A microwave-enzymatic coupled method for degrading chitosan

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

Active Publication Date: 2016-07-06
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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  • A microwave-enzymatic coupled method for degrading chitosan
  • A microwave-enzymatic coupled method for degrading chitosan
  • A microwave-enzymatic coupled method for degrading chitosan

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Prepare 100ml of pH=5 buffer solution with acetic acid and sodium acetate, and add 1g of high molecular chitosan (relative molecular weight (hereinafter abbreviated as Mw) 1.8×10 to the buffer solution 6 Da), the number-average molecular weight is 5.0×105Da-5.0×106Da) by HPLC, and the chitosan solution is prepared by stirring thoroughly; then 0.03g cellulase is added to the chitosan solution, and the chitosan is mixed evenly after stirring. Pour the glycan solution into the reaction flask, then put the reaction flask into the microwave reactor, and pour nitrogen into the reaction flask, control the temperature at 55℃, the microwave power 600W for degradation reaction, and the microwave will be heated to 100℃ after 10 minutes. Enzyme inactivation, the inactivation reaction time is 10-15min; adjust the pH value of the solution after the reaction to 7-8 with 1mol / L sodium hydroxide solution, and then filter to remove the precipitate, after the filtrate is dialyzed to remove s...

Embodiment 2

[0019] Prepare 100ml of pH=5 buffer solution with acetic acid and sodium acetate, add 1g high molecular chitosan (Mw=1.8×10) to the buffer solution 6 Da, number average molecular weight 5.0×105Da-5.0×106Da), stir well to prepare a chitosan solution; then add 0.03g cellulase to the chitosan solution, stir evenly, pour the solution into the reaction flask, and then The reaction flask was put into a microwave reactor, and nitrogen was introduced into the reaction flask. The temperature was controlled at 55°C and the microwave power was 600W for degradation reaction. After 60 minutes, the microwave was heated to 100°C to inactivate the enzyme. The inactivation reaction time was 10- 15min; Adjust the pH value of the reacted solution to 7-8 with 1mol / L sodium hydroxide solution, then filter to remove the precipitate, the filtrate is dialysis to remove salt ions and freeze-dried to obtain low molecular weight chitosan, yield Is 82%; measured by high performance liquid chromatography, t...

Embodiment 3

[0021] The steps of the method for degrading chitosan in this example are the same as those in Example 1 and Example 2. 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] Among them, the low molecular weight chitosan in Example 10 was measured by high performance liquid chromatography for its degree of polymerization. It mainly contained trisaccharides to hexasaccharides. image 3 As shown; the comparison between the control example and the example shows that the yield of enzymatic chitosan at room temperature is significantly lower than the yield of enzymatic chitosan under microwave heating conditions, so the microwave-enzymatic coupling degraded shell designed by the present invention The glycan method has higher degradation efficiency.

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Abstract

The invention belongs to the technical field of chitosan preparation, and relates to a microwave-enzymatic coupled method for degrading chitosan: first, dissolving high molecular weight chitosan in an acetic acid-sodium acetate buffer solution with a pH value of 4-6 , fully stirred to make a chitosan solution; secondly, in the prepared chitosan solution, add the enzyme whose quality is 0.5-3% of the chitosan quality, stir the chitosan solution into the reaction bottle, Then put the reaction bottle into the microwave reactor, and pass nitrogen into the reaction bottle, control the temperature below 65 ° C, and carry out the degradation reaction under the condition of microwave power 100-1000W for 10-180min; after the degradation reaction is completed, microwave the temperature to 100 ℃ to inactivate the enzyme, the inactivation reaction time is 10-15min; finally, adjust the pH value of the reaction solution to 7-8 with an alkaline solution, and then filter to remove the precipitate, and use the obtained filtrate to remove the salt ion by dialysis Low molecular weight chitosan is prepared by freeze-drying; the method is simple to operate, easy to control, mild in condition and friendly to the environment.

Description

Technical field: [0001] The invention belongs to the technical field of chitosan preparation, and specifically relates to a microwave-enzymatic coupling degradation method of chitosan. A buffer solution with a pH value of 4-6 is selected as a solvent, and the chitosan is degraded by microwave-enzymatic coupling. Preparation of low molecular weight chitosan. Background technique: [0002] Chitosan, also known as chitosamine, chitin, and chitin, is obtained from the deacetylation of chitin, which is widely present in nature, and is the only natural alkaline polysaccharide found so far. Chitosan has a large relative molecular weight and can be dissolved in acidic solutions, but is insoluble in water and general organic solvents, which limits its application to a large extent. The low molecular weight chitosan obtained by degradation is not only soluble in water , And has physiological activities such as lowering cholesterol, lowering blood lipid, antibacterial, anti-tumor, etc., an...

Claims

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Application Information

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Patent Type & Authority Patents(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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