东亚钳蝎毒透明质酸酶的纯化和部分性质的研究

用CM-SephadexC50,CM-SephadexC25和SephadexG-75凝胶过滤,从东亚钳蝎毒中提纯蝎毒透明质酸酶,应用低pH系统不连续聚丙烯酰胺凝胶圆盘电泳,SDS-不连续聚丙烯酰胺凝胶垂直板电泳鉴定均为单一条带,活力提高34倍,产率为12％,纯品无出血活性,无神经毒性。用凝胶过滤法和SDS电泳法测得分子量为54000,PAS染色证实为糖蛋白。 纯化的透明质酸酶的最适pH为4.5～6.5,最适温度为37℃,该酶对热的稳定性比蛇毒透明质酸酶高一些,但在碱性环境中也易失活。0.15MNaCl对酶活性有明显稳定作用,Fe~(2+)、Fe~(3+)及肝素对酶活性有明显的抑制作用,Cu~(2+)对酶活力也有一定影响。

ISOLATION, PURIFICATION AND SOME PROPERTIES OF HYALURONIDASE FROM SCORPION VENOM OF Buthus martensii Karsch

A hyaluronidase was isolated and purified from scorpion venom o?Buthus martensii Karsch by ion-exchange chromatography on CM-Sephadex C-50 and CM-Sephadex C-25, gel filtration on Sephadex G-75. The homogenity of the purified enzyme was confirmed by polyacrylamide gel disc-electrophoresis at pH 4.3 and SDS-polyacrylmide gel slab electrophoresis, At least a 34-fold purified preparation was obtained. The hyaluronidase was a glycoprotein ( positive PAS staining ) with a molecular weight of 54,000 which was determined by gel filtration on Sephadex G-75 and SDS-discontinuous polyacrylamide gel slab electrophoresis. None of the hemorrhagic or neurotoxic activity was found. The hyaluronidase had an optimum pH of 4.5-6.5 and an optimum temperature of 37℃. After purification, hyaluronidase became very labile. It was heat sensitive, stable in acid and lost its activity in alkali. The enzyme was inhibited by Fe++, Fe+++, heparin and obviously affected by Cu++.