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Gamma-polyglutamic acid producing bacterium and method for efficiently synthesizing gamma-polyglutamic acid

A polyglutamic acid and bacteria-producing technology, applied in the field of microbial fermentation, can solve the problems of long production cycle and low production efficiency, etc.

Inactive Publication Date: 2019-01-08
南京卉之源生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although the fermentation production of γ-PGA has made great progress, there are still problems of long production cycle and low production efficiency.

Method used

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  • Gamma-polyglutamic acid producing bacterium and method for efficiently synthesizing gamma-polyglutamic acid
  • Gamma-polyglutamic acid producing bacterium and method for efficiently synthesizing gamma-polyglutamic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Slant medium components: peptone 10g / L, yeast powder 5g / L, sodium chloride 10g / l, agar 15g / L, pH 7.0, sterilized at 121°C for 20 minutes.

[0078] Preparation of seed medium: peptone 10g / L, yeast powder 5g / L, sodium chloride 10g / L, pH7.0, sterilized at 121°C for 20 minutes.

[0079] Fermentation medium preparation: glucose 40g / L, peptone 5 g / L, sodium glutamate 30 g / L, K2HPO4 3H2O 2g / L, MgSO4 0.25 g / L, ammonium sulfate 3 g / L, manganese sulfate 0.1g / L L, pH7.0, sterilized at 115°C for 15 minutes (glucose is sterilized separately from other components).

[0080] The cryopreserved Bacillus subtilis (Bacillus Subtilis) W84 strain was inoculated on the slant medium for activation, and cultured at 37°C for 20h. Inoculate the strain activated on the slant into the seed medium, and culture at 37° C. for 12 h with a shaker rotating at 200 rpm.

[0081] Put the seed solution activated in the previous step into the fermentation medium of the shake flask, the inoculum amount is 2...

Embodiment 2

[0084] Slant-plane activation of strains: the same as in Example 1.

[0085] Seed medium preparation: glucose 20 g / L; sodium glutamate 5 g / L; dipotassium hydrogen phosphate 2 g / L; magnesium sulfate 2.5 g / L; yeast extract 5 g / L; manganese sulfate 0.2 g / L, pH 7.0. Sterilize at 115°C for 15 minutes (glucose is sterilized separately from other components).

[0086] Preparation of fermentation medium: glucose 40g / L, peptone 5 g / L, sodium glutamate 50 g / L, dipotassium hydrogen phosphate 8g / L, magnesium sulfate 0.5 g / L, ammonium sulfate 3 g / L, manganese sulfate 0.1g / L, pH7.0, sterilized at 115°C for 15 minutes (glucose is sterilized separately from other components).

[0087]The cryopreserved Bacillus subtilis (Bacillus Subtilis) W84 strain was inoculated on the slant medium for activation, and cultured at 37°C for 20h. Inoculate the strain activated on the slant into the seed medium, and culture at 37° C. for 12 h with a shaker rotating at 200 rpm.

[0088] Put the seed solution ...

Embodiment 3

[0091] Slant-plane activation of strains: the same as in Example 1.

[0092] Seed medium preparation: glucose 20 g / L; sodium glutamate 5 g / L; dipotassium hydrogen phosphate 2 g / L; magnesium sulfate 2.5 g / L; yeast extract 5 g / L; manganese sulfate 0.2 g / L, pH 7.0. Sterilize at 115°C for 15 minutes (glucose is sterilized separately from other components).

[0093] Preparation of fermentation medium: sucrose 40g / L, peptone 5 g / L, sodium glutamate 50 g / L, dipotassium hydrogen phosphate 8 g / L, magnesium sulfate 0.5 g / L, ammonium sulfate 3 g / L, sulfuric acid Manganese 0.1g / L, pH7.0, sterilized at 115°C for 15 minutes.

[0094] The cryopreserved Bacillus subtilis (Bacillus Subtilis) W84 strain was inoculated on the slant medium for activation, and cultured at 37°C for 20h. Inoculate the strain activated on the slant into the seed medium, and culture at 37° C. for 12 h with a shaker rotating at 200 rpm.

[0095] Put the seed solution activated in the previous step into the fermentat...

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Abstract

The invention discloses a gamma-polyglutamic acid producing bacterium. The gamma-polyglutamic acid producing bacterium is classified and named as Bacillus Subtilis hzy-1, has been deposited in the Chinese Industry Culture Collection Center with the accession number of CCTCC M 2018226 in May 3, 2018. Meanwhile, the invention further discloses a method for efficiently synthesizing gamma-polyglutamicacid. Compared with an existing technology for producing gamma-polyglutamic acid, the method has the advantages that by a used bacillus subtilis strain, the gamma-polyglutamic acid can be effectivelyfermented and produced, the molecular weight of the produced gamma-polyglutamic acid is 1000 KDa or above, by optimization of bacillus subtilis strain culture conditions, the content of the gamma-polyglutamic acid in fermentation liquor reaches 40-60 g / L, and therefore, a method for effectively preparing the gamma-polyglutamic acid at low price is provided. The produced gamma-polyglutamic acid can be applied to the fields of agriculture, cosmetics and the like.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation, in particular to a synthetic strain of gamma-polyglutamic acid and a method for efficiently preparing macromolecule gamma-polyglutamic acid. Background technique [0002] γ-polyglutamic acid (γ-polyglutamic acid, hereinafter referred to as γ-PGA) is a type of homopolyamino acid formed by connecting D-glutamic acid and L-glutamic acid monomers through γ-amide bonds. Fermentation synthesis. Because γ-PGA has excellent physical, chemical and biological properties such as film-forming, fibroblast, plasticity, adhesion, moisture retention, and degradability, it has great application potential in the fields of industry, agriculture, cosmetics, and medicine. . [0003] There are three methods for preparing polyglutamic acid: chemical synthesis, extraction and microbial fermentation. Compared with the previous two methods, the microbial fermentation method has the advantages of lower pr...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P13/02C12R1/125
CPCC12P13/02C12N1/205C12R2001/125
Inventor 殷志敏黄正才
Owner 南京卉之源生物科技有限公司
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