title: beta-Glycosyl azides as substrates for alpha-glycosynthases: preparation of efficient alpha-L-fucosynthases.
abstract: Fucose-containing oligosaccharides play a central role in physio-pathological events, and fucosylated oligosaccharides have interesting potential applications in biomedicine. No methods for the large-scale production of oligosaccharides are currently available, but the chemo-enzymatic approach is very promising. Glycosynthases, mutated glycosidases that synthesize oligosaccharides in high yields, have been demonstrated to be an interesting alternative. However, examples of glycosynthases available so far are restricted to a limited number of glycosidases families and to only one retaining alpha-glycosynthase. We show here that new mutants of two alpha-L-fucosidases are efficient alpha-L-fucosynthases. The approach shown utilized beta-L-fucopyranosyl azide as donor substrate leading to transglycosylation yields up to 91%. This is the first method exploiting a beta-glycosyl azide donor for alpha-glycosynthases; its applicability to the glycosynthetic methodology in a wider perspective is presented.
authors.affiliation: Institute of Protein Biochemistry - Consiglio Nazionale delle Ricerche, 80131 Naples, Italy.
authors.forename: Beatrice
authors.initials: B
authors.lastname: Cobucci-Ponzano
authors.forename: Fiorella
authors.initials: F
authors.lastname: Conte
authors.forename: Emiliano
authors.initials: E
authors.lastname: Bedini
authors.forename: Maria Michela
authors.initials: MM
authors.lastname: Corsaro
authors.forename: Michelangelo
authors.initials: M
authors.lastname: Parrilli
authors.forename: Gerlind
authors.initials: G
authors.lastname: Sulzenbacher
authors.forename: Alexandra
authors.initials: A
authors.lastname: Lipski
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Dal Piaz
authors.forename: Laura
authors.initials: L
authors.lastname: Lepore
authors.forename: Mosè
authors.initials: M
authors.lastname: Rossi
authors.forename: Marco
authors.initials: M
authors.lastname: Moracci
mesh_terms: D019943:Amino Acid Substitution; D001386:Azides / Q000737:chemistry* / Q000494:pharmacology; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D006031:Glycosylation; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D009844:Oligosaccharides / Q000096:biosynthesis; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D005644:alpha-L-Fucosidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D001386:Azides; D009844:Oligosaccharides; D011994:Recombinant Proteins; D005644:alpha-L-Fucosidase