少棘蜈蚣活性蛋白提取及纤溶作用性质研究

Study on Extraction of Active Protein from Scolopendra subspinipes mutilans and its Fibrinolytic Activity

目的研究少棘蜈蚣活性蛋白的纤溶性及其性质。方法通过硫酸铵分段盐析、DEAE纤维素柱和Sephadex G-75柱层析从少棘蜈蚣Scolopendra subspinipes mutilans L. Koch组织分离纯化出纤溶性活性蛋白,纤维蛋白平板法测定其纤溶性,SDS-PAGE测定样品纯度和分子量,温度、pH改变对少棘蜈蚣纤溶性蛋白活性的影响。结果分离纯化到一种相对分子质量约为48.3 kD的纤溶性蛋白,水解纤维蛋白的比活力为42.36 u/mg。结论少棘蜈蚣活性蛋白具有纤溶性,该成分在40℃下基本稳定,最适温度35℃,最适pH8.0。

Objective To study the fibrinolytic activity of the active protein of Scolopendra sulspinipes multicans. Methed A novel of fibrinolytic protein was separated and purified by ammonium suplhate sub -precipitation, DEAE -cellulose and SephadexG - 75 column chromatography from the tissue of the Scolopendra subspinipes rnutilans L. Koch. Result The protein showed an apparent molecular weight of about 48.3kD in sodium dodecyl sulfate - polyacrylamide gel electrophoresis analysis. Its specific activity to hydrolyze fibrin was 42.36 u/mg. The activity of enzyme was inhibited by serine protease inhibitor such as phenylmethanesulfonyl fluoride （PMSF）, but wasn＇t inhibited by ethylenedianfinotetraacetic acid （EDTA）. Corclusion The protein is stable under 40℃, the optimum temperature is 35℃, and the optimum pH is 8.0.