Pichia surface display: display of proteins on the surface of glycoengineered Pichia pastoris strains.
Biotechnol Lett
; 30(12): 2173-81, 2008 Dec.
Article
em En
| MEDLINE
| ID: mdl-18679585
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.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Pichia
/
Proteínas Recombinantes
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Glicoproteínas de Membrana
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Engenharia Genética
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Proteínas de Saccharomyces cerevisiae
Limite:
Humans
Idioma:
En
Ano de publicação:
2008
Tipo de documento:
Article