A bioreactor system operating in a continuous mode was designed to generate biofilms on polished and as-received surfaces of AISI 316 stainless steel coupons exposed for 36 d to a pure culture of marine Pseudomonas NCIMB 2021. Scanning electron microscopy (SEM) and atomic force microscopy were employed to determine the degree of surface colonisation and to examine corrosion damage of the steel. X-ray photoelectron spectroscopy analysis was carried out to characterise the chemistry of the passive layers on polished steel stored for a period of time, freshly re-polished coupons, and as-received steel. The effect of biofilms on the composition of layers formed on the steel specimens was evaluated. SEM revealed that the surfaces of polished and stored steel appeared to accumulate more biofilm compared to as-received specimens. Micropitting of steel occurred underneath the biofilm, regardless of surface finish. The concentration of elements in the passive layers differed significantly between freshly re-polished and as-received or polished and stored coupons. In the presence of Pseudomonas NCIMB 2021 biofilm, the composition of the passive layer on the as-received steel surface was considerably altered compared to unexposed steel or steel exposed to abiotic medium.