【摘要】 对土鳖虫水浸醇沉提取物2号样品进行了离子交换柱层析,得组分Ⅰ、Ⅱ、Ⅲ。实验结果表明,组分Ⅲ的溶栓活性明显高于组分Ⅰ和组分Ⅱ,其蛋白质含量为88.9%,分子量约为38,018,效价为313 UK/mg,比活力为352 UK/mg蛋白。将组分Ⅲ再进行凝胶过滤柱层析,得组分Ⅳ、Ⅴ、Ⅵ。组分Ⅵ的生物活性高于组分Ⅳ和组分Ⅴ,蛋白质含量为89.3%,电泳呈现两条带,分子量约为34,623和39,811,效价为77 UK/mg,比活力为86 UK/mg蛋白。将组分Ⅵ进行反相高效液相色谱柱层析,收集保留时间为17 m in的洗脱峰,得到土鳖虫纯蛋白质,呈白色絮状,极易溶于水。再一次用反相高效液相色谱检查其纯度,保留时间为17.073 m in,无杂质峰。纤溶活性实验结果表明,组分Ⅵ既有直接降解纤维蛋白的作用,同时也有纤溶酶原激活剂样作用。更多还原

【Abstract】 The sample has been gotten from Eupolyphaga sinensis Walker by means of water extracting and(alcohol) sinking,which has been purified by ion exchange column,and then,three fractions(Ⅰ,Ⅱ and Ⅲ) have been obtained.The experimental results demonstrate that the biological activity of the fraction Ⅲ is(higher) than that of the fractionⅠorⅡ,in which the content of protein is 88.9%,molecular weight(MW) is about 38.018,the effective value is 313 UK/mg and the comparative activity is 352 UK/mg.The fractions Ⅳ,Ⅴ and Ⅵ have been gotten,when the fraction Ⅲ has been filtrated through gel filtration column.And the experimental results show that the biological activity of the fraction Ⅵ is higher than that of the fraction Ⅳ or Ⅴ,in which the content of protein is 89.3%,it has been found that the fraction Ⅵ gave two strips on the(sodium)dodecyl-sulphonate-polyacrylamide gel electrophoresis(SDS-PAGE) film,their MW data are about 34.623 and 39.811,respectively.The effective value and comparative activity of the fraction Ⅵ are(77 UK/mg) and 86 UK/mg,respectively,which has been purified by the reversed phase high performance(liquid) chromatography(HPLC),the peak,whose retention time is about 17 min,has been collected as the pure protein,which is white,catkin like and easily resolved in water.The purity of the protein has been(secondly) inspected by the reversed phase HPLC,the experimental results show that the retention time of the protein is 17.073 min and the impurity peak is not found.The experimental results of activity indicate that the fraction Ⅵ has the ability to degrade fibrin directly and to active the plas minogen.更多还原