title: Cell Surface Display and Characterization of Rhizopus oryzae Lipase in Pichia pastoris Using Sed1p as an Anchor Protein.
abstract: It has been investigated to conduct the surface displaying of lipase from Rhizopus oryzae onto the cells of Pichia pastoris yeast using Sed1p as an anchor protein. A yeast cell surface display plasmid pPICZαA-rol-histag-sed1p was constructed by fusing rol and sed1p gene fragments into the plasmid pPICZαA, followed by introducing recombinant plasmid into P. pastoris cells. Surface display levels were monitored by Western Blot and immunofluorescence microscopy. The activity of displaying lipase obtained from recombinant mutS reached at 60 U/g-dry cell. In addition, the displaying lipase was stable in broad ranges of temperatures and pH, with the optimum temperature at 40 °C and pH 7.5. These results indicate that the P. pastoris displaying lipase may have potential in whole-cell biocatalyst.
authors.affiliation: College of Chemical Engineering, Nanjing Forestry University, Nanjing, China.
authors.forename: Wenqian
authors.initials: W
authors.lastname: Li
authors.forename: Hao
authors.initials: H
authors.lastname: Shi
authors.forename: Huaihai
authors.initials: H
authors.lastname: Ding
authors.forename: Liangliang
authors.initials: L
authors.lastname: Wang
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
authors.forename: Xun
authors.initials: X
authors.lastname: Li
authors.forename: Fei
authors.initials: F
authors.lastname: Wang
mesh_terms: D015153:Blotting, Western; D061505:Cell Surface Display Techniques; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012233:Rhizopus / Q000201:enzymology* / Q000235:genetics; D013696:Temperature
chemical_list: D011994:Recombinant Proteins; D008049:Lipase