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Process of preparing gamma-poly glutaminic acid

A technology of polyglutamic acid and glutamic acid, which is applied in the field of preparation of γ-polyglutamic acid, can solve the problems of many by-products, small molecular weight of the product, and high production cost, achieve good hygroscopic performance and moisturizing performance, and reduce separation and purification The effect of optimization of process and strain culture conditions

Inactive Publication Date: 2007-07-18
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the chemical synthesis method has a long synthesis route, many by-products, low yield, and the product has a small molecular weight, which is difficult to meet the requirements as a new drug carrier material, and has no industrial application value.
However, due to the low concentration of γ-PGA in the extraction method, and with different conditions, the content varies greatly
Therefore, the extraction process is very complicated, the production cost is very high, and it is also difficult to carry out large-scale industrial production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 1) Bacillus subtilis (Bacillus subtills) zju-7 was activated on a slant at 37°C, and the slant medium: glucose 1.5%, peptone 1%, glutamic acid 3%, MgSO 4 0.1%, NaCl 1%, K 2 HPO 4 0.01%, make up volume with water; activate for 8 hours.

[0035] 2) Liquid medium: glucose 2%, peptone 1.5%, MgSO 4 0.1%, NaCl 1%, K 2 HPO 40.01%, at the same time, dilute the final fermentation broth of Corynebacterium glutamicum that was centrifuged to remove the thalline according to the concentration of glutamic acid to a concentration of 3% of glutamic acid in the shake flask culture medium, make up the volume with water, pH7, pack Liquid volume 50ml / 500ml shake flask, sterilized at 115°C for 20 minutes.

[0036] After the sterilization, cool down, inoculate the above-mentioned Bacillus subtilis (Bacillus subtills) zju-7, the inoculum amount is 3%, culture in a shake flask at 37° C. with a rotation speed of 200 rpm, carry out a shake flask fermentation experiment under the above con...

Embodiment 2

[0039] Same as in Example 1, the gamma-polyglutamic acid production strain was replaced by Bacillus natto (Bacillus natto), activated on a slant at 37°C, the slant medium: glucose 1.5%, peptone 1%, glutamic acid 3%, MgSO 4 0.1%, NaCl 1%, K 2 HPO 4 0.01%, make up volume with water, activate for 8 hours.

[0040] Liquid medium: glucose 2%, peptone 2%, glutamic acid 5%, MgSO 4 0.2%, NaCl 2%, K 2 HPO 4 0.05%, pH7, liquid volume 30 / 250ml shake flask, sterilized at 115°C for 20 minutes. Cool after sterilization, inoculate with an inoculum size of 3%, make up the volume with water, cultivate at 37°C for 24 hours, and shake the flask at a rotational speed of 200rpm.

[0041] After the fermentation, the γ-polyglutamic acid was separated and purified as in Example 1, and the yield could reach 10.3 g / L.

Embodiment 3

[0043] As in Example 1, the crystallization mother liquor was used as a substitute for the glutamic acid precursor, and cultured in a shake flask at 37° C. at a rotation speed of 200 rpm for 24 hours.

[0044] After the fermentation is over, centrifuge to remove the bacteria in the fermentation broth, adjust the pH of the supernatant to 4 with hydrochloric acid, then add 4 times the volume of methanol to precipitate, redissolve the precipitate in 100 times the volume of water, and filter to remove insolubles , and then dialysis to remove small molecular substances, and then freeze-dry the solution to obtain γ-polyglutamic acid, and the yield can reach 12g / L.

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Abstract

The present invention discloses poly-gamma-glutamic acid preparing process. The process includes the steps of culturing different kinds of poly-gamma-glutamic acid producing bacterial strains in culture medium containing carbon source, nitrogen source, NaCl and intermediate to obtain poly-gamma-glutamic acid fermenting liquor, precipitating in organic solvent, dialysis and drying to obtain poly-gamma-glutamic acid product. The process has the intermediate as the precursor for preparing poly-gamma-glutamic acid, and is simple, low in cost and high in yield.

Description

technical field [0001] The invention relates to a method for preparing gamma-polyglutamic acid. Background technique [0002] γ-polyglutamic acid (Polyγ-glumatic acid, γ-PGA) is a homo-amino acid compound that can be synthesized by microorganisms. As a biopolymer material, γ-PGA has the advantages of good biodegradability, edible, non-toxic to human body and environment. Therefore, γ-PGA and its derivatives are widely used in food, cosmetics, medicine and water treatment. [0003] At present, the methods for synthesizing γ-PGA mainly include chemical synthesis, extraction and microbial synthesis. However, the chemical synthesis method has a long synthesis route, many by-products, low yield, and the product has a small molecular weight, which is difficult to meet the requirements as a new type of drug carrier material, and has no industrial application value. However, due to the low concentration of γ-PGA in the extraction method, the content varies greatly with different ...

Claims

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Application Information

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IPC IPC(8): C12P13/00C12P1/04C12R1/125C12R1/10C12R1/07C08G69/02
Inventor 徐志南石峰岑沛霖
Owner ZHEJIANG UNIV
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