The nano-biocomposite electrodes composed of carbon nanotube (CNT), polypyrrole (PPy), and E. coli-bacteria were investigated for electrochemical supercapacitors. For this purpose, PPy/CNT–E. coli was successfully synthesized through oxidative polymerization. The PPy/CNT–E. coli electrode exhibited a high specific capacitance of 173 F∙g−1 at the current density of 0.2 A∙g−1, which is much higher than that (37 F∙g−1) of CNT. Furthermore, it displayed sufficient stability after 1000 charge/discharge cycles. The CNT, PPy/CNT, and PPy/CNT–E. coli composites were characterized by x-ray diffraction, scanning electron microscopy, and surface analyzer (Brunauer–Emmett–Teller, BET). In particular, the pyrrole monomers were easily adsorbed and polymerized on the surface of CNT materials, as well as E. coli bacteria enhanced the surface area and porous structure of the PPy/CNT–E. coli composite electrode resulting in high performance of devices.