A novel intracellular 66,500 Da monomeric enzyme was purified from the Staphylococcus sp. Strain S3/C cells cultured in Pseudomonas Mineral Salts (PMS) medium containing dibenzothiophene as sole carbon and sulfur source. The enzyme had the ability to degrade dibenzothiophene and 4,6-dimethyl dibenzothiophene in the presence of organic solvents e.g. toluene, iso-octane and n hexadecane under aqueous – hydrocarbon biphasic conditions as well as under complete non-aqueous conditions at pH 6.0 when incubated at 30°C. Dibenzothiophene was the inducer of the enzyme and NADH acted as an essential coenzyme for the oxidation of dibenzothiophene. Km of DBT for the enzyme was 1.7 ´ 10-5 M and the Vmax was 0.34 μM min-1.
