扩展青霉FS1884脂肪酶基因的定点诱变及其活性分析  

Site-directed Mutagenesis on the Lipase Gene of Penicillium expansum FS1884 and the Activity Analysis of the Mutant

作　　者：李强[1] 张建军[1] 蔡容华[1] 吴伟斌[1] 施碧红[1] 施巧琴[1] 

机构地区：[1]福建师范大学生命科学学院,福建福州350108

出　　处：《福建师范大学学报（自然科学版）》2009年第4期105-108,共4页Journal of Fujian Normal University：Natural Science Edition

基　　金：福建省自然科学基金资助项目(B0610027)

摘　　要：为提高扩展青霉FS1884脂肪酶活性,采用一步突变法对脂肪酶(PEL)的基因进行体外M220V定点诱变,然后将其连接到表达载体pPIC3.5K中,构建重组表达载体pPIC3.5K-M220V-PEL,电转化毕赤酵母GS115中,经甲醇诱导后脂肪酶成功分泌表达.重组脂肪酶的活性分析表明:突变体在毕赤酵母中得到活性表达,获得脂肪酶表达产物PEL-M220V-GS,在35℃下具有最高酶活为3099U/mg,比野生型提高了5%.To increase the activity of Penicillium expansum FS1884 lipase （PEL）, onestep site-directed mutagenesis method was used to introduce a mutation site of Met220Val into the lipase encoding gene. The recombinant vector of pPIC3.5K-M220V-PEL was constructed and transformed into Pichia pastoris GS115 by electroporation. The recombinant mutant was expressed in P. pastoHs and can secret the mutant lipase of PEL-M220V-GS into the culture medium induced by Methanol. The specific activity of the mutant lipase was 3 099 U/mg, which was increased by 5% to that of wild type lipase PEL-GS.

关 键 词：扩展青霉 碱性脂肪酶 毕赤酵母 一步突变法 

分 类 号：Q78[生物学—分子生物学] TS210.9[轻工技术与工程—粮食、油脂及植物蛋白工程]