The optimization of biomass and oil productivities in heterotrophic cultures of Auxenochlorella protothecoides was achieved using a non-linear model-based approach. A 10-fold increase in the average biomass productivity, and a 16-fold increase in the maximum productivity, was observed with respect to batch cultures as a result of the proposed optimization strategy. Final cell density in the optimized culture was 144 g/L (dry weight), with 49.4%w/w oil content. Maximum lipid productivity was 20.16 g/L d, achieved during the exponential growth phase at an average cell density of 86 g/L. Lipid productivity in the optimized microalgal culture was higher than previously reported values for other oleaginous microorganisms. Oil composition analysis showed that the oil has a high quality as biodiesel precursor. The higher productivity and excellent lipid profile of the optimized microalgal culture make A. protothecoides an exceptional source for biodiesel production and a potential source of single cell oil for other applications. (C) 2011 Elsevier Ltd. All rights reserved.