Oxidative microbial degradation of 2,4,6-trinitrotoluene via 3-methyl-4,6-dinitrocatechol

A novel pathway for biodegradation of 2,4,6-trinitrotoluene (TNT) was investigated where TNT was the sole carbon, nitrogen, and energy source. Results showed the ability of microorganismsto metabolize TNT through removal of a nitro-group, oxygenation of the aromatic ring, and production of a metabolite that is typically a precursor to oxygenolytic ring cleavage. Nitrite production was observed in active systems, and TNT degradation activity was repeatable and transferable. The metabolic intermediate, 3-methyl-4,6-dinitrocatechol, was positively identified through stable isotope mass spectrometry and tandem mass spectrometry. Experimentation with 14C-TNT showed >3% 14C-labeled CO2 in active systems after 30 d exposure to microorganisms. An increasing fraction of 14C-labeled material was associated with biomass with time, where 11.41 +/- 2.91% and 17.09 +/- 1.49% of 14C was associated with biomass in active systems after 20 and 30 d, respectively, as compared with 5.68 +/- 1.33% and 6.08 +/- 1.27% in inactive systems. Parallel degradation of TNT and production of organic metabolites and nitrite were observed in shake flasks constructed with soil from historically contaminated sites, indicating that the novel pathway identified herein is disturbed in the environment. Therefore, results presented provide evidence of a previously unreported pathway for oxidative degradation of TNT.