基于抗凝血活性的不同基原水蛭酶解工艺对比研究  

Comparative Study on Enzymatic Hydrolysis Processes of Different Radicals Based on Anticoagulant Activity

作　　者：张学涛 庄滢潭 李震[1] 吴家劲 邹婧 高鹏[1] 代龙 ZHANG Xue-tao;ZHUANG Ying-tan;LI Zhen;WU Jia-jin;ZOU Jing;GAO Peng;DAI Long(College of Pharmacy,Shandong University of Traditional Chinese Medicine,Shandong Jinan 250355,China;College of Pharmacy,Binzhou Medical University,Shandong Yantai 264003,China)

机构地区：[1]山东中医药大学药学院,山东济南250355 [2]滨州医学院药学院,山东烟台264003

出　　处：《当代化工》2020年第3期549-554,共6页Contemporary Chemical Industry

基　　金：山东省重点研发计划项目(重大科技创新工程),项目号:2019JZZY020906;山东中医药大学大学生创新创业训练计划项目,项目号:S201910441027.

摘　　要：以抗凝血活性作为评价指标,优选出抗凝活性最佳的水蛭品种和最佳的酶解工艺。采用了单因素试验法确定三种基原水蛭的酶解工艺,筛选出具有最佳抗凝血活性的品种,对活性最强的日本医蛭进行响应面优化设计,确定蛋白酶用量、酶解温度、酶解pH值三个影响因素的最优水平。结果三种水蛭抗凝血活性由高到低顺序为:水蛭>柳叶蚂蟥>蚂蟥;酶解因素影响的顺序为:酶用量>酶解温度>酶解pH值;优选的最佳酶解工艺为:最佳蛋白酶是胰蛋白酶,蛋白酶最佳用量为7657.28 u/g,最适宜温度为53.20℃,最佳酶解pH值为8.30,以此最优条件对日本医蛭进行酶解,得到的产物抗凝血活性达到558u/g。模型可靠性较高,通过对比试验优选了抗凝血活性最强的水蛭品种和最佳的酶解工艺,可以提高水蛭的抗凝血活性。Using anticoagulant activity as an evaluation index,optimum leeches with the best anticoagulant activity and the best enzymatic hydrolysis process were screened out.Single factor test was used to determine the enzymatic hydrolysis process and screen out the varieties with the best anticoagulant activity,optimized design of response surface for the most active Japanese medicine was carried out,the optimal protease dosage,temperature and pH were determined.The order of anticoagulant activity of the three leeches was as follows:Hirudonipponica Whitman>Whitmaniaacranulata Whitman>Whitmaniapigra Whitman,the order of influence of enzymatic hydrolysis factors was as follows:enzyme dosage>enzymatic hydrolysis temperature>enzymatic hydrolysis pH;The optimal enzymatic process was as follows:using trypsin digestion,enzyme dosage 7757.28 u/g,enzymatic hydrolysis temperature 53.20℃,enzymatic pH 8.30.The anticoagulant activity of the Japanese medical entrapment product obtained under this optimal condition reached 558 u/g.The model established in this study has high reliability,through comparison experiments,the species of leech with the best anticoagulant activity and the best enzymatic hydrolysis technology were selected.The research method is scientific,the results are accurate.

关 键 词：水蛭 酶解 抗凝血活性 响应面分析 工艺优选 

分 类 号：R285[医药卫生—中药学]