This work assessed the effect of an enriched culture medium and synthetic seawater on the growth and production of exopolymeric substances (EPS) of a Desulfovibrio sp. strain, isolated from a Mexican oil well. The EPS (mainly consisting of proteins) growth was only achieved after exposing sulfate-reducing bacteria to culture media under dissimilative conditions that predominantly promoted the growth of the biofilm and a small concentration of microorganisms. Once this EPS film was obtained, the evolution of SAE 1018 carbon steel/biofilm/synthetic seawater (VNNS medium) interface was further studied using electrochemical impedance spectroscopy technique (EIS). This study revealed strong adhesion of the biofilm during the formation of iron sulfide (pirrotite) on carbon steel surface. The biofilm inhibits the accelerated damage of the steel for some time exhibiting impedance values of 30000Ω. However, at longer times the chemical environment around the biofilm, as a result of microbial metabolism, may become quite corrosive to steel.