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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
【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%.
【Abstract】 To increase the activity of Penicillium expansum FS1884 lipase (PEL),one-step 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. pastoris 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.
【Key words】 Penicillium expansum； lipase； Pichia pastoris； one-step site-directed mutagenesis；
- 【文献出处】 福建师范大学学报(自然科学版) ,Journal of Fujian Normal University(Natural Science Edition) , 编辑部邮箱 ,2009年04期
- 【分类号】Q93
- 【被引频次】7
- 【下载频次】157