解淀粉芽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

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 gone 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 swain GS 115 by electroporation. 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 72h, the enzyme activity can reach to 2330 U/L.