Microorganisms were cultured in the R-2 A medium with inoculum from biofilm in a reclaimed wastewater distribution system and then extracellular polymeric substances (EPS) were extracted from the culture. Characterization of EPS and their effects on the corrosion of cast iron were examined. EPS extracted from different culturing stages contained different proportions of protein and polysaccharide but with similar functional groups. All types of EPS could inhibit cast iron corrosion and the EPS from the stationary stage had the highest inhibition efficiency. The inhibition efficiency was increased with addition of a small amount of EPS while decreased with excessive amount of EPS. EPS formed a protective film on the metal surface, which retarded the cathodic reduction of oxygen. Excessive amount of EPS promoted anodic dissolution through EPS-Fe binding. The C=O and C-(O, N) in EPS could be the anodic electrochemical sites with possible products of C-(C, H). (C) 2014 Elsevier Ltd. All rights reserved.