Extracellular polymeric substances (EPS) are an important medium for communication and material exchange between iron-oxidizing bacteria and the external environment and could induce the iron (oxyhydr) oxides production which reduced arsenic (As) availability. The main component of EPS secreted by iron-oxidizing bacteria (Ochrobactrum EEELCW01) was composed of polysaccharides (150.76-165.33 mg/g (DW⁻¹)) followed by considerably smaller amounts of proteins (16.12-12.98 mg/g (DW⁻¹)). Low concentrations of As (100 or 500 μmol·L⁻¹) promoted the amount of EPS secretion. FTIR results showed that EPS was composed of polysaccharides, proteins, and a miniscule amount of nucleic acids. The functional groups including -COOH, -OH, -NH, -C=O, and -C-O played an important role in the adsorption of As. XPS results showed that As was bound to EPS in the form of As³⁺. With increasing As concentration, the proportion of As³⁺ adsorbed on EPS increased. Ferrihydrite with a weak crystalline state was only produced in the system at 6 h during the mineralization process of Ochrobactrum sp. At 8 d, the minerals were composed of goethite, galena, and siderite. With the increasing mineralization time, the main mineral phases were transformed from weakly crystalline hydrous iron ore into higher crystallinity siderite (FeCO₃) or goethite (α-FeOOH), and the specific surface area and active sites of minerals were reduced. It can be seen from the distribution of As elements that As is preferentially adsorbed on the edges of iron minerals. This study is potential to understand the biomineralization mechanism of iron-oxidizing bacteria and As remediation in the environment. Copyright © 2022 The Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences.