Among the different types of secondary post-consumption wastes, E-wastes or waste electrical and electronic equipment represent the fastest growing and most problematic waste stream with printed circuit boards (PCBs) constituting its major ingredient. Results from the extraction of Cu and Ni from PCBs using biogenic Fe2(SO4)3 obtained from the original isolate Acidithiobacillus ferrooxidans 61 (KM819692) are presented. At. ferrooxidans 61 was grown at a temperature of 30 °C in a modified 9 K medium supplemented with ferrous iron. Two-stage bioleaching was carried out at 600 rpm and 40 °C. Experiments were performed at 10% of pulp density (PD) with 48-h duration (each stage of 24 h), under pH 1 and 20 g/L Fe3+. Under these conditions, overall recovery of Cu and Ni of 95% and 87% respectively was achieved. The obtained results indicate that non-ferrous metals in PCBs may be efficiently leached within two-stage bioleaching coupling bio-oxidation to subsequent redoxolysis. Scanning electron microscope (SEM) images acquisition and elemental mapping were performed to assess the liberation degree of essential phases after size-reduction steps and their implication on bioleaching efficiency.