Variations in protein glycosylation in Hansenula polymorpha depending on cell culture stage.
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.
Kim
So-Young
SY
Systems Microbiology Research Center, KRIBB, Daejeon 305-333, Korea.
Sohn
Jung-Hoon
JH
Pyun
Yu-Ryang
YR
Choi
Eui-Sung
ES
D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000378:metabolism; D001709:Biotechnology / Q000379:methods*; D018929:Cell Culture Techniques / Q000379:methods; D002480:Cellulase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D005949:Glucose Oxidase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism*; D006031:Glycosylation; D010843:Pichia / Q000201:enzymology / Q000254:growth & development / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism
D006023:Glycoproteins; D011994:Recombinant Proteins; D000429:Alcohol Oxidoreductases; D005949:Glucose Oxidase; C029293:methanol oxidase; D002480:Cellulase; C019826:carboxymethylcellulase