Pentachlorophenol-degrading Sphingomonas sp. UG30 and Sphingomonas chlorophenolica strains RA2 and ATCC 39723 can transform p-nitrophenol in either mineral salts-glutamate or mineral salts-glucose medium after an initial lag period. However, mineralization of p-nitrophenol by these bacterial strains was observed only in mineral salts-glucose medium. When p-nitrophenol was the sole nitrogen source in the growth medium, UG30 mineralized 32% of 140 mM [14C]p-nitrophenol which was 10% higher than the amount of [14C]p-nitrophenol mineralized in mineral salts-glucose medium. UG30 did not transform or mineralize p-nitrophenol (in a growth medium) in the absence of glucose or glutamate. All three strains released nitrite during p-nitrophenol degradation in mineral salts-glucose medium and mineral salts-glutamate medium. The transformation rate of p-nitrophenol by UG30 was dependent on the initial p-nitrophenol concentration, with the optimal rate being found at 310 μM of p-nitrophenol and inhibition observed at ≥1100 μM of p-nitrophenol. Pre-exposure of UG30 cells to p-nitrophenol eliminated the initial lag phase of p-nitrophenol transformation. However, pre-growth of UG30 cells on pentachlorophenol did not reduce the lag period for p-nitrophenol transformation. Both p-nitrophenol- and pentachlorophenol-induced UG30 cells degraded pentachlorophenol without any lag phase. Thin layer chromatographic analysis of the reaction mixture suggested 4-nitrocatechol was an intermediate of p-nitrophenol transformation by UG30.