解淀粉芽孢杆菌中性植酸酶基因的克隆及其在毕赤酵母中的表达  

Cloning and Expression of Neutral Phytase Gene from Bacillus Amyloliquefaciens in Pichia Pastoris

作　　者：张建云[1] 崔树军[1] 谷立坤[1,2] 武秀琴[1] 

机构地区：[1]河南工程学院资源与环境工程系,河南郑州451191 [2]中国科学院研究生院,北京100049

出　　处：《河南科技大学学报（自然科学版）》2010年第2期69-72,共4页Journal of Henan University of Science And Technology:Natural Science

基　　金：河南省科技攻关项目(092102210212);河南工程学院青年基金项目(Y2007031)

摘　　要：采用PCR法从解淀粉芽孢杆菌BA11中克隆到一个中性植酸酶phyc基因,将该基因克隆到毕赤酵母表达载体pPIC9K上并电转化至宿主细胞GS115后进行诱导表达。SDS-PAGE试验表明:该重组中性植酸酶在毕赤酵母宿主细胞中实现了高效分泌性表达。植酸酶活性测定结果显示阳性克隆子在诱导72h酶活性达到最高值,活性为2330U/L。Phyc gene was amplified from Bacillus amyloliquefaciens genomic DNA by PCR.The Phyc gene was inserted into the expression vector pPIC9K and transformed into the Pichia pastoris strain GS115 by elect-roporation.Then the Phyc gene was expressed by induction of methyl alcohol.SDS-PAGE analysis showed that recombinant neutral phytase was successfully produced at a high level in Pichia pastoris.And after induced for 72 h,the enzyme activity can reach 2 330 U/L.

关 键 词：中性植酸酶 毕赤酵母 解淀粉芽孢杆菌 异源表达 

分 类 号：Q939.124[生物学—微生物学] Q556