鲎血胶原蛋白酶解条件及其体外活性的研究

Study on enzymatic hydrolysis conditions of collagen from blood in Limulus and its vitro activities

为了考察鲎血胶原蛋白(HXJ)酶解条件及其体外活性,以酶解后的肽得率为考察指标,在单因素的基础上,采用正交设计试验对酶解p H、酶解温度和底物浓度进行筛选,寻找HXJ的最佳酶解条件;对酶解后的HXJ进行体外α-葡萄糖苷酶和乙酰胆碱酶抑制活性测试,同时对其总抗氧化力和对DPPH自由基、羟自由基、超氧阴离子清除率进行测试。结果表明,HXJ的最佳酶解条件为:p H为7.0、温度为70℃、底物质量浓度为0.38 mg/m L、胰酶质量浓度为2.20 mg/m L、酶解时间为5 h,此时肽得率为36.50%;对α-葡萄糖苷酶和乙酰胆碱酯酶的最大抑制率分别为47.9%和97.5%;HXJ具有一定总抗氧化能力,对DPPH自由基、羟自由基和超氧阴离子的最大清除率分别为29.3%、97.3%和25.9%。HXJ能有效抑制α-葡萄糖苷酶和乙酰胆碱酯酶,具有较强的抗氧化活性。

To explore the enzymatic hydrolysis conditions of collagen from blood in Limulus（HXJ） and its vitro activities,the optimal hydrolysis conditions for HXJ are screened through selecting enzymatic hydrolysis p H value,enzymatic hydrolysis temperature and substrate concentration by means of the orthogonal test based on single factor test with the peptide yield after enzymatic hydrolysis as evaluating index.Subsequently,the inhibitive activities of hydrolyzed HXJ are determined in vitro against α-glycosidases and acetylcholinesterase,and the hydrolyzed HXJ＇s total antioxidant capacity and scavenging rates on DPPH free radicals,hydroxyl radicals and superoxide anion are studied too.The best enzymatic hydrolysis conditions for HXJ are as follows：p H is 7.0,enzymolysis temperature is 70℃,substrate concentration is 0.38 mg/mL,pancreatin concentration is 2.20 mg/mL and enzymolysis lasts for 5 h.Under the best conditions,the peptide yield can reach 36.50%,HXJ＇s inhibitive rates on a-glucosidase and acetylcholinesterase are47.9% and 97.5%,respectively,HXJ has certain total antioxidant capacity,and its scavenging rate on DPPH free radicals,hydroxyl radicals and superoxide anion are 29.3%,97.3% and 25.9% respectively in the experimental range.To sum up,HXJ can inhibit effectively a-glucosidase and acetylcholinesterase and has strong antioxidant activities.