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Method for degrading chitosan through microwave-enzyme method coupling

A technology of chitosan and chitosan solution, which is applied in the field of chitosan preparation, can solve the problems of high cost, poor product uniformity, and long degradation reaction cycle, and achieve the effect of low cost, low cost, and easy control

Active Publication Date: 2014-10-15
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the disadvantages of long enzymatic degradation reaction cycle, high cost and poor product uniformity during physical degradation when chitosan is degraded in the prior art, and provide a combination of physical and enzymatic degradation of chitosan. method to increase the degradation efficiency

Method used

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  • Method for degrading chitosan through microwave-enzyme method coupling
  • Method for degrading chitosan through microwave-enzyme method coupling
  • Method for degrading chitosan through microwave-enzyme method coupling

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Prepare buffer solution 100ml of pH=5 with acetic acid and sodium acetate, add 1g polymer chitosan (relative molecular mass (hereinafter abbreviated as Mw) 1.8 * 10 6 Da), its number-average molecular weight is 5.0×105Da-5.0×106Da) through high performance liquid phase measurement, fully stirred to make chitosan solution; then add 0.03g cellulase to chitosan solution, after stirring evenly, shell Pour the polysaccharide solution into the reaction bottle, put the reaction bottle into the microwave reactor, and pass nitrogen gas into the reaction bottle, control the temperature at 55°C, and microwave power 600W to carry out the degradation reaction. After 10 minutes, the microwave is heated to 100°C for Enzyme inactivation, the inactivation reaction time is 10-15min; use 1mol / L sodium hydroxide solution to adjust the pH value of the reaction solution to 7-8, then filter to remove the precipitate, and use the obtained filtrate to remove salt ions by dialysis Low molecular ...

Embodiment 2

[0019] Prepare buffer solution 100ml of pH=5 with acetic acid and sodium acetate, add 1g polymer chitosan (Mw=1.8×10 6 Da, number-average molecular weight 5.0×105Da-5.0×106Da), fully stirred to make chitosan solution; then add 0.03g cellulase in the chitosan solution, pour the solution into the reaction bottle after stirring evenly, and then Put the reaction bottle into a microwave reactor, and pass nitrogen gas into the reaction bottle, control the temperature at 55°C, and microwave power 600W to carry out the degradation reaction. After 60 minutes, the microwave temperature is raised to 100°C to inactivate the enzyme. The inactivation reaction time is 10- 15min; with 1mol / L sodium hydroxide solution, the pH value of the solution after the reaction is adjusted to 7-8, and then the precipitate is removed by filtration, and the obtained filtrate is lyophilized to obtain low molecular weight chitosan after removing salt ions by dialysis, and the yield is It is 82%; Measured by h...

Embodiment 3

[0021] The method step of degrading chitosan in the present embodiment is identical with embodiment 1 and embodiment 2, and the number average molecular weight of high molecular weight chitosan is 5.0 * 10 5 Da-5.0×10 6 Da, other relevant experimental data are shown in Table 1:

[0022]

[0023]

[0024] Table 1

[0025] Wherein the product low-molecular-weight chitosan in example 10, surveys its degree of polymerization with high performance liquid chromatography, mainly contains trisaccharide to hexasaccharide, specifically as image 3 Shown; Know that compared with example by contrast example, the productive rate of enzymolytic chitosan under normal temperature is obviously lower than the productive rate of enzymatic chitosan under microwave heating condition, so the microwave-enzymatic method coupling degradation shell of the present invention design The glycan method has a high degradation efficiency.

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Abstract

The invention belongs to the technical field of chitosan preparation and relates to a method for degrading chitosan through microwave-enzyme method coupling. The method comprises the steps as follows: firstly, dissolving the high-molecular-weight chitosan in an acetic acid-sodium acetate buffer solution with the pH value in a range of 4-6, and fully mixing the chitosan and the buffer solution to prepare a chitosan solution; then adding an enzyme with the mass accounting for 0.5-3% of the mass of the chitosan into the prepared chitosan solution, pouring the chitosan solution into a reaction bottle after even mixing, then placing the reaction bottle into a microwave reactor, feeding a nitrogen gas into the reaction bottle, and controlling the temperature to be below 65 DEG C and the microwave power in a range of 100-1000 W to have a degradation reaction for 10-180 min; after the degradation reaction is finished, heating a microwave to 100 DEG C to inactivate the enzyme for 10-15 min; finally, adjusting the pH value of the reacted solution to be in a range of 7-8 with an alkaline solution, removing sediment through filtration, removing saline ions from an obtained filtrate with a dialysis method, and then freezing and drying the filtrate to prepare the low-molecular-weight chitosan. The method is simple to operate, easy to control, mild in condition and environment-friendly.

Description

Technical field: [0001] The invention belongs to the technical field of chitosan preparation, and specifically relates to a method for degrading chitosan through microwave-enzymatic coupling. A buffer solution with a pH value of 4-6 is selected as a solvent to degrade chitosan through microwave-enzymatic coupling. Preparation of low molecular weight chitosan. Background technique: [0002] Chitosan, also known as chitosamine, deacetylated chitin, and chitin sugar, is obtained by deacetylation of chitin, which exists widely in nature, and is the only natural alkaline polysaccharide found so far. Chitosan has a relatively large molecular weight and can be dissolved in acidic solutions, but it is insoluble in water and general organic solvents, which limits its application to a large extent; and the low molecular weight chitosan obtained after degradation can not only dissolve in water , and has physiological activities such as lowering cholesterol, lowering blood fat, antibac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/08
Inventor 刘松周苗苗李克成李鹏程邢荣娥于华华秦玉坤
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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