Method for removing protein of Brazil mushroom crude polysaccharide

A crude polysaccharide and Agaricus blazei technology is applied in the field of Agaricus blazei crude polysaccharide deproteinization, which can solve the problems of polysaccharide loss, long operation period and high cost, and achieve the effects of high protein removal rate, short operation time and little polysaccharide loss.

Inactive Publication Date: 2006-08-23
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problems of severe loss of polysaccharides, huge consumption of organic solvents, high cost, great environmental pollution and damage to operators and long ope...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Example 1 The cation passes through the column once

[0015] Pack the treated ion exchange resin into the chromatographic column, the specification of the cation resin chromatographic column: 2.6×55cm, and the height of the ion exchanger packing is 45cm;

[0016] Sample loading: cationic column deproteinization, sample volume is 10ml, polysaccharide content 13.00mg / ml in Agaricus blazei crude polysaccharide, protein content 6.23mg / ml during sample loading, crude polysaccharide feed solution pH is natural pH (=6.4 during extraction ).

[0017] Elution: After loading the cation exchange resin, elute with a deionized water peristaltic pump at a flow rate of 1-4mL / min, detect the sugar content in the effluent by the phenol-sulfuric acid method, and elute until no sugar is precipitated, about 400-800ml, and then regenerate ,reuse. The separation results are shown in Table 1.

Embodiment 2

[0018] Example 2 Anions pass through the column once

[0019] Pack the treated ion exchange resin into the chromatographic column, the specification of the anion exchange resin: 2.6×55 cm, and the height of the ion exchange material is 45 cm.

[0020] Sample loading: anion passes through the column to deproteinize, and the loading amount is 10ml. When loading the sample, the polysaccharide content is 13.00mg / ml in the Agaricus blazei crude polysaccharide, and the protein content is 6.23mg / ml. The pH of the crude polysaccharide feed liquid is the natural pH when extracting (= 6.4).

[0021] Elution: After loading the anion exchange resin, elute with a deionized water peristaltic pump at a flow rate of 1-4mL / min, detect the sugar content in the effluent by the phenol-sulfuric acid method, and elute until no sugar is precipitated, about 400-800ml. The separation results are shown in Table 1.

Embodiment 3

[0022] Example 3 Anions pass through the column once

[0023] The anion exchange resin described in Example 2 was eluted with 4% NaOH solution at 3 times the column volume, dynamic for 1 hour, and then eluted with pure water instead of NaOH until neutral.

[0024] Sample loading: anion passes through the column to deproteinize, and the loading amount is 10ml. When loading the sample, the polysaccharide content is 13.00mg / ml in the Agaricus blazei crude polysaccharide, and the protein content is 6.23mg / ml. The pH of the crude polysaccharide feed liquid is the natural pH when extracting (= 6.4).

[0025] Elution: After loading the anion exchange resin, elute with a deionized water peristaltic pump at a flow rate of 1-4mL / min, detect the sugar content in the effluent by the phenol-sulfuric acid method, and elute until no sugar is precipitated, about 400-800ml. The separation results are shown in Table 1.

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PUM

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Abstract

The present invention relates to the method of removing protein from crude Brazil mushroom polysaccharide. The crude Brazil mushroom polysaccharide is column chromatography separated with at least one kind of weak acid cation resin or weak alkali anion resin to separate free protein from polysaccharide. The method of the present invention has short operation time of 3-5 hr, high protein eliminating rate, less polysaccharide loss, no use of organic solvent. less environmental pollution, low cost and reusablility of ion exchange resin.

Description

(1) Technical field [0001] The invention relates to a deproteinization method of Agaricus blazei crude polysaccharide. (2) Background technology [0002] Fungal polysaccharides are a type of biologically active products. Generally, they need to be separated and purified to obtain relatively pure polysaccharides to play a better role. Especially when they are to be developed into high-purity drugs, the impurities contained in crude polysaccharides must be removed. An essential purification step in the deproteinization process. The method currently applied to the deproteinization of polysaccharides mainly includes the addition of a certain proportion of organic solvents, such as trichloroacetic acid, trifluoroacetic acid method, tannic acid, Sevag reagent (a mixture of trichloroacetic acid and n-butanol in a certain proportion), and can also be used Enzyme method, enzyme-Sevag combination method, etc. Among them, the Sevag method is a classic method and is the most widely us...

Claims

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Application Information

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IPC IPC(8): C08B37/00C07K1/14
Inventor 孙培龙杨开何荣军吴学谦
Owner ZHEJIANG UNIV OF TECH
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