Production method of low-endotoxin gamma-polyglutamic acid

A technology of polyglutamic acid and production method, applied in the field of microbial fermentation, can solve problems such as complicated process and secondary pollution, and achieve the effects of good stability, pollution prevention and reduction of endotoxin production.

Pending Publication Date: 2022-07-22
SHANDONG FREDA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the problems of secondary pollution and complicated process in the existing endotoxin removal method, the present invention provides a production method containing low endotoxin γ-polyglutamic acid, which is not easy to be contaminated by microorganisms in the endotoxin treatment process, and the process is simple and suitable Industrialized production, effectively reducing the content of endotoxin in the production of γ-polyglutamic acid

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The production method of low-endotoxin-containing γ-polyglutamic acid according to the present invention comprises the following steps:

[0028] S1. Preparation of γ-polyglutamic acid fermentation broth

[0029] (1) Seed culture: Bacillus subtilis FRD518 was inoculated in a conical flask equipped with a seed medium, and the seed medium was composed of the following concentrations: glucose 5g / L, yeast powder 3g / L, ammonium sulfate 2g / L , MgSO 4 0.1g / L, the pH of the medium is 7.2, sterilized at a temperature of 115°C for 30min, the inoculum amount is 1%, and shaken at 35°C, 180r / min shaker culture until the OD600 reaches 1.6;

[0030] (2) Fermentation culture: the seed liquid prepared in step (1) is inoculated in a fermentation medium, and the fermentation medium is a component according to the following concentrations: peptone 5g / L, yeast extract 5g / L, ammonium sulfate 5g / L , Sodium glutamate 50g / L, citric acid 2g / L, MgSO 4 0.1g / L, K 2 HPO 4 1g / L, glucose 100g / L, m...

Embodiment 2

[0038] The production method of low-endotoxin-containing γ-polyglutamic acid according to the present invention comprises the following steps:

[0039] S1. Preparation of γ-polyglutamic acid fermentation broth

[0040] (1) Seed culture: Bacillus subtilis FRD518 was inoculated in a conical flask equipped with a seed medium, and the seed medium was the following components: glucose 7.5g / L, yeast powder 4g / L, ammonium sulfate 6g / L L. MgSO 4 0.5g / L, the pH of the medium is 7.3, sterilized at a temperature of 115°C for 30min, the inoculum volume is 1%, and shaken at 36°C, 200r / min shaker culture until the OD600 reaches 2.0;

[0041] (2) Fermentation culture: the seed liquid prepared in step (1) is inoculated into a fermentation medium, and the fermentation medium is composed of the following concentrations: peptone 15g / L, yeast extract 15g / L, ammonium sulfate 15g / L , sodium glutamate 80g / L, citric acid 3g / L, MgSO 4 0.3g / L, K 2 HPO 4 2.0g / L, glucose 150g / L, medium pH 7.3, steri...

Embodiment 3

[0049] The production method of low-endotoxin-containing γ-polyglutamic acid according to the present invention comprises the following steps:

[0050] S1. Preparation of γ-polyglutamic acid fermentation broth

[0051] (1) Seed culture: Bacillus subtilis FRD518 was inoculated in a conical flask equipped with a seed medium, and the seed medium was the following components: glucose 10g / L, yeast powder 5g / L, ammonium sulfate 10g / L , MgSO 4 1.0g / L, the pH of the medium is 7.5, sterilized at a temperature of 115°C for 30min, the inoculum amount is 1%, and shaken at 38°C, 220r / min shaker culture until the OD600 reaches 2.2;

[0052] (2) Fermentation culture: the seed liquid prepared in step (1) is inoculated in a fermentation medium, and the fermentation medium is a component according to the following concentrations: peptone 30g / L, yeast extract 30g / L, ammonium sulfate 20g / L , Sodium glutamate 100g / L, citric acid 5g / L, MgSO 4 0.5g / L, K 2 HPO 4 4.0g / L, glucose 200g / L, medium pH...

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Abstract

The invention relates to the technical field of microbial fermentation, in particular to a production method of low-endotoxin gamma-polyglutamic acid, which comprises the following steps: S1, preparing gamma-polyglutamic acid fermentation liquor; s2, carrying out acid treatment on the gamma-polyglutamic acid fermentation liquor to adjust the pH value to 3.0-5.0, carrying out high-temperature treatment, cooling, decolorizing and filtering to obtain filtrate; s3, adding acid into the filtrate to adjust the pH value to 1.0-3.0, standing to separate out precipitate, and pumping away supernatant liquid; and S4, adding an organic solvent into the bottom precipitate, stirring and precipitating, pumping away the organic solvent on the upper layer, and drying the precipitate. The method is not easily polluted by microorganisms in the endotoxin treatment process, the process is simple and suitable for industrial production, and the content of the endotoxin in gamma-polyglutamic acid production is effectively reduced.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation, in particular to a production method containing low endotoxin γ-polyglutamic acid. Background technique [0002] γ-Polyglutamic acid is water-soluble, biodegradable, edible, non-toxic to human body, and non-polluting to the environment. Today, with emphasis on environmental protection and sustainable development, this biosynthetic degradable functional material is favored by people, and is gradually being used in pharmaceutical manufacturing, food processing, antifreeze and preservation of vegetables, fruits, and seafood. It can also be developed and applied in many fields such as cosmetics industry, tobacco, leather manufacturing and plant seed protection. It is a new multifunctional biological product with great development value and broad prospects. [0003] In order to use γ-polyglutamic acid in the cosmetic industry and pharmaceutical manufacturing, it is necessary to ensure ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/02C08G69/46C12R1/125
CPCC12P13/02C08G69/46
Inventor 王庆波李海军张英华马双双贾开钰胡红涛李艳艳徐波张杰
Owner SHANDONG FREDA BIOTECH
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