毕赤酵母表达罗氏耶尔森氏菌(Yersinia rohdei)植酸酶和酶学特性研究  

Expression of phytase from Yersinia rohdei in Pichia pastoris and its enzymatic characteristics

作　　者：陈磊[1,2] 许晶[2] 韩红娟[2] 邱瑾[1,2] 孙百慧[1,2] 彭日荷[2] 姚泉洪[2] 

机构地区：[1]上海海洋大学食品学院,上海201306 [2]上海市农业科学院生物技术研究所,上海201106

出　　处：《上海农业学报》2016年第6期18-25,共8页Acta Agriculturae Shanghai

基　　金：上海市科委农业成果转化项目(143919NO300);上海市种业发展项目[沪农科种字(2016)第1-2号]

摘　　要：按毕赤酵母偏爱密码子,将来自Yersinia rohdei的植酸酶基因(YrAPPA)进行了化学合成,并成功在毕赤酵母GS-115中异源分泌表达,重组植酸酶分泌量能达到72μg/mL。酶学特性研究结果表明:酶比活达到1 500 U/mg,重组植酸酶的最适反应温度和pH分别为55℃和4.5;纯化的重组植酸酶能耐受60℃高温,且pH 3—7.5范围内重组植酸酶有良好的稳定性;Cu^(2+)、Zn^(2+)、Fe^(3+)对植酸酶活性有抑制作用;重组植酸酶有抗胃蛋白酶和胰蛋白酶水解特性。Yersinia rohdei phytase gene （YrAPPA） was chemically synthesized according to the codon usage of Pichia pastoris and successfully expressed in P. pastoris GS-115, with the secretion yield of recombinant phytase being up to about 72μg/mL. The enzymatic characterization showed that the specific activity of enzyme is 1 500 U/rag, and the optimal reacting temperature and pH of recombinant phytase are respectively 55℃ and 4.5;The purified recombinant phytase can tolerate high temperature at 60℃ and has a good stability within pH 3-7.5 ;The phytase whose activity is inhibited by Cu^2＋, Zn^2＋ and Fe^3＋ has characteristics resistant to trypsin and pepsin.

关 键 词：植酸酶 酶学特性 毕赤酵母 电击转化 异源表达 

分 类 号：Q78[生物学—分子生物学]