Poly(3-thiopheneacetic acid-co-3-methylthiophene) conductive polymer was electrosynthesized with ferrocene and used for an enzymatic fuel cell including glucose oxidase and bilirubin oxidase enzymes. The system was operated in a single-compartment and membrane-less cell by using glucose as fuel. Detailed optimization ensured to achieve considerable power output to generate sustainable energy from municipal wastewater as a renewable fuel source. Maximum power density of 1 mu W/cm(2) was generated at a cell voltage of +0.56 V in 100 mM, pH 7.4 phosphate buffer with the addition of 10 mM synthetic glucose. The working electrodes could harvest glucose readily found in the municipal wastewater of Zonguldak City in Turkey by generating a power density of 4 mu W/cm(2) for the municipal wastewater sample. In this way, the organic pollutants in wastewater could be evaluated by converting them into the electrical energy using an enzymatic fuel cell for the first time. (C) 2015 American Institute of Chemical Engineers Environ Prog, 35: 859-866, 2016