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Nanozymes (NZs) are nanomaterials that mimic enzyme-like catalytic activity. They have attracted substantial attention due to their inherent physicochemical properties for use as promising alternatives to natural enzymes (NEs) in a variety of research fields. Particularly, in biosensing and bioassays, NZs have opened a new horizon to eliminate the intrinsic limitations of NEs, including their denaturation at extreme pH values and temperatures, poor reusability and recyclability, and high production costs. Moreover, the catalytic activity of NZs can be modulated in the preparation step by following an appropriate synthesis strategy. This review aims to gain insight into the potential substitution of NEs by NZs in biosensing and bioassays while considering both the pros and cons. The physicochemical and catalytic properties of nanozymes and natural enzymes are compared to obtain insight into their application in biosensing and bioassays. [Display omitted] • Nanozymes (NZs) are the nanomaterials mimicking the enzyme-like catalytic activity. • NZs and their application in biosensing and bioassay are summarized. • Natural enzymes (NEs) vs NZs in biosensing and bioassay are discussed. • Compositions, modifications, and immobilization strategies and are highlighted. [ABSTRACT FROM AUTHOR]