Single-cell oil from M. circinelloides was analyzed as a potential feedstock for biodiesel production. The total oil obtained was 44% of dried biomass after 96 h of cultivation. Microwave-assisted eth- anol extraction is a fast and sustainable method for recovering high lipid concentrations from fungi. The lipid profile of M. circinelloides showed high concentrations of saturated and monounsaturated fatty acids, predominantly C16:0 and C18:1. The enzymatic transe- sterification of the single-cell oil with ethanol achieved yields of ethyl esters of 93%. The transesterification of the single-cell oil from M. circinelloides is a suitable approach to completely fulfill the standard specifications for renewable diesel.