Microbial electrolysis is a new technology for the production of value-added products, such as gaseous biofuels, from waste organic substrates. This study describes the performance of a methane-producing microbial electrolysis cell (MEC) operated at ambient temperature with a Geobacter sulfurreducens microbial bioanode and a methanogenic microbial biocathode. The cell was initially operated at a controlled cathode potential of -850 mV (vs. standard hydrogen electrode, SHE) in order to develop a methanogenic biofilm capable of reducing carbon dioxide to methane gas using abiotically produced hydrogen gas or directly the polarized electrode as electron donors. Subsequently, G. sulfurreducens was inoculated at the anode and the MEC was operated at a controlled anode potential of +500 mV, with acetate serving as electron donor. The rate of methane production at the cathode was found to be primarily limited by the acetate oxidation kinetics and in turn by G. sulfurreducens concentration at the anode of the MEC. Temperature had also a main impact on acetate oxidation kinetics, with an apparent activation energy of 58.1 kJ mol(-1). (C) 2011 Elsevier B.V. All rights reserved.