The "Digestive Enzymes in Health and Disease" Special Issue Reprint explores the diverse roles of digestive enzymes in physiological processes, their implications in disease mechanisms, and their therapeutic potential in clinical applications. Digestive enzymes are essential to maintaining metabolic homeostasis, aiding in nutrient digestion and absorption, and protecting against gastrointestinal dysfunction. Disruptions in digestive enzyme activity are associated with the development of disorders such as malabsorption syndromes and inflammatory bowel disease and metabolic conditions such as obesity and diabetes. Recent advances in the research field offer novel insights into the functions of these digestive enzymes, many of which include "extra-digestive" roles, paving the way for possible innovative therapeutic strategies. The Special Issue Reprint will thus deepen our understanding of digestive enzymes and their clinical relevance.