title: Endo-β-N-acetylglucosaminidase catalysed glycosylation: tolerance of enzymes to structural variation of the glycosyl amino acid acceptor.
abstract: Endo-β-N-Acetylglucosaminidases (ENGases) are highly useful biocatalysts that can be used to synthetically access a wide variety of defined homogenous N-linked glycoconjugates in a convergent manner. The synthetic efficiency of a selection of family GH85 ENGases was investigated as the structure of the acceptor substrate was varied. Several different GlcNAc-asparagine acceptors were synthesised, and used in conjunction with penta- and decasaccharide oxazoline donors. Different enzymes showed different tolerances of modification of the GlcNAc acceptor. Whilst none tolerated modification of either the 4- or 6-hydroxyl, both Endo M and Endo D tolerated modification of OH-3. For Endo D the achievable synthetic efficiency was increased by a factor of three by the use a 3-O-benzyl protected acceptor. The presence of a fucose at position-3 was not tolerated by any of the enzymes assayed.
authors.affiliation: Department of Chemistry, University of Canterbury, Private Bag 4800, Christchurch, 8140, New Zealand. antony.fairbanks@canterbury.ac.nz.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Tomabechi
authors.forename: Marie A
authors.initials: MA
authors.lastname: Squire
authors.forename: Antony J
authors.initials: AJ
authors.lastname: Fairbanks
mesh_terms: D000596:Amino Acids / Q000096:biosynthesis* / Q000737:chemistry; D055162:Biocatalysis; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D006027:Glycosides / Q000096:biosynthesis* / Q000737:chemistry; D006031:Glycosylation; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / Q000378:metabolism*; D008969:Molecular Sequence Data
chemical_list: D000596:Amino Acids; D006027:Glycosides; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase