This study explored the potential of caffeine being utilized as the fuel for the microbes to produce electrons for electricity generation in up-flow constructed wetland-microbial fuel cell (UFCW-MFC). The effect of caffeine concentration was investigated to identify the availability of UFCW-MFC in the conversion of caffeine to electrons for electricity production; and the effect of operating conditions (circuit connection, supplementary aeration, and plant) was studied to determine their significance in the treatment of caffeine containing wastewater. The UFCW-MFC achieved about 98% of decaffeination efficiency regardless of caffeine concentration; while a decrease of efficiency was observed when UFCW-MFC operated without supplementary aeration and plant (~93%). COD removal efficiency decreased correspondingly to the increase of caffeine concentration, which could be contributed by the higher concentration of caffeine and its intermediates. The degradation pathway of caffeine in UFCW-MFC was explored in this study. It was remarkable that ammonia was produced and converted to ammonium ions during caffeine catabolism. Supplementary aeration and macrophyte play a crucial role in removing excess caffeine, intermediates as well as accumulated ammonium ions. The toxicity assessment revealed that caffeine was degraded to less toxic products. The closed circuit connection not only contributed to electricity generation but also enhanced the caffeine and COD removal efficiency by 4.6 and 5.4% in the anaerobic region, respectively. The increase of voltage and maximum power density from phase I to phase IV indicated that caffeine could be converted to electrons by the anaerobes for electricity production.
