In vivo synthesis of complex N-glycans by expression of human N-acetylglucosaminyltransferase I in the filamentous fungus Trichoderma reesei.
The human N-acetylglucosaminyltransferase I gene was introduced in the genome of Trichoderma reesei strain VTT-D-80133. Expression was studied after induction from the cellobiohydrolase I promoter. Successful in vivo transfer of GlcNAc was demonstrated by analyzing the neutral N-glycans which were synthesized on cellobiohydrolase I. Final proof of the formation of GlcNAcMan5GlcNAc2 was obtained by NMR analysis.
Maras
M
M
Department of Molecular Biology, Flanders Interuniversity Institute for Biotechnology and University of Gent, Belgium.
De Bruyn
A
A
Vervecken
W
W
Uusitalo
J
J
Penttilä
M
M
Busson
R
R
Herdewijn
P
P
Contreras
R
R
D002240:Carbohydrate Sequence; D002480:Cellulase / Q000235:genetics; D043366:Cellulose 1,4-beta-Cellobiosidase; D003001:Cloning, Molecular; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics / Q000378:metabolism*; D009844:Oligosaccharides / Q000737:chemistry; D011134:Polysaccharides / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000378:metabolism; D013172:Spores, Fungal; D014242:Trichoderma / Q000378:metabolism / Q000502:physiology*
D009844:Oligosaccharides; D011134:Polysaccharides; D011994:Recombinant Proteins; D017351:N-Acetylglucosaminyltransferases; C024850:alpha-1,3-mannosyl-glycoprotein beta-1,2-N-acetylglucosaminyltransferase I; D002480:Cellulase; D043366:Cellulose 1,4-beta-Cellobiosidase