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Method for producing gamma-polyglutamic acid through adding sodium chloride (NaCl) in fermentation process

A technology of polyglutamic acid and fermentation process, which is applied in the field of high-efficiency fermentation and production of γ-polyglutamic acid, which can solve the problems of high production cost, restrictions on the industrial production and application of γ-polyglutamic acid, and low yield, and achieve The effect of increased output, good application prospects, and simple operation

Inactive Publication Date: 2015-02-04
DONGGUAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are mainly four kinds of γ-PGA producing bacteria reported so far: Bacillus subtilis (Natto) (BaciLLus subtiLis), Bacillus licheniformis (BaciLLus Licheniformis), Bacillus thermostable (BaciLLus thermotoLerant), Bacillus anthracis (BaciLLus.anthracis ), although many researchers have done a lot of research on the production of γ-polyglutamic acid by microbial fermentation, the yield is generally still at a low level and the production cost is high, which limits the production of γ-polyglutamic acid to a large extent. industrial production and application

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Embodiment 1: Shake flask fermentation

[0015] (1) Preparation of seed solution

[0016] The Bacillus subtilis natto used in the production of γ-PGA comes from the China Industrial Microorganism Collection Center (CICC), and the strain number is CICC 20643. The composition of the seed medium is: peptone 10g / L, beef extract 5g / L, NaCl 10g / L, pH 7.0, the rest is distilled water or tap water, the liquid volume is 50mL / 250mL triangular flask, sterilized at 121°C for 30min, after cooling, Take a ring of Bacillus natto preserved in the slant medium in advance and inoculate it into the seed medium, and culture it in a shaker flask at 37°C and 120r / min for 24h to form the Bacillus natto seed solution.

[0017] (2) shake flask culture

[0018] Composition of fermentation medium: 25g / L, yeast extract 5g / L, sodium glutamate 40g / L, Na 2 HPO 4 1g / L, NaH 2 PO 4 1g / L, MgSO 4 0.5g / L, the rest is distilled water; the initial pH value is 6.5, the liquid volume is 70mL / 250mL Erlen...

Embodiment 2

[0021] Embodiment 2: liquid submerged fermentation

[0022] (1) Preparation of seed solution

[0023] Same as example 1.

[0024] (2) Liquid submerged culture

[0025] The composition of the fermentation medium: glucose 20g / L, yeast powder 10g / L, sodium glutamate 20g / L, disodium hydrogen phosphate 1g / L, disodium hydrogen phosphate 1g / L, magnesium sulfate 0.5g / L, the rest is Tap water; the initial pH value is 7.0, add 3L fermentation medium in a 5L microbial fermentation tank, insert Bacillus natto seed liquid with 5% inoculation amount after sterilization and cooling, cultivate at 37°C, and the aeration ratio is 0.5~0.8(v / v), the stirring speed is 200r / min, 4% NaCl is added after culturing for 24 hours, and NaCl is added as a concentrated solution after being sterilized by aseptic operation. After continuing to cultivate at 37°C for 36 hours, the aeration ratio was reduced to 0.3-0.5 (v / v) during the cultivation process.

[0026] (3) Extraction of γ-PGA

[0027] The extr...

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Abstract

The invention provides a method for producing gamma-polyglutamic acid through adding sodium chloride (NaCl) in a fermentation process. According to the method, bacillus subtilis natto is used as a producing bacterium for the gamma-polyglutamic acid. The method comprises the following main three fermentation processing steps: performing aerobic culture on the producing bacterium in a liquid fermentation medium for a period of time in advance; then, adding the NaCl so as to increase the osmotic pressure of a fermentation substrate; and then, continuing performing the aerobic culture to a fermentation end and the like. Compared with the yield of the gamma-polyglutamic acid produced through adding the NaCl before inoculation or not adding the NaCl, the yield of the gamma-polyglutamic acid produced by the method for producing gamma-polyglutamic acid through adding the NaCl in the fermentation process can be obviously increased. Moreover, the method for producing gamma-polyglutamic acid through adding the NaCl in the fermentation process is simple in operation, is convenient for large-scale production and has a good application prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for efficiently fermenting and producing gamma-polyglutamic acid. Background technique [0002] γ-polyglutamic acid (poly-γ-glutamic acid, γ-PGA for short) is a water-soluble biopolymer formed by combining L-glutamic acid or D-glutamic acid through γ-amide bonds. . Because γ-polyglutamic acid has many characteristics such as water absorption, degradability, and hydrolysis, it is widely used in various fields such as medicine, food processing, agriculture, greening, and water treatment, and has great development value and application prospect. [0003] γ-polyglutamic acid was originally discovered in Bacillus anthracis, and it was found in high CO 2 Under the induction of concentration, Bacillus anthracis can produce γ-D-polyglutamic acid, and the produced γ-D-polyglutamic acid is attached to the cell wall and forms a capsule outside the microbial cell, the toxicity limit of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P13/02C12R1/07
Inventor 邹水洋朱丹
Owner DONGGUAN UNIV OF TECH
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