The influence of the organic loading rate (also known as active anodic chamber volume) on bioelectricity generation in a continuous, two-chamber microbial fuel cell for the treatment of synthetic wastewater, with glucose as the only carbon source, was examined. Ten sets of experiments with different combinations of hydraulic retention times (0.24-1.14 d) and influent chemical oxygen demand concentrations were performed to verify the impact of organic loading rate on the voltage generation capacity of a simple dual-chamber microbial fuel cell working in continuous mode. We found that there is an optimal hydraulic retention time value at which the maximum voltage is generated: 0.41 d. However, there were no similar effects, in terms of voltage generation, when a constant hydraulic retention time with different influent chemical oxygen demand of wastewater was used. The obtained maximal voltage value (600 mV) has also been compared to literature data. Computational fluid dynamics (CFD) was used to calculate the fluid flow and the exit age distribution of fluid elements in the reactor to explain the obtained experimental results and identify the crucial parameters for the design of bioreactors on an industrial scale.