Glycerol-3-phosphate oxidase is primarily utilized in biochemical research and diagnostic applications. It plays a critical role in enzymatic assays for measuring glycerol-3-phosphate levels, which are important in studying metabolic pathways, particularly glycolysis and lipid metabolism. This enzyme is also employed in biosensors for detecting and quantifying glycerol-3-phosphate in biological samples, aiding in the diagnosis of metabolic disorders. Additionally, it is used in industrial biotechnology for the production of dihydroxyacetone phosphate, a key intermediate in various biochemical processes. Its application extends to the study of oxidative stress and mitochondrial function, providing insights into cellular energy metabolism.