title: Chemoenzymatic Synthesis of a Type 2 Blood Group A Tetrasaccharide and Development of High-throughput Assays Enables a Platform for Screening Blood Group Antigen-cleaving Enzymes.
abstract: A facile enzymatic synthesis of the methylumbelliferyl β-glycoside of the type 2 A blood group tetrasaccharide in good yields is reported. Using this compound, we developed highly sensitive fluorescence-based high-throughput assays for both endo-β-galactosidase and α-N-acetylgalactosaminidase activity specific for the oligosaccharide structure of the blood group A antigen. We further demonstrate the potential to use this assay to screen the expressed gene products of metagenomic libraries in the search for efficient blood group antigen-cleaving enzymes.
authors.affiliation: Centre for High-Throughput Biology Department of Chemistry, University of British Columbia, Vancouver, BC, Canada V6T 1Z1.
authors.forename: David H
authors.initials: DH
authors.lastname: Kwan
authors.affiliation: Institute of Pathology Helmholtz Institute for Biomedical Engineering, Biointerface Group, RWTH Aachen University, Pauwelsstraße 30, 52074 Aachen, Germany.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Ernst
authors.affiliation: Department of Chemistry, University of British Columbia, Vancouver, BC, Canada V6T 1Z1.
authors.forename: Miriam P
authors.initials: MP
authors.lastname: Kötzler
authors.affiliation: Centre for High-Throughput Biology Department of Chemistry, University of British Columbia, Vancouver, BC, Canada V6T 1Z1 withers@chem.ubc.ca.
authors.forename: Stephen G
authors.initials: SG
authors.lastname: Withers
mesh_terms: D000017:ABO Blood-Group System / Q000737:chemistry* / Q000378:metabolism; D001681:Biological Assay; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005453:Fluorescence; D015870:Gene Expression; D015723:Gene Library; D006027:Glycosides / Q000096:biosynthesis / Q000138:chemical synthesis*; D057166:High-Throughput Screening Assays; D006801:Humans; D006923:Hymecromone / Q000138:chemical synthesis* / Q000378:metabolism; D054892:Metagenome; D009844:Oligosaccharides / Q000096:biosynthesis / Q000138:chemical synthesis*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D048809:alpha-N-Acetylgalactosaminidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001616:beta-Galactosidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D000017:ABO Blood-Group System; D006027:Glycosides; D009844:Oligosaccharides; D011994:Recombinant Proteins; D006923:Hymecromone; D001616:beta-Galactosidase; D048809:alpha-N-Acetylgalactosaminidase
keywords: blood group antigen; enzymatic oligosaccharide synthesis; glycosyltransferase; metagenomics