解脂耶氏酵母AS 2.1216中两种脂肪酶的表达与纯化
Expression and purification of two lipases from Yarrowia lipolytica AS 2.1216.
作者信息
Song Hui-Ting, Jiang Zheng-Bing, Ma Li-Xin
机构信息
College of Chemistry and Material Science, Hubei University, Wuhan, PR China.
出版信息
Protein Expr Purif. 2006 Jun;47(2):393-7. doi: 10.1016/j.pep.2006.02.007. Epub 2006 Mar 10.
We isolated two lipase genes LIPY7, LIPY8 from Yarrowia lipolytica CGMCC (China general microbiological culture collection center) AS 2.1216. The LIPY7 and LIPY8 genes encode a 366 and a 371-amino acid protein, respectively. The lipase genes with 6 x His tag sequence were cloned into expression vector pPIC9K and successfully integrated into a heterologous fungal host Pichia pastoris KM71, respectively. The recombinants were induced by methanol to secrete active lipases into cultural medium. The recombinant lipases were also purified and characterized.
我们从解脂耶氏酵母CGMCC（中国普通微生物菌种保藏管理中心）AS 2.1216中分离出两个脂肪酶基因LIPY7和LIPY8。LIPY7和LIPY8基因分别编码含366个和371个氨基酸的蛋白质。将带有6×组氨酸标签序列的脂肪酶基因分别克隆到表达载体pPIC9K中，并成功整合到异源真菌宿主巴斯德毕赤酵母KM71中。重组体用甲醇诱导，将活性脂肪酶分泌到培养基中。对重组脂肪酶也进行了纯化和特性分析。
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