Journal
ORGANIC & BIOMOLECULAR CHEMISTRY
Volume 19, Issue 4, Pages 775-784Publisher
ROYAL SOC CHEMISTRY
DOI: 10.1039/d0ob02412k
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Funding
- Vanderbilt University Discovery Award grant
- National Science Foundation [HRD 1618408]
- Indiana Academy of Science Senior Research Grant Program [IASSG-F20-01]
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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.
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.
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