We explore for the first time the ability of a three-dimensional polyacrylonitrile/gold material prepared by a low-cost and scalable synthesis method, based on the combination of electrospinning and sputtering, as a new material with large surface area to provide high loadings of enzymes to enhance the electrochemical performances of enzyme electrodes in biofuel cells (BFCs). An ethanol/O-2 BFC has been developed based on enzymatic reactions performed at both the cathode and anode with immobilization of the respective enzymes and mediators on the three-dimensional nanostructured electrodes. The power density delivered is 1.6 mW cm(-2) at 0.75 V, which is five times the power density delivered by the BFC built on flat bioelectrodes. The greatly improved performance of these synthesized nanostructured electrodes makes them exciting materials for their implantation in biofuel cell applications.