Reactive oxygen species (ROS) are the weapons of neutrophiles against bacterial pathogens, and also the central effectors in reactive oxygen therapy for skin and soft tissue infection. Nanozymes that spontaneously generate ROS under physiological conditions are new antibacterials that hold promise towards multidrug resistant pathogens. The clinical use of the nanozymes is however limited by their low biocompatibility and toxicity in vivo. Here, we develop an oleic acid (OA) nanoemulsion template method for the one-pot synthesis of OA-manganese dioxide (MnO2) nanozyme. The OA-MnO2 nanozyme showed high stability and biocompatibility under physiological conditions with marked oxidase-like activity. The ROS generated by the OA-MnO2 nanozyme effectively kill the Gram-positive Staphylococcus aureus and the Gram-negative Escherichia coli strains. Moreover, the OA-MnO2 nanozyme shows promising abilities to prevent and destruct biofilm formation by Staphylococcus aureus, and result in superior in vivo antibacterial performance as compared to vancomycin. The reactive oxygen therapy based on OA-MnO2 nanozyme cures the infected skin and promotes wound healing in mice, manifesting its potential use in skin and soft tissue infection.