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Method for separating purified gamma-polyglutamic acid from fermentation liquor

A polyglutamic acid, separation and purification technology, applied in the field of separation and purification of γ-polyglutamic acid, can solve the problems of low yield of γ-polyglutamic acid, high cost of separation and purification, adverse environmental impact, etc. Conducive to environmental protection, cost reduction, low cost effect

Active Publication Date: 2010-10-13
浙江赛灵特医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The above-mentioned purification method has the disadvantage of many times of freeze-drying, which leads to high cost of separation and purification, and the yield of γ-polyglutamic acid product is not high. At the same time, the use of methanol also brings certain environmental problems and has adverse effects on the environment.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] 100mL γ-polyglutamic acid fermentation broth was diluted 3 times with water to adjust the pH to 3.0; the diluted fermentation broth was filtered and sterilized with a high-efficiency plate airtight filter, the filter aid used was perlite, and the filter membrane pore size used was 0.2 micron, get filtrate;

[0028] Adjust the pH of the above filtrate to 7.1, precipitate with 3 times the volume of ethanol, and let stand in ethanol for 180 minutes to remove the water in the precipitate; centrifuge to separate the precipitate, and the resulting precipitate is dried under reduced pressure at 0.2Mpa to obtain γ -Crude polyglutamic acid;

[0029] Dissolve the crude γ-polyglutamic acid with an equal volume of deionized water, adjust the pH to 7.0, add 2 g of an industrial protease preparation with a titer of 1000 U / ml, and incubate at 37°C for 60 minutes to remove impurities; adjust the pH to 3.0, Ultrafiltration removes pigments and small molecular impurities; then adjust the pH ...

Embodiment 2

[0032] 100mL γ-polyglutamic acid fermentation broth was diluted 3 times with water to adjust the pH to 3.2; the diluted fermentation broth was filtered and sterilized with a high-efficiency plate-type airtight filter, the filter aid used was perlite, and the membrane pore size used was 0.2 micron, get filtrate;

[0033] Adjust the pH of the above filtrate to 7.3, use 3 times the volume of ethanol for precipitation, let stand in ethanol for 180 minutes to remove the water in the precipitate; centrifuge to separate the precipitate, and the resulting precipitate is dried under reduced pressure at 0.2Mpa to obtain γ -Crude polyglutamic acid;

[0034] Dissolve the crude γ-polyglutamic acid with an equal volume of deionized water, adjust the pH to 7.1, add 2g of an industrial protease preparation with a titer of 1000 U / ml, and incubate at 37°C for 60 minutes to remove impurities; adjust the pH to 3.1, Ultrafiltration removes pigments and small molecular impurities; then adjust the pH to...

Embodiment 3

[0037] 100mL γ-polyglutamic acid fermentation broth was diluted 3 times with water to adjust the pH to 3.0; the diluted fermentation broth was filtered and sterilized with a high-efficiency plate airtight filter, the filter aid used was perlite, and the filter membrane pore size used was 0.2 micron, get filtrate;

[0038] Adjust the pH of the above filtrate to 7.1, precipitate with 3 times the volume of ethanol, and let stand in ethanol for 170 minutes to remove the moisture in the precipitate; centrifuge to separate the precipitate, and the obtained precipitate is dried under reduced pressure at 0.2Mpa to obtain γ -Crude polyglutamic acid;

[0039] Dissolve the crude γ-polyglutamic acid with an equal volume of deionized water, adjust the pH to 7.0, add 2 g of an industrial protease preparation with a titer of 1000 U / ml, and incubate at 37°C for 60 minutes to remove impurities; adjust the pH to 3.0, Ultrafiltration removes pigments and small molecular impurities; then adjust the p...

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PUM

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Abstract

The invention discloses a method for separating purified gamma-polyglutamic acid from fermentation liquor. The method comprises the following steps of: firstly, diluting the fermentation liquor by using water and regulating the pH value to 2.8-3.2; carrying out plate filter for degerming, and regulating the pH value of filter liquor to 6.9-7.3; carrying out alcohol precipitation and removing water in the precipitate; centrifuging to separate out the precipitate as well as decompressing and drying to obtain a crude gamma-polyglutamic acid product; dissolving again and regulating the pH value to 6.9-7.1; adding protease, preserving heat and removing hybrid protein; regulating the pH value to 2.9-3.1 and ultra-filtering to remove pigment and micromolecule impurities; regulating the pH value to 6.9-7.1 and carrying out alcohol precipitation; and decompressing and drying to obtain a finished high-purity gamma-polyglutamic acid product. In the invention, the purposes of separating and purifying high-purity gamma-polyglutamic acid are achieved through regulating the pH values, centrifuging, precipitating and filtering in multiple steps. The method has the advantages of high purity, high yield, low cost and environmental protection.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and particularly relates to a method for separating and purifying γ-polyglutamic acid from fermentation broth. Background technique [0002] γ-Polyglutamic acid (γ-PGA) is an extracellular high molecular polymer secreted by bacteria of the genus Bacillus. It is formed by connecting L- or / and D-glutamic acid through γ-polyglutamyl bonds. Its molecular weight is generally between 100 and 1000KDa, with excellent water solubility, super adsorption and biodegradability, and the degradation product is pollution-free glutamic acid. It is an excellent environmentally friendly polymer material that can be used as Water retention agents, heavy metal ion adsorbents, flocculants, slow-release agents and drug carriers have great commercial and social value in cosmetics, environmental protection, food, medicine, agriculture, desert management and other industries. [0003] At present, the production method of γ-polyglut...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08G69/10
Inventor 汤瑜刘金明王震黄云战
Owner 浙江赛灵特医药科技有限公司
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