Abstract

Enzymology, the study of enzymes and their catalytic mechanisms, has evolved from a discipline characterized by the pursuit of complexity to one focused on unravelling the mechanistic underpinnings of physiological processes and disease. This transformation has been facilitated by advances in single molecule studies, which allow researchers to directly observe the behaviour of individual enzyme molecules in real-time. By circumventing the limitations of traditional bulk assays, single molecule enzymology offers unprecedented insights into the dynamic interplay between enzymes, substrates, and cofactors. This article traces the journey from masochistic enzymology to mechanistic physiology and disease, highlighting the transformative impact of single molecule studies on our understanding of enzyme function and regulation. Furthermore, it explores the application of enzymological principles to the study of physiological processes and disease, showcasing the role of enzymology in driving advances in biomedical research and therapeutics.