Chlorella protothecoides, a lipid-producing microalga, was grown heterotrophically and autotrophically in separate reactors, the off-gases exiting the former being used to aerate the latter. Autotrophic biomass productivity with the two-reactor association, 0.0249 g L-1 h(-1), was 2.2-fold the value obtained in a control autotrophic culture, aerated with ambient air. Fatty acid productivity was 1.7-fold the control value. C. protothecoides heterotrophic biomass productivity was 0.229 g L-1 h(-1). This biomass' fatty acid content was 34.5% (w/w) with a profile suitable for biodiesel production, according to European Standards. The carbon dioxide fixed by the autotrophic biomass was 45 mg CO2 L-1 h(-1) in the symbiotic arrangement, 2.1 times the control reactor value. The avoided CO2 atmospheric emission represented 30% of the CO2 produced in the heterotrophic stage, while the released O-2 represented 49% of the oxygen demand in that stage. Thus, an increased efficiency in the glucose carbon source use and a higher environmental sustainability were achieved in microalgal biodiesel production using the proposed assembly. (C) 2014 Elsevier B.V. All rights reserved.