title: Surface display of active lipase in Saccharomyces cerevisiae using Cwp2 as an anchor protein.
abstract: Lipase Lip2 from Yarrowia lipolytica was displayed on the cell surface of Saccharomyces cerevisiae using Cwp2 as an anchor protein. Successful display of the lipase on the cell surface was confirmed by immunofluorescence microscopy and halo assay. The length of linker sequences was further examined to confirm that the correct conformation of Lip2 was maintained. The results showed that the displayed Lip2 exhibited the highest activity at 7.6 +/- 0.4 U/g (dry cell) when using (G(4)S)(3) sequence as the linker, with an optimal temperature and pH at 40 degrees C and pH 8.0. The displayed lipase did not lose any activity after being treated with 0.1% Triton X-100 and 0.1% Tween 80 for 30 min, and it retained 92% of its original activity after incubation in 10% DMSO for 30 min. It also exhibited better thermostability than free Lip2 as reported previously.
authors.affiliation: Key Laboratory of Molecular Biophysics, The Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, 430074 Wuhan, China.
authors.forename: Wenshan
authors.initials: W
authors.lastname: Liu
authors.forename: Heyun
authors.initials: H
authors.lastname: Zhao
authors.forename: Bin
authors.initials: B
authors.lastname: Jia
authors.forename: Li
authors.initials: L
authors.lastname: Xu
authors.forename: Yunjun
authors.initials: Y
authors.lastname: Yan
mesh_terms: D002462:Cell Membrane / Q000378:metabolism*; D004789:Enzyme Activation; D008049:Lipase / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000502:physiology*; D015202:Protein Engineering / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000502:physiology*
chemical_list: C093811:CWP2 protein, S cerevisiae; D008562:Membrane Glycoproteins; D029701:Saccharomyces cerevisiae Proteins; D008049:Lipase