ORIGINAL ARTICLE High lipids accumulation in Rhodosporidium toruloides by applying single and multiple nutrients limitation in a simple chemically defined medium Yolanda González-García 1 & L. Melisa Rábago-Panduro 2 & Todd French 3 & David Isidoro Camacho-Córdova 2 & Porfirio Gutiérrez-González 4 & Jesús Córdova 2 Received: 8 February 2017 /Accepted: 27 June 2017 /Published online: 18 July 2017 # Springer-Verlag GmbH Germany and the University of Milan 2017 Abstract Microbial oil produced by the oleaginous yeast Rhodosporidium toruloides ATCC 204091 (formerly referred to as Rhodotorula glutinis) has a similar fatty acid composi- tion 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 nutri- ent’ 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 accu- mulation by R. toruloides was investigated. Biomass produc- tion 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 de- sign 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 me- dium composed of (g/L): glucose, 100; KH 2 PO 4 , 4.6; NaNO 3 , 13.4; MgSO 4 . 7H 2 O, 0.2; and CaCl 2 . 2H 2 O, 0.11. For the sec- ond culture step, the biomass was transferred to lipid accumu- lation media. A 2 3 factorial experimental design was conduct- ed to investigate the effect of N, P and S limitations (individ- ually 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’ limita- tions, respectively. Keywords Microbial oil . Nutrient limited medium . Oleaginous yeast . Rhodosporidium Electronic supplementary material The online version of this article (doi:10.1007/s13213-017-1282-2) contains supplementary material, which is available to authorized users. * Yolanda González-García yolacea@yahoo.com L. Melisa Rábago-Panduro meli_rplm@hotmail.com Todd French french@che.msstate.edu David Isidoro Camacho-Córdova davidicaco@yahoo.com.mx Porfirio Gutiérrez-González pgutierrezglez@gmail.com Jesús Córdova jesuscordovaudg@yahoo.com.mx 1 Departamento de Madera, Celulosa y Papel, Universidad de Guadalajara, Km 15.5 Carretera Guadalajara-Nogales, 45010 Zapopan, Jalisco, Mexico 2 Departamento de Química, Universidad de Guadalajara, Blvd. Marcelino García Barragán 1421, 44430 Guadalajara, Jalisco, Mexico 3 Dave C. Swalm School of Chemical Engineering, Mississippi State University, Starkville, MS 39762, USA 4 Departamento de Matemáticas, Universidad de Guadalajara, Blvd. Marcelino García Barragán 1421, 44430 Guadalajara, Jalisco, Mexico Ann Microbiol (2017) 67:519–527 DOI 10.1007/s13213-017-1282-2