Journal
BIOTECHNOLOGY LETTERS
Volume 30, Issue 12, Pages 2173-2181Publisher
SPRINGER
DOI: 10.1007/s10529-008-9807-1
Keywords
N-glycosylation; Pichia pastoris; Surface display; Yeast
Categories
Funding
- Institute for the Promotion of Innovation through Science and Technology in Flanders (IWT-Vlaanderen)
- Research Corporation Technologies (Tucson, AZ)
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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.
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