芽孢杆菌L_4所产角蛋白酶的纯化工艺、理化性质及其应用研究  

PURIFICATION, CHARACTERIZATION AND APPLICATION OF KERATINASE FROM Bacillus sp. L_4 STAIN

作　　者：张奇[1] 孙丹[2] 杨文博[3] 刘茜[3] 白芳[1] 白钢[1,3] 

机构地区：[1]南开大学药学院,天津300071 [2]南开大学学报编辑部,天津300071 [3]南开大学生命科学学院,天津300071

出　　处：《离子交换与吸附》2009年第5期448-455,共8页Ion Exchange and Adsorption

基　　金：天津市应用基础及前沿技术研究计划(08JCYBJC04400)

摘　　要：根据芽孢杆菌L4菌株发酵产生的角蛋白酶的分子量和等电点等特征,分别采用硫酸铵盐析沉淀、SephadexG-75凝胶过滤、DEAE-cellulose52离子交换和Sepharose4B凝胶过滤等色谱层析对其进行分离纯化,经SDS-PAGE检测呈现单一条带,分子量为149kD;变性电泳结果显示,该酶由4个分子量约为37.5kD的亚基组成;采用等电聚焦电泳测得其等电点为7.76,并通过纸层析法初步确认该酶为高甘露糖型糖蛋白;最后通过羊毛水解实验确认该酶具有水解羊毛角蛋白的能力,在羊毛仿羊绒等领域具有一定的应用价值。Keratinase from Bacillus sp. L4 strain was purified by salting out, Sephadex G-75 gel filtration, DEAE-cellulose 52 ion-exchange and Sepharose 4B gel filtration chromatography, which showed one single band using SDS-PAGE analysis, and the molecular weight was 149kD. Denaturing gel electrophoresis results showed that the enzyme consists of four subunits molecular weight of about 37.5kD. Through IEF appropriately measured isoelectric point of 7.76, and using paper chromatography the enzyme was confirmed the initial high-mannose type glycoprotein. Finally, the experimental of wool hydrolysis confirmed that the enzyme owned the hydrolytic ability of wool keratin, which could be used in the fields of wool imitation cashmere and has a certain value.

关 键 词：芽孢杆菌 角蛋白酶 离子交换层析 凝胶过滤层析 

分 类 号：TQ92[轻工技术与工程—发酵工程]