Method for extracting and purifying antler active oligopeptide and chondroitin sulfate by using enzymatic hydrolysis method
A chondroitin sulfate and enzymatic hydrolysis method is applied in the field of purifying antler oligopeptide, continuous extraction, and chondroitin sulfate, which can solve the problems of discontinuous extraction, low extraction rate and long time, and improve bone quality and extraction yield. High, immune-boosting effect
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Embodiment 1
[0020] Fresh velvet antler was peeled, weighed 1g of velvet antler after peeling, washed, added 20mL distilled water to make velvet antler aqueous solution, adjusted the pH of the solution to 9.5 with 0.1mol / L NaOH, and heated at 80°C for 25min. Adjust pH 10, add 0.8g alkaline protease at 45℃, enzyme activity ≥ 200000U / g, hydrolyze for 3h, then adjust pH 7 with 0.1mol / L HCL, add 0.8g papain at 65℃ , Enzyme activity 50000U / g, enzymolysis 3h. Inactivated after enzymatic digestion. The deer antler solution after enzymatic hydrolysis and inactivation was centrifuged, and the supernatant and precipitate were separated. Add 10 mL of distilled water to the precipitate to make an aqueous solution, add 0.4 g of alkaline protease, adjust the pH to 10, and enzymatically hydrolyze for 40 min. After inactivation by enzymatic hydrolysis, the supernatant was collected by centrifugation. The supernatants obtained after the second centrifugation were combined, and vacuum rotary evaporation ...
Embodiment 2
[0022] Fresh velvet antler was peeled, weighed 1g of velvet antler after peeling, washed, added 40mL of distilled water to make velvet antler aqueous solution, adjusted the pH of the solution to 10 with 0.1mol / L NaOH, and heated at 75°C for 40min. Adjust pH 9, add 1.2g alkaline protease at 35℃, enzyme activity ≥ 200000U / g, hydrolyze for 2h, then adjust pH 6.5 with 0.1mol / L HCL, add 1.2g papain at 55℃ , Enzyme activity 50000U / g, enzymolysis 2h. Inactivated after enzymatic digestion. The deer antler solution after enzymatic hydrolysis and inactivation was centrifuged, and the supernatant and precipitate were separated. Add 10 mL of distilled water to the precipitate to make an aqueous solution, add 0.4 g of alkaline protease, adjust the pH to 9, and enzymatically hydrolyze for 30 min. After inactivation by enzymatic hydrolysis, the supernatant was collected by centrifugation. The supernatants obtained after the second centrifugation were combined, and vacuum rotary evaporatio...
Embodiment 3
[0024] Peel fresh antler, weigh 10g of peeled antler, wash, add 300mL distilled water to make antler aqueous solution, adjust the pH of the solution to 10 with 1mol / L NaOH, and heat at 85°C for 30min. Adjust the pH to 9.5, add 9g of alkaline protease at a temperature of 45°C, the enzyme activity is ≥200,000U / g, and hydrolyze for 2 hours, then adjust the pH to 7 with 1mol / L HCL, add 9g of papain at a temperature of 60°C, and the enzyme activity 50000U / g, enzymatic hydrolysis for 2h. Inactivated after enzymatic digestion. The deer antler solution after enzymatic hydrolysis and inactivation was centrifuged, and the supernatant and precipitate were separated. Add 100mL of distilled water to the precipitate to make an aqueous solution, add 4g of alkaline protease, adjust the pH to 10, and enzymatically hydrolyze for 30min. After inactivation by enzymatic hydrolysis, the supernatant was collected by centrifugation. The supernatants obtained after the second centrifugation were co...
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