D-Glucosamine is an important building block of major structural components of the fungal cell wall, namely chitin, chitosan and mannoproteins. Other amino sugars, such as D-mannosamine and D-galactosamine, relatively abundant in higher eukaryotes, rarely occur in fungal cells and are actually absent from yeast and yeast-like fungi. The glucosamine-containing sugar nucleotide UDP-GlcNAc is synthesized in yeast cells in a four-step cytoplasmic pathway. This article provides a comprehensive overview of the present knowledge on the enzymes catalysing the particular steps of the pathway in Candida albicans and Saccharomyces cerevisiae, with a special emphasis put on mechanisms of the catalysed reactions, regulation of activity and perspectives for exploitation of enzymes participating in UDP-GlcNAc biosynthesis as potential targets for antifungal chemotherapy. Copyright © 2006 John Wiley & Sons, Ltd.