Method for extracting collagen type II by taking agkistrodon as raw material
A technology of collagen and viper, which is applied in the field of extracting type II collagen, can solve the problems of complex operation, difficult chemical methods, and differences in results, and achieve the effect of simple operation and high purity
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[0029] Methods as below:
[0030] 1. Crush Viper (purchased from Huangshan Snake Science Research Institute) into a powder with a particle size of 2 mm;
[0031] 2. Suspend 150 grams of Viper powder in 2000 mL of 4 M guanidine hydrochloride-50 mM Tris-HCl mixed solution (pH7.5), control the temperature at 4 ° C, stir for 24 h; 4 ° C, 14000 × Centrifuge at g for 1 h, remove the supernatant, wash the precipitate with 50 mM Tris-HCl solution to remove guanidine hydrochloride, centrifuge at 14,000×g for 1 h at 4°C, and remove the supernatant;
[0032] 3. The precipitate obtained in the previous step was resuspended with 20 times the volume of the precipitate in 0.5 M acetic acid, and the pH was adjusted to 2.8 with 70% formic acid, and 5 g of pepsin (purchased from Wuxi Zhongtian Food Additives Co., Ltd., unit Enzyme activity 15000U / g), stirred gently at 4°C for 48 h;
[0033] 4. Centrifuge to remove the precipitate: 4°C, 14000×g, centrifuge for 1 h, discard the precipitate (if...
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