CAJ | 학술논문

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1598．5 U/mL.
종해지야씨효모중극륭출지방매기인Lip2，장기련접도표체재체pPIC9K，연후전격전화도필적효모GS115균주중，통과사선，획득일주표체활성교고적공정균주HL10.통과대발효조건화발효삼수적우화，재요병중용1．5％적갑순유도3천，표체적지방매적활력체도1598．5 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 (α-factor) under the control of the methanol inducible promoter of the alcohol oxidase 1 gene.