应用细胞透性化技术快速提取氧化葡萄糖杆菌胞内右旋糖酐糊精酶  

Method for determinating dextran dextrinase from Gluconobacter oxydans by cell permeabilization

作　　者：王舒[1] 毛相朝[1,2] 张鲁嘉[3] 邢艳陇[1] 王华磊[1] 魏东芝[1,3] 

机构地区：[1]中国科学院青岛生物能源与过程研究所,中国科学院生物燃料重点实验室,青岛266101 [2]中国海洋大学食品科学与工程学院,青岛266003 [3]华东理工大学生物反应器工程国家重点实验室,鲁华生物技术研究所,上海200237

出　　处：《生物加工过程》2010年第3期35-39,共5页Chinese Journal of Bioprocess Engineering

基　　金：国家自然科学基金资助项目(30900025);国家重点基础研究发展计划(973计划)资助项目(2009CB724703)

摘　　要：将细胞透性化技术应用到右旋糖酐酶的提取检测过程中,根据单因素实验和正交试验设计确定了从氧化葡萄糖杆菌（Gluconobacter oxydans）细胞中提取胞内右旋糖酐糊精酶（DDase）的最佳方法：甘氨酸（Gly）质量分数15%,Triton X-100体积分数1%,菌悬液OD600为0.5~6.0,pH为6.81,冰浴处理时间1 h,透性化试剂的提取效率可达到酶活最大值的90%以上。与超声波细胞破碎法相比,该方法条件温和,酶的释放率较高并易于大量试样的平行实验操作。The study was focused on the application of disrupting Gluconobacter oxydans to release dextran dextrinase（DDase） by cell permeabilization.Kind of agents,solution concentration,solution pH and the disruption time were studied,which affected the disruption efficiency.A rapid method for extracting and determinating of the DDase with glycine and Triton X-100 was presented.The optimal conditions for extraction were as follows： glycine concentration 15%,Triton X-100 concentration 1%,OD600 of Gluconobacter oxydans suspension 0.5 to 6,pH 6.81,extraction time 1 h.The extraction efficiency of permeability reagents could achieve over 90%.The method could obtain the higher breaking efficiency and be easier to scale up compared with the ultrasonic disruption.

关 键 词：细胞透性化 氧化葡萄糖杆菌 右旋糖酐糊精酶 甘氨酸 

分 类 号：Q555.4[生物学—生物化学]