In this study, a portable and disposable paper-based microbial fuel cell (MFC) was fabricated. The MFC was powered by Rhodopseudomonas palustris bacteria (R. palustris). An activated carbon sheet-based anode pre-loaded organic matter (starch) and R. palustris was used. By using starch in the anode, R. palustris-loaded on the anode could be preserved for a long time in dry conditions. The MFC could generate electricity on-demand activated by adding water to the anode. The activated carbon sheet anode was treated by UV-ozone treatment to remove impurities and to improve its hydrophilicity before being loaded with R. palustris. The developed MFC could generate the maximum power density of 0.9 μW/cm2 and could be preserved for long-term usage with little performance degradation (10% after four weeks).