Abstract
Cytochrome P450s are ubiquitous monooxygenase enzymes that are responsible for the drug metabolism and biosynthesis of bioactive organic small molecules. For many bacterial species, P450 monooxygenases play substantial roles in the biosynthesis of useful secondary metabolites such as antibiotics. To date, thousands of P450 genes have been found in the genomes of most organisms, but substrates of many of their product enzymes have not yet been identified and thus need to be clarified. Moreover, characterizing the substrate specificity for natural and engineered P450 enzymes of interest is thought to be important for the production of useful organic small molecules and expansion of the library. Therefore, technology identifying substrates and/or characterizing the substrate specificity for P450 enzymes of interest would be very important not only for P450 chemistry and biology but also for the pharmaceutical discovery and production process. Toward this end, we have developed a droplet array format on an NADH-TEG-SC-slide, and an NAD^+ detection system using biotinylated acetophenone derivative 3. By using this platform, the substrate of P450cam was successfully identified on the array format.
