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Mycopremna generating gamma- polyglutamic acid and culturing method thereof

A technology of polyglutamic acid and cultivation method, which is applied in the production field of γ-polyglutamic acid, can solve the problems of high production cost and low output of γ-PGA, achieve low price, reduce production cost, and simple and easy cultivation method line effect

Active Publication Date: 2010-06-16
领先生物农业股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the production of γ-PGA by fermentation of bacteria belonging to the genus Bacillus has low yield and high production cost, so large-scale industrial production has not been realized.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Mutagenesis and screening of high-yield γ-PGA strains

[0029] Pick out a single colony of Bacillus natto CICC 10023 (purchased from China Industrial Microbial Culture Collection and Management Center), connect it to a sterile 250ml Erlenmeyer flask containing glass beads, add 30ml sterile water, and place it in HZQ- Shake on an X100 shaker (Erbin Donglian Electronics Co., Ltd.) at 170 r / min for 20 minutes to break the colonies to make a bacterial suspension. Place a plate containing 30ml of bacterial suspension on a 78-1 magnetic stirrer (Guohua Electric Co., Ltd.), irradiate it with a 40-watt violet lamp for 1 min, 2 min, and 3 min, and then separate the three bacterial suspensions with different doses Make 10 -5 And 10 -3 Diluted solution of, coated plate (for the formula of the fermentation medium solid plate, see the technical solution section), cultivated in the dark for 24 hours, and screened out mutants with altered colony morphological characteristics. ...

Embodiment 2

[0031] Example 2 Cultivation of Bacillus subtillis CGMCC No. 2108 and detection of γ-PGA

[0032] (1) Strain activation: pick Bacillus subtillis (Bacillus subtillis) CGMCC No. 2108 and place it on the slant medium. The components of the slant medium are: potato 200g / l, glucose 20g / l, agar 15g / l, pH The value is natural. Put the inoculated slope into a constant temperature incubator at 28°C for 48 hours.

[0033] (2) Seed culture: Take the activated Bacillus subtillis CGMCCNo.2108 and connect it to the seed culture medium (500ml Erlenmeyer flask with 100ml culture solution). The components of the seed culture medium are: glucose 20g / l, yeast Cream 5g / l, sodium glutamate 20g / l, dipotassium hydrogen phosphate 2g / l, magnesium sulfate 0.25g / l. The inoculated seed culture solution was placed in a constant temperature shaker at 37°C for 24 hours at 218r / min.

[0034] (3) Fermentation culture: Take 3ml of the cultured seed culture liquid and put it into the fermentation medium (500ml tria...

Embodiment 3

[0037] Example 3 Cultivation of Bacillus subtillis CGMCC No. 2108 and γ-PGA detection

[0038] Pick a ring of Bacillus subtillis (Bacillus subtillis) CGMCC No. 2108 on the slant medium, put it in a constant temperature incubator at 28°C for 48 hours, then take a ring and connect it to the seed medium, and place the inoculated seed culture solution Into the 37 ℃ constant temperature shaker 218r / min culture for 24 hours.

[0039] Take 3 ml of the cultured seed culture solution and put it into the fermentation medium. Among them, the fermentation medium composition is: citric acid 40-60g / l, and the remaining components are the same as in Example 2. Put the connected fermentation medium on a constant temperature shaker at 37°C for 36-72 hours at 218r / min.

[0040] Test result: The yield of γ-PGA produced by fermentation with the above medium components and culture method is 8.2g / l.

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PUM

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Abstract

The invention provides a high-productive strain Bacillus subtillis CGMCC No.2108 of poly-Gamma -glutamic-acid; the invention also provides a culture method and an applicable culture medium of the strain. The poly-Gamma -glutamic-acid provided by the invention is has the simple requirement of strain nutrient, stable passage, maximum output of 34.1g per liter, simple and practical culture method, wide culture medium raw material source and low cost, thus effectively reducing the production cost and being suitable for large-scale industrial production.

Description

Technical field [0001] The present invention relates to the production field of γ-polyglutamic acid, in particular to a strain of γ-polyglutamic acid producing strain and its culture method. Background technique [0002] γ-polyglutamic acid (γ-PGA) is a high molecular polymer formed by the condensation of α-amino and γ-carboxyl groups between glutamic acid monomers. The degree of polymerization is generally greater than 300,000. γ-PGA can be synthesized by microorganisms, has good water solubility, strong water absorption, and is biodegradable, and has no toxic effects on the human body. Therefore, it is widely used in food, cosmetics, water treatment, agriculture and medicine. For example, γ-PGA is an ideal drug carrier. The highly active carboxyl group (-COOH) on the γ-PGA molecular chain can combine with some drugs to form a more stable complex, which can be degraded into Endogenous glutamate and release the drug. Another example is the twice repeated water absorption of γ-P...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P13/00C12R1/125
Inventor 李颖孟永宏孙杉杉
Owner 领先生物农业股份有限公司
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