珊邱晓颖黄小茉
Owner 广东迪美生物技术有限公司
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