L-Tryptophan is a nutritionally essential amino acid for monogastric animals and preweaning ruminants because it cannot be synthesized in the body. Besides serving as a building block for proteins, tryptophan is a critical nutrient for the functions of nervous and immune systems. Over the past decades, much attention has been directed to study the role of tryptophan as a limiting amino acid in mammalian and avian nutrition. However, emerging evidence from recent studies shows that tryptophan and its metabolites (e.g., serotonin (5-hydroxytryptamine, 5-HT) and melatonin)) can regulate feed intake, reproduction, immunity, neurological function, and anti-stress responses. Additionally, tryptophan may modulate gene expression and nutrient metabolism to impact whole-body homeostasis in organisms. Thus, adequate intake of this amino acid from the diet is crucial for growth, development, and health of animals and humans.