Glycerol degradation with electricity production by a pure culture of Bacillus subtilis in a single-chamber air cathode of microbial fuel cell (MFC) has been demonstrated. Steady state polarization curves indicated a maximum power density of 0.06 mW/cm(2) with an optimal external resistance of 390 Omega Analysis of the effect of pH on MFC performance demonstrated that electricity generation was sustained over a long period of time under neutral to alkaline conditions. Cyclic voltammetry exhibited the increasing electrochemical activity with the increase of pH of 7, 8 and 9. Voltammetric studies also demonstrated that a two-electron transfer mechanism was occurring in the reactor. The low Coulombic efficiency of 23.08% could be attributed to the loss of electrons for various activities other than electricity generation. This study describes an application of glycerol that could contribute to transformation of the biodiesel industry to a more environmentally friendly microbial fuel cell-based technology. (C) 2010 Elsevier Ltd. All rights reserved.