The production of bioethanol was studied using recombinant Escherichia coil with glycerol as a carbon source. Glycerol is an attractive feedstock for biofuels production since it is generated as a major byproduct in biodiesel industry; therefore, we investigated the conversion of glycerol to bioethanol using E. coil BL21 (DE3) which harbors several genes in ethanol production pathway of Enterobacter aerogenes KCTC 2190. Fermentation was carried out at 34 degrees C for 42 h, pH 7.6, using defined production medium. Under optimal conditions, bioethanol production by the recombinant E. coil BL21 (DE3), strain pEB, was two-fold (3.01 g/L) greater than that (1.45 g/L) by the wild-type counterpart. The results obtained in this study will provide valuable guidelines for engineering bioethanol producers. (C) 2013 Elsevier Inc. All rights reserved.