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- Title
Cell Surface Display and Characterization of Rhizopus oryzae Lipase in Pichia pastoris Using Sed1p as an Anchor Protein.
- Authors
Li, Wenqian; Shi, Hao; Ding, Huaihai; Wang, Liangliang; Zhang, Yu; Li, Xun; Wang, Fei
- 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.
- Subjects
RHIZOPUS oryzae; PICHIA pastoris; LIPASES; WESTERN immunoblotting; IMMUNOFLUORESCENCE
- Publication
Current Microbiology, 2015, Vol 71, Issue 1, p150
- ISSN
0343-8651
- Publication type
Article
- DOI
10.1007/s00284-015-0835-5