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Method for extracting gamma-polyglutamic acid by inorganic salt/organic solvent coprecipitation effect

A polyglutamic acid and organic solvent technology, applied in the field of biological synthesis of biodegradable polymer materials, can solve the problems of extremely high requirements for fiber membranes and difficult scale-up of industrial production, so as to improve extraction yield and avoid environmental problems , the effect of reducing viscosity

Inactive Publication Date: 2011-08-17
天津北洋百川生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method has extremely high requirements on the fiber membrane, and it is difficult to scale up production in industrial production.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Extraction of gamma-polyglutamic acid:

[0025] Preparation of the first medium

[0026] (1) Strain activation medium: LB medium: yeast powder 5g / L, tryptone 10g / L, NaCl 10g / L, agar 20g / L, pH 7.0, sterilized by high pressure steam at 121°C for 15min;

[0027] (2) Seed medium: K 2 HPO 4 0.5g / L, MgSO 4 ·7H 2 O 0.5g / L, glucose 30g / L, yeast powder 5g / L, tryptone 10g / L, pH 7.0; 121℃ high pressure steam sterilization for 15min

[0028] (3) Fermentation medium: glucose 80g / L, MgSO 4 ·7H 2 O 0.5g / L, yeast extract 20g / L, NaCl 10g / L, CaCl 2 1g / L, sodium glutamate 8g / L (sterilized separately), pH 7.0, 121℃ high pressure steam sterilization for 15min;

[0029] The second step of bacterial activation

[0030] Transfer the slant strain - Bacillus licheniformis (B. licheniformis) to LB slant medium, activate and cultivate at 37°C for 12h;

[0031] The third step of seed culture Inoculate the activated slant bacteria in the second step into a 500mL conical flask containing 50...

Embodiment 2

[0039] In this example, in the fifth step (2) fermentation filtrate added three times the volume of absolute ethanol according to the method of Example 1 to make the final concentration in the solution reach 75% (v / v), 2% (m / v) v) of sodium chloride, at 40°C, slowly stirred and placed overnight, the obtained precipitate was γ-polyglutamic acid, and the supernatant was poured to obtain polyglutamic acid precipitate, which was dissolved in 1:5 (m:v) In distilled water, centrifuge again to remove insoluble small molecules, and the supernatant is freeze-dried to obtain crude γ-polyglutamic acid. The γ-polyglutamic acid crude product obtained in the fifth step (2) was dissolved in distilled water at 1:5 (m:v), 2% (m / v) sodium chloride was added, and dehydrated alcohol was added to make it in the distilled water. The final concentration in the solution reached 30% (v / v), slowly stirred to obtain γ-polyglutamic acid precipitation, dissolved in distilled water at 1:5 (m:v), and filter...

Embodiment 3

[0041] In this example, in the fifth step (2) fermentation filtrate according to the method of Example 1, add absolute ethanol to make the final concentration in the supernatant reach 80% (v / v), 1% (m / v) v) potassium chloride, at 30 ° C, slowly stirred and placed overnight, the obtained precipitate was γ-polyglutamic acid, and the supernatant was poured to obtain polyglutamic acid precipitate, which was dissolved in 1:10 (m:v) In distilled water, centrifuge again to remove insoluble small molecules, and the supernatant is freeze-dried to obtain crude γ-polyglutamic acid. The γ-polyglutamic acid crude product obtained in the fifth step (2) was dissolved in distilled water at 1:10 (m:v), 1% (m / v) potassium chloride was added, and dehydrated alcohol was added to make it in distilled water. The final concentration in the solution reaches 30% (v / v), slowly stir to obtain γ-polyglutamic acid precipitation, dissolve in distilled water at 1:10 (m:v), and filter through ultrafiltration...

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Abstract

The invention discloses a method for extracting gamma-polyglutamic acid. The scheme of the method comprises the following steps: filtering out thallus by pumping fermentation liquor; adding organic solvent into fermentation supernate, adding sodium chloride, potassium chloride or ammonium sulfate, and putting for one night to obtain precipitate which is a gamma-polyglutamic acid coarse product; dissolving the gamma-polyglutamic acid coarse product into distilled water, filtering out undissolved micromolecule substance in a centrifuging way, adding sodium chloride, potassium chloride or ammonium sulfate and organic solvent into the supernate, stirring to obtain a gamma-polyglutamic acid precipitate, dissolving the gamma-polyglutamic acid precipitate into the distilled water according to 1: (5-10) (m : v), and filtering out impurity by an ultrafiltration membrane; and performing the freeze drying to ultrafiltration peritoneal effluent so as to obtain white gamma-polyglutamic acid powder. After the method is used, the product extraction yield can be effectively improved, the usage amount of the organic solvent can be reduced, the production cost can be lowered, the environment problem caused by a large number of organic solvent during extracting can be avoided, and the environment pollution can be reduced.

Description

technical field [0001] The technical scheme of the invention belongs to the research field of biological synthesis of biodegradable polymer materials. Background technique [0002] γ-Polyglutamic acid [γ-ployglutamic acid, referred to as γ-PGA] is a copolymer of glutamic acid as the only monomer. γ-PGA was first discovered in 1937. The researchers found γ-PGA in the cell capsules of Bacillus anthracis and Bacillus mesentericus, which is one of the main components of some microbial capsules. Later Japanese researchers also found γ-PGA in Bacillus subtilis and Bacillus natto. So far, foreign units represented by Ajinomoto Co., Ltd., Meiji Seika Co., Ltd. and Hirosaki University have conducted in-depth research on the properties, synthesis and application of γ-PGA, and γ-polyglutamic acid has commercial products. However, there are relatively few studies in this area in our country. Only in recent years have scholars made basic research on the synthesis and properties of pol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/02C08G69/10C12R1/10
Inventor 乔长晟楼鹏李政李雪蒋磊
Owner 天津北洋百川生物技术有限公司
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