Method for removing protein in polysaccharide extraction liquid by dialdehyde cellulose

A dialdehyde cellulose and protein technology, applied in the innovative field of deproteinization method, can solve the problems of affecting efficacy, destroying the structure of proteoglycan complex, and weakening the resolubility of polysaccharides, so as to avoid waste and residue, improve deproteinization rate, The effect of low degradability

Active Publication Date: 2017-01-25
江苏灵源沂岸科技股份有限公司
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AI Technical Summary

Problems solved by technology

The reagents used for Sevage deproteinization are chloroform and n-butanol, and chloroform is a toxic substance, which is likely to cause a decrease in the activity of polysaccharides and solvent residues. This method is time-consuming to operate, consumes a lot of organic solvents, has high costs, and has great environmental pollution and harm to operators. more damage
The precipitation phenomenon in the deproteinization operation of the trichloroacetic acid method is more obvious than that of the Sevage method, but more polysaccharides are easily lost during deproteinization, and the resolubility of polysaccharides after deproteinization is significantly weakened, which has a greater impact on the structure of polysaccharides. Therefore, the deproteinization effect of this method poor
Although the effect of hydrochloric acid deproteinization is better, if the experimental conditions are not strictly controlled, the loss rate of polysaccharides is high, and there are disadvantages of polysaccharide hydrolysis
The enzymatic method is to use protease to hydrolyze the protein in the polysaccharide extract under the catalysis of protease, thereby removing the protein. The specificity of the protease method makes the effect unstable, and it may also destroy the structure of the proteoglycan complex and affect its efficacy. , and proteases are expensive, resulting in a higher cost for this method

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  • Method for removing protein in polysaccharide extraction liquid by dialdehyde cellulose
  • Method for removing protein in polysaccharide extraction liquid by dialdehyde cellulose
  • Method for removing protein in polysaccharide extraction liquid by dialdehyde cellulose

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preparation example Construction

[0026] (1) Preparation of dialdehyde cellulose: Weigh coniferous wood or broad-leaved wood cellulose powder, add sodium periodate and deionized water, adjust pH=3, and react in a water bath at 30°C for a certain period of time in the dark, and the degree of oxidation is different The dialdehyde cellulose is dried and ground into powder for later use.

[0027] (2) According to the procedures of hot water extraction, concentration, alcohol precipitation, filtration and drying, the crude polysaccharides of yam, ganoderma lucidum and astragalus are obtained;

[0028] (3) All three polysaccharides were deproteinized by Sevage method, and compared with dialdehyde cellulose method. Sevage method: Add 10 mL of Sevage reagent (chloroform:n-butanol=4:1) to the crude polysaccharide solution at a volume ratio of 5:1, shake for 20 min, and centrifuge to remove denatured protein at the junction of the water layer and the reagent layer. This step was repeated 7 times.

Embodiment 1

[0031] 1) Move the yam crude polysaccharide into a reaction tank, add purified water to prepare a polysaccharide solution with a mass fraction of 1%, and prepare dialdehyde cellulose prepared from coniferous wood for use;

[0032] 2) Add 20g of dialdehyde cellulose with an oxidation degree of 78% to 1L of yam polysaccharide solution, control the range of solid-liquid ratio to 4:200 (g / mL), adjust the pH value to 8, mix well, and react at a constant temperature of 40°C for 4 hours, after the reaction was completed, the supernatant was obtained by cooling and centrifugation; the above operation was repeated 3 times to obtain the polysaccharide solution after deproteinization. Taking the deproteinization rate as an index, compared with the Sevage method, the deproteinization rate reached 88.35% after the dialdehyde cellulose deproteinization for 4 times; and the deproteinization rate reached 74.38% after the Sevage method 4 times.

Embodiment 2

[0034] 1) Move the crude polysaccharide of yam into a reaction tank, add purified water to prepare a polysaccharide solution with a mass fraction of 2%, and prepare dialdehyde cellulose prepared from hardwood as a raw material for use;

[0035] 2) Add 5g of dialdehyde cellulose with an oxidation degree of 23% to 1L of yam polysaccharide solution, control the range of solid-liquid ratio to 1:200 (g / mL), adjust the pH value to 7, mix well, and react at a constant temperature of 20°C for 8 Hours, after the reaction is completed, the polysaccharide solution after deproteinization is obtained by cooling and centrifuging. Taking the deproteinization rate as an index, compared with the Sevage method, the deproteinization rate reached 49.87% after one deproteinization by the dialdehyde cellulose method; and 47.93% after one deproteinization by the Sevage method.

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Abstract

The invention provides a novel method for removing protein in the polysaccharide separation and purification process. The method comprises the following steps of firstly preparing a polysaccharide solution at a certain weight percentage; then, adding dialdehyde cellulose at different oxidization degrees in the polysaccharide solution; controlling the material-liquid ratio (g / mL) and the reaction temperature; regulating the pH value; performing reaction for a certain time at the specified temperature; after the reaction is completed, performing cooling and centrifugal dispersion; taking supernate; repeating the operation for many times to obtain the protein removal polysaccharide solution. The method provides a new idea for the protein removal method; in addition, the process is simple; the operation is easy; meanwhile, the raw material cellulose has the advantages of rich resources, environment-friendly performance and the like; the method belongs to a green and environment-friendly protein removal method and is favorable for the scaled separation and purification of crude polysaccharide.

Description

technical field [0001] The invention relates to a new method for removing protein in the process of separating and purifying polysaccharides. It particularly relates to a deproteinization method that does not use traditional chemical or enzymatic methods, but uses dialdehyde cellulose to react with protein to generate Schiff base for deproteinization, which belongs to the innovative field of deproteinization methods. Background technique [0002] Polysaccharides widely exist in various animal, plant and microbial tissues, and have become a research hotspot in natural medicine due to their diverse biological activities. However, impurity proteins are often contained in polysaccharide solutions, so how to effectively remove proteins has become a very meaningful research content. Wang Shan and Huang Shengyang reviewed the methods of deproteinization in plant polysaccharide extracts in "Research Progress on Deproteinization Methods of Plant Polysaccharide Extracts", and compreh...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/00
Inventor 张黎明许萌郑意强吴嘉乘王金艳葛欢欢申蕊王劲峰郭然戴玉杰
Owner 江苏灵源沂岸科技股份有限公司
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