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Title 

Characterization of N-linked oligosaccharides assembled on secretory recombinant glucose oxidase and cell wall mannoproteins from the methylotrophic yeast Hansenula ploymorpha

Authors 

Moo Woong KimSang Ki RheeJ Y KimY ShimmaY ChibaY JigamiHyun Ah Kang

Publisher 

Oxford University Press (OUP)

Issue Date 

2004

Citation 

Glycobiology, vol. 14, no. 3, pp. 243-251

Keywords 

cell wall mannoproteinsglycan profilinghansenula polymorphan-linked oligosaccharidesrecombinant glucose oxidasealpha mannosidaseantibodyasparagine linked oligosaccharideglucose oxidaseglycoprotein

Abstract 

Presently almost no information is available on the oligosaccharide structure of the glycoproteins secreted from the methylotrophic yeast Hansenula polymorpha, a promising host for the production of recombinant proteins. In this study, we analyze the size distribution and structure of N-linked oligosaccharides attached to the recombinant glycoprotein glucose oxidase (GOD) and the cell wall manno-proteins obtained from H. polymorpha. Oligosaccharide profiling showed that the major oligosaccharide species derived from the H. polymorpha-secreted recombinant GOD (rGOD) had core-type structures (Man8-12GlcNAc2. Analyses using anti-α1,3-mannose antibody and exoglycosidases specific for α1,2- or ,6-mannose linkages revealed that the mannose outer chains of N-glycans on the rGOD have very short α1,6 extensions and are mainly elongated in α1,2-linkages without a terminal α1,3-linked mannose addition. The N-glycans released from the H. polymorpha mannoproteins were shown to contain mostly mannose in their outer chains, which displayed almost identical size distribution and structure to those of H. polymorpha-derived rGOD. These results strongly indicate that the outer chain processing of N-glycans by H. polymorpha significantly differs from that by Saccharomyces cerevisiae, thus generating much shorter mannose outer chains devoid of terminal α1,3-linked mannoses.

ISSN 

0959-6658

Link 

http://dx.doi.org/10.1093/glycob/cwh030

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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