In biohydrometallurgical processes, a mineral exhibits different oxidation phases due to the system's heterogeneity, especially in heap leach pads. Oxidation chemically modifies the mineral surface altering its interface, and thereby affecting the bacteria-mineral interaction, mineral reactivity and leaching velocity. Given that the mineral can be found in different oxidation states in heap bioleaching processes, three oxidation conditions of FeS2, in which iron and/or sulfur related compounds are formed on the mineral surface were assayed. This paper studies the interaction between the modified surface of a sulfide mineral, FeS2, and 0 K culture medium, typically used in biomining processes. Chemical and electrochemical changes on surfaces were characterized by subjecting them to weathering in an acidic culture medium (pH 1.8), without applying potential or current. The chemical species formed were identified by Raman spectroscopy. The modified pyrite surfaces showed significant interfacial transformations upon immersion in the culture medium, and the formation of passive chemical species, such as elemental sulfur, jarosite, phosphates and oxides, were identified. These interfacial modifications are correlated with changes in the open circuit potential (OCP) values during immersion of pyrite and surface modified pyrites in 0 K culture medium. Electrochemical characterization showed a decrease in mineral oxidation capacity, which directly affects the extent of leaching and possibly, of the interaction with other elements participating in the process, such as microorganisms. To study the interactions among bacteria and the pyrite mineral suffering different surface modifications, the attachment of the bioleaching bacterium Leptospirillum sp. was evaluated.