Enzymatic route to preparative-scale synthesis of UDP-GlcNAc/GalNAc, their analogues and GDP-fucose.
Enzymatic synthesis using glycosyltransferases is a powerful approach to building polysaccharides with high efficiency and selectivity. Sugar nucleotides are fundamental donor molecules in enzymatic glycosylation reactions by Leloir-type glycosyltransferases. The applications of these donors are restricted by their limited availability. In this protocol, N-acetylglucosamine (GlcNAc)/N-acetylgalactosamine (GalNAc) are phosphorylated by N-acetylhexosamine 1-kinase (NahK) and subsequently pyrophosphorylated by N-acetylglucosamine uridyltransferase (GlmU) to give UDP-GlcNAc/GalNAc. Other UDP-GlcNAc/GalNAc analogues can also be prepared depending on the tolerance of these enzymes to the modified sugar substrates. Starting from L-fucose, GDP-fucose is constructed by one bifunctional enzyme L-fucose pyrophosphorylase (FKP) via two reactions.
Zhao
Guohui
G
Department of Chemistry, The Ohio State University, Columbus, Ohio, USA.
Guan
Wanyi
W
Cai
Li
L
Wang
Peng George
PG
D006031:Glycosylation; D016695:Glycosyltransferases / Q000378:metabolism; D006154:Guanosine Diphosphate Fucose / Q000096:biosynthesis*; D009844:Oligosaccharides / Q000096:biosynthesis / Q000737:chemistry; D055601:Organic Chemistry Phenomena; D010766:Phosphorylation; D011134:Polysaccharides / Q000096:biosynthesis / Q000737:chemistry; D014536:Uridine Diphosphate N-Acetylgalactosamine / Q000031:analogs & derivatives / Q000096:biosynthesis*; D014537:Uridine Diphosphate N-Acetylglucosamine / Q000031:analogs & derivatives / Q000096:biosynthesis*
D009844:Oligosaccharides; D011134:Polysaccharides; D006154:Guanosine Diphosphate Fucose; D014537:Uridine Diphosphate N-Acetylglucosamine; D014536:Uridine Diphosphate N-Acetylgalactosamine; D016695:Glycosyltransferases