解脂耶氏酵母脂肪酶基因Lip2在毕赤酵母中高活力表达  

High Level Expression of Yarrowia lipolytica Lipase Gene Lip2 in Pichia pastoris

作　　者：解复红[1,2] 权淑静[1,2] 马焕[1,2] 刘德海[1,2] 程雁[2] 

机构地区：[1]河南省工业酶工程技术研究中心,郑州450008 [2]河南省科学院生物研究所有限责任公司,郑州450008

出　　处：《河南科学》2013年第5期589-593,共5页Henan Science

基　　金：河南省自然科学基金项目(122102213117;122102210110)

摘　　要：从解脂耶氏酵母中克隆出脂肪酶基因Lip2,将其连接到表达载体pPIC9K,然后电击转化到毕赤酵母GS115菌株中,通过筛选,获得一株表达活性较高的工程菌株HL10.通过对发酵条件和发酵参数的优化,在摇瓶中用1.5%的甲醇诱导3天,表达的脂肪酶的活力达到1 598.5 U/mL.The extracellular lipase gene from Yarrowia lipolytica（Lip2） was cloned, ligated into vector pPIC9K. The recombinant plasmid pPIC9K-Lip2 was linearized by restriction enzyme SalI, then integrated into the genome of the methylotrophic yeast Pichia pastoris GS115. A clone named HL10 with higher lipase was selected. When the medium and fermentation condition were optimized,the lipase activity reached 1598.5 U/mL in a shaken flask cultivation with an apparent molecular weight of 36.0 kDa using Saccharomyces cerevisiae secretion signal peptide （a-factor） under the control of the methanol inducible promoter of the alcohol oxidase 1 gene.

关 键 词：解脂耶氏酵母 毕赤酵母 脂肪酶 表达 

分 类 号：Q556.2[生物学—生物化学]