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Method for extracting Pleurotus tuber-regium cell wall glycoprotein

An extraction method and cell wall technology, which is applied in the field of extracting glycoproteins from the cell walls of tiger milk mushrooms, can solve the problems that glycoproteins are rarely involved

Active Publication Date: 2021-07-30
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there have been sufficient studies on polysaccharides, proteins, amino acids, and lectins of tiger milk mushroom, but few studies on its glycoproteins

Method used

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  • Method for extracting Pleurotus tuber-regium cell wall glycoprotein
  • Method for extracting Pleurotus tuber-regium cell wall glycoprotein
  • Method for extracting Pleurotus tuber-regium cell wall glycoprotein

Examples

Experimental program
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Effect test

Embodiment 1

[0037] (1) After harvesting, the dried sclerotia of tiger milk mushroom is pulverized by a mill, and passed through a 50-mesh sieve to obtain dried sclerotia powder of tiger milk mushroom.

[0038] (2) At room temperature, wash and centrifuge the dried sclerotia powder of tiger milk mushroom after sieving with deionized water for 3 times to remove impurities, then wash and centrifuge 3 times with PBS solution with pH 6.5 in turn, the sclerotia powder and PBS material The liquid ratio is 1g:(10~20)mL, remove the soluble components in the cells, remove the filtrate by suction filtration with a vacuum degree of 0.05~0.1MPa, wash the obtained residue with ultrapure water 8 times, and further remove the soluble components and residual components in the cells of PBS to obtain sclerotia cell wall fragments.

[0039] (3) According to the solid-liquid ratio of 1:5 (w / v), add detergent-reductant mixed solution (DRAS) to the cell wall fragments obtained in step (2) for extraction; after ...

Embodiment 2

[0046] (1) After harvesting, the dried sclerotia of tiger milk mushroom is pulverized by a mill, and passed through a 50-mesh sieve to obtain dried sclerotia powder of tiger milk mushroom.

[0047] (2) At room temperature, wash and centrifuge the dried sclerotia powder of tiger milk mushroom after sieving with deionized water for 3 times to remove impurities, then wash and centrifuge 3 times with PBS solution of pH 7.0 successively, the sclerotia powder and the material of PBS The liquid ratio is 1g:(10~20)mL, remove the soluble components in the cells, remove the filtrate by suction filtration with a vacuum degree of 0.05~0.1MPa, wash the obtained residue with ultrapure water 8 times, and further remove the soluble components and residual components in the cells of PBS to obtain sclerotia cell wall fragments.

[0048] (3) According to the solid-liquid ratio of 1:10 (w / v), add DRAS to the cell wall fragments obtained in step (2) for extraction; after mixing evenly, extract at ...

Embodiment 3

[0054] (1) Pour the harvested fresh sclerotium of tiger milk mushroom into liquid nitrogen to freeze, and grind to obtain fresh sclerotia powder of tiger milk mushroom.

[0055] (2) Under normal temperature, wash and centrifuge the fresh sclerotia powder 3 times with deionized water to remove impurities, then wash and centrifuge 3 times with the PBS solution of pH 6.5 successively, the solid-liquid ratio of sclerotia powder and PBS is 1g: (10 ~20) mL, remove intracellular soluble components, remove the filtrate by suction filtration with a vacuum degree of 0.05-0.1 MPa, wash the obtained residue with ultrapure water 10 times, further remove intracellular soluble components and residual PBS, and obtain fresh sclerotia Cell wall fragments.

[0056] (3) According to the solid-liquid ratio of 1:5 (w / v), add DRAS to the cell wall fragments obtained in step (2) for extraction; after mixing evenly, extract at 90°C for 10 minutes, and then centrifuge at room temperature at a speed of ...

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Abstract

The invention discloses an extraction method of Pleurotus tuber-regium cell wall glycoprotein, and belongs to the technical field of biology. The Pleurotus tuber-regium sclerotium cell wall glycoprotein is obtained by crushing dry or fresh Pleurotus tuber-regium sclerotium, cleaning with PBS, leaching with a detergent-reducing agent mixed solution, precipitating with trichloroacetic acid, cleaning the material with acetone and dialyzing the liquid to obtain the product, wherein the glycoprotein accounts for 4.93-5.36% of the dry weight of the sclerotium cell wall, the total sugar content in the glycoprotein is about 32%, the total protein content is about 67%, and the protein content is about 67.7%, there is no free polysaccharide and macromolecular protein residues; and column separation is performed to obtain pure glycoprotein products with different molecular weights. The method is simple in process, high in operability, easy to implement and relatively high in purity, and has a wide industrial application prospect.

Description

technical field [0001] The invention relates to a method for extracting cell wall glycoprotein of tiger milk mushroom, belonging to the field of biotechnology. Background technique [0002] Glycoprotein (Glycoprotein) is a complex sugar composed of branched oligosaccharide chains and covalently linked proteins. It exists widely in nature and has various functions and activities such as anti-cancer, anti-oxidation, anti-fatigue, and anti-tumor. Its structure And the particularity of the function makes it attract more and more attention. [0003] Edible fungus is a kind of large-scale fungus with wide sources, low price, safety and harmlessness, and its cultivation technology is relatively mature, so it has great potential in the development of glycoprotein resources. However, there are certain difficulties in the efficient extraction and separation of glycoproteins in edible fungi, and the methods for purifying proteins that are commonly used now have low product purity. Th...

Claims

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Application Information

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IPC IPC(8): C07K1/14C07K1/36C07K1/34C07K1/30C07K1/18C07K1/16
CPCC07K1/145C07K1/36C07K1/34C07K1/30C07K1/18C07K1/16
Inventor 朱晨曦杨茜茹陈磊林演汤晴严快陈献忠
Owner JIANGNAN UNIV
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