Pyrite ash, a waste by-product formed during roasting of pyrite ores, is a good source of valuable metals. The waste is associated with several environmental issues due to its dumping in sea and/or land filling. Although several other management practices are available for its utilization, the waste still awaits and calls for an eco-friendly biotechnological application for metal recovery. In the present study, chemolithotrophic meso-acidophilic iron and sulphur oxidisers were evaluated for the first time towards simultaneous mutli-metal recovery from pyrite ash. XRD and XRF analysis indicated higher amount of Hematite (Fe₂O₃) in the sample. ICP–OES analysis indicated concentrations of Cu > Zn > Co> As that were considered for bioleaching. Optimization studies indicated Cu – 95%, Co – 97%, Zn – 78% and As – 60% recovery within 8 days at 10% pulp density, pH – 1.75, 10% (v/v) inoculum and 9 g/L Fe²⁺. The productivity of the bioleaching system was found to be Cu – 1696 ppm/d (12% dissolution/d), Co – 338 ppm/d (12.2% dissolution/d), Zn k 576 ppm/d (9.8% dissolution/d) and As – 75 ppm/d (7.5% dissolution/d). Synergistic actions for Fe²⁺ – S° oxidation by iron and sulphur oxidisers were identified as the key drivers for enhanced metal dissolution from pyrite ash sample.