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Compositions and methods for enzymatic decontamination by inactivation of Hazardous agents are provided. Hazardous agents of microbial and chemical origin can be neutralized by H2O2. The methods described herein provide for enzymatic production of H2O2 in situ using oxidoreductase enzymes that use oxygen as an acceptor and their alcohol substrates. The enzymatically produced H2O2 and corresponding aldehydes have potent antimicrobial properties. The enzymatically produced H2O2 also can detoxify chemical agents in situ. The decontaminating power of the oxidoreductase enzymes that use oxygen as an acceptor may be amplified by addition of reagents, such as acetyl donors or base catalysts that, in the presence of H2O2, yield peroxy acid derivatives and hydroperoxy anions. Such derivatives can neutralize biological and chemical agents, thus providing a broadly applicable decontamination method. In addition, catalytic production of H2O2 in situ results in controlled synthesis of decontamination reagents at their point of use, mitigating the need to store, transport and dispose of hydrogen peroxide solutions in the field.