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Bacillus subtilis for high-yield gamma-polyglutamic acid and fermentation method of bacillus subtilis

A technology of Bacillus subtilis and polyglutamic acid, applied in the field of microorganisms, can solve the problems of high cost, low yield and purity of γ-PGA, and achieve the effects of low production cost, narrow product molecular weight distribution and high purity

Inactive Publication Date: 2019-06-28
ZHEJIANG HUIJIA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the microbial production of γ-PGA still has the disadvantages of high cost, low yield and low purity of γ-PGA

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Isolation and identification of Bacillus subtilis HJBA058

[0024] The strain was isolated from the gut of chicks. The specific implementation steps are as follows:

[0025] 1. Preparation of bacterial suspension: After dissecting healthy chicks of a certain age (4-5 days), scrape the surface mucus and contents of the ileum or cecum and place them in a triangular flask filled with sterile saline. Stir magnetically for 30 minutes to make a bacterial suspension and treat at 80°C for 15 minutes;

[0026] 2. Bacterial suspension dilution and colony picking: The above-mentioned bacterial suspension was diluted into bacterial suspensions of different concentrations using a 10-fold ratio dilution method, and the dilution of 10 3 、10 4 、10 5 、10 6 Each 100 μL of bacterial suspension was spread on a solid medium plate; after 2 days of constant temperature cultivation at 37°C, pick a single colony whose surface was rough, opaque, viscous, and could be picked up with a toothp...

Embodiment 2

[0041] The isolation and identification of Bacillus subtilis HJBA058 in this example is the same as in Example 1.

[0042] Utilize bacillus subtilis HJBA058 fermentation to produce the detection of gamma-polyglutamic acid and fermentation product, its specific operation steps are as follows:

[0043] 1. Strain activation: inoculate Bacillus subtilis HJBA058 into LB solid medium (its components and concentrations thereof are as follows: peptone 10g / L, yeast powder 5g / L, sodium chloride 5g / L, agar 2%, the The pH value in the LB solid medium is 7.2), and cultivated at a constant temperature of 37°C for 24 hours;

[0044] 2. For seed cultivation, take an appropriate amount of the strain slope prepared in the above step 1. Under aseptic conditions, each slope is cleaned with 5 milliliters of sterile water, and inserted into the seed culture with an inoculum of 1.5%. Liquid (its components and concentrations are as follows: maltose 50g / L, yeast powder 10g / L, sodium glutamate 30g / L,...

Embodiment 3

[0050] The isolation and identification of Bacillus subtilis HJBA058 in this example is the same as in Example 1.

[0051] Utilize bacillus subtilis HJBA058 fermentation to produce the detection of gamma-polyglutamic acid and fermentation product, its specific operation steps are as follows:

[0052] 1. Strain activation: inoculate Bacillus subtilis HJBA058 into LB solid medium (its components and concentrations thereof are as follows: peptone 10g / L, yeast powder 5g / L, sodium chloride 5g / L, agar 2%, the The pH value in the above-mentioned LB solid medium is 7.0), and cultivated at a constant temperature of 37° C. for 22 hours;

[0053] 2. For seed culture, take an appropriate amount of the slanted surface of the bacteria prepared in the above step 1. Under aseptic conditions, each slanted surface is cleaned with 5 ml of sterile water, and the inoculum of 3% is inserted into the seed for cultivation. Liquid (its components and concentrations are as follows: maltose 50g / L, yeas...

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Abstract

The invention discloses bacillus subtilis for high-yield gamma-polyglutamic acid and a fermentation method of the bacillus subtilis. The preservation number of the bacillus subtilis is CGMCC NO: 9383,and the bacillus subtilis is preserved in China General Microbiological Culture Collection Center on June 25th, 2014. The gamma-polyglutamic acid is fermented and extracted through the four steps ofstrain activation, seed culture, tank loading fermentation and extraction of the gamma-polyglutamic acid, the fermentation method of the bnacillus subtili is simple, the production cost is low, the fermentation condition is easy to control, the purity of the fermentation product is high, the yield of the gamma-polyglutamic acid is as high as 56.5 g / L, and the bacillus subtilis can be convenientlyapplied to the production of the gamma-polyglutamic acid.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a high-yield γ-polyglutamic acid bacillus subtilis and a fermentation method thereof. Background technique [0002] γ-polyglutamic acid (Poly(γ-glutamic acid), γ-PGA) is a unique biopolymer formed by the polymerization of L-glutamic acid and D-glutamic acid through α-amino and γ-carboxyl groups . It 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 free hydrophilic carboxyl groups, which can form hydrogen bonds inside or between molecules, and have extremely high water solubility and water absorption and moisture retention; these free carboxyl groups At the same time, it provides a cation-bonding group, which makes it have good adsorption to metal ions. In addition, γ-polyglutamic acid, as an environmentally friendly...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P13/02C12R1/125
Inventor 杨彩梅郗洪生曾新福李慧刘金松
Owner ZHEJIANG HUIJIA BIOTECH
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