Pichia surface display: display of proteins on the surface of glycoengineered Pichia pastoris strains
| Autor: | Roland Contreras, Steven Geysens, Pieter P. Jacobs, Stefan Ryckaert, Kristof De Vusser, Nico Callewaert |
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| Rok vydání: | 2008 |
| Předmět: |
Glycosylation
Saccharomyces cerevisiae Proteins Blotting Western Genetic Vectors Bioengineering Applied Microbiology and Biotechnology Pichia Pichia pastoris chemistry.chemical_compound N-linked glycosylation Gene Expression Regulation Fungal Humans Cloning Molecular chemistry.chemical_classification Membrane Glycoproteins biology Wild type General Medicine biology.organism_classification Flow Cytometry Molecular biology Yeast Recombinant Proteins Epitope mapping chemistry Biochemistry Microscopy Fluorescence Glycoprotein Genetic Engineering Biotechnology |
| Zdroj: | Biotechnology letters. 30(12) |
| ISSN: | 1573-6776 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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