In situ generation of hydrogen peroxide and its use for enzymatic degradation of 2,4,6-trinitrotoluene

A novel electroenzymatic method hybridizing both electrochemical and enzymatic reactions and using lignin peroxidase (Lip) for treatment of TNT (2,4,6-trinitrotoluene) waste was studied. In the presence of Phanerochaete chrysosporium lignin peroxidase, TNT was degraded in an electrochemical reactor using hydrogen peroxide, produced by an electrode reaction. The effectiveness of the electroenzymatic method was examined in this study. The efficiency of removal of TNT was greater than that of biochemical methods under optimal conditions. The effects of reaction conditions on TNT degradation and denitrification, TOC removal efficiency, and powder consumption were also investigated. A potential (vs Ag/AgCl) of +0.1 V was selected as being optimal for the electroenzymatic reaction. TNT degradation was significantly improved in the combined veratryl alcohol-lignin peroxidase oxidation procedure, showing complementary effects of veratryl alcohol (VA) in the TNT degradation reaction. Denitrification was found to be proportional to the amount of TNT degraded. One intermediate, 2,4-diamino-6-nitrotoluene, derived from the oxidative degradation of TNT, was eluted on an HPLC chromatogram and was detected using mass spectrometry. The electroenzymatic method had a lower power requirement than electrochemical oxidation at -0.2 and -0.4 V. The electroenzymatic method may be easily applied to biodegradation systems and provide added benefit for highly recalcitrant chemicals since the system would not be susceptible to the toxicity of the chemical. Also H₂O₂ instability and decomposition in the samples after preparation would not be of concern.