The total amount of lipids produced in Rhodotorula glutinis is a subject which has attracted increasing attention due to the potential biodiesel conversion from these microbial oils. The effects of the dissolved oxygen (DO) level in lipid accumulation were examined in this study. Variations of different medium volumes (30, 40 and 50 ml) and shaking speed (60, 150 and 210 rpm) in the flask trials were adopted to explore the DO effects on lipid production. All of the results revealed that a low DO could retard cell growth, while enhancing lipid accumulation. The 5 l-fermentor results also confirm that a low DO (25 +/- 10%) batch could have higher lipid content than that of high DO batch (60 +/- 10%). Nevertheless, the DO level would not obviously affect the lipid composition profile. Oleic acid (C18:1) was the primary fatty acid in both batches. Due to the slow biomass growth rate resulting from the low DO, a two-stage DO controlled strategy (consisting of a high DO stage and following a low DO stage) was performed to improve the cell growth and lipid accumulation simultaneously. However, the strategy was not successful on the enhancement of total lipid production as compared to other batches. Conclusively, even a low DO could retard cell growth: the total production of lipids in the batch with low DO was higher that of the high DO batch due to the enhancement of lipid accumulation. Therefore, the batch operation of R. glutinis at the low DO was suggested for the purpose of lipid production. (C) 2011, The Society for Biotechnology, Japan. All rights reserved.