芳香黄杆菌弹性蛋白酶的纯化及性质研究

报道了弹性蛋白亲和层析分离纯化芳香黄杆菌产胞外弹性蛋白酶。经一步柱层析可获聚丙烯酰胺凝胶电泳均一的酶制品，收率可达５０％，并制备了酶的结晶。该酶在ＳＤＳ－ＰＡＧＥ上测得分子量为２１３８０，ＩＥＦ－ＰＡＧＥ测得等电点８．９，最适作用温度为５０℃，最适作用ｐＨ为７．４，在４０℃以下热稳定性良好，ｐＨ４．５－９．５范围内稳定，重金属离子Ｆｅ３＋、Ｚｎ２＋、Ｃｒ３＋、Ｃｏ２＋、Ｈｇ２＋、Ｎｉ２＋、Ａｇ＋、Ｃｕ２＋等严重抑制酶活性，黄杆菌弹性蛋白酶除了特异水解弹性蛋白外，对干酪素、血纤维蛋白、白蛋白、明胶、血红蛋白等多种蛋白质均能水解。

Purification and Properties of the Elastase from Flavobacterium odoratum

The purification of the elastase from Flavobacterium odoratum17-87 was achieved by the combination of (NH4)2SO4 fractionation and affinity chromatography on insoluble substrate.PAGE-homogenous of preparation elastase was obtained and crystalized.The molecular weight estimated by SDS-PAGE was 21380,and the isoelectric point determined by IEF-PAGE was 8.9. The elastiolytic activity was optimal at PH7. 4 and 50℃. The enzyme was stable over the PH range of 4. 5-9.5 at temperature below 40℃, but was completely inhibited by Fe3+、Cu2+、Zn2+、Cr3+、Co2+、Ni2+、Hg2+、Ag+. The elastase from F.odoratum 17-87 had a broad action spectrum, it could not only degrade insoluble elastin specifically, but also hydrolyze casein, fibrin, bovin serum albumin, gelatin and hemoglobin.