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Compositions and methods for catalytic buffering of enzymatic decontamination reactions are provided. Enzymatic decontamination of organophosphorus or organohalogen compounds generates acidic reaction products that precipitously reduce the pH of the medium, thus impairing activity of the decontaminating enzymes. Catalytic buffering, that is, the use of an enzyme to produce ions from a substrate to modulate pH, can provide effective pH control. The compositions provided here include urease enzymes with mutations in the alpha subunit of the urease holoenzyme. These mutant ureases maintain urease activity in the presence of fluoride ions, which are organophosphorus and organohalogen hydrolysis products that otherwise inhibit urease activity. The fluoride-resistant ureases act as effective catalytic buffers during organofluorophosphorus hydrolysis reactions. Methods for using the fluoride-resistant ureases in enzymatic decontamination are also provided. Catalytic buffering afforded by fluoride-resistant ureases facilitates the application of safe and effective enzymatic methods for decontamination of personnel, equipment and the environment.