嗜热土芽孢杆菌GSEY01及其高温蛋白酶的初步研究  

Study on A Thermophilic Geobacillus sp.GSEY01 and Its Thermophilic Protease

作　　者：廖艳江[1] 季秀玲[1] 魏云林[1] 林连兵[1] 

机构地区：[1]昆明理工大学生物工程技术研究中心,昆明650224

出　　处：《生物技术通报》2010年第6期189-194,共6页Biotechnology Bulletin

基　　金：中国科学院微生物研究所微生物资源前期开发国家重点实验室开放课题(SKLMR-080605);国家自然科学基金项目(30660009;30960022)

摘　　要：从云南腾冲热海热泉中分离出一株产高温蛋白酶的菌株GSEY01。该菌株最适生长温度为60℃,16S rRNA基因序列分析表明,该菌株为土芽孢杆菌属(Geobacillus)的耐热菌株。该菌株所产高温蛋白酶可以通过超滤浓缩,硫酸铵分级沉淀和强阴离子交换层析获得纯酶。此高温蛋白酶分子量约为42kD,最适催化温度为80℃,最适催化pH7.5,Mg2+能增强该酶活力,Fe3+,Cd2+和Ni2+对其活性则有抑制作用。PMSF对该酶影响较小,乙二胺四乙酸(EDTA)和十二烷基磺酸钠(SDS)则对其有强烈的抑制作用,此高温蛋白酶和其他土芽孢杆菌所产蛋白酶有较大差异,可以应用于相关的高温催化环境。A thermostable extracellular protease producing strain GSEY01 was isolated from a hot spring in Rehai,Tengchong,Yunnan Province.The optimal growth temperature of strain GSEY01 was 60℃.16S rRNA gene phylogenetic analysis showed that this strain belonged to thermophilic Geobacillus sp.Protease secreted by strain GSEY01 was purified to homogeneity by ultra filtration,ammonium sulfate precipitation and ion exchange chromatography.Molecular weight of this proteinase was 42 kD.The protease showed the highest activity at 80℃and pH7.5.The activity of the protease could be activated by Mg 2＋,but inhibited partially by Fe3＋,Cd2＋and Ni2＋.In addition,the activity of the enzyme was inhibited by EDTA and SDS strongly,but not by PMSF.Protease from strain GSEY01was greatly different from those proteinase isolated from other strains of Geobacillus,and could be applied in industrial processes that are performed under high temperature.

关 键 词：土芽孢杆菌 高温蛋白酶 纯化 酶学性质 

分 类 号：Q936[生物学—微生物学]