In the Ball State Laboratory for Biocatalysis Research, we strive to develop new green-chemical tools through enzyme engineering that can contribute to alleviating the chemical industry’s reliance on fossil fuels.<br/> <br/>With the recent advances in Synthetic Biology and Directed Evolution, the potential for engineering enzymes as robust, selective, and environmentally benign chemical catalysts has exploded. Rieske dioxygenases, with their unique ability to perform oxidative dearomatization to produce chiral <i>cis</i>-diol metabolites, have long been utilized as green-chemical catalysts in bioremediation efforts and by synthetic chemists, although their utility has been limited by their substrate scope and selectivity. In the Ball State Laboratory for Biocatalysis Research, we have developed a novel periodate-based reactive assay system that has demonstrated the capacity to screen for the cis-dihydroxylation activity of large Rieske dioxygenase variant libraries, and to identify Rieske dioxygenase variants with significantly altered reactivity. By applying our novel assay system, we are developing new catalysts based on Rieske dioxygenases, which have expanded the chemical utility of this class of enzymes. To this end, we have developed new dioxygenase catalysts with significantly increased activity in the production of valuable chiral synthons through rational mutagenesis and high-throughput screening. Through our continued mutagenesis and screening efforts, we are endeavoring to alleviate the constraints on the substrate scopes and activities of Rieske dioxygenases, enabling their application in a much wider variety of synthetic contexts.

Expanding the utility of Rieske dioxygenases through enzyme engineering

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