Cloning, purification and possible use of a Bacillus nitroreductase in biotechnological applications

), respectively. The enzymatic reduction of CB1954 analyzed by LC-ESI-MS gave prominent molecular ions at the expected m/z values of CB1954 (M + H⁺: m/z 253), primary hydroxylamine intermediates (M + H⁺: m/z 239), and amino end-products (M + H⁺: m/z 223). The purified enzyme was also found to biotransform TNT to the product 2-amino 4,6-DNT. In addition, a proof-of-concept electrochemical detection platform based on immobilized nitroreductase produced a measurable cathodic response toward nitrofurazone, demonstrating the enzyme's electrochemical applicability. ANOVA identified significant differences in activity levels among multiple groups, while regression analysis enabled prediction of enzyme responses and characterization of the underlying kinetic patterns in statistical analysis.