<p>Enzymes are biological macromolecules with highly efficient catalytic activity, playing essential roles in metabolic reactions within living organisms. Although natural enzymes exhibit excellent catalytic performance, their high production cost, low stability, and susceptibility to deactivation restrict their broader application. As a new generation of artificial enzymes, nanozymes exhibit the lower costs to produce, and the higher catalytic stability, which are also easier to be modified when compared to natural enzymes. Among the numerous nanomaterials, two-dimensional (2D) nanozymes demonstrate unique advantages, such as the large specific surface area, excellent biocompatibility, outstanding electron transport capabilities, and simple methods of surface modification, which satisfy the ideal properties for electrochemical sensing, biosensing, and electrocatalysis. Herein, this article reviewed the research progress of various 2D nanozymes, including graphene nanosheets (GNSs), transition metal sulfide nanosheets (TMS NSs), 2D metal-organic frameworks (MOFs), and 2D transition metal carbides/nitrides (MXenes). Furthermore, this article also systematically introduced the modification of various 2D functional nanozymes, the simulated catalytic mechanism of catalysis of peroxidase, as well as an overview of the state-of-the-art advances of the 2D hybrid nanozymes in various biomedical applications.</p> Graphical Abstract <p></p>