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OALib Journal期刊
ISSN: 2333-9721
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大肠杆菌植酸酶在毕赤酵母中的表达与纯化
DOI: 10.3724/SP.J.1145.2012.00678, PP. 678-681
Keywords: 大肠杆菌植酸酶,毕赤酵母,甲醇浓度,表达与纯化,圆二色谱
Abstract:
为探讨植酸酶的结构与功能，研究了来源于大肠杆菌的植酸酶基因在酵母中的表达和纯化条件.将含有大肠杆菌植酸酶基因的毕赤酵母工程菌在不同甲醇浓度和不同诱导时间下培养，检测植酸酶的表达情况.结果表明：诱导培养基中一次性添加2.5%的甲醇，诱导96h后，蛋白浓度达到1.36mgml-1，酶活力达到6530uml-1；将在毕赤酵母中表达的植酸酶粗酶液经过硫酸铵盐析、resources柱和superdex-75三步纯化，得到单峰纯的蛋白，比活力为137280umg-1.用圆二色谱分析纯化后的蛋白在ph5.0和8.0环境中的结构变化，结果显示ph8.0环境使大肠杆菌植酸酶发生了变性.图3参16
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