Pichia surface display: display of proteins on the surface of glycoengineered Pichia pastoris strains

被引：22
作者：
Jacobs, Pieter P. [2 ,3 ]
Ryckaert, Stefan [2 ,3 ]
Geysens, Steven [2 ,3 ]
De Vusser, Kristof [2 ,3 ]
Callewaert, Nico [1 ,4 ]
Contreras, Roland [2 ,3 ]
机构：
[1] Univ Ghent, Lab Prot Biochem & Biomol Engn, Dept Biochem Physiol & Microbiol, B-9000 Ghent, Belgium
[2] VIB, Dept Mol Biomed Res, Unit Fundamental & Appl Mol Biol, B-9052 Ghent, Belgium
[3] Univ Ghent, Dept Mol Biol, B-9052 Ghent, Belgium
[4] VIB, Dept Mol Biomed Res, Unit Mol Glycobiol, B-9052 Ghent, Belgium
来源：
BIOTECHNOLOGY LETTERS | 2008年 / 30卷 / 12期
关键词：
N-glycosylation; Pichia pastoris; Surface display; Yeast;
D O I：
10.1007/s10529-008-9807-1
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
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.
引用
收藏
页码：2173 / 2181
页数：9
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