Glycobiology Advance Access originally published online on December 23, 2003
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Glycobiology vol 14 no 3 pp. 243-251, 2004
Glycobiology vol. 14 no. 3 © Oxford University Press 2004; all rights reserved.
Characterization of N-linked oligosaccharides assembled on secretory recombinant glucose oxidase and cell wall mannoproteins from the methylotrophic yeast Hansenula polymorpha
2 Metabolic Engineering Laboratory, Korea Research Institute of Bioscience and Biotechnology, Oun-dong 52, Yusong-gu, Daejeon 305-600, Korea; 3 Department of Microbiology, Chungnam National University, Daejeon 305-764, Korea; and 4 Research Center for Glycoscience, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-4 Higashi, Tsukuba, Ibaraki, 305-8566, Japan
Received on August 27, 2003; revised on October 27, 2003; accepted on October 30, 2003
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 (Man8–12GlcNAc2). Analyses using anti-
1,3-mannose antibody and exoglycosidases specific for 1,2- or 1,6-mannose linkages revealed that the mannose outer chains of N-glycans on the rGOD have very short 1,6 extensions and are mainly elongated in 1,2-linkages without a terminal 1,3-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 1,3-linked mannoses.
1 To whom correspondence should be addressed; e-mail: hyunkang{at}mail.kribb.re.kr
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