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Bacterial adsorption on mineral surface is one of the key steps in bioleaching process. The bacteria adsorb on the mineral surface via the extracellular polymeric substances (EPS) layer. In this paper, the behavior of glucuronic acid, one of the key substances in EPS layer, adsorbed on the pyrite surface is studied using DFT and electrochemical methods. Adsorption capacity of glucuronic acid is stronger than that of water. Glucuronic acid adsorbs on pyrite surfaces and it follows a mixed type of interactions (physisorption and chemisorption). Adsorption of glucuronic acid on pyrite surface followed Langmuir’s adsorption isotherm with adsorption standard free energy of –27.67kJ mol–1. The structural and electronic parameters were calculated and discussed.