Fifteen eukaryotic microorganisms were tested for their ability to assimilate biodiesel derived waste glycerol and convert it into value-added metabolic products. For this purpose yeast and Zygomycetes strains were cultivated in nitrogen-limited raw glycerol-based media (initial glycerol concentration 30 g/L). Yeasts tested accumulated restricted lipid quantities (up to similar to 22%, wt/wt, in the case of Rhodotorula sp), while differentiations in their fatty acid composition were recorded in relation to the yeast strains employed and the fermentation time. On the contrary, fungi accumulated higher quantities of lipid inside their mycelia (ranging between 18.1 and 42.6%, wt/wt, of dry biomass) that contained in variable amounts the medically important CIA (gamma-linolenic acid). Moreover, Yarrowia lipolytica, Pichia membranifaciens and Thamnidium elegans were further studied in media having increased initial glycerol concentrations. In these conditions Y. lipolytica secreted significant amounts of acetic acid (29.2 g/L), as well as mannitol (19.4 g/L) while P. membramfaciens reached 28.4 g/L of biomass at glycerol concentration 90 g/L T elegans produced 11.6 g/L of oil, with 71.1%, wt/wt, of fat in biomass, while the maximum concentration of GLA was 371 mg/L Detailed analysis of T. elegans lipids indicated that the phospholipids fraction was particularly rich in polyunsaturated fatty acids. (C) 2010 Elsevier Ltd. All rights reserved.