The culture media for glycerol to ethanol biotransformation by Escherichia coli under anaerobic conditions was optimized. A Plackett-Burman screening design was used to determine which factors were significant with a 95% confidence interval. A full factorial 2(2) and response surface model were employed to determine the optimum conditions for the selected response variable. The response variable considered in this work was biomass productivity (Q(x)). Profiles for biomass growth, glycerol consumption and ethanol production were obtained at optimum conditions and fermentation parameters were calculated. Glycerol to ethanol yield and ethanol specific productivity were determined to be 845 g kg (1) of glycerol and 212 g kg (1) h (1) of cell mass respectively. Optimized culture media is presented along with the main results and experimental profiles obtained from this work. (C) 2011 Elsevier Ltd. All rights reserved.