Acid invertases (Ac-Invs) are crucial enzymes in plant physiology, regulating sucrose metabolism and hydrolyzing sucrose into glucose and fructose. These sugars serve not only as energy sources and structural components but also as signaling molecules, influencing diverse developmental processes, including seed and fruit growth, flowering, and stress responses. Ac-Invs are classified into cell wall invertases (CWINs) and vacuolar invertases (VINs) based on their subcellular localization, with both playing distinct roles in sucrose unloading, osmotic regulation, and sugar accumulation. Recent studies have also highlighted their involvement in abiotic stress adaptation and hormonal regulation, emphasizing their central role in plant resilience and productivity. However, gaps remain in understanding their regulatory mechanisms, particularly their interactions with plant hormones, defective invertases, and responses to environmental stresses. This review summarizes the biochemical characteristics, functions, and regulatory mechanisms of Ac-Invs, providing insights into their evolutionary significance and potential applications in crop improvement. Future research directions are proposed to elucidate unresolved questions and leverage Ac-Invs for enhancing agricultural sustainability.