This paper reports the studies on mechanism of arsenic tolerance and bioremoval of arsenic ions (arsenite or arsenate) by Acidithiobacillus ferrooxidans. Exposure of cells to arsenic ions resulted in increased cell surface hydrophobicity, decreased electrophoretic mobility and stronger adsorption affinity towards arsenopyrite. The mechanism of tolerance to arsenic ions were specific and could be attributed to the changes in specific protein expression in the outer membrane and cytosolic membrane fractions. Biosorption studies showed decrease in solution arsenic concentration only with ferrous–grown cells indicating that presence of ferric ions in the EPS was necessary for binding or entrapment of arsenic ions in the EPS. Bacterial EPS of ferrous–grown wild cells were able to uptake arsenate ions due to the strong affinity of ferric ions towards arsenate ions. Neither cells nor the ferric ions were capable of precipitating or oxidizing arsenite ions directly. Both arsenate ions and arsenite ions were co–precipitated with ferric ions formed during the growth of the bacteria.