Development of a novel rDNA based plasmid for enhanced cell surface display on Yarrowia lipolytica

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dc.contributor.author Bulani, Siyavuya Ishmael
dc.contributor.author Moleleki, Lucy N.
dc.contributor.author Albertyn, Jacobus
dc.contributor.author Moleleki, Ntsane
dc.date.accessioned 2012-12-03T13:02:46Z
dc.date.available 2012-12-03T13:02:46Z
dc.date.issued 2012-05-20
dc.description.abstract In this study, a novel rDNA based plasmid was developed for display of heterologous proteins on the cell surface of Yarrowia lipolytica using the C-terminal end of the glycosylphosphatidylinositol (GPI) anchored Y. lipolytica cell wall protein 1 (YlCWP1). mCherry was used as a model protein to assess the efficiency of the constructed plasmid. Y. lipolytica transformants harbouring the expression cassettes showed a purple colour phenotype on selective YNBcasamino plates as compared to control cells indicating that mCherry was displayed on the cells. Expression of mCherry on cells of Y. lipolytica was confirmed by both fluorescent microscopy and flow cytometry. Furthermore, SDS-PAGE analysis and matrix-assisted laser desorption/ionization (MALDI)-time-of (TOF)-mass spectrometry (MS) peptide mass fingerprinting (PMF) confirmed that the protein cleaved from the yeast cells using enterokinase was mCherry. Efficient cleavage of mCherry reported in this work offers an alternative purification method for displayed heterologous proteins on Y. lipolytica cells using the plasmid constructed in this study. The developed displaying system offers great potential for industrial production and purification of heterologous proteins at low cost. en_US
dc.description.sponsorship CSIR en_US
dc.description.uri http://www.amb-express.com/content/2/1/27 en_US
dc.identifier.citation Bulani et al.: Development of a novel rDNA based plasmid for enhanced cell surface display on Yarrowia lipolytica. AMB Express 2012 2:27. en_US
dc.identifier.issn 2191-0855
dc.identifier.other 10.1186/2191-0855-2-27
dc.identifier.uri http://hdl.handle.net/2263/20629
dc.language.iso en en_US
dc.rights © 2012 Bulani et al.; licensee Springer. This is an Open Access article distributed under the terms of the Creative Commons Attribution License en_US
dc.subject mCherry en_US
dc.subject rDNA vector en_US
dc.subject YlCWP1 en_US
dc.subject Yarrowia lipolytica en_US
dc.subject Cell surface display en_US
dc.title Development of a novel rDNA based plasmid for enhanced cell surface display on Yarrowia lipolytica en_US
dc.type Article en_US


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