Chinese wolfberry polysaccharide protein removal method based on cloud point extraction

A technology of wolfberry polysaccharide and protein removal, which is applied in the field of separation and purification of biomacromolecules, can solve the problems of long purification process flow, large pollution, high material consumption and energy consumption, and achieve short process flow, low production cost and high biocompatibility Effect

Inactive Publication Date: 2015-11-04
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the defects of long process flow, high material consumption and energy consumption, and large pollution of the current Lycium barbarum polysaccharide purification process, and provide a separation and purification method of Lycium barbarum pure polysaccharide with simpler process flow, low cost, and environmental protection

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] (1) The novel thermosensitive block copolymer polyethylene oxide polypropylene oxide polyethylene oxide (EOPOEO) was mixed with distilled water to prepare 8% EOPOEO aqueous solution.

[0028] (2) Add crude polysaccharide solution of Lycium barbarum to the solution obtained in step (1), so that the concentration of Lycium barbarum polysaccharide reaches 4 mg / ml. Adjust the pH value of the system to 5, shake and mix, separate the phases at 70°C and stand still to form a two-phase aqueous system. After 12 hours, separate the upper and lower phases.

[0029] (3) Concentrate the upper phase obtained in step (2) under reduced pressure, mix it with 5 times the volume of 95% ethanol, and let it stand at 4°C for 12 hours to obtain purified Lycium barbarum polysaccharide. The recovery rate of polysaccharide was 81%, and the removal rate of protein was 87%

[0030] (4) Let the lower phase obtained in step (2) stand overnight at 4°C to precipitate the protein, centrifuge to get th...

Embodiment 2

[0032] (1) A 12% EOPOEO aqueous solution was prepared by mixing the novel temperature-sensitive block copolymer polyethylene oxide polypropylene oxide polyethylene oxide (EOPOEO) with distilled water.

[0033] (2) Add crude polysaccharide solution of Lycium barbarum to the solution obtained in step (1), so that the concentration of Lycium barbarum polysaccharide reaches 4 mg / ml. Adjust the pH value of the system to 5, shake and mix, separate the phases at 70°C and stand still to form a two-phase aqueous system. After 12 hours, separate the upper and lower phases.

[0034] (3) Concentrate the upper phase obtained in step (2) under reduced pressure, mix it with 5 times the volume of 95% ethanol, and let it stand at 4°C for 12 hours to obtain purified Lycium barbarum polysaccharide. The recovery rate of polysaccharide was 71%, and the removal rate of protein was 64%

[0035] (4) Let the lower phase obtained in step (2) stand overnight at 4°C to precipitate the protein, centrifug...

Embodiment 3

[0037] (1) A 4% EOPOEO aqueous solution was prepared by mixing the novel temperature-sensitive block copolymer polyethylene oxide polypropylene oxide polyethylene oxide (EOPOEO) with distilled water.

[0038] (2) Add crude polysaccharide solution of Lycium barbarum to the solution obtained in step (1), so that the concentration of Lycium barbarum polysaccharide reaches 4 mg / ml. Adjust the pH value of the system to 5, oscillate and mix, and separate the phases at 70° C. to form a two-phase aqueous system. After 12 hours, separate the upper and lower phases.

[0039] (3) Concentrate the upper phase obtained in step (2) under reduced pressure, mix it with 5 times the volume of 95% ethanol, and let it stand at 4°C for 12 hours to obtain purified Lycium barbarum polysaccharide. The recovery rate of polysaccharide was 65%, and the removal rate of protein was 72%

[0040] (4) Let the lower phase obtained in step (2) stand overnight at 4°C to precipitate the protein, centrifuge to ge...

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Abstract

The invention discloses a Chinese wolfberry polysaccharide protein removal method based on cloud point extraction, relating to the technical field of bioengineering. The method comprises the following basic steps: adding a prepared temperature-sensitive block copolymer polyethylene oxide-polypropylene oxide-polyethylene oxide (EOPOEO) solution, mixing with a Chinese wolfberry crude polysaccharide water solution, and carrying out liquid-liquid-phase separation on the system under the induction of temperature, wherein the impurity proteins in the Chinese wolfberry polysaccharide are successfully removed under the action of the polymer phase; after the phase separation, mixing the upper phase of the system with ethanol, and precipitating to obtain a polysaccharide finished product; and meanwhile, cooling the lower phase with enriched temperature-sensitive polymer to precipitate most proteins, and carrying out comprehensive operation (comprising the steps of pH value regulation, electrolyte addition, heating and the like) to form a secondary cloud point extraction system, thereby successfully recovering the temperature-sensitive polymer in the lower phase. The invention provides a separation purification method of Chinese wolfberry pure polysaccharide, which has the advantages of simpler technical process, low cost and environment friendliness.

Description

technical field [0001] The invention relates to the technical field of separation and purification of biological macromolecules, in particular to a method for removing protein from Lycium barbarum polysaccharides based on cloud point extraction. Background technique [0002] Lycium barbarum, as a traditional precious Chinese herbal medicine, has good health care and medical value. As one of the main active ingredients in Lycium barbarum, Lycium barbarum polysaccharide has attracted much attention for its special effects. As a macromolecular substance with a molecular weight between 80 and 120 kDa, Lycium barbarum polysaccharides have been shown to enhance immune function, protect the liver and anti-aging, lower blood sugar and blood lipids, anti-tumor, sensitize radiotherapy, and increase the efficacy and detoxification of chemotherapy. good effect. To sum up, Lycium barbarum polysaccharide shows broad commercial value due to its application prospect in food and medicine. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/00
Inventor 王赟胡晓玮韩娟唐旭倪良陈桐王蕾
Owner JIANGSU UNIV
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