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Oleochemical activities (e.g. biodiesel production, fat saponification) generate annually very high amounts of concentrated glycerol‐containing waters (called crude glycerol) as the principal residues of these processes. Crude glycerol is an industrial residue the valorization of which attracts remarkable and constantly increasing interest. In the current investigation, biodiesel‐derived glycerol was employed as substrate for yeast and fungal strains cultivated under nitrogen‐limited conditions in shake flasks. Glucose was employed as reference substrate. Several yeasts (Candida diddensiae, Candida tropicalis, Pichia ciferrii, Williopsis saturnus, Candida boidinii, and Candida oleophila) rapidly assimilated glucose and converted it into ethanol, despite aerobic conditions imposed, and were Crabtree‐positive. None of these yeasts produced ethanol during growth on glycerol or accumulated significant quantities of lipid …