High butanol accumulation is due to feedback inhibition which leads to the low butanol productivity observed in acetone-butanol-ethanol (ABE) fermentation. The aim of this study is to use biodiesel as an extractant for the in situ removal of butanol from the broth. The results indicate that adding biodiesel as an extractant at the beginning of fermentation significantly enhances butanol production. No significant toxicity of biodiesel on the growth of Clostridium acetobutylicum is observed. In the fed-batch operation with glucose feeding, the maximum total butanol obtained is 31.44 g/L, as compared to the control batch (without the addition of biodiesel) at 9.85 g/L. Moreover, the productivity obtained is 0.295 g/L h in the fed-batch, which is higher than that of 0.185 g/L h for the control batch. The in situ butanol removal by the addition of biodiesel has great potential for commercial ABE production. ? 2012 Elsevier Ltd. All rights reserved.