A method for extracting and separating and purifying polysaccharides from the leftovers of Cordyceps militaris culture medium

A technology of separation and purification and culture medium, which is applied in bone diseases, antibacterial drugs, antiviral agents, etc. It can solve the problems of complex operation process, inability to obtain pure polysaccharide components, high cost, etc., achieve good repeatability, and benefit human metabolism , low cost effect

Active Publication Date: 2021-10-26
SOUTH CHINA NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] 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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  • A method for extracting and separating and purifying polysaccharides from the leftovers of Cordyceps militaris culture medium
  • A method for extracting and separating and purifying polysaccharides from the leftovers of Cordyceps militaris culture medium
  • A method for extracting and separating and purifying polysaccharides from the leftovers of Cordyceps militaris culture medium

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Embodiment 1

[0045] A method for extracting and separating and purifying polysaccharides from the leftovers of Cordyceps militaris culture medium, comprising the following steps:

[0046](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;

[0047] (2) Decolorization of the polysaccharide of the Cordyceps militaris medium: the ...

Embodiment 2

[0056] The polysaccharides of the Cordyceps militaris culture medium obtained in Example 1 were 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.

[0057] figure 2 is the UV spectrum of P1, image 3 It is the ultraviolet spectrogram of P2. The results show that P1 has weak absorption peaks at 260nm and 280nm, indicating that P1 contains a small amount of protein and nucleic acid; P2 has no obvious absorption peaks at 260nm and 280nm, indicating that P2 does not contain protein and nucleic acid. Nucleic acid substances.

Embodiment 3

[0059] Carry out polysaccharide molecular weight analysis to the polysaccharide of Cordyceps militaris culture medium that embodiment 1 obtains, concrete experimental method is as follows:

[0060] 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.

[0061] 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),

[0062] TSKG-4000K gel column (300mm×7.8mm) and TSKG-2500K gel column (300mm×7.8mm); Waters2414 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...

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Abstract

The invention discloses a method for extracting and separating and purifying polysaccharides from the leftovers of the Cordyceps militaris culture medium. The method comprises the following steps: crushing the leftovers of the Cordyceps militaris rice culture medium and then adding water to reflux for extraction, concentrating the extract, adding ethanol to statically Place overnight, centrifuge and get the precipitate to be the polysaccharide extract of Cordyceps militaris culture medium; add the polysaccharide extract to H 2 o 2 Decolorize, then add papain for enzymatic hydrolysis, then add Sevage reagent, centrifuge to get the supernatant, which is crude polysaccharide; put it on an ion exchange chromatography column, and use NaCl solution for gradient elution to obtain multiple polysaccharide components. The method of the invention can complete a large-flux polysaccharide extraction operation, and is suitable for large-scale industrial production. The DEAE ion-exchange chromatographic column has good separation effect and good repeatability, and the obtained polysaccharide components have high purity and stable properties.

Description

technical field [0001] The invention relates to a method for extracting, separating and purifying polysaccharides from leftovers of culture medium of Cordyceps militaris. Background technique [0002] 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, Northwest China and other regions . Cordyceps militaris can parasitize on the larvae or pupae of insects such as Lepidoptera, Coleoptera, and Diptera, and can be artificially cultivated in batches using silkworm pupae and rice culture medium. [0003] Due to the limited resources of natural Cordyceps militaris, there are many artificial cultivation methods at present. There are mainly three cultivation methods: (1) collecting wild Cordyceps militaris, carrying out the separation and purification of strains, and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/00A61P19/06A61P39/06A61P31/04
CPCA61P19/06A61P31/04A61P39/06C08B37/0003
Inventor 黄儒强张竞雯王静辉高林林王倩
Owner SOUTH CHINA NORMAL UNIVERSITY
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