Ozone-based processes gained much attention in recent years. However, due to low oxidative stability and utilization rate, single ozonation process (SOP) is insufficient for complete mineralization of pollutants. As a result, the single ozonation process is performed in the presence of a catalyst, a process known as catalytic ozonation process (COP). A promising catalyst (Fe/BC) was prepared by impregnating iron on biochar surface to remove methylene blue from aqueous solution via heterogeneous catalytic ozonation process (HCOP). The prepared Fe/BC features were characterized using Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM) and Brunauer–Emmett–Teller method (BET) before and after HCOP. Furthermore, the effect of various operating parameters such as ozone dose, catalyst dose, initial dye concentration, initial pH on the efficiency of SOP and HCOP were compared. In comparison to single ozonation process, the experimental study found that heterogeneous catalytic ozonation process has the highest efficiency. At pH 7.0, approximately 76% of methylene blue is removed during single ozonation process in 60 min. Heterogeneous catalytic ozonation process showed 95% methylene blue elimination from aqueous solution. The efficiency of heterogeneous catalytic ozonation process was decreased by 52% in the presence of hydroxyl radical (●OH) scavenger, indicating that hydroxyl is the major oxidant during heterogeneous catalytic ozonation process for the removal of methylene blue from aqueous solution. Fe/BC catalyst appears to have a lot of industrial promise, as well as the ability to remove methylene blue from aqueous solution via heterogeneous catalytic ozonation process.