In this study, nine heterotrophic yeast isolates were tested for their ability to assimilate crude glycerol and consecutive conversion to triacylglycerides (TGAs). All the organisms were initially screened on crude glycerol-based selection media, and those producing lipid globules were further evaluated for lipid production. Sudan Black B staining of eight isolates showed lipid globules. These strains were further studied at different C/N ratio. The molecular identification revealed that the isolates belonged to the genera of Yarrowia and Candida. Among these isolates, SKY7 (Yarrowia lipolytica) produced up to 42.04 ± 0.11 % of lipid w/w) with a C/N ratio of 100 and fermentation time of 72 h. The other strains produced 5.82 ± 0.4 to 34.57 ± 0.44 % lipid (w/w). The GC-flame ionization detector (FID) lipid profile showed that the lipid produced by the strains had close resemblance with vegetable oil and could serve as a feedstock for biodiesel production. Biolog test of the isolates revealed a wide spectrum of carbon utilization.