title: Development of novel yeast cell surface display system for homo-oligomeric protein by coexpression of native and anchored subunits.
abstract: Streptavidin derived from Streptomyces avidinii was displayed on the cell surface of the yeast Saccharomyces cerevisiae by cell-surface engineering using two types of plasmid for the expression of a native subunit and an anchored subunit fused with the C-terminus of 318 amino acids of Flo1p containing a glycosylphosphatidylinositol anchor attachment signal. The displayed streptavidin had the binding ability for biotinylated compounds. This was confirmed by fluorescence microscopy after the adsorption of yeast cells displaying streptavidin and biotinylated fluorescein isothiocyanate. On the other hand, streptavidin produced by cells harboring only the plasmid for the expression of the anchored subunit showed a very low binding activity for biotinylated compounds. Cells displaying streptavidin may constitute novel whole-cell affinity adsorbents widely used for immunoassay and biosensing. This coexpression method will ensure that proteins, such as homo- and hetero-oligomeric proteins, are displayed on the cell surface in an active form.
authors.affiliation: Yokohama Research Center, Chisso Corporation, Kanazawa-ku, Japan.
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Furukawa
authors.forename: Takanori
authors.initials: T
authors.lastname: Tanino
authors.forename: Hideki
authors.initials: H
authors.lastname: Fukuda
authors.forename: Akihiko
authors.initials: A
authors.lastname: Kondo
mesh_terms: D000327:Adsorption; D019921:Biotinylation; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D005240:Feasibility Studies; D015870:Gene Expression; D006735:Horseradish Peroxidase / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019809:Streptavidin / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D013499:Surface Properties
chemical_list: D019809:Streptavidin; D006735:Horseradish Peroxidase