扩展青霉脂肪酶ep8与R182K叠加突变体的构建  

R182K Double-Mutagenesis of Penicillium expansum Lipase

作　　者：王彩梅[1] 蔡少丽[1] 吴义真[1] 林琳[1] 

机构地区：[1]福建师范大学生命科学学院,福州350007

出　　处：《生物技术通报》2007年第4期165-168,共4页Biotechnology Bulletin

基　　金：国家自然科学基金资助项目(No.30270033);福建省自然科学基金资助项目(No.B0120001和C0410009);福建省工业微生物研发平台建设(2006H0085)项目资助

摘　　要：利用重叠延伸PCR法对扩展青霉碱性脂肪酶(PEL)基因进行体外定点突变,并将含突变基因的重组质粒pAO815-ep8-R182K在毕赤酵母(Pichiapastoris)GS115中进行表达。叠加突变体PEL-ep8-R182K表达产物与野生型PEL、PEL-ep8比较实验表明:叠加突变体表达蛋白PEL-ep8-R182K最适反应温度与野生型PEL、PEL-ep8一致,均为40℃;热稳定性与野生型相似,比PEL-ep8降低2.25℃。但是,在比活上,PEL-ep8-R182K与PEL-ep8、野生型PEL相比,其比酶活分别提高了14.03%和3.86%。Double-Mutagenesis using megaprimer PCR was applied to the gene of Penicillium expansum lipase, and the mutant recombinant plasmid was expressed in Piehia pastoris GSll5.The comparision experiments of PEL-ep8- R182K with PEL and PEL-ep8 showed that the optimum temperature of them was 40℃ and its thermostability was similar with the wild type and 2.25℃ lower than the random mutant, However,its activity was increased by 14.03% than the wild type and 3.86% than the random mutant.

关 键 词：扩展青霉脂肪酶 叠加突变 最适温度 热稳定性 

分 类 号：Q78[生物学—分子生物学]