Abstract Objective To compare physiological and process parameters, as well as lipid accumulation capacity, of six strains of Yarrowia lipolytica and two strains of Rhodosporidium toruloides in media containing glycerol as the main carbon and energy source. Results The strains Y. lipolytica IMUFRJ 50678, Po1g, W29 and CCT 5443 displayed very similar physiological parameters, with µ max , and average values of 0.31 h −1 , 0.53 (g DW/g S), 0.60 (g S/g DW.h) and 0.46 (g DW/g S), respectively. The two strains of R. toruloides presented physiological and process parameters with no significant difference, with average values of 0.084 h −1 , 0.53 (g DW/g S), 0.17 (g S/g DW.h) and 0.44 (g DW/g S). Among all Y. lipolytica strains, Y. lipolytica CCT 5443 strain presented the highest Y Lip/S , 0.054 (g Lip/g S), and P Lip of 0.040 (g Lip/l.h). Among all investigated strains ( Y. lipolytica and R. toruloides ), the yeast R. toruloides CCT 7815 displayed the highest lipid accumulation capacity, with Y Lip/S equal to 0.11 (g Lip/g S) and P Lip equal to 0.10 (g Lip/l.h). Conclusion Among all strains investigated in our study, the yeast strain R. toruloides CCT 7815 presents the most promising characteristics for industrial single cell oil production. List of abbreviations DW dry weight (g/l) Lip lipid (g/l) P Lip lipid productivity (g Lip/l.h) maximum specific substrate consumption rate (g S/g DW.h) S substrate (g/l) Y Lip/S lipid yield on substrate (g Lip/g S) biomass yield on substrate during the exponential growth phase (g DW/g S) global biomass yield on substrate (g DW/g S) µ max maximum specific growth rate (h −1 )