Bacterial biofilms play a crucial role in the emergence of antibiotic resistance and the persistence of chronic infections. The challenge of effectively eradicating bacterial biofilms while ensuring minimal toxicity to normal cells persists. Carbon-based artificial nanoenzymes have attracted considerable attention as emerging nanotheranostic agents, owing to their biocompatibility, cost-effectiveness, and straightforward synthesis. In this study, we have developed a multifunctional carbon dots (CDs) system, specifically CDs functionalized with 1-(3-aminopropyl) imidazole (API), termed CDs-API. This system demonstrates acid-activated antibiofilm activity. The CDs-API were synthesized from chlorogenic acid (ChA), a bioactive compound naturally occurring in coffee, and subsequently functionalized with API to achieve charge-switchable properties under acidic conditions. This distinctive feature enables CDs-API to efficiently penetrate bacterial biofilms and selectively target the colonized bacteria. The enzyme-like activity of CDs-API effectively consumes high levels of glutathione (GSH) within the biofilm, leading to the accumulation of reactive oxygen species (ROS). Consequently, this process degrades the extracellular polymeric substance (EPS) matrix, damages bacterial DNA and protein structures, and disrupts the redox balance, ultimately leading to bacterial cell death. Experimental results demonstrated that CDs-API effectively inhibited the growth of methicillin-resistant