SCREENING METHOD FOR THE DISCOVERY AND DIRECTED EVOLUTION OF OXYGENASE ENZYMES.

摘要

The present invention relates to screening methods for oxidation enzymes, particularly mono and dioxygenases. According to the methods of the invention, a product of an oxidation reaction is converted into a phenol or a catechol, which is easily detected by a Gibbs assay. This conversion allows for a sensitive and efficient assay. Both high throughput liquid phase and solid phase methods using these principles are provided. Also described are methods for detecting phenolic ether products and sulfhydryl products from oxidation reactions, also using a Gibbs assay. Figure (1) shows this athway for a generic, monosubstituted benzene being degraded by the toluene dioxygenase system.