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Single‐atom (SA) nanoparticles exhibit considerable potential in terms of photothermal properties for bactericidal applications. Nevertheless, the restricted efficacy of their targeted and controlled antibacterial activity has hindered their practical implementation. This study aims to overcome this obstacle by employing chemical modifications to tailor SAs, thereby achieving targeted and light‐controlled antimicrobial effects. By conducting atomic‐level modifications on palladium SAs using glutathione (GSH) and mercaptophenylboronic acid (MBA), their superior targeted binding capabilities toward Escherichia coli cells are demonstrated, surpassing those of SAs modified with cysteine (Cys). Moreover, these modified SAs effectively inhibit wound bacteria proliferation and promote wound healing in rats, without inducing noticeable toxicity to major organs under 808 nm laser irradiation. This study highlights the significance of chemical engineering in tailoring the antibacterial properties of SA nanoparticles, opening avenues for combating bacterial infections and advancing nanoparticle‐based therapies.
By chemically tailoring single atoms at the atomic level, targeted ability is achieved in palladium SAs while maintaining excellent photothermal properties in a bio‐environment. SA nanoparticles modified with GSH and MBA, rather than Cys, demonstrate remarkable in vivo wound disinfection without noticeable toxicity under 808 nm laser irradiation, highlighting the importance of chemical engineering in controlling antibacterial capabilities.