Nanozymes, a class of nanomaterials mimicking the function of enzymes, have aroused much attention as the candidate in diverse fields with the arbitrarily tunable features owing to the diversity of crystalline nanostructures, composition, and surface configurations. However, the uncertainty of their active sites and the lower intrinsic deficiencies of nanomaterial‐initiated catalysis compared with the natural enzymes promote the pursuing of alternatives by imitating the biological active centers. Single‐atom nanozymes (SAzymes) maximize the atom utilization with the well‐defined structure, providing an important bridge to investigate mechanism and the relationship between structure and catalytic activity. They have risen as the new burgeoning alternative to the natural enzyme from in vitro bioanalytical tool to in vivo therapy owing to the flexible atomic engineering structure. Here, focus is mainly on the three parts. First, a detailed overview of single‐atom catalyst synthesis strategies including bottom‐up and top‐down approaches is given. Then, according to the structural feature of single‐atom nanocatalysts, the influence factors such as central metal atom, coordination number, heteroatom doping, and the metal–support interaction are discussed and the representative biological applications (including antibacterial/antiviral performance, cancer therapy, and biosensing) are highlighted. In the end, the future perspective and challenge facing are demonstrated.

Comprehensive Summary Oxygen reduction reaction (ORR) is the pivotal portion in many electrochemical energy conversion and storage technologies. However, the complex mechanisms and sluggish kinetics of ORR have also become one of the key issues hindering the development and application of these technologies. Recently, single‐atom catalysts (SACs) with well‐defined atomic active centers and theoretical 100% atomic utilization have attracted broad interests in the area of electrocatalytic ORR. The electrocatalytic ORR performance of SACs is fundamentally determined by the intrinsic activity of the single‐atom active site and increasing the number of active sites involved in the ORR can further enhance the ORR activity of SACs. In this review, advances in atomically dispersed metal electrocatalysts for ORR in the last three years are summarized. Three main regulation strategies for achieving SACs with excellent intrinsic activity in the context of the ORR mechanism are involved including modulation of coordination environments, the construction of intrinsic defects, and the introduction of dual active sites. Moreover, discussions on improving the loading and utilization of active sites are given. Finally, the current challenges and opportunities for the development of SACs for ORR are presented.