As a result of ubiquitous excretion by micro-organisms, extracellular polymeric substances are reported in high concentrations in marine systems. The majority of this material is exopolysaccharide (EPS). Despite previous studies showing that EPS can affect carbon as well as trace metal cycling, little is known about the effect on Fe – a critical nutrient limiting primary productivity in up to 40% of the ocean. Here, we have characterised an EPS purified from bacteria isolated from the pelagic Southern Ocean (Pseudoalteromonas sp.) and investigated its role in Fe chemical speciation, solubility, as well as bioavailability for two keystone Southern Ocean phytoplankton strains. This EPS has an average molecular weight of 4.6 MDa, exhibiting mainly –OH, COO– and –NH2 functional groups. An asymmetrical flow field-flow fractionation coupled online with UV-spectrophotometer, differential refractive index, and multiangle laser light scattering (aFlFFF-UV-DRI-MALS) demonstrates that this EPS is polydisperse with three, not well resolved, size populations having molar masses in the range from 0.57 to 15.8 MDa. Fe was exclusively associated with the medium size fraction of this EPS and was the most abundant trace metal with 2.2 nM Fe per nM EPS. Only a third of this Fe was chemically labile, and the strength of Fe-EPS complexes increased with equilibration time. 1 nM EPS is efficient to retain Fe in solution, mainly in the colloidal phase (0.02–0.2 μm). Fe bound to the EPS was highly bioavailable (25% as much as for inorganic Fe). Due to combined effect of EPS on Fe solubility and bioavailability, it can increase the residence time of bioavailable Fe in the euphotic zone, therefore possibly sustaining and controlling primary productivity in sensitive oceanic regions, such as the Southern Ocean.