Abstract: Bacterial resistance and the limitations of conventional antibacterial strategies have posed severe threats to public health, agriculture and ecological safety, making it urgent to develop novel and highly efficient antibacterial materials. Copper single-atom nanozymes feature high atom utilization efficiency, excellent enzyme-mimetic catalytic activity and favorable biosafety. They can exert efficient antibacterial effects through reactive oxygen species (ROS)-mediated oxidative damage and effectively circumvent bacterial drug resistance. This paper reviews three mainstream preparation methods including pyrolysis, metal exchange strategy and coordination-driven self-assembly, elaborates the mechanisms of ROS-based sterilization and multi-mode synergistic antibacterial effects, and summarizes the application progress in plant disease control, fruit preservation, antibacterial textiles and bacterial infection therapy. Current challenges in catalytic mechanism elucidation, in vivo metabolism and large-scale preparation are discussed, and future prospects in precision antibacterial therapy, green agriculture and translational medicine are proposed, providing theoretical references for the design of high-performance single-atom antibacterial materials.