Taxonomic group: fungi / Ascomycota
(Phylum: Ascomycota)
The structure was elucidated in this paperNCBI PubMed ID: 34939126Publication DOI: 10.1093/glycob/cwab127Journal NLM ID: 9104124Publisher: IRL Press at Oxford University Press
Correspondence: K. Jensen <khjn
novozymes.com>
Institutions: Department of Science and Environment, Roskilde University, Universitetsvej 1, Building 28, Roskilde 4000, Denmark, Novozymes A/S, Biologiens Vej 2, Kongens Lyngby 2800, Denmark, Complex Carbohydrate Research Center, 315 Riverbend Rd. University of Georgia, Athens, GA, USA USA, Department of Biotechnology and Biomedicine, Technical University of Denmark, Building 224, Kongens Lyngby 2800, Denmark
Glycoengineering ultimately allows control over glycosylation patterns to generate new glycoprotein variants with desired properties. A common challenge is glycan heterogeneity, which may affect protein function and limit the use of key techniques such as mass spectrometry. Moreover, heterologous protein expression can introduce nonnative glycan chains that may not fulfill the requirement for therapeutic proteins. One strategy to address these challenges is partial trimming or complete removal of glycan chains, which can be obtained through selective application of exoglycosidases. Here, we demonstrate an enzymatic O-deglycosylation toolbox of a GH92 α-1,2-mannosidase from Neobacillus novalis, a GH2 β-galactofuranosidase from Amesia atrobrunnea and the jack bean α-mannosidase. The extent of enzymatic O-deglycosylation was mapped against a full glycosyl linkage analysis of the O-glycosylated linker of cellobiohydrolase I from Trichoderma reesei (TrCel7A). Furthermore, the influence of deglycosylation on TrCel7A functionality was evaluated by kinetic characterization of native and O-deglycosylated forms of TrCel7A. This study expands structural knowledge on fungal O-glycosylation and presents a ready-to-use enzymatic approach for controlled O-glycan engineering in glycoproteins expressed in filamentous fungi.
O-glycosylation, glycoside hydrolase, Aspergillus oryzae, cellobiohydrolase, fungal glycoproteins
Structure type: oligomer
Location inside paper: Fig. 4A
Aglycon: (1->3)Ser/Thr
Trivial name: O-glycan
Compound class: glycoprotein
Contained glycoepitopes: IEDB_130701,IEDB_136104,IEDB_137485,IEDB_143632,IEDB_144983,IEDB_152206,IEDB_983930,SB_136,SB_196,SB_44,SB_67,SB_72
Methods: methylation, SDS-PAGE, kinetics assays, enzyme assay, HPAEC-PAD, LC-MS, cloning, glycoengineering, site-directed mutagenesis, expression, purification, reductive β-elimination
Enzymes that release or process the structure: cellobiohydrolase from Trichoderma reesei (TrCel7A)
Comments, role: Possible O-glycan structures from TrCel7A expressed in A.oryzae; major glycan structure 81% DP2
Related record ID(s): 51766, 51767, 51768, 51769, 51770
NCBI Taxonomy refs (TaxIDs): 5062
Show glycosyltransferases
There is only one chemically distinct structure:
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