This study presents an enzymatic bioanode fabricated with fructose dehydrogenase and a polyaniline film to construct a single-compartment fructose biofuel cell. The enzymatic bioanode provided fructose oxidation current, which accompanied the electron transfer between the heme c moiety of fructose dehydrogenase and polyaniline. Characterization of the bioanode at a pH of 4.5 indicated an onset potential of -0.1 V (vs. Ag/AgCl) with respect to the redox potential corresponding to heme c of fructose dehydrogenase as well as high current densities for fructose oxidation of 1.0 +/- 0.1 mA/cm(2) at + 0.50 V (vs. Ag/AgCl). A single compartment fructose biofuel cell was constructed by use of the bioanode together with an enzymatic biocathode fabricated with laccase and polythiophene copolymer film. The fructose biofuel cell possessed an open circuit potential of 0.55 V with an associated short-circuit current of 1.4 +/- 0.2 mA/cm(2). In addition, the maximum power density of the biofuel cell was 0.36 +/- 0.04 mW/cm(2) at a cell voltage of 0.3 V.