In this study, Y. lipolytica NBRC 1658 and a novel domestic strain Y. lipolytica 57 were used for citric acid production from two different substrates; glycerol and mannitol, in a batch system. In each substrate media, the growth was defined by the non-competitive substrate inhibition model for both of the strains. Maximum citric acid concentration obtained by the domestic strain was higher than that of NBRC 1658 in glycerol medium. The best results for productivity and maximum specific citric acid production rate were obtained at 120 g/L initial glycerol concentration for NBRC 1658, and at 160 g/L initial glycerol concentration for the domestic strain. In the medium containing mannitol, maximum citric acid concentration and productivity were obtained at 120 g/L initial mannitol concentration for both of the strains. The study gave promising data for gaining a novel citric acid producer, Y. lipolytica 57, determining substrate profiles of the used strains for citric acid production.