The present report describes the isolation of oleaginous yeast, Pichia kudriavzevii MTCC 5493, from rotten
fruits and the production of biodiesel from the yeast oil. Optimization of physiological and nutritional
parameters was carried out for improved biomass production using cheap substrates such as crude glycerol,
yeast autolysate and corn steep liquor. Fed-batch fermentation yielded a dry biomass and oil yield of
up to 33 g L1 and 19% (w/w) respectively and was scaled up successfully to 26 L. Optimization of the C/N
ratio further increased the oil yield to 23% (w/w). The oil extraction process was also optimized using
environmentally safe solvents. The oil profile indicated a high oleic acid content followed by palmitic, linoleic
acid, and stearic acid. The presence of increased levels of saturated and monounsaturated fatty acids
in the oil make it ideal for biodiesel.