Evaluation of Pseudoenantiomeric Mixed Carbonates as Efficient Fluorogenic Probes for Enantioselectivity Screening

We report mixed carbonates as enzyme substrates and demonstrate their application as fluorogenic probes for lipase and esterase enantiopreference screening. By the application of pseudoenantiomers with distinct fluorescence behaviors, it is possible to evaluate the activity and enantiopreference of hydrolytic enzymes. In order to validate our method, enantioselectivities calculated from fluorometric measurements were compared with the results obtained from larger‐scale kinetic resolution. Pseudoenantiomeric mixed carbonates with distinct fluorogenic properties are shown to be very useful probes for screening lipase and esterase enantioselectivity. The simultaneous use of pseudoenantiomeric substrates overcomes conventional assays with single enantiomer substrates, as the effects of competitive binding are taken into account.