影响广地龙粗提液蚓激酶活力因素的研究  

Study on the factors on the activity of lumbrokinase in the crude extracts of lumbricus

作　　者：徐兴[1] 索绪斌[1] 向兰[2] 岳静静[1] 游杨杨[1] 张涵 

机构地区：[1]广东药科大学中药学院,广东广州510006 [2]广东药科大学生命科学与制药学院,广东广州510006 [3]广州市茗川生物科技有限公司,广东广州510006

出　　处：《广东药学院学报》2016年第2期159-163,共5页Academic Journal of Guangdong College of Pharmacy

基　　金：国家自然科学基金项目(81303288);国家科技创新基金项目(国科发计[2013]583号);广东省教育厅创新强校特色创新项目(粤教科函[2015]3号)

摘　　要：目的以广地龙粗提液中的蚓激酶为研究对象,研究影响其酶活力的影响因素,并计算其酶学反应动力学常数。方法将蚓激酶与酪蛋白溶液一起培养,通过检测溶液中反应得到的酪氨酸含量,计算蚓激酶的酶活力;分别研究溶液pH值,不同的金属离子、吐温-80、EDTA的浓度,及乙醇的体积分数等条件对蚓激酶酶活性的影响,用米氏方程拟合蚓激酶降解酪蛋白过程,计算动力学参数。结果蚓激酶的活力受pH的影响明显,当pH7.0时酶活力最高;随着钙离子浓度的增加酶活力降低,Na^+、Mg^(2+)、Ba^(2+)对酶的活力影响不大,K^+、Fe^(2+)对酶的活力有一定的促进作用;乙醇对蚓激酶的活力具有抑制作用,体积分数为60%时蚓激酶完全失活;EDTA会降低蚓激酶的活力;吐温-80能提高蚓激酶的活力;蚓激酶降解酪蛋白的活化能Ea=-19.4 k J/mol。结论研究结果可为蚓激酶的提取分离及制剂研究提供参考。Objective To study the factors on the activity of lumbrokinase in the crude extracts of lumbricus.The enzymatic kinetic constants such as Michaelis constant and degradation activation free energy had been calculated. Methods Lumbrokinase and casein had been cultured together in solution,and the lumbrokinase activity had been calculated according to the rise of the tyrosine content which obtained from the reaction by the alteration of the conditions of lumbrokinase solution,such as pH,surfactant,Tween-80,EDTA and ethanol concentration. Michaelis＇ Menten equation was used to fit the lumbrokinase protein degradation process. Results Lumbrokinase activity significantly influenced by pH,which was highest in pH7. 0. The enzyme activity decreased with the increase of calcium concentration; Na~＋,Mg^（2＋）,Ba^（2＋） had little influence on lumbrokinase activity while K~＋ and Fe^（2＋）might promote the activity of enzyme. Ethanol had inhibitory effect on the activity of lumbrokinase and its activity completely lost in ethanol concentration of 60%. EDTA might decrease the activity of lumbrokinase activity,and Tween-80 could improve the activity of lumbrokinase. The activation energy（ Ea） of lumbrokinase degrade casein was-19.4 k J/mol. Conclusion The results will provide some reference for the separation and purification of lumbrokinase and its preparations.

关 键 词：蚓激酶 米氏方程 动力学参数 活化能 

分 类 号：R284[医药卫生—中药学]