嗜热真菌Thermomyces lanuginosus热稳定蛋白酶的纯化及特性  

PURIFICATION AND PROPERTIES OF A THERMOSTABLE PROTEASE FROM THE THERMOPHILIC FUNGUS THERMOMYCES LANUGINOSUS

作　　者：朱俊华[1] 李多川[1] 陈静[1] 

机构地区：[1]山东农业大学植保学院,山东泰安271018

出　　处：《山东农业大学学报（自然科学版）》2002年第1期19-22,共4页Journal of Shandong Agricultural University：Natural Science Edition

基　　金：山东省自然科学基金项目

摘　　要：嗜热真菌Thermomyceslanuginosus在液体培养基中于 5 0℃振荡培养 1周后 ,培养液经硫酸铵分级沉淀、DEAE-Sepharose离子交换层析、Phenythl-Sepharose疏水层析 ,得到了凝胶电泳均一的蛋白酶。用SDS -PAGE测定分子量为 5 5kD ,该酶的最适反应温度为 70℃ ,最适作用pH值为 6 .0。该酶具有很好的耐热性 ,在pH值 6 .0 5 0℃条件下酶是稳定的 ;6 0℃保温 1h ,仍保留 92 %的原酶活性 ;90℃时酶活半衰期为 6min。抑制剂试验证明 。A thermostable protease from the culture supernatant from 7-day-old shake culture grown on a modified medium(YPS) of the thermophilic fungus Thermomyces lanuginosus was purified to SDS-PAGE homogenous by ammonium sulfate fraction , DEAE-Sepharose chromatography, and Phenythl- Sepharose chromatography. The molecular weight of the enzyme estimated with SDS-PAGE is approx. 55 kD.The optimum condition for activity is 60℃,pH6.0.The protease is thermostable at 50℃.The half life time of the enzyme is 6 min at 90℃.It is indicated to be a serine protease by inhibition test.

关 键 词：嗜热真菌 THERMOMYCES LANUGINOSUS 蛋白酶 纯化 热稳定 

分 类 号：Q51[生物学—生物化学]