Resorufin β-D-glucopyranoside is widely used as a substrate in enzymatic assays, particularly for detecting β-glucosidase activity. It is highly sensitive and specific, making it a valuable tool in biochemical research and diagnostics. When cleaved by β-glucosidase, it releases resorufin, a fluorescent compound that can be easily measured, enabling the quantification of enzyme activity. This property is utilized in high-throughput screening for drug discovery, where it helps identify potential inhibitors or activators of β-glucosidase. Additionally, it is employed in microbiology to study enzyme kinetics and in environmental monitoring to assess microbial activity in soil or water samples. Its fluorescence-based detection method offers a rapid and reliable approach for various analytical applications.