Method for preparing chitooligosaccharide with snailase and application thereof
A technology of snail enzyme and chitosan oligosaccharide, which is applied in the direction of medical preparations, applications, and pharmaceutical formulas containing active ingredients, and can solve the problems of limited source of specific enzymes, many steps in chemical degradation methods, and low efficiency of single enzyme degradation, etc. problems, achieve good industrial promotion practicability and value, improve idiopathic pulmonary fibrosis, and improve the effect of lung fibroblast activation
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[0036] Example 1. A method for preparing chito-oligosaccharides with snail enzyme
[0037] S1: Take the chitosan raw material and dry it to a constant weight, dissolve it in a 0.2mol / L acetic acid-sodium acetate buffer with a pH of 4.5, stir to dissolve, and prepare a chitosan solution with a mass concentration of 1.0%;
[0038] S2 Add the snail enzyme solution with a mass concentration of 1.0% to the chitosan solution obtained in step S1, the snail enzyme and the chitosan substrate mass ratio is 1:10, after enzymatic hydrolysis in a 40°C water bath for 2.5 hours, Inactivate in a water bath at 100°C for 10 minutes, cool to room temperature, continue stirring, add NaOH solution with a mass concentration of 5%, adjust the pH to 4.5, filter, and collect the filtrate;
[0039] S3 Put the filtrate obtained in step S2 into the inner layer equipped with a 3000Da regenerated cellulose dialysis bag, and the outer layer is covered with a dialysis device with a 1000Da regenerated cellulose dial...
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[0041] Example 2. A method for preparing chito-oligosaccharides with snailase
[0042] S1: Take the chitosan raw material and dry it to a constant weight, dissolve it in a 0.2mol / L acetic acid-sodium acetate buffer with a pH of 4.5, stir to dissolve, and prepare a chitosan solution with a mass concentration of 1.0%;
[0043] S2 Add the snail enzyme solution with a mass concentration of 1.0% to the chitosan solution obtained in step S1, and the mass ratio of the snail enzyme to the chitosan substrate is 1:10. After enzymatic hydrolysis in a 40°C water bath for 2 hours, 100 Inactivate in a water bath at ℃ for 15 minutes, cool to room temperature, continue to stir, add dropwise NaOH solution with a mass concentration of 5%, adjust the pH to 6, filter, and collect the filtrate;
[0044] S3 Put the filtrate obtained in step S2 into the inner layer equipped with a 3000Da regenerated cellulose dialysis bag, and the outer layer is covered with a dialysis device with a 1000Da regenerated cell...
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