An Optimized System for the Study of Rieske Oxygenase‐catalyzed Hydroxylation Reactions <i>In vitro</i>

Rieske non-heme iron oxygenases (ROs) are primarily known for their ability to catalyze the stereoselective formation of vicinal cis-diols in a single step, endowing valuable products pharmaceutical and chemical applications. In addition, ROs can several other oxidation reactions with high regio- stereoselectivity typically broad substrate scope. Owing dependence on multicomponent electron transfer, majority synthetic applications relies recombinant whole-cell catalysts. this context, important properties system that determine catalytic efficiency, including transfer via redox partner proteins, stability uncoupling, have been investigated lesser extent recent years. Here, we show one most prominent ROs, cumene dioxygenase from Pseudomonas fluorescens IP01 (CDO) by developing optimizing an efficient vitro system, activities be achieved. highlight continuous supplementation electrons oxygenase is required sustain activity, while uncoupling major limitation CDO efficiency stability.