The objective of this study was to prepare bifunctional catalysts based on iron and CaO and test them in the biodiesel production using waste cooking oil (WCO) as feedstock. Two iron precursors were studied, Fe2O3 and Fe(NO3)3·9H2O. The identified crystalline phases were Ca2Fe2O5 and CaFeO3. Surface morphology and textural properties (distribution of active species, specific surface area, size, and pore volume) were also analyzed. Additionally, thermal stability was studied and 800 °C was established as the optimum calcination temperature. The density of both acidic and basic sites was higher with the catalyst prepared with Fe2O3 than with that prepared with Fe(NO3)3·9H2O. The latter, however, leads to reach equilibrium in half of the time than with the former. This was ascribed to the ratio of acidic to basic sites, which is higher with the catalyst prepared with the precursor salt. This ratio not only affects the overall cost of the process by affecting the time at which equilibrium is reached but also by dictating the methanol/oil molar ratio at which the equilibrium is reached sooner. The prepared bifunctional catalyst allowed us to produce biodiesel with 90% of methyl ester content at atmospheric pressure, reaction temperature of 60 °C, reaction time of 2 h, with 12:1 M ratio of methanol/WCO, 10 wt % of Fe over CaO, and a catalyst loading of 5 wt %. This catalyst can be used at least 3 times. The so-obtained biodiesel met the European norm EN-14214 regarding viscosity and density.