Specificity of Donor Structures for endo-β-N-Acetylglucosaminidase-Catalyzed Transglycosylation Reactions

To demonstrate the structural specificity of the glycosyl donor for the transglycosylation reaction by using endo-β-N-acetylglucosaminidase from Mucor hiemalis (endo-M), a series of tetrasaccharide oxazoline derivatives was synthesized. These derivatives correspond to the core structure of an asparagine-linked glycoprotein glycan with a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety. The transglycosylation activity of wildtype (WT) endo-M and two mutants, N175Q and N175A, was examined by using these tetrasaccharide donors with p-nitrophenyl N-acetylglucosaminide (GlcNAc-pNp). The essential configuration of the hydroxy group for the transglycosylation reaction was determined. On the basis of these results, the transglycosylation reaction was investigated by using chemically modified donors, and transglycosylated products were successfully obtained.