黑蚂蚁纤溶活性蛋白的分离纯化及其性质研究  

Purification and Biochemical Characterization of a Fibrinolytic Protein from Black Ants

作　　者：冼永权 谭竹钧[1] 韩雅莉[1] 

机构地区：[1]广东工业大学轻工化工学院,广州510006

出　　处：《基因组学与应用生物学》2015年第2期284-289,共6页Genomics and Applied Biology

基　　金：国家自然科学基金项目(No.30772739);海口市重点科技计划项目(2011-0044)共同资助

摘　　要：通过硫酸铵分段盐析、DEAE-52离子交换和Sephacryl S-100分子筛层析从黑蚂蚁（Polyrhachis vicina Roger）中纯化到一种纤溶活性蛋白,再通过SDS-PAGE凝胶电泳测定其分子质量,Braford法测定蛋白浓度,蒽酮硫酸法测定其含糖量,纤维蛋白平板法测定其酶活性。并研究温度、pH改变及不同金属离子对其活性的影响。结果显示,分离得到的活性蛋白具有纤溶活性,其分子质量约为25 kD,蛋白浓度为2.8041 mg/mL,糖含量为8.171 4%,酶活力为67.455 2 U/g。研究表明,该活性蛋白最适温度为45℃,最适pH为3.0,pH在4.0-8.0基本稳定,Na^＋、K^＋对其酶活性影响较小,Mg^2＋对该酶纤溶活性有强烈的激活作用,而Zn^2＋、Ba^2＋、Mn^2＋、Ca^2＋等对此酶活力有没明显的抑制作用。A fibrinolytic protein was isolated and purified by ammonium sulfate fractionation, DEAE-52ion-exchange and Sephacryl S-100 HR gel-filtration chromatography from the black ants, and then determined molecular weight by SDS-PAGE gel-electrophoresis analyze, determined the protein concentration by Braford,measured the sugar content by anthrone sulfuric acid method, determined its fibfinolytic activity by fibrin plate. To study the effects of temperature, pH changes and different metal ions on the fibrinolytic protein. The results showed that the active protein showed an molecular weight of about 25 kD, the protein concentration was 2.804 1 mg/mL, the sugar content was 8.171 4%, the enzyme activity was 67.455 2 U/g. Studies have shown that the fibrinolytic protein from black ants has fibrinolytic activity, the optimal temperature and pH of the fibrinolytic enzyme were 45℃ and3.0. This enzyme was stable at pH from 4.0 to 8.0, Na^＋, K^＋had little effect on its activity, Mg^2＋had activation apparently, but was significantly inhibited by Zn^2＋, Ba^2＋, Mn^2＋, Ca^2＋.

关 键 词：黑蚂蚁 纤溶活性蛋白 分离纯化 纤溶活性 

分 类 号：R284.2[医药卫生—中药学]