Method for separating and purifying ultra-high molecular weight poly-gamma-glutamic acid from high-viscosity fermentation liquor

An ultra-high molecular weight, separation and purification technology, which is applied in the field of separation and purification of ultra-high molecular weight poly-γ-glutamic acid, can solve the problems of lower PGA recovery rate, low processing efficiency, and increased production costs, and achieve easy removal of low molecular weight impurities, Product purity and recovery rate are high, and the effect of reducing production cost

Pending Publication Date: 2021-03-12
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The main problems of these three methods are: (1) all will bring in different degrees of foreign substances such as inorganic salts and organic solvents in the separation system, thereby increasing the process operation unit to remove these foreign substances; (2) these Exogenous substances may lead to the reduction or denaturation of PGA molecular weight, thereby reducing the recovery rate of PGA; (3) The use of these foreign substances not only increases production costs, but also poses a greater safety hazard in organic solvents during the production process
But there are still some problems to be solved: (1) The molecular weight of the isolated and purified PGA is lower than 1×10 6 , which limits the scope of application of PGA; (2) The separation and purification process includes fermentation liquid viscosity reduction, filtration sterilization, heating method to remove protein and other macromolecular impurities, ion exchange resin method to remove metal ions, activated carbon decolorization, nanofiltration to remove small molecule impurities , ultrafiltration concentration and drying, the operation unit is cumbersome, and the processing efficiency is low, which makes industrial application more difficult

Method used

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  • Method for separating and purifying ultra-high molecular weight poly-gamma-glutamic acid from high-viscosity fermentation liquor
  • Method for separating and purifying ultra-high molecular weight poly-gamma-glutamic acid from high-viscosity fermentation liquor

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Embodiment 1

[0037] A method for separating and purifying ultra-high molecular weight poly-γ-glutamic acid from high-viscosity fermentation broth, comprising the steps of:

[0038] (1) Dilute and acidify the fermentation broth

[0039] Use viscosity > 3000mpa s, molecular weight of poly-γ-glutamic acid contained in the fermentation broth > 2×10 6 The high-viscosity fermentation broth of poly-γ-glutamic acid was diluted to 2 times the original volume with distilled water in the fermenter, and then the pH was adjusted to 3 with 3 M HCl, at 37°C and 150r / min. Acidify for 30 minutes;

[0040] (2) Plate and frame filter

[0041] Add commercially available vermiculite powder filter aid to the diluted and acidified fermentation broth at a mass percentage of 1%, filter through a 400-mesh filter cloth, and filter with a plate and frame to remove bacteria and pigments at a pressure of 0.2 MPa;

[0042] (3) Cyclic ultrafiltration

[0043] The molecular weight cut-off is 3×10 5 The ultrafiltratio...

Embodiment 2

[0047] A method for separating and purifying ultra-high molecular weight poly-γ-glutamic acid from high-viscosity fermentation broth, comprising the steps of:

[0048] (1) Dilute and acidify the fermentation broth

[0049] Bacillus subtilis ( Bacillus subtilis ) GXA-28 (preservation number CCTCC NO: M2012347, depository unit: China Type Culture Collection Center) is a production strain, a high-viscosity fermentation broth containing ultra-high molecular weight poly-γ-glutamic acid obtained through liquid fermentation in a fermenter, The viscosity of the fermentation broth is >3000mpa·s, and the molecular weight of poly-γ-glutamic acid contained in the fermentation broth is >3×10 6 ; Dilute the fermentation broth to 5 times the original volume with distilled water in the fermenter, then adjust the pH to 5 with 3M HCl, and acidify for 90 minutes at 45°C and 250r / min;

[0050] (2) Plate and frame filter

[0051] Add the commercially available vermiculite powder filter aid to t...

Embodiment 3

[0057] A method for separating and purifying ultra-high molecular weight poly-γ-glutamic acid from high-viscosity fermentation broth, comprising the steps of:

[0058] (1) Dilute and acidify the fermentation broth

[0059] Use viscosity > 3000mpa s, molecular weight of poly-γ-glutamic acid contained in the fermentation broth > 2×10 6 The high-viscosity fermentation broth of poly-γ-glutamic acid was diluted to 2.5 times the original volume with distilled water in the fermenter, and then adjusted to pH 4 with 3 M HCl, at 42°C and 200r / min. Acidify for 60min;

[0060] (2) Plate and frame filter

[0061] Add the commercially available vermiculite powder filter aid to the diluted and acidified fermentation broth at a mass percentage of 2%, filter through a 600-mesh filter cloth, and filter with plate and frame to remove bacteria and pigments at a pressure of 0.2 MPa;

[0062] (3) Cyclic ultrafiltration

[0063] The molecular weight cut-off is 4×10 5 The ultrafiltration membran...

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Abstract

The invention belongs to the technical field of separation and purification, and particularly relates to a method for separating and purifying ultra-high molecular weight (more than 2*10<6>) poly-gamma-glutamic acid from high-viscosity (more than 3000 mpa.s) fermentation liquor. The refined white amorphous poly-gamma-glutamic acid is obtained through four-step unit operation of fermentation liquordilution acidification, plate-frame filtration, circulating ultrafiltration and low-temperature drying, the molecular weight of the obtained poly-gamma- glutamic acid product is larger than 3*10<6>,the purity is higher than 94.8%, and the total recovery rate is higher than 57%. The method has the advantages of simple process, low cost, ultrahigh molecular weight of the product, high purity of the product and high recovery rate, and has a great industrialization potential.

Description

technical field [0001] The invention belongs to the technical field of separation and purification, and in particular relates to a method for separating and purifying ultrahigh molecular weight poly-γ-glutamic acid from high-viscosity fermentation broth. Background technique [0002] Poly-γ-glutamic acid (PGA) is a microorganism composed of L-glutamic acid and (or) D-glutamic acid linked by amide bonds through α-amino and γ-carboxyl groups. source polymer. Due to the differences in production strains, medium components and fermentation conditions, the molecular weight of PGA shows diversity, generally in the range of 1×10 5 ~8×10 6 . PGA has excellent water solubility, biocompatibility, biodegradability, and chemical modification, and can be widely used in food, medicine, cosmetics, agriculture, and environmental protection. However, as a high molecular weight polymer, the molecular weight is the main factor affecting the application range of PGA. For example, low molec...

Claims

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Application Information

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IPC IPC(8): C08G69/10
CPCC08G69/10
Inventor 曾伟梁智群陈桂光张璇
Owner GUANGXI UNIV
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