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J. Microbiol. Biotechnol. 2007; 17(12): 1949-1954

Published online December 28, 2007

Copyright © The Korean Society for Microbiology and Biotechnology.

Variations in Protein Glycosylation in Hansenula polymorpha Depending on Cell Culture Stage

Kim, So-Young , Jung-Hoon Sohn , Yu-Ryang Pyun 1 and Eui-Sung Choi 1*

Systems Microbiology Research Center, KRIBB, Daejeon 305-333, Korea, 1Department of Biotechnology, Yonsei University, Seoul 120-749, Korea

Abstract

A simple way to prevent protein hyperglycosylation in Hansenula polymorpha was found. When glucose oxidase from Aspergillus niger and carboxymethyl cellulase from Bacillus subtilis were expressed under the control of an inducible methanol oxidase (MOX) promoter using methanol as a carbon source, hyperglycosylated forms occurred. In contrast, MOX-repressing carbon sources (e.g., glucose, sorbitol, and glycerol) greatly reduced the extent of hyperglycosylation. Carbon source starvation of the cells also reduced the level of glycosylation, which was reversed to hyperglycosylation by the resumption of cell growth. It was concluded that the proteins expressed under actively growing conditions are produced as hyperglycosylated forms, whereas those under slow or nongrowing conditions are as short-glycosylated forms. The prevention of hyperglycosylation in the Hansenula polymorpha expression system constitutes an additional advantage over the traditional Saccharomyces cerevisiae system in recombinant production of glycosylated proteins.

Keywords: Glycosylation, glucose oxidase, Hansenula polymorpha, CMCase