A new application of a combined solvent extraction and two-phase biodegradation processes using two-liquid phase partitioning bioreactor (TLPPB) technique was proposed and developed to enhance the cleanup of high concentration of crude oil from aqueous phase using acclimated mixed culture in an anaerobic environment. Silicone oil was used as the organic extractive phase for being a water-immiscible, biocompatible and non-biodegradable. Acclimation, cell growth of mixed cultures, and biodegradation of crude oil in aqueous samples were experimentally studied at 30±2ºC. Anaerobic biodegradation of crude oil was examined at four different initial concentrations of crude oil including 500, 1000, 2000, and 5000 mg/L. Complete removal of crude oil was achieved biphasic bioreactor after 3 weeks compared to 73-82% in the monophasic bioreactor for the same time period.
