Gamma-polyglutamic acid producing bacteria and method for breeding and preparing gamma-polyglutamic acid

A technology for producing polyglutamic acid and bacteria, which is applied in the field of γ-polyglutamic acid-producing bacteria and breeding, to prepare γ-polyglutamic acid, which can solve the shortage of γ-polyglutamic acid production technology, γ-polyglutamic acid The production capacity of glutamic acid is not strong, etc., to achieve the effect of low pollution, easy process and process control

Active Publication Date: 2019-08-16
汉肽生物医药集团有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] An object of the present invention is to provide a γ-polyglutamic acid-producing bacterium, which solves the defect that most of the existing γ-polyglutamic acid-producing bacteria products are soil-grade crude products, and provides a γ-polyglutamic acid-producing bacterium Breeding method
[0008] Another object of the present invention is to provide a kind of method that γ-polyglutamic acid producing bacteria prepare γ-polyglutamic acid, solve the deficiency of the production technology of γ-polyglutamic acid and the insufficient production capacity of domestic γ-polyglutamic acid strong question

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] (1) Bacterial culture: Bacillus licheniformis) BL (screened from soil) was inoculated in a Erlenmeyer flask filled with seed medium, and cultured with shaking at 37±1°C for 22 hours to obtain a seed solution.

[0054] Seed shake flask medium formula: glycerin 15g / L, citric acid 2g / L, ferric ammonium citrate 0.5g / L, L-sodium glutamate 4g / L, potassium dihydrogen phosphate 0.5g / L, magnesium sulfate 0.5g / L, the balance is water, PH 7.0±1.

[0055] (2) Preparation of bacterial suspension: Take the above seed liquid in a sterile centrifuge tube, centrifuge at 3000r / min for 10min, discard the supernatant, add sterile water to shake and wash, centrifuge for 10min, discard the supernatant, and then add sterile water, shake evenly.

[0056] (3) Mutagenesis: Put the bacterial suspension in a sterile petri dish, irradiate for 1.2 minutes under a 254nm ultraviolet lamp in a UV mutagenesis ultra-clean bench, at a distance of 20cm; put the bacterial suspension that has been subjecte...

Embodiment 2

[0063] (1) Bacterial culture: take Bacillus licheniformis) BL (screened from soil), inoculate in a Erlenmeyer flask filled with seed medium, shake and culture at 37±1°C for 16 hours, and obtain seed liquid.

[0064] Seed shake flask medium formula: glycerin 10g / L, citric acid 0.8g / L, ferric ammonium citrate 0.1g / L, L-sodium glutamate 2g / L, potassium dihydrogen phosphate 0.3g / L, magnesium sulfate 0.3 g / L, the balance is water, PH 7.0±1.

[0065] (2) Preparation of bacterial suspension: Take the above seed liquid in a sterile centrifuge tube, centrifuge at 2000r / min for 5min, discard the supernatant, add sterile water to shake and wash, centrifuge for 5min, discard the supernatant, and then add sterile water, shake evenly.

[0066] (3) Mutagenesis: Put the bacterial suspension in a sterile petri dish, irradiate for 0.5 min under a 254nm ultraviolet lamp in an ultraviolet mutagenesis ultra-clean bench, at a distance of 15cm; put the bacterial suspension that has been subjected t...

Embodiment 3

[0073] (1) Bacterial culture: Bacillus licheniformis) BL (screened from soil) was inoculated in a Erlenmeyer flask filled with seed medium, and cultured with shaking at 37±1°C for 32 hours to obtain a seed solution.

[0074] Seed shake flask medium formula: glycerin 20g / L, citric acid 3g / L, ferric ammonium citrate 1g / L, L-sodium glutamate 7g / L, potassium dihydrogen phosphate 0.8g / L, magnesium sulfate 0.8g / L L, the balance is water, pH 7.0±1.

[0075] (2) Preparation of bacterial suspension: Take the above seed liquid in a sterile centrifuge tube, centrifuge at 5000r / min for 30min, discard the supernatant, add sterile water to shake and wash, centrifuge for 30min, discard the supernatant, and then add sterile water, shake evenly.

[0076] (3) Mutagenesis: put the bacterial suspension in a sterile petri dish, irradiate for 1.5 min under a 254nm ultraviolet lamp in an ultraviolet mutagenesis ultra-clean bench, at a distance of 25cm; put the bacterial suspension that has been sub...

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Abstract

The invention belongs to the technical field of bioengineering fermentation, in particular to gamma-polyglutamic acid producing bacteria and a method for breeding and preparing gamma-polyglutamic acid. The Bacillus licheniformis BL1 is deposited in the China General Microbiological Culture Collection Center (CGMCC for short); the address of the deposit institution is: Institute of Microbiology, Chinese Academy of Sciences, No.3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing; the postal code is 100101; the registered number is CGMCC NO. 17508; and the date of deposit is April 1, 2019. This bacterium is used as a production strain. The preparation method of gamma-polyglutamic acid includes the steps of strain screening, fermentation, disinfection, microfiltration sterilization,ultrafiltration concentration and precipitation drying. The gamma-polyglutamic acid prepared by the method has high purity and no impurities, and is more suitable for cosmetics and medicines.

Description

[0001] Technical field: [0002] The invention belongs to the technical field of bioengineering fermentation, and in particular relates to a γ-polyglutamic acid-producing bacterium and a method for breeding and preparing γ-polyglutamic acid. [0003] Background technique: [0004] γ-polyglutamic acid (γ-PGA) is an unusual anionic polyamide compound consisting of d- and l-glutamic acid units linked by amide bonds between amino and carboxylic acid groups. According to the current glutamic acid residues, γ~PGA can be divided into γ~l~PGA (only l~glutamic acid residues), γ~d~PGA (only d~glutamic acid residues) and γ~ld~ PGA (l~ and d~ glutamic acid residues). Currently, there are three methods for producing γ-PGA: chemical synthesis, extraction and microbial fermentation. Compared with other methods, microbial fermentation is the most cost-effective and has many advantages, including cheap raw materials, minimal environmental pollution, high natural product purity, and mild react...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02C12P13/02C12R1/10
CPCC12N1/02C12N1/20C12P13/02C12N1/205C12R2001/10
Inventor 朱永真李艳琪李同金赵传海
Owner 汉肽生物医药集团有限公司
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