贵阳腐霉Pr1蛋白酶的初步纯化及其酶学特性研究  

A primary study on purification of subtilisin-like protease(Pr1) from Pythium guiyangense and its enzymological properties

作　　者：王荣新[1] 雷大卫[1] 苏晓庆[1] 

机构地区：[1]贵阳医学院基础医学院,贵阳550004

出　　处：《安徽农业大学学报》2007年第4期505-508,共4页Journal of Anhui Agricultural University

基　　金：国家自然科学基金(30160003);贵州省省长基金;贵州省高层次人才特助经费共同资助

摘　　要：将Pr1粗酶液通过凝胶过滤层析、SDS-PAGE等方法进行了初步的纯化。在凝胶过滤层析后,发现两种分子量不同的Pr1同工酶(Pr1E1,Pr1E2)。以Suc-(Ala)2-Pro-Phe-PNA为底物时,Pr1酶的Km值为0.0176μmol.L-1,50μL粗酶液反应10 min时的Vmax为0.1446;Pr1E1和Pr1E2最适反应温度均为50℃;Pr1E1和Pr1E2最适反应pH均为8.2。Pr1酶的最佳保存温度在-20℃与-80℃。Subtilisin-like protease （ Prl ） of Pythium guiyangense was primarily purified by gel filtration chro- matography and SDS-PAGE, and two Prl isoenzymes （Prl El, PrlE2 ） were found. In the crude enzyme solution, Km value and V m= for substrate of Suc-（ Ala）2-Pro-Phe-PNA were 0. 0176 μmol · L^-1 and 0. 1446/50 μL · 10 min,respectively. The optimum temperature and pH for both PrlE1 and PrlE2 were 50℃ and pHS. 2, respectively. The optimum preservation temperature for Prl were both -20℃ and -80℃.

关 键 词：贵阳腐霉 Prl蛋白酶 纯化 特性 

分 类 号：Q55[生物学—生物化学] S476.12[农业科学—农业昆虫与害虫防治]