Environmentally friendly alternative energy sources are needed due to the global energy crises and to mitigate the environmental issues related to fossil fuels. Among alternative energy sources, biodiesel has several advantages, especially when microbial lipids are used as biodiesel raw materials. Efforts to reduce microbial lipid production costs are currently being made. Candida lipolytica, an oleaginous yeast, was grown in nitrogen-limited media. Biomass production, lipid yield, and lipid content were evaluated. The impacts of inoculum size, pH, temperature, agitation speed, and incubation time, as well as variations in glucose concentration and different nitrogen sources were investigated. The yeast was grown in two-stage fermentation by combining the different optimized parameters together. In the first stage of fermentation, the lipid content was 29.35 � 0.5%, whereas in the second stage, it was 41.6 � 0.1% (on Day 10). Although the use of only glucose in the second stage increased the lipid content by 32%, this approach limited the cost-efficiency. In this study, to reduce the raw material cost and achieve a high lipid yield from a yeast that can eventually be used to produce biofuels, expired and rejected soft drinks (ERSD) and chey whey were used. The maximum lipid content (56.6 %) and lipid yield (88 %)were obtained with using only ERSD as carbon-rich medium. In addition, the use of ERSD instead of glucose in the second stage was cost-effective. Chemical oxygen demand (COD) was decreased by 48 %, indicating that C. lipolytica could effectively remove COD from the mixtures.
The dominant fatty acids, in order of their relative abundance (%w/w), were oleic, palmitic, linoleic, and stearic acids. Biodiesel production and wastewater treatment were performed simultaneously with high efficiency.