Rhodococcus erythropolis USTB-03 is a promising bacterial strain for the biodesulfurization of dibenzothiophene (DBT) via a sulfurspecific pathway in which DBT is converted to 2-hydroxybiphenyl (2HBP) as an end product. The effects of nicotinamide and riboflavin on the sulfur specific activity (SA) of DBT biodesulfurization by R. erythropolis USTB-03 were investigated. Both nicotinamide and riboflavin were found to enhance the expression of SA, which was not previously reported. When R. erythropolis USTB-03 was grown on a medium containing nicotinamide of 10. 0 mmol or riboflavin of 50. 0 μnol, SA was raised from 68. 0 or so to more than 130 mmol 2HBP/(kg dry cells. h). When R. erythropolis USTB-03 was grown in the presence of both nicotinamide of 5. 0 mmol and riboflavin of 25. 0 μmol, SA was further increased to 159. 0 mmol 2HBP/(kg dry cells. h). It is suggested that the biological synthesis of reduced form of flavin mononucleotide (FMNH2), an essential coenzyme for the activities of biodesulfurization enzyme Dsz C and A, might be enhanced by nicotinamide and riboflavin, which was responsible for the increased SA of R. erythropolis USTB-03.