title: Pichia surface display: display of proteins on the surface of glycoengineered Pichia pastoris strains.
abstract: Expression of proteins on the surface of yeasts has a wide range of applications in biotechnology, such as directed evolution of proteins for increased affinity and thermal stability, screening of antibody libraries, epitope mapping, and use as whole-cell biocatalysts. However, hyperglycosylation can interfere with overall protein accessibility on the surface. Therefore, the less elaborate hyperglycosylation in wild type Pichia pastoris and the availability of glycoengineered strains make this yeast an excellent alternative for surface display of glycoproteins. Here, we report the implementation of the well-established a-agglutinin-based yeast surface display technology in P. pastoris. Four heterologous proteins were expressed on the surface of a wild type and a glycoengineered strain. Surface display levels were monitored by Western blot, immunofluorescence microscopy, and FACS analysis. The availability of glycoengineered strains makes P. pastoris an excellent alternative for surface display of glycoproteins and paves the way for new applications.
authors.affiliation: Department for Molecular Biomedical Research, Unit for Fundamental and Applied Molecular Biology, VIB, Technologiepark 927, 9052 Ghent, Belgium.
authors.forename: Pieter P
authors.initials: PP
authors.lastname: Jacobs
authors.forename: Stefan
authors.initials: S
authors.lastname: Ryckaert
authors.forename: Steven
authors.initials: S
authors.lastname: Geysens
authors.forename: Kristof
authors.initials: K
authors.lastname: De Vusser
authors.forename: Nico
authors.initials: N
authors.lastname: Callewaert
authors.forename: Roland
authors.initials: R
authors.lastname: Contreras
mesh_terms: D015153:Blotting, Western; D003001:Cloning, Molecular; D005434:Flow Cytometry; D015966:Gene Expression Regulation, Fungal; D005818:Genetic Engineering; D005822:Genetic Vectors; D006031:Glycosylation; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; C069939:a-agglutinin protein, S cerevisiae