Tropane alkaloids (TAs) are a class of specialized metabolites with significant pharmaceutical importance, including compounds like hyoscyamine, anisodamine, and scopolamine. These molecules, primarily derived from Solanaceae plants, exhibit potent biological activities, including anticholinergic, analgesic, and sedative properties. However, the natural production of TAs is often constrained by low yields from plants, limited availability of source plants, and environmental variability. Metabolic engineering provides a promising avenue to address these challenges by enhancing TA biosynthesis in plants or heterologous systems. This chapter highlights recent advances in metabolic engineering, focusing on boosting the production of specific metabolites by targeting bottleneck steps or blocking competing pathways. Furthermore, it discusses the strategies employed to engineer primary metabolic flux and enhance the expression of pathway-specific genes.