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Anti-oxidation polysaccharide in boletus subsplendidus as well as preparation method and application thereof

A technology for tawny boletus and oxidized polysaccharide, which is applied in the field of extraction and purification of antioxidant polysaccharides in chrysanthemum boletus, can solve problems such as research difficulties, achieves simple preparation, is suitable for large-scale production, and has good safety Effect

Active Publication Date: 2017-01-11
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, the separation, purification and determination of biological activity of polysaccharides have been relatively mature, but the methods are still being developed and improved. Sexual constraints, this part of the research is still very difficult

Method used

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  • Anti-oxidation polysaccharide in boletus subsplendidus as well as preparation method and application thereof
  • Anti-oxidation polysaccharide in boletus subsplendidus as well as preparation method and application thereof
  • Anti-oxidation polysaccharide in boletus subsplendidus as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1. Preparation of antioxidant polysaccharide of the present invention

[0037] (1) Dry the fruiting bodies of Boletus edulis, pulverize with a high-speed grinder, add 30 times volume of distilled water, shake in a water bath at 80℃ for 5h, centrifuge at 8000r for 10min, take the supernatant, and repeat the extraction 2-3 times , Combine the centrifugal supernatants, use a rotary evaporator for rotary evaporation, concentrate to a certain volume, add 4 times the volume of absolute ethanol and let stand overnight at 4°C. After centrifugation, the precipitate was reconstituted to obtain a polysaccharide solution. To remove protein by sevage method, Sevage reagent was prepared according to the ratio of chloroform: n-butanol=4:1, and Sevage reagent was added according to the ratio of crude solution: Sevage reagent=4:1, vigorously shaken for 8-10min, and left to stand. When the interface between the aqueous and organic phases appears white, open the separatory funnel to...

Embodiment 2

[0040] Example 2. Determination of the purity and molecular weight of the antioxidant polysaccharide of the present invention

[0041] Prepare standard dextran (Dextran T-10, T-50, T-70, T-100, T-200) of known molecular weight with double-distilled water to prepare a standard solution with a concentration of 10 mg / mL. After the filter membrane was filtered, the sample was loaded on the SUGAR KS-804 sugar column, the injection volume was 20μl, the molecular weight Mw of different dextran was used as the ordinate, and the retention time t of the standard dextran was used as the abscissa to draw a standard curve ( Figure 4 ).

[0042] Prepare a solution with a concentration of 10mg / mL with double distilled water, filter it through a 0.22μm filter membrane and load the sample, collect the spectrum, and determine the purity according to the peak situation. image 3 Shows a single narrow symmetrical peak, it can be judged that the polysaccharide is pure product, the retention time is 6.0...

Embodiment 3

[0043] Example 3. The DPPH free radical scavenging effect of the antioxidant polysaccharide of the present invention

[0044] (1) Use anhydrous methanol as a solvent for the antioxidant polysaccharide to prepare a solution with a concentration of 0.5 mg / mL, and use anhydrous methanol for DPPH to prepare a final concentration of 1×10 -4 mol / L solution.

[0045] (2) Take 0.06, 0.12, 0.18, 0.3, and 0.6 mL of the above polysaccharide solution in a 1.5 mL centrifuge tube, make up 1.2 mL with DPPH solution, and use an equal volume of anhydrous methanol to replace the sample solution as a blank control, and ascorbic acid as a positive control , Placed in the dark for 30 minutes.

[0046] (3) Measure the absorbance of each treatment at 517nm.

[0047] (4) Press "Removal rate=[1-(A s -A b ) / A i ]×100%" formula to calculate DPPH clearance rate, where A i Is the absorbance value of DPPH solution without sample, A s Is the absorbance of the sample after reaction with DPPH, A b Is the absorbance o...

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Abstract

The invention discloses an anti-oxidation polysaccharide in boletus subsplendidus as well as a preparation method and application thereof. The anti-oxidation polysaccharide is extracted from boletus subsplendidus and is named HNG2. The anti-oxidation polysaccharide is obtained by grinding boletus subsplendidus sporocarp into powder and then carrying out hot water extraction, ethanol precipitation, protein removal, dialysis, freeze-drying, ion exchange and molecular sieving. The HNG2 is yellowish powder, is a polysaccharide containing alpha- and beta-glycosidic bond configurations simultaneously, has a molecular weight of 177KDa, is easily soluble in hot water, and is insoluble in organic solvents, such as ethanol, ethyl ether and acetone. The anti-oxidation polysaccharide is composed of fucose, xylose, glucose, galactose and mannitose in a ratio of 0.36:0.8:30:2:1.6. The polysaccharide has a relatively strong effect on eliminating DPPH, superoxide anion and hydroxyl radical. The anti-oxidation polysaccharide in boletus subsplendidus is a natural extract and is simple in preparation method, relatively low in cost and high in purity.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a method for extracting and purifying antioxidant polysaccharides from Boletus chrysogenum. Background technique [0002] my country is rich in fungi resources, and many fungi have received widespread attention as precious medicinal and edible fungi. Edible fungus is rich in nutrients, rich in vitamins and minerals, high in dietary fiber, low in energy, high in protein, and low in fat. The protein content in edible fungi is about 18%-35%, which is about 1.6 times that of beef. It contains 8 kinds of amino acids necessary for human body, and the ratio is close to that of human body, so it is easily absorbed. In addition, edible fungus is called "plant meat" because it contains a variety of vitamins and is rich in VB 1 , VB 2 , VC, folic acid, VB 12 Wait. With the continuous in-depth development and research on fungal resources, many active ingredients have been isolated and ob...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/00A61K31/715A61P39/06
CPCA61K31/715C08B37/0003
Inventor 刘方张可人王一君
Owner NANKAI UNIV
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