Solid-phase oligosaccharide and glycopeptide synthesis using glycosynthases.
Enzymatic approaches for the preparation of oligosaccharides are interesting alternatives to traditional chemical synthesis, the main advantage being the regio- and stereoselectivity offered without the need for protecting groups. The use of solid-phase techniques offers easy workup procedures and the prospect of automatability. Here, we report the first application of glycosynthases to solid-phase oligosaccharide synthesis by use of the 51 kDa serine and glycine mutants of Agrobacterium sp. beta-glucosidase, Abg E358S and E358G. Acceptors were linked to PEGA resin through a backbone amide linker (BAL), and using these mutated enzymes, a galactose moiety was transferred from a donor sugar, alpha-D-galactosyl fluoride, with high efficiency (>90%) together with excellent recovery of material. Furthermore, it was demonstrated that a resin-bound model glycopeptide was also an acceptor for the glycosynthase.
Tolborg
Jakob F
JF
Department of Chemistry, Building 201, Kemitorvet, Technical University of Denmark, DK-2800, Kgs. Lyngby, Denmark.
Petersen
Lars
L
Jensen
Knud J
KJ
Mayer
Christoph
C
Jakeman
David L
DL
Warren
R Antony J
RA
Withers
Stephen G
SG
D000107:Acetylation; D000586:Amination; D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D005998:Glycine / Q000737:chemistry; D006020:Glycopeptides / Q000138:chemical synthesis*; D006031:Glycosylation; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D015394:Molecular Structure; D009154:Mutation; D009844:Oligosaccharides / Q000138:chemical synthesis*; D010455:Peptides / Q000138:chemical synthesis; D012116:Resins, Plant / Q000138:chemical synthesis; D012231:Rhizobium / Q000201:enzymology*; D012694:Serine / Q000737:chemistry; D001617:beta-Glucosidase / Q000378:metabolism*
D006020:Glycopeptides; D009844:Oligosaccharides; D010455:Peptides; D012116:Resins, Plant; D012694:Serine; D001617:beta-Glucosidase; D005998:Glycine