Kinetic isotopic fractionation of carbon and nitrogen during RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine) biodegradation was investigated with pure bacterial cultures under aerobic and anaerobic conditions. Relatively large bulk enrichments in 15 ؊ production were much larger than, but systematically related to, the bulk RDX N isotope effects with different bacteria. Apparent intrinsic 15 N-NO 2 ؊ values were consistent with an initial denitration pathway in the aerobic experiments and more complex processes of NO 2 ؊ formation associated with anaerobic ring cleavage. These results indicate the potential for isotopic analysis of residual RDX for the differentiation of degradation pathways and indicate that further efforts to examine the isotopic composition of potential RDX degradation products (e.g., NO x ) in the environment are warranted.
IMPORTANCEThis work provides the first systematic evaluation of the isotopic fractionation of carbon and nitrogen in the organic explosive RDX during degradation by different pathways. It also provides data on the isotopic effects observed in the nitrite produced during RDX biodegradation. Both of these results could lead to better understanding of the fate of RDX in the environment and help improve monitoring and remediation technologies.
Explosives manufacturing, testing, and training range activities have resulted in the release of explosive compounds, including the nitramine explosive hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), in soils at numerous U.S. Department of Defense (DoD) installations. Because RDX is mobile in soils and persistent, it has also been detected in groundwater or drinking water at some of these sites (1, 2).There has been extensive research examining the biological degradation of explosive compounds by pure cultures of bacteria as well as mixed consortia (for a review, see reference 3). RDX biodegradation has been observed under a range of redox conditions (4-9), and at least three predominant degradation pathways have been elucidated (Fig. 1). Typically, RDX is degraded anaerobically as an electron acceptor, requiring a carbon source as an electron donor, and entails either successive reduction of the three nitro groups via successive two-electron (2e Ϫ ) transfers followed by ring cleavage (pathway A) or an earlier opening of the ring structure (pathway B1), with or without initial denitration (pathway B2). The specific mechanism of ring destabilization and the steps that lead to ring cleavage may be variable and remain subject to debate (6,7,10). A denitration reaction that converts hexahydro-1-nitroso-3,5-dinitro-1,3,5-triazine (MNX) (in pathway A) through multiple intermediates to methylenedinitramine (MEDINA) (in pathway B) under anaerobic conditions has also been proposed (7,11,12). Aerobic degradation proceeds via two single-electron transfer steps resulting in sequential denitration (2 NO 2 Ϫ removed) followed by ring cleavage and production of 4-nitro-2,4-diazabutanal (NDAB) (pathway C) (7). Some bacteria are able to utilize R...