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
李强;张建军;蔡容华;吴伟斌;施碧红;施巧琴;
摘要(Abstract):
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%.
关键词(KeyWords): 扩展青霉;碱性脂肪酶;毕赤酵母;一步突变法
基金项目(Foundation): 福建省自然科学基金资助项目(B0610027)
作者(Authors): 李强;张建军;蔡容华;吴伟斌;施碧红;施巧琴;
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