In tank bioleaching, decreasing particle size of milled concentrates has been shown to improve leaching performance owing to increased mineral surface area and mechanical activation effects of fine mineral particles. However, evidence suggests a critical lower limit of particle size distribution exists below which the performance of the thermophilic iron and sulfur oxidizer Sulfolobus metallicus is compromised and complete culture death may result. This paper proposes an explanation for these observations at fine fractions through identifying a relationship between mineral composition of six sulfide concentrates, their extent of milling and the generation of the reactive oxygen species (ROS) hydrogen peroxide (H2O2) and hydroxyl radicals (•OH) in slurries. The effect of oxidative stress induced in the absence of minerals on growth and bioleaching performance of S. metallicus is studied.