11 research outputs found
Development of a S-cerevisiae whole cell biocatalyst for in vitro sialylation of oligosaccharides
In Vivo Synthesis of Mammalian-Like, Hybrid-Type N-Glycans in Pichia pastoris
The Pichia pastoris N-glycosylation pathway is only partially homologous to the pathway in human cells. In the Golgi apparatus, human cells synthesize complex oligosaccharides, whereas Pichia cells form mannose structures that can contain up to 40 mannose residues. This hypermannosylation of secreted glycoproteins hampers the downstream processing of heterologously expressed glycoproteins and leads to the production of protein-based therapeutic agents that are rapidly cleared from the blood because of the presence of terminal mannose residues. Here, we describe engineering of the P. pastoris N-glycosylation pathway to produce nonhyperglycosylated hybrid glycans. This was accomplished by inactivation of OCH1 and overexpression of an α-1,2-mannosidase retained in the endoplasmic reticulum and N-acetylglucosaminyltransferase I and β-1,4-galactosyltransferase retained in the Golgi apparatus. The engineered strain synthesized a nonsialylated hybrid-type N-linked oligosaccharide structure on its glycoproteins. The procedures which we developed allow glycan engineering of any P. pastoris expression strain and can yield up to 90% homogeneous protein-linked oligosaccharides
Pichia surface display: display of proteins on the surface of glycoengineered Pichia pastoris strains
Dependence of neurotrophic factor activation of Trk tyrosine kinase receptors on cellular sialidase
Trypanosome trans-sialidase targets TrkA tyrosine kinase receptor and induces receptor internalization and activation
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Immunization with an engineered mutant trans-sialidase highly protects mice from experimental Tryponosoma cruzi infection: A vaccine candidate
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