梅花鹿鹿角盘胶原肽制备及其活性作用研究

Study on preparation and activities of collagen peptide from Sika antlerbase

目的研究梅花鹿鹿角盘胶原肽的护肤作用并优化酶解法提取胶原肽的工艺。方法以热水法提取的胶原蛋白为原料,胶原蛋白水解度为指标,采用胰蛋白酶酶解梅花鹿鹿角盘胶原蛋白。借助单因素试验、Box-Behnken设计试验来优化制备鹿角盘胶原肽的工艺。在10、20、30、40、50 g/L浓度范围内探究胶原肽的抗氧化能力及还原能力。将40只昆明小鼠随机均分为空白组、对照组及胶原肽溶液低、中、高剂量组(0.1 g/L、0.2 g/L、0.4 g/L),每天涂抹小鼠皮肤2次,6周结束实验并测定SOD活性、Hyp含量、MDA含量水平变化。结果鹿角盘胶原蛋白含量为22.46%。鹿角盘胶原肽最优制备工艺为酶解时间2.2 h、pH值为8.0、温度50℃、加酶量1 200μ/g,此条件下的蛋白水解度为15.97%。浓度达到50 g/L时,鹿角盘胶原肽具有最强的抗氧化及还原能力。与空白组比较,胶原肽低、中、高剂量组均有显著的护肤作用,SOD和Hyp水平显著升高(P<0.01),MDA水平显著降低(P<0.01)。结论试验优化的酶解提取工艺条件稳定可行,鹿角盘胶原肽具有较好的护肤作用。

Objective To study the skincare effect of collagen peptide from Sika antlerbase and to optimize the process of extracting the collagen peptide by enzymatic hydrolysis. Methods The collagen extracted by hot water method as the raw material was digested with trypsin. The degree of collagen hydrolysis was used as an index. The single-factor test and Box-Behnken design test were used to optimize the preparation process of collagen peptide from the antlerbase. The antioxidation and reducing ability of the collagen peptide were investigated in the concentration range of 10,20,30,40 and 50 g/mL. Forty Kunming mice were randomly divided into blank group,control group,low,middle and high dose collagen peptide solution groups(0.1 g/L,0.2 g/L,0.4 g/L).The skin of mice was smeared with the peptide twice a day. At the end of 6 weeks,the activity of SOD,the content of hydroxyproline(Hyp) and the level of MDA were measured. Results The antlerbase collagen comtent is 22.46%. The optimal preparation process of the antlerbase collagen peptide was enzymatic hydrolysis time 2.2 h,pH 8.0,temperature 50 ℃,enzyme amount 1 200 μ/g. Under this condition,the degree of protein hydrolysis is 15.97%.The collagen peptide of the antlerbase had the strongest antioxidation and reducing ability when the concentration reached 50 mg/mL. Compared with the blank group,the low,middle and high dose groups of collagen peptide had significant skincare effect as SOD and Hyp significantly increased(P<0.01) and MDA significantly decreased(P<0.01). Conclusion The optimized enzymatic extraction conditions are stable and feasible,and the antlerbase collagen peptide has a good skincare effect.