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Cordyceps militaris culture medium polysaccharide and its separation and purification method and application

A technology of separation, purification and culture medium, which is applied in the direction of medical preparations containing active ingredients, pharmaceutical formulas, organic active ingredients, etc., can solve the problems of inability to obtain pure polysaccharide components, high cost, complicated operation process, etc., and achieve beneficial Human metabolism, low cost, good repeatability

Active Publication Date: 2021-06-01
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] Chinese patent application (application number 201110086808.2) discloses a method for extracting polysaccharides from Cordyceps militaris medium, using different enzyme solutions to remove protein and starch in the Cordyceps militaris medium, and then obtaining the crude Cordyceps militaris medium through alcohol precipitation. Polysaccharide; the operation process is complicated, the cost is high, and the polysaccharide cannot be further separated and purified, and the polysaccharide component with higher purity cannot be obtained

Method used

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  • Cordyceps militaris culture medium polysaccharide and its separation and purification method and application
  • Cordyceps militaris culture medium polysaccharide and its separation and purification method and application
  • Cordyceps militaris culture medium polysaccharide and its separation and purification method and application

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Embodiment 1

[0046] A method for extracting and separating and purifying polysaccharides from the leftovers of Cordyceps militaris culture medium, comprising the following steps:

[0047](1) Extraction of polysaccharide from Cordyceps militaris culture medium: Weigh 55g of Cordyceps militaris rice culture culture leftovers, fully dry and crush, pass through a 40-mesh sieve, weigh 50g of dry powder, add 800ml of distilled water, ultrasonic for 30min, and reflux extraction at 70°C for 1.5h , extract the combined filtrate for 3 times, vacuum filter and concentrate to 100ml at 55°C to obtain the polysaccharide concentrate; add 400ml of 95% ethanol to the polysaccharide concentrate, stir continuously to make the polysaccharide evenly precipitate, and stand overnight at 4°C ; Centrifuge at 5000r / min for 15min and dry the precipitate to obtain the polysaccharide extract of Cordyceps militaris culture medium;

[0048] (2) Decolorization of the polysaccharide of the Cordyceps militaris medium: the ...

Embodiment 2

[0057] The polysaccharide P2 of the Cordyceps militaris culture medium obtained in Example 1 was analyzed by ultraviolet spectrum, and 1 mg of polysaccharide samples were weighed to prepare a 1 mg / mL polysaccharide solution, and the ultraviolet spectrum was scanned in the range of 200-800 nm.

[0058] figure 2 It is the ultraviolet spectrogram of P2, and the results show that P2 has no obvious absorption peak at 260nm and 280nm, indicating that P2 does not contain protein and nucleic acid substances.

Embodiment 3

[0060] Carry out polysaccharide molecular weight analysis to the Cordyceps militaris culture medium polysaccharide P2 that embodiment 1 obtains, concrete experimental method is as follows:

[0061] Molecular weights were determined using gel permeation chromatography (GPC). Weigh 2 mg of the freeze-dried polysaccharide sample, add 0.02M phosphate buffer to dissolve, prepare a 2.0 mg / mL solution, filter it with a 0.22 μm sterile filter membrane, and take the filtrated liquid for later use.

[0062] Chromatographic conditions: column temperature 35°C; 0.02mol / L phosphate buffer (pH 7.0) as mobile phase, flow rate 0.6ml / min, injection volume 20μL; TSK gel guard column (40mm×6.0mm), TSKG- 4000K gel column (300mm×7.8mm) and TSKG-2500K gel column (300mm×7.8mm); Waters 2414 differential refraction detector detection. Prepare a series of dextran solutions with different molecular weights (700, 400, 200, 100, 50, 30, 10, 5kD) as standard samples, and make a standard curve. The molecu...

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Abstract

The invention discloses a cordyceps militaris culture medium polysaccharide and its separation and purification method and application. The polysaccharide is composed of the following molar percentages of monosaccharides: 0.26% ribose, 0.11% rhamnose, 2.01% arabinose, 0.34% xylose, 29.62% mannose, 67.19% glucose, 0.47% galactose. The extraction method of the present invention does not affect the biological activity of the polysaccharide P2 of the Cordyceps militaris culture medium, and the obtained pure polysaccharide P2 has high purity and stable properties, has remarkable effects in anti-oxidation, uric acid reduction, and antibacterial aspects, and is beneficial to human metabolism; The cost is low, and the pure polysaccharide P2 can be further used in the development of health care products, medicines and cosmetics.

Description

technical field [0001] The invention relates to a cordyceps militaris culture medium polysaccharide, a separation and purification method and application thereof. Background technique [0002] Polysaccharide, also known as polysaccharide, is a linear or branched chain polymer connected by aldose or ketose through glycosidic bonds. It is a polar complex macromolecule with a degree of polymerization greater than 10, and its molecular weight is generally tens of thousands. The above is one of the four basic substances that constitute life activities. In addition to existing in a free state, polysaccharides in organisms also combine with proteins or fats to form proteoglycans and lipopolysaccharides. [0003] Cordyceps militaris (C.militaris), also known as Cordyceps militaris, belongs to the subphylum Ascomycota, Sclerotinia, and Spheroides in taxonomy. It belongs to the genus Ergotaceae with Cordyceps militaris. It is mainly distributed in Northeast my country, North China, No...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/00A61K31/715A61P39/06A61P31/04A61P19/06
CPCA61K31/715A61P19/06A61P31/04A61P39/06C08B37/0003C08B37/006
Inventor 黄儒强张竞雯王静辉高林林王倩
Owner SOUTH CHINA NORMAL UNIVERSITY
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