大豆过氧化物酶基因在毕赤酵母中的表达  

Expression of Soybean Peroxidase Gene in the Pichia Pastoris

作　　者：曾家豫[1] 刘芙蓉[1] 廖世奇[2] 王黎[2] 袁红霞[1] 王宇仙[1] 张渭波[1] 

机构地区：[1]西北师范大学生命科学学院,甘肃兰州730070 [2]甘肃省医学科学研究院,甘肃兰州730050

出　　处：《大豆科学》2010年第5期733-737,共5页Soybean Science

基　　金：甘肃省教育厅资助项目(0601-28;0501B-16);甘肃省自然科学基金资助项目(096RJZA035);甘肃省高分子材料重点实验室开放课题(KF-06-01)

摘　　要：大豆过氧化物酶(Soybean peroxidase,SBP)是一类主要催化过氧化氢的氧化还原酶,具有耐热性高和pH适用范围宽等优点。利用已构建好的重组表达载体pPICZα-A-sbp,通过氯化锂化学转化法将重组质粒转化进入毕赤酵母GS115中,并在甲醇诱导下进行表达。利用SDS-PAGE法和愈创木酚法测定了重组菌株的SBP表达活性。结果显示,重组毕赤酵母表达了分子量约为40 KD的SBP蛋白,同时发酵24 h后SBP表达活性开始迅速增高,发酵72 h SBP表达活性达到最大,并筛选出表达活性高的菌株。Soybean peroxidase （SBP） is a kind of oxidoreductase, which mainly catalyzed the hydrogen peroxide. The SBP has the advantages of good heat resistance and wide range of pH application and so on. Used the reeombinants expression vector pPICα-A-sbp transformed into Pichia pastoris GS115 by chemical transformation with lithium chloride. The transformants were induced by methanol to express a protein, which was verified by SDS-PAGE and guaiacol. The results showed that the recombinants express approximately 40 KD protein in size, fermentation began rapidly increasing activities of SBP ex- pression after 24 h, maximum in 72 h and the high expressions of sbp gene were filtered.

关 键 词：大豆过氧化物酶 毕赤酵母 基因表达 

分 类 号：S565.1[农业科学—作物学]