Preparation Of Red Deer Antler Collagen Peptide With Skin Fibroblast-Promoting Activity By Dual-Enzyme Method
Keywords
Cervus Elaphus Antler Collagen Peptide The Proliferation Of Cell
Abstract
Collagen, as the main component of the extracellular matrix, is closely related to cell proliferation and differentiation. In order to explore the appropriate process conditions for preparing red deer antler collagen peptides with the function of promoting skin fibroblast proliferation, this study used red deer antler as raw material, enzymatic hydrolysis using Alcalase and trypsin, and compared the hydrolysis degree, peptide yield and normal human skin. Single-factor experiments and orthogonal experiments were carried out to determine the appropriate process conditions for the target value of the proliferation rate of fibroblasts (NHDF cells). Experimental results show that the suitable preparation conditions for red deer antler collagen peptide are: first use Alcalase for enzymatic hydrolysis at a substrate concentration of 1%, an enzyme-to-base ratio of 2%, pH 9.0, and a temperature of 60¡ãC; then, at a substrate concentration of 1%, Trypsin was used for enzymatic hydrolysis at an enzyme-base ratio of 6%, pH 8.0, and a temperature of 42¡ãC. The final hydrolysis degree of the enzymatic hydrolyzate was 21.7%, the peptide yield was 86%, and the cell proliferation rate was 22.75%. Alcalase-trypsin dual enzymes hydrolyze red deer antler collagen step by step to obtain collagen peptides that promote a high proliferation rate of normal human skin fibroblasts. As the main component of the extracellular matrix, collagen was closely related to the proliferation and differentiation of the cells. In order to explore the suitable preparation conditions of the collagen peptides from red deer (Cervus elaphus) antler with the function of promoting the proliferation of the skin fibroblasts, the antler was used as the raw material and hydrolyzed by alcalase and trypsin. The degree of hydrolysis (DH), peptides yield and the proliferation rate of normal human dermal fibroblasts (NHDF) were used as the target to perform the single factor experiments and orthogonal experiments and determine the appropriate process conditions. The results showed that the optimal preparation conditions were as follows: alcalase was used for enzymolysis under the conditions of substrate concentration of 1%, enzyme substrate ratio of 2%, pH 9.0 and temperature of 60 ¡æ, and then trypsin was used for enzymolysis under the conditions of substrate concentration of 1%, enzyme substrate ratio of 6%, pH 8.0 and temperature of 42 ¡æ. The final hydrolysis degree of the hydrolyzate, the yield of peptides and proliferation The rate of cells were 21.7%, 86% and 22.75%, respectively. Collagen peptide with promoting the proliferation rate of NHDF could be obtained from red deer antler by alcalase-trypsin bi-enzyme hydrolysis.
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Original research was done by Yang Congshan
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