Zymostenol is primarily used in biochemical research and pharmaceutical development due to its role as a steroidal intermediate. It serves as a key precursor in the synthesis of various biologically active steroids, including hormones with anti-inflammatory, metabolic, and reproductive effects. Its structure allows for selective modifications, making it valuable in the production of customized steroid analogs for drug discovery. In industrial settings, Zymostenol is employed in enzymatic transformation studies, where specific enzymes are used to modify its functional groups, enabling the production of complex steroid derivatives with high stereoselectivity. This makes it useful in green chemistry approaches, reducing the need for harsh chemical reagents in steroid synthesis. Additionally, Zymostenol finds application in metabolic pathway studies, helping researchers understand steroid biosynthesis and enzyme kinetics in mammalian and microbial systems. Its use in assay development supports the screening of enzyme inhibitors or activators relevant to endocrine disorders and cancer research.