High lipids accumulation in Rhodosporidium toruloides by applying single and multiple nutrients limitation in a simple chemically defined medium

Abstract

Microbial oil produced by the oleaginous yeast Rhodosporidium toruloides ATCC 204091 (formerly referred to as Rhodotorula glutinis) has a similar fatty acid composition to the vegetable oils and represents a potential alternative for biodiesel production. Finding strategies to improve the oil production by this yeast is desirable, as it is one of this nutrient’s limitations during the accumulation phase, as well as one of the main factors influencing the process. Therefore, the effect of single or combined nutrient limitation on lipid accumulation by R. toruloides was investigated. Biomass production and lipid accumulation by R. toruloides was improved using experimental designs in a two-step batch culture on a chemically-defined culture medium with high initial glucose concentration. For the first culture step, a Box–Behnken design was applied to optimize the main medium components’ concentrations, while maintaining a high biomass production. A biomass concentration of 44.3 g/L was reached with a medium composed of (g/L): glucose, 100; KH2PO4, 4.6; NaNO3, 13.4; MgSO4 .7H2O, 0.2; and CaCl2 .2H2O, 0.11. For the second culture step, the biomass was transferred to lipid accumulation media. A 23 factorial experimental design was conducted to investigate the effect of N, P and S limitations (individually or jointly) on lipid production from glucose (100 g/L). Lipid accumulation on dry cell mass was 77.04, 65.42, 70.13 and 69.84% for N, P, S and simultaneous nutrients’ limitations, respectively.