Functional expression of Escherichia coli enzymes synthesizing GDP-L-fucose from inherent GDP-D-mannose in Saccharomyces cerevisiae

Glycobiology. 2000 Oct;10(10):1041-7. doi: 10.1093/glycob/10.10.1041.


Fucosylation of glycans on glycoproteins and -lipids requires the enzymatic activity of relevant fucosyltransferases and GDP-L-fucose as the donor. Due to the biological importance of fucosylated glycans, a readily accessible source of GDP-L-fucose would be required. Here we describe the construction of a stable recombinant S.cerevisiae strain expressing the E.coli genes gmd and wcaG encoding the two enzymes, GDP-mannose-4,6-dehydratase (GMD) and GDP-4-keto-6-deoxy-D-mannose-3,5-epimerase/4-reductase (GMER(FX)) respectively, needed to convert GDP-mannose to GDP-fucose via the de novo pathway. Taking advantage of the rich inherent cytosolic GDP-mannose pool in S.cerevisiae cells we could easily produce 0.2 mg/l of GDP-L-fucose with this recombinant yeast strain without addition of any external GDP-mannose. The GDP-L-fucose product could be used as the fucose donor for alpha1,3fucosyltransferase to synthesize sialyl Lewis x (sLex), a glycan crucial for the selectin-dependent leukocyte traffic.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbohydrate Epimerases / genetics
  • Carbohydrate Epimerases / metabolism
  • Escherichia coli / enzymology*
  • Escherichia coli Proteins*
  • Genes, Bacterial
  • Genetic Engineering / methods
  • Guanosine Diphosphate Fucose / biosynthesis*
  • Guanosine Diphosphate Fucose / chemistry
  • Guanosine Diphosphate Mannose / metabolism*
  • Hydro-Lyases / genetics
  • Hydro-Lyases / metabolism
  • Ketone Oxidoreductases*
  • Multienzyme Complexes*
  • Recombinant Proteins / metabolism
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Stereoisomerism
  • Sugar Alcohol Dehydrogenases / genetics
  • Sugar Alcohol Dehydrogenases / metabolism
  • Transformation, Genetic


  • Escherichia coli Proteins
  • Multienzyme Complexes
  • Recombinant Proteins
  • wcaG protein, E coli
  • Guanosine Diphosphate Fucose
  • Guanosine Diphosphate Mannose
  • Sugar Alcohol Dehydrogenases
  • Ketone Oxidoreductases
  • Hydro-Lyases
  • GDPmannose 4,6-dehydratase
  • Carbohydrate Epimerases