Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Mutagenized strain Bacillus licheniformis TKPG091 for generating great amount of gamma-poly glutamic acid

A technology of TKPG091 and Bacillus licheniformis, applied in the field of fermentation engineering

Inactive Publication Date: 2010-09-22
天津北洋百川生物技术有限公司
View PDF0 Cites 16 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Extraction method Due to the inherent characteristics of this product, the extraction process is complicated, the cost is high, and it cannot be mass-produced
The preparation method of γ-PGA mainly focuses on microbial fermentation and synthesis. Biosynthesis of γ-PGA has outstanding advantages, such as high molecular weight, mild production conditions and high product purity. However, the yield of γ-PGA produced by microbial fermentation is not high, and the The production cost failed to achieve large-scale industrial production, which affected the application of the substance

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 The bacterial strain Bacillus licheniformis NXTK0007 screened out from the soil for the production of γ-polyglutamic acid

[0032] 1. Bacterial morphology: The thallus is short rod-shaped, with obvious capsules formed around the thallus, pili growing around it, and wrapped by a mucus layer. Bacteria divides by binary fission, gradually forming a septum from the middle, and then splits into two daughter cells of the same shape, size and structure. Spores grow in the middle or on one side of the fungus.

[0033] 2. Colony morphology: On the plate, when the medium is wet, the colony is generally round, with a smooth surface, relatively neat surroundings, and opaque colonies; when the medium is dry, the colonies are rough and wrinkled, with a concave center and irregular edges .

[0034] 3. Culture characteristics: This bacterium is a highly oxygen-consuming bacterium, the optimum growth temperature is 28 ℃ ~ 40 ℃, the optimum growth pH is 6-8; the rotation spee...

Embodiment 2

[0036] Example 2 Using Bacillus licheniformis NXTK0007 as the starting strain to screen strain TKPG091 by multiple compound mutagenesis techniques

[0037] 1. Obtaining of mutagenized strains

[0038] (1) Select the well-grown gamma-polyglutamic acid production strain NXTK0007 slant strain, wash the spores with sterile saline, and make 10 6 Each / mL spore suspension was irradiated under 30W UV lamp at 30cm for 10s, 20s, 30s, 40s, 50s, 60s, 70s, 80s respectively, the bacterial suspension was diluted and spread on a plate, and cultured at 37°C in the dark. Calculate the fatality rate;

[0039] (2) Select the irradiation time of three different doses of 40s, 60s, and 80s to carry out ultraviolet irradiation treatment to the spore suspension of bacterial strain NXTK0007 respectively;

[0040] (3) Then the bacterial suspensions of three different irradiation times were mixed evenly, and serially diluted 10 times for 10 -1 ~10 -6 , take 10 -4 、10 -5 、10 -6 Three dilutions of t...

Embodiment 3

[0054] The cultivation of embodiment 3 bacillus licheniformis TKPG091

[0055] 1. Preparation of culture medium

[0056] Preservation and separation medium: LB medium: trypsin 10g / L, yeast extract 5g / L, NaCl 10g / L, agar 15-20g / L, distilled water, use 1mol / L NaOH to adjust the pH to 7.2, steam at 121°C Sterilize for 20 minutes.

[0057] Seed medium composition: K 2 HPO 4 0.5, ferric ammonium citrate 0.5, MgSO 4 ·7H 2 O 0.5, glycerin 20.0, citric acid 2.0, L-glutamic acid 4.0, distilled water 1.0L, initial pH 7.4, sterilized at 121°C for 15 minutes, culture temperature 37°C.

[0058] Fermentation medium: L-glutamic acid 20.0, citric acid 12.0, glycerol 80.0, NH 4 Cl 7.0, K 2 HPO 4 0.5, MgSO 4 ·7H 2 O 0.5, FeCl 3 ·6H 2 O 0.04, CaCl 2 0.075, MnSO 4 ·H 2 O 0.104, 1.0L of distilled water, adjust the pH to 7.4 with 1mol / L NaOH, sterilize at 121°C for 15min, and cultivate at 37°C.

[0059] 2. Activation of strains

[0060] Transfer Bacillus licheniformis to TKGA091 ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention obtains a gamma-poly glutamic acid high-yield strain TKPG091, namely Bacillus licheniformis, by screening through composite mutagenesis technology. (Bacillus licheniformis is preserved in a general microbial center of China microbial strain preservation and management committee, and has a stain number of CGMCC No. 3336). The strain is prepared by taking Bacillus licheniformis stains NXTK0007 screened from fermented soya beans produced in Sichuan province in China as original stains, and mutagenizing the original stains by using ultraviolet rays. The yield of gamma-poly glutamic acid reaches 17.5+ / -2.1 g / L under optimized conditions.

Description

technical field [0001] The invention belongs to the technical field of fermentation engineering, and in particular relates to a mutagenic strain Bacillus licheniformis TKPG091 capable of producing a large amount of γ-polyglutamic acid and a cultivation method thereof. Background technique [0002] γ-polyglutamic acid [γ-poly(glutamic acid), γ-PGA] is formed by combining L-glutamic acid [L-Glu] and D-glutamic acid [D-Glu] through γ-amide bonds A high molecular weight amino acid polymer. The degradation product of this kind of material is non-toxic and harmless, and will not cause secondary pollution to the environment. In recent years, the development and research of this kind of polymer material has been developed rapidly. As a high molecular polymer, γ-polyglutamic acid has some unique physical, chemical and biological properties, such as biodegradability, good biocompatibility, strong water retention, and non-toxic to the human body. These characteristics determine the w...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P13/02C12R1/10
Inventor 乔长晟陈笑徐勇虎蒋磊李雪
Owner 天津北洋百川生物技术有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com