title: Development of a new yeast surface display system based on Spi1 as an anchor protein.
abstract: Yeast surface display is a powerful tool widely used for many biotechnological and biomedical applications. It consists in exposing peptides and proteins of interest on the surface of Saccharomyces cerevisiae and other yeasts. These molecules are fused to the amino or carboxy terminus of an appropriate cell wall protein, usually bound by glycosylphosphatidylinositol. Several systems for this purpose have been reported to date. In this work, we describe a new yeast surface display strategy based on cell wall protein Spi1 as an anchor, which is expressed in centromeric and episomal plasmids under the control of its own promoter or that corresponding to the PGK1 glycolytic gene. Exposure efficiency was demonstrated by western blot, flow cytometry analyses, and fluorescence microscopy by taking advantage of including the V5 epitope. We also demonstrated the ability of this new strategy for the exposure of several peptides and proteins of different sizes. The regulation of the SPI1 promoter by the stationary phase and other stress conditions reveals interesting potential applications of systems based on it for industrial and biotechnological processes.
authors.affiliation: Departament de Química Orgànica, Facultat de Farmàcia, Universitat de València, Vicent Andrés Estellés s/n E-46100 Burjassot, Valencia, Spain.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Andreu
authors.affiliation: Departament de Bioquímica i Biologia Molecular, Facultat de Biologia, Universitat de València, Dr. Moliner 50, E-46100 Burjassot, Valencia, Spain. m.del.olmo@uv.es.
authors.forename: Marcel Lí
authors.initials: ML
authors.lastname: Del Olmo
mesh_terms: D015153:Blotting, Western; D061505:Cell Surface Display Techniques / Q000379:methods*; D005434:Flow Cytometry; D005822:Genetic Vectors; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism*
chemical_list: D008562:Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; C516345:SPI1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins
keywords: Saccharomyces cerevisiae; Spi1; Stationary phase; Stress conditions; Yeast surface display