Method for extracting polysaccharide from cordyceps militaris medium

A technology for Cordyceps polysaccharide and Cordyceps militaris, which can be applied to non-central analgesics, metabolic diseases, cardiovascular system diseases, etc., can solve the problems of low extraction rate and affect the quality of Cordyceps polysaccharides, and achieve no waste discharge and reasonable process. Effect

Active Publication Date: 2012-10-17
DALIAN POLYTECHNIC UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the extraction of Cordyceps polysaccharides in Cordyceps militaris medium reported publicly at home and abroad is limited to hot water extraction, or using auxiliary physical methods such as ultrasonic waves, the extraction rate is relatively low, and there is a large amount of starch that has not been removed. Affect the quality of Cordyceps polysaccharide

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0077] Example 1 (alkaline extraction method)

[0078] Take 100g of Cordyceps militaris culture medium dry powder, add 2000mL of water in weight, mix well, leaching with 0.5mol / L NaOH at pH 9 at 60°C for 4 hours, then centrifuge to obtain supernatant and precipitate. Add 1000mL of water to the precipitation, adjust the pH to 9, repeat the above operation 1 to 2 times, combine the obtained supernatant, adjust the pH to neutral, add 2g of medium temperature amylase, 2mL of 2% Cacl 2 After hydrolysis at 70°C for 60 minutes, inactivate the enzyme at 90°C for 5 minutes, cool down, and centrifuge at high speed for 10 minutes. The supernatant was concentrated to a polysaccharide content of 4% (detected by the phenol-sulfuric acid method); 3 times the volume of the concentrated solution was added with 95% ethanol, precipitated at 4°C for 12 hours, and then centrifuged at 4000 rpm for 10 minutes to obtain a centrifugal precipitate. The precipitate was vacuum freeze-dried at -50°C to o...

example 2

[0079] Example 2 (single enzyme method)

[0080] Cordyceps militaris medium dry powder 100g, add water 8000mL, adjust the pH value to 2.0, add pepsin 2g, hydrolyze at 37°C for 4 hours, adjust the pH value to 7.0, inactivate the enzyme at 90°C for 10 minutes, cool, and centrifuge at 4000 rpm Obtain the supernatant and precipitate in 10 minutes, add 100 mL of water to the precipitate and repeat the above operation 1 or 2 times, and combine the supernatant. Add 0.2 g of medium-temperature amylase to the supernatant, hydrolyze at 70°C for 60 minutes, inactivate the enzyme at 90°C for 5 minutes, cool, and centrifuge at 4000 rpm for 10 minutes. Concentrate the supernatant to 80mL, add 240mL of 95% ethanol, precipitate at 4°C for 12 hours, then centrifuge at 4000 rpm for 10 minutes to obtain a centrifugal precipitate -50°C and vacuum-dry to obtain 15.8g of crude polysaccharide, which is diluted with water to 50mL , add Sevag reagent (V chloroform: V n-butanol = 5: 1) 10mL, shake vig...

example 3

[0081] Example 3 (double enzyme method)

[0082] Take 100g of fresh Cordyceps militaris medium, mash it, add 200mL of water, add 1800mL of water after homogenization, adjust the pH to 2.0 with 6mol / L HCL, 1g of pepsin, stir at 38°C for 1-4 hours, hydrolyze Keep the pH value at 2.0; then adjust the pH to 8.0 with 0.5mol / L NaOH solution, add trypsin with a weight of 1g of the raw material, stir the enzymolysis in a water bath at 37°C for 4 hours, and keep the pH value at 8.0 during hydrolysis; Adjust the pH to neutral, inactivate at 90-98°C for 5-10 minutes, and centrifuge after cooling to obtain the supernatant and precipitate; the supernatant is set aside, add 500 mL of water to the precipitate and repeat the above steps twice, and combine the supernatant , adjust the pH to neutral, add 1g of medium-temperature amylase, hydrolyze at 70°C for 60 minutes, inactivate the enzyme at 90°C for 5 minutes, cool, and centrifuge at high speed for 10 minutes. After the supernatant is con...

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Abstract

The invention discloses a method for extracting polysaccharide from a cordyceps militaris medium. The cordyceps militaris is crushed and mixed with water, is extracted in an enzyme liquor and an alkali liquor under 30 DEG C to 80 DEG C, and an obtained solution is centrifugalized. An insoluble substance is extracted repeatedly, and supernatants are combined. Amylase enzymolysis is performed on the obtained product under 50 DEG C to 75 DEG C. After centrifugation, a supernatant is condensed to be with a sugar content of 1.5 percent to 3.0 percent. The supernatant is added with ethanol and alcohol precipitation is performed on the obtained product under 0 DEG C to 4 DEG C, a product is obtained by drying the obtained product after centrifugation. After deproteinization and dialysis, crude cordyceps polysaccharide is obtained. A single enzyme method, a double enzyme method, a compound enzyme method and a supersonic wave enzyme method can be adopted, and protease, trypsin, etc. can be adopted. The method optimizes the technology route for extracting the cordyceps polysaccharide, enhances an obtaining rate and an extraction efficiency of the cordyceps polysaccharide. Extraction, separation and purification of the cordyceps polysaccharide are carried out effectively. The method provides a solution approach for the utilization of agricultural wastes, and provides a technology base for further exploitation of the cordyceps polysaccharide.

Description

technical field [0001] The invention relates to the field of biological products, in particular to a method for extracting polysaccharides from Cordyceps militaris culture medium. Background technique [0002] Cordyceps militaris (L.) Link, also known as Cordyceps militaris, belongs to Asconlycotina, Pyrenomycetes, Sphaeriales, Claviciptiaceae, Cordyceps ( Cordyceps) and Cordyceps sinensis (Cordyceps Sinensis (Berk) Sacc) belong to different species. Can produce a variety of biologically active substances: polysaccharides, mannitol, cordycepin, steroids, etc. In recent years, the research on the artificial cultivation of Cordyceps militaris and its active ingredients has been particularly active. [0003] Cordyceps polysaccharide is a kind of polymer compound, it is one of the main active components of Cordyceps militaris, and it is the pharmacological active substance with the highest content in Cordyceps militaris. A large number of pharmacological experiments have show...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/00A61P35/00A61P37/02A61P29/02A61P13/12A61P9/10A61P3/10A61P3/06
Inventor 朱蓓薇安冬董秀萍李冬梅杨静峰
Owner DALIAN POLYTECHNIC UNIVERSITY
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