Biotransformation products and mineralization potential for hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) in abiotic versus biological degradation pathways with anthraquinone-2,6-disulfonate (AQDS) and Geobacter metallireducens

This study investigated extracellular electron shuttle-mediated RDX biodegradation and the distribution of ring cleavage metabolites generated by biological degradation (cells) versus the products formed by abiotic degradation (reduced electron shuttles), and when the two pathways were acting simultaneously. All pathways were influenced by pH. Buffered suspensions (pH 6.8/7.9/9.2) were performed with cell-free anthrahydroquinone-2,6-disulfonate as the sole electron donor, cells (Geobacter metallireducens) + acetate, or cells/acetate + anthraquinone-2,6-disulfonate as an electron shuttle. The metabolites identified included methylenedinitramine, formaldehyde, nitrous oxide, nitrite, ammonium and carbon dioxide. As pH increased, the rates of RDX reduction by AH₂QDS also increased. Cells alone reduced RDX faster at the lower pH values. However, at all pH the rates of the electron shuttle-mediated pathways were consistently the fastest, and the proportion of carbon present as formaldehyde, which is a precursor to mineralization, was highest in the presence of electron shuttles. Formaldehyde accounted for 45/51/54% of the carbon in electron shuttle amended cell suspensions as opposed to 13/42/45% of carbon without shuttles at the pH 6.8/7.9/9.2, respectively. Approximately 7-20% of RDX was mineralized to CO₂ in the presence of cells at all pH tested; AQDS increased the extent of ¹⁴CO₂ produced. Nitrous oxide and nitrite were end products in the strictly abiotic pathway, but nitrite was depleted in the presence of cells to form ammonium. Understanding the different products formed in the abiotic versus biological pathways and the influence of pH is critical to developing mixed biotic-abiotic remediation strategies for RDX.