Development and application of a high throughput assay system for the detection of Rieske dioxygenase activity

Herein we report the development of a new periodate-based reactive assay system for the fluorescent detection of the cis-diol metabolites produced by Rieske dioxygenases. This sensitive and diastereoselective assay system successfully evaluates the substrate scope of Rieske dioxygenases and determines the relative activity of a rationally designed Rieske dioxygenase variant library. The high throughput capacity of the assay system enables rapid and efficient substrate scope investigations and screening of large dioxygenase variant libraries.

Automated summary

A new periodate-based reactive assay system for fluorescent detection of cis-diol metabolites produced by Rieske dioxygenases has been developed. This sensitive and diastereoselective system effectively evaluates substrate scope and relative activity of a designed Rieske dioxygenase variant library. Its high throughput capacity allows for rapid and efficient substrate scope investigations and screening of large dioxygenase variant libraries.