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Preparing gamma-poly glutamic acid, glutamate, glutathione and its precursor with bacillus subtilis NX-2

A technology of Bacillus subtilis and polyglutamic acid, applied in the direction of bacteria, fermentation, etc., can solve the problems of high consumption of raw materials, long production cycle, low productivity, etc., and achieve the effect of simplifying the process route

Inactive Publication Date: 2003-06-25
WUHAN ELITE CHEM FERTILIZER
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, it has been found that many bacteria can produce γ-PGA, such as B.licheniformic ATCC9945, B.subtilis IFO3335, B.subtilisF02-1, etc., but there are still problems such as large consumption of raw materials, long production cycle, low productivity, and high production cost. Therefore, Efficient and low-cost production of γ-polyglutamic acid still needs to be developed

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Glucose 1.5%, yeast extract 0.5%, glutamic acid 3%, KH 2 PO 4 0.5%, MgSO 4 .7H 2 O0.05%, the ratio of making culture medium, the pH before sterilization is 7.0~7.5. Fill liquid 50ml in the Erlenmeyer flask of 500ml capacity, sterilize at 121 ℃ for 20 minutes, cool down.

[0033] The above-mentioned Bacillus subtilis NX-2 strain was inoculated in the culture medium, cultured at 37° C. for 24 hours, and the rotation speed of the shaker flask was 200 r / ml. The content of gamma-polyglutamic acid in the fermented liquid is 30g / l, and the productivity is 1.25g h -1 l -1 .

Embodiment 2

[0035] The medium is glucose 1.5%, yeast extract 1.0%, corn steep liquor 1.0%, MgSO 4 0.01%, K 2 HPO 4 0.01%, pH7, filling volume 50ml / 500ml Erlenmeyer flask, 120°C for 20 minutes. After the sterilization is finished, cool down and inoculate. The bacterial species is Bacillus subtilis NX-2 strain, the inoculum amount is 1%, and the shake flask is cultivated at 37° C. for 24 hours, and the rotation speed is 180 r / min. The activity of γ-glutamyl transpeptidase in the culture medium was up to 3.0U / ml.

Embodiment 3

[0037] The L-cysteinyl L-glycine prepared by chemical synthesis is prepared to a concentration of 20mmol / l with 50ml of water, and then added with L-glutamic acid to make the concentration 20mmol / l, mixed as the enzyme reaction substrate, filled with N 2 stand-by. The fermentation broth obtained in Example 2 was added to the reaction substrate with an enzyme amount of 0.5 U / ml, placed in a constant temperature water bath at 37°C for 2 to 10 hours, and samples were taken every 30 minutes for HPLC analysis. After reacting for 10 hours, 6 mmol / l of glutathione was produced, and the conversion rate of L-cysteinyl L-glycine was 30%.

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Abstract

The bacillus subtilis NX-2, or CGMCC NO.0833 is one bacillus spawn obtained through screening and is used for the production of gamma-polyglutamic acid and its salt as well as glutathione and its precursor. In optimized condition, gamma-polyglutamic acid may be accumulated to 30-50 g / L and produced in the production efficinecy of 0.8-2.5 g / L. When bacillus subtilis NX-2 is used in fermenting to produce gamma-glutamyl transpeptidase in optimized condition, the fermented liquid may reach enzyme activity of 1-5 U / ml; and the fermented liquid or separation obtained enzyme or the immobilized enzyme is used in the production of glutathione and its precursor with substrate dipeptide or dipeptide precursor converting rate of 30-80% being reached.

Description

technical field [0001] The present invention relates to a microbial strain of high-yielding gamma-polyglutamic acid (gamma-PGA) and high-yield gamma-glutamyl transpeptidase (gamma-GTP), and using it for gamma-polyglutamic acid and salts thereof and production of glutathione and its precursors. Background technique [0002] γ-polyglutamic acid (γ-PGA) and its salts are biodegradable polymer substances that are harmless to humans and the environment, and are expected to be widely used in the fields of food, cosmetics, agriculture, medicine, and water treatment . At present, it has been found that many bacteria can produce γ-PGA, such as B.licheniformic ATCC9945, B.subtilis IFO3335, B.subtilisF02-1, etc., but there are still problems such as large consumption of raw materials, long production cycle, low productivity, and high production cost. Therefore, Efficient and low-cost production of γ-polyglutamic acid remains to be developed. [0003] Glutathione is N-(N-L-γ-glutamyl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P21/02
Inventor 徐虹欧阳平凯李霜韦萍
Owner WUHAN ELITE CHEM FERTILIZER
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