With the aim of providing new methods for the regioselective protection at the 2,3 and 4 positions of monosaccharides, we have studied the acetylation of a class of rigid sugars: the 1,6-anhydro-β-d- andl-hexopyranoses (hexopyranosanesD-1toD-5andL-1toL-5), using vinyl acetate as an acyl donor and two common lipases,Candida rugosaandPseudomonas cepacia, as catalysts. Our results indicate that the relative orientation of the hydroxyls governs the regioselectivity of acetylation. In thed-series, when the 3-OH is in the axial position, acetylation occurs mainly at the 4-axial OH, while the 2-axial OH is preferred when the 4-OH is equatorial. Conversely, when the 3-OH is equatorial, a strong selectivity affects the equatorial 2-OH. Compounds of thel-series were shown to be poor substrates for the lipasePseudomonas cepaciaexcept forl-galactosane for which the 2-monoacetyl ester was obtained in good yield. An attempt to rationalize the results by means of molecular modelling is also made to account for the catalytic activity of theCandida rugosalipase on hexopyranosanes1–3.