In recent years, gold nanoparticles have demonstrated excellent enzyme-mimicking activities which resemble those of peroxidase, oxidase, catalase, superoxide dismutase or reductase. This, merged with their ease of synthesis, tunability, biocompatibility and low cost, makes them excellent candidates when compared with biological enzymes for applications in biomedicine or biochemical analyses. Herein, over 200 research papers have been systematically reviewed to present the recent progress on the fundamentals of gold nanozymes and their potential applications. The review reveals that the morphology and surface chemistry of the nanoparticles play an important role in their catalytic properties, as well as external parameters such as pH or temperature. Yet, real applications often require specific biorecognition elements to be immobilized onto the nanozymes, leading to unexpected positive or negative effects on their activity. Thus, rational design of efficient nanozymes remains a challenge of paramount importance. Different implementation paths have already been explored, including the application of peroxidase-like nanozymes for the development of clinical diagnostics or the regulation of oxidative stress within cells via their catalase and superoxide dismutase activities. The review also indicates that it is essential to understand how external parameters may boost or inhibit each of these activities, as more than one of them could coexist. Likewise, further toxicity studies are required to ensure the applicability of gold nanozymes in vivo. Current challenges and future prospects of gold nanozymes are discussed in this review, whose significance can be anticipated in a diverse range of fields beyond biomedicine, such as food safety, environmental analyses or the chemical industry.