摘要
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。
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.
关键词
芳香黄杆菌
弹性蛋白酶
亲和层析
提纯
生化药物
Flavobacterium odoratumelastase
Purification
Affinity chromatography
Property
