Unravelling an Efficient O2/Reductant-Dependent Catalytic Route in Heme Peroxygenases for Sustainable Applications

Heme peroxygenases are attractive biocatalysts for incorporating oxygen into the organic molecules using H2O2 as oxygen source under mild conditions. However, their practical applications are hindered by irreversible oxidative inactivation caused by exogenous H2O2 usage. Herein, we report a novel catalytic pathway in heme peroxygenases that relies on O2 and small-molecule reductants such as ascorbate acid (AscA), dehydroascorbic acid (DHA), gallic acid (GA), or pyrogallol (PA) to drive reactions. For reactions of unspecific peroxygenase (UPO) with either AscA or DHA, experimental and computational studies revealed that DHAA (the hydrated form of DHA) is the actual co-substrate responsible for activating oxygen to generate oxyferryl heme (compound I, Cpd I) as the oxygenation species. Subsequently, we demonstrate the universality of this O2/reductant-dependent route across various heme peroxygenases, highlighting its biological significance as monooxygenases. Compared to the conventional H2O2-dependent process, this innovative route can efficiently eliminate the excessive production of H2O2, thereby preventing the heme destruction and related enzyme inactivation. Finally, scale-up reactions were performed for the preparations of chiral, value-added products with unprecedented productivity, underscoring the great synthetic capabilities of the developed peroxygenase technology, which paves the way for sustainable and practical applications in various chemical transformations.