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Method for producing gamma-polyglutamic acid though hot fermentation of bacillus subtilis

A technology of Bacillus subtilis and polyglutamic acid, applied in the field of microbial fermentation, can solve the problems of long fermentation period, low production efficiency, complex fermentation medium composition and the like

Active Publication Date: 2013-02-06
广西多得乐生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0013] The patents found above have the following deficiencies: the fermentation temperature must be strictly controlled below 40°C, the requirements for fermentation conditions are high, the composition of the fermentation medium is complicated, the cost of the fermentation process is high, the fermentation cycle is long, and the production efficiency is low. Restricted the process of γ-polyglutamic acid fermentation industrialization

Method used

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  • Method for producing gamma-polyglutamic acid though hot fermentation of bacillus subtilis
  • Method for producing gamma-polyglutamic acid though hot fermentation of bacillus subtilis
  • Method for producing gamma-polyglutamic acid though hot fermentation of bacillus subtilis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1~5

[0041] (1) Activation and preservation of strains: Inoculate CCTCC NO: M2012347 on solid slant medium, culture at 45°C for 16 hours, and store at 4°C for short-term; the concentration of solid slant medium is composed of: glucose 10g / l, yeast extract 5g / l, sodium glutamate 5g / l, MgSO 4 ·7H 2 O 0.1g / l, KH 2 PO 4 0.5g / l, agar 15g / l, pH 7.0, prepared with distilled water;

[0042] (2) Seed liquid preparation: put 1.0cm on the above slope 2 The bacterial strains were inserted into a 250ml Erlenmeyer flask containing 30ml of liquid seed medium, the shaking table rotated at 180rpm, and cultured at 45°C for 12h to the mid-logarithmic growth stage of the strain; the concentration of the liquid seed medium was composed of: glucose 10g / l, yeast extract 5g / l l, sodium glutamate 5g / l, MgSO 4 ·7H 2 O 0.1g / l, KH 2 PO 4 0.5g / l, pH 7.0, prepared with distilled water;

[0043] (3) Liquid shake flask fermentation: put the seed liquid into the sterilized liquid fermentation medium, th...

Embodiment 6~12

[0049] (1) Activation and preservation of strains: Inoculate CCTCC NO: M2012347 on solid slant medium, culture at 45°C for 16 hours, and store at 4°C for short-term; the concentration of solid slant medium is composed of: glucose 10g / l, yeast extract 5g / l, sodium glutamate 5g / l, MgSO 4 ·7H 2 O 0.1g / l, KH 2 PO 4 0.5g / l, agar 15g / l, pH 7.0, prepared with distilled water;

[0050] (2) Seed liquid preparation: put 1.0cm on the above slope 2 The bacterial strains were inserted into a 250ml Erlenmeyer flask containing 30ml of liquid seed medium, the shaking table rotated at 180rpm, and cultured at 45°C for 12h to the mid-logarithmic growth stage of the strain; the concentration of the liquid seed medium was composed of: glucose 10g / l, yeast extract 5g / l l, sodium glutamate 5g / l, MgSO 4 ·7H 2 O 0.1g / l, KH 2 PO 4 0.5g / l, pH 7.0, prepared with distilled water;

[0051] (3) Liquid shake flask fermentation: put the seed liquid into the sterilized liquid fermentation medium, th...

Embodiment 13

[0057] (1) Strain activation: Inoculate CCTCC M2012347 on a solid slant medium, culture at 45°C for 16 hours, and store at 4°C for short-term storage; the concentration of the solid slant medium is composed of: glucose 10g / l, yeast extract 5g / l, Sodium glutamate 5g / l, MgSO 4 ·7H 2 O 0.1g / l, KH 2 PO 4 0.5g / l, agar 15g / l, pH 6.5, prepared with distilled water;

[0058] (2) Seed liquid preparation: put 1.0cm on the above slope 2 The bacterial lawn was inserted into a 250ml Erlenmeyer flask equipped with 30ml liquid seed culture medium, the shaking table rotated at 180rpm, and cultivated at 45°C for 12h to the mid-logarithmic growth stage of the bacterial strain; wherein the liquid seed culture medium concentration components were: glucose 10g / l , yeast extract 5g / l, sodium glutamate 5g / l, MgSO 4 ·7H 2 O 0.1g / l, KH 2 PO 4 0.5g / l, pH7.0, prepared with distilled water.

[0059] (3) Liquid shake flask fermentation: put the seed liquid into the sterilized liquid fermentatio...

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Abstract

The invention discloses a method for producing gamma-polyglutamic acid though hot fermentation of bacillus subtilis. A bacterial strain of bacillus subtilis (Bacillussubtilis) GXA-28 with a preservation serial number of CCTCCNO:M2012347 is used for preparation of the gamma-polyglutamic acid through actication of culture, preservation, seed liquid preparation, liquid shake flask fermentation and gamma-polyglutamic acid extraction and purification. the method can widely use low-cost carbon nitrogen sources, such as molasses and inorganic ammonium salt, wherein the inorganic ammonium salt can serve as a sole nitrogen source, the gamma-polyglutamic acid can be efficiently produced at a 40-50 DEG C hot fermentation condition, fermentation time is 18-25h, highest output can achieve 20-30g / l, a production rate can achieve 0.9-1.3g / l.h, and the method has the advantages of being high in efficiency and low in cost.

Description

technical field [0001] The invention belongs to the field of microbial fermentation, and in particular relates to a method for producing gamma-polyglutamic acid by high-temperature fermentation of bacillus subtilis. Background technique [0002] γ-polyglutamic acid (Poly(γ-glutamic acid), γ-PGA) is a unique biologically high molecular. It was first discovered in the capsule of Bacillus anthracis by Ivánovics et al. in 1937 (Ivánovics and Bruckner 1937; Ivánovics and Erdōs 1937); and it was confirmed in 1942 that it could be secreted by Bacillus subtilis into liquid medium as a fermentation product (Bovamick 1942). Therefore, Bacillus subtilis and Bacillus licheniformis have been extensively studied as the main γ-polyglutamic acid producing bacteria. [0003] γ-polyglutamic acid is usually polymerized by 500-5000 glutamic acid monomers, and its molecular weight is between 100-1000kDa. The glutamic acid residues in the γ-polyglutamic acid molecule have a large number of fre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/02C12R1/125
Inventor 梁智群曾伟陈桂光张云开
Owner 广西多得乐生物科技有限公司
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