title: Cloning, expression and characterization of a new lipase from Yarrowia lipolytica.
abstract: Bioinformatic analysis of the Yarrowia lipolytica CLIB122 genome has revealed 18 putative lipase genes all of which were expressed in Escherichia coli and screened for hydrolyzing activities against p-nitrophenyl-palmitate. One positive transformant containing an ORF of 1,098 bp encoding a protein of 365 amino acids was obtained. To characterize its enzymatic properties, the lipase gene was functionally expressed in Pichia pastoris. The resulting lipase exhibited the highest activity towards p-NP-decanoate at pH 7 and 35 °C. In addition, the new lipase had a lower optimal temperature and pH compared to other Y. lipolytica lipases. It was noticeably enhanced by Ca(2+), but was inhibited by PMSF, Hg(2+) and Ni(2+). The new lipase displayed the 1,3-specificity for triolein.
authors.affiliation: Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China. xiaohehc@gmail.com
authors.forename: Heyun
authors.initials: H
authors.lastname: Zhao
authors.forename: Lina
authors.initials: L
authors.lastname: Zheng
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Wang
authors.forename: Yun
authors.initials: Y
authors.lastname: Liu
authors.forename: Li
authors.initials: L
authors.lastname: Xu
authors.forename: Yunjun
authors.initials: Y
authors.lastname: Yan
mesh_terms: D003001:Cloning, Molecular; D004789:Enzyme Activation; D004795:Enzyme Stability; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D008049:Lipase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D025062:Yarrowia / Q000201:enzymology* / Q000235:genetics
chemical_list: D008049:Lipase