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Gamma-polyglutamic acid high-yield strain and method for preparing gamma-polyglutamic acid by liquid fermentation by using strain

A technology of polyglutamic acid and high-yielding strains, applied in the field of bioengineering

Active Publication Date: 2019-04-12
CHAMBROAD CHEM IND RES INST CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the maximum concentration of γ-polyglutamic acid produced by large-scale (500L fermenter) fermentation reported in the literature is generally around 35g / L. The concentration can reach more than 50g / L or higher, but there are no reports of practical application in production. Therefore, the highest concentration of gamma-polyglutamic acid in the existing industrial production is generally around 35g / L

Method used

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  • Gamma-polyglutamic acid high-yield strain and method for preparing gamma-polyglutamic acid by liquid fermentation by using strain
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Embodiment 1

[0088] A method for preparing gamma-polyglutamic acid by liquid fermentation, the specific steps are as follows:

[0089] (1) Activation of strains: move the strains on the slant of the test tube stored at 4°C on the LB medium to room temperature for activation for 4 hours;

[0090] (2) Preparation of liquid seeds: On an aseptic operating table, use a sterile inoculation loop to inoculate 2 single colonies into a triangular flask containing sterilized seed medium, culture at 30°C, shaker speed at 170rpm, and culture for 24 hours ;

[0091] Seed medium composition: glucose 20g / L, yeast extract 7g / L, sodium glutamate 19g / L, K 2 HPO 4 ·3H 2O 2g / L, MgSO 4 0.20g / L, distilled water 1000mL, pH 7.3, sterilization conditions 121°C, 0.15Mpa, sterilize for 20min.

[0092] (3) Fermentation, regulation and control process: the liquid seed is received in the fermentation medium with the inoculation amount of 7% (v / v), and the 500L fermenter fermentation medium filling liquid volume is...

Embodiment 2

[0100] A method for preparing gamma-polyglutamic acid by liquid fermentation, the specific steps are as follows:

[0101] (1) Activation of strains: move the strains on the slant of the test tube stored at 4°C on the LB medium to room temperature for activation for 4 hours;

[0102] (2) Preparation of liquid seeds: On an aseptic operating table, use a sterile inoculation loop to inoculate 2 single colonies into a triangular flask containing sterilized seed medium, cultivate at a temperature of 32°C, rotate at a shaker speed of 180rpm, and cultivate for 20 hours ;

[0103] Seed medium composition: glucose 20g / L, yeast extract 8g / L, sodium glutamate 20g / L K 2 HPO 4 ·3H 2 O 2g / L, MgSO 4 0.20g / L, distilled water 1000mL, pH 7.5, sterilization condition 121℃, 0.15Mpa, sterilization 20min;

[0104] (3) Fermentation, regulation and control process: the liquid seed is received in the fermentation medium with the inoculum size of 5% (v / v), and the liquid filling capacity of the fe...

Embodiment 3

[0112] A method for preparing gamma-polyglutamic acid by liquid fermentation, the specific steps are as follows:

[0113] (1) Activation of strains: move the strains on the slant of the test tube stored at 4°C on the LB medium to room temperature for activation for 4 hours;

[0114] (2) Preparation of liquid seeds: On an aseptic operating table, use a sterile inoculation loop to inoculate 3 single colonies into a triangular flask containing sterilized seed medium, cultivate at a temperature of 35°C, rotate at a shaker speed of 190rpm, and cultivate for 16 hours ;

[0115] Seed medium composition: glucose 18g / L, yeast extract 8g / L, sodium glutamate 20g / L K 2 HPO 4 ·3H 2 O 2g / L, MgSO 4 0.20g / L, distilled water 1000mL, pH 7.0, sterilization condition 121℃, 0.15Mpa, sterilization 20min;

[0116] (3) Fermentation and regulation process: the liquid seed is received in the fermentation medium with the inoculum size of 5% (v / v), and the liquid filling capacity of the fermentatio...

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Abstract

The invention belongs to the field of bioengineering technology, and provides a gamma-polyglutamic acid high-yield strain and a method for preparing gamma-polyglutamic acid by liquid fermentation by using the strain. The strain produced by the gamma-polyglutamic acid is bacillus, and is bacillus subtilis identified as by 16S rDNA. The strain code is YJY18-11, and the preservation number of the strain preserved in the China General Microbiological Culture Collection Center is CGMCC No.16159. The strain can be applied to produce the gamma-polyglutamic acid, and the content of the gamma-polyglutamic acid obtained fermented by the method can reach 40 g / L or above.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and provides a high-yield gamma-polyglutamic acid strain and a method for preparing gamma-polyglutamic acid by liquid fermentation using the strain. Background technique [0002] γ-polyglutamic acid is an anionic polyamino acid formed by the polymerization of glutamic acid monomers through γ-amide bonds, and its molecular weight is generally between 10-2000kDa. It is a multifunctional and environmentally friendly biopolymer material. The water solubility, biocompatibility, and biodegradability of γ-polyglutamic acid benefit from the large number of free carboxyl groups contained in its main chain. These active groups are prone to reactions such as crosslinking, chelation, and derivatization. It has broad application prospects in agriculture, environmental governance, cosmetics, medicine, food and other fields. [0003] Since Ivanovic and others discovered that the cell capsule of Bacill...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P13/02C12R1/125
CPCC12P13/02C12R2001/125C12N1/205
Inventor 刘元元杨传伦徐泽平吴文雷刘海玉傅英旬和富明张心青
Owner CHAMBROAD CHEM IND RES INST CO LTD