Even though great progress has been achieved in mimicking natural enzyme engineering, few artificial enzymes with efficient catalytic performance and multifunction have been reported. In this study, novel manganese-iron dual single-atom catalysts (Mn/Fe SACs) were synthesized via a hydrothermal/pyrolysis recipe. Iron atoms inside the Mn/Fe SACs adequately exerted the peroxidase (POD)-like activity, its Michaelis-Menten constant, and maximum initial velocity superior to the horseradish peroxidase. Manganese atoms sufficiently catalyzed the H2O2 to generate oxygen (O2), which alleviated the challenge of the continued lack of O2 in the infected wound. In addition, Mn/Fe SACs possess a glutathione oxidase-like activity that further enhanced POD-like activity in the therapeutic process. The antibacterial rates of Mn/Fe SACs were 95 and 94.5% for Escherichia coli and Staphylococcus aureus, respectively. In vitro anti-inflammatory experiments demonstrated that Mn/Fe SACs could regulate the polarization of macrophages into the anti-inflammatory M2 subtype. In vivo wound healing experiments suggested that the combination therapy of Mn/Fe SACs and chemodynamic therapy presented a great promotion of the recovery rate. Moreover, the O2 generated by the catalase-like process contributed to the catalysts permeating the interior of the infected wounds and achieved preferable abscess elimination ability. This work revealed the potential of Mn/Fe SACs as broad-spectrum antimicrobial materials, which provided a novel strategy for treating infected and abscess wounds.