Enzymatic route to preparative-scale synthesis of UDP-GlcNAc/GalNAc, their analogues and GDP-fucose

Nat Protoc. 2010 Apr;5(4):636-46. doi: 10.1038/nprot.2010.3. Epub 2010 Mar 11.


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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Glycosylation
  • Glycosyltransferases / metabolism
  • Guanosine Diphosphate Fucose / biosynthesis*
  • Oligosaccharides / biosynthesis
  • Oligosaccharides / chemistry
  • Organic Chemistry Phenomena
  • Phosphorylation
  • Polysaccharides / biosynthesis
  • Polysaccharides / chemistry
  • Uridine Diphosphate N-Acetylgalactosamine / analogs & derivatives
  • Uridine Diphosphate N-Acetylgalactosamine / biosynthesis*
  • Uridine Diphosphate N-Acetylglucosamine / analogs & derivatives
  • Uridine Diphosphate N-Acetylglucosamine / biosynthesis*


  • Oligosaccharides
  • Polysaccharides
  • Guanosine Diphosphate Fucose
  • Uridine Diphosphate N-Acetylglucosamine
  • Uridine Diphosphate N-Acetylgalactosamine
  • Glycosyltransferases