REDUCTION OF 2,4,6-TRINITROTOLUENE BY CLOSTRIDIUM-ACETOBUTYLICUM THROUGH HYDROXYLAMINO-NITROTOLUENE INTERMEDIATES

Studies were conducted to isolate and identify intermediates of 2,4,6- trinitrotoluene (TNT) transformation by Clostridium acetobutylicum and to quantify their concentrations in active whole cell cultures. Only two intermediates of TNT reduction were detected in cell cultures and were identified as 4-hydroxylamino-2,6-dinitrotoluene and 2,4-dihydrox ylamino-6-nitrotoluene. Structures were confirmed with H-1-NMR, C-13-N MR, and desorption chemical ionization mass spectroscopy. When cells w ere suspended in a non-growth saline medium, both hydroxylamine forms accumulated. In media capable of supporting cell growth, the 2,4-dihyd roxylamino-6-nitrotoluene accumulated with concentrations of 4-hydroxy lamino-2,6-dinitrotoluene remaining near detection limits. Studies usi ng purified 2,4-dihydroxylamino-6-nitrotoluene confirmed that its biot ransformation rate in active cultures greatly exceeded abiotic decompo sition in aqueous medium.