Characterization of N-linked oligosaccharides assembled on secretory recombinant glucose oxidase and cell wall mannoproteins from the methylotrophic yeast Hansenula polymorphaopen access
- Authors
- Kim, Moo Woong; Rhee, Sang Ki; Kim, Jeong-Yoon; Shimma, Yoh-ichi; Chiba, Yasunori; Jigami, Yoshifumi; Kang, Hyun Ah
- Issue Date
- Mar-2004
- Publisher
- OXFORD UNIV PRESS INC
- Keywords
- Cell wall mannoproteins; Glycan profiling; Hansenula polymorpha; N-linked oligosaccharides; Recombinant glucose oxidase
- Citation
- GLYCOBIOLOGY, v.14, no.3, pp 243 - 251
- Pages
- 9
- Journal Title
- GLYCOBIOLOGY
- Volume
- 14
- Number
- 3
- Start Page
- 243
- End Page
- 251
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/57023
- DOI
- 10.1093/glycob/cwh030
- ISSN
- 0959-6658
1460-2423
- 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 mannoproteins 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 (Man(8-12)GlcNAc(2)). Analyses using anti-alpha1,3-mannose antibody and exoglycosidases specific for alpha1,2- or alpha1,6-mannose linkages revealed that the mannose outer chains of N-glycans on the rGOD have very short alpha1,6 extensions and are mainly elongated in alpha1,2-linkages without a terminal alpha1,3-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 alpha1,3-linked mannoses.
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