C−H Amination via Nitrene Transfer Catalyzed by Mononuclear Non‐Heme Iron‐Dependent Enzymes

Abstract Expanding the reaction scope of natural metalloenzymes can provide new opportunities for biocatalysis. Mononuclear non‐heme iron‐dependent enzymes represent a large class of biological catalysts involved in the biosynthesis of natural products and catabolism of xenobiotics, among other processes. Here, we report that several members of this enzyme family, including Rieske dioxygenases as well as α‐ketoglutarate‐dependent dioxygenases and halogenases, are able to catalyze the intramolecular C−H amination of a sulfonyl azide substrate, thereby exhibiting a promiscuous nitrene transfer reactivity. One of these enzymes, naphthalene dioxygenase (NDO), was further engineered resulting in several active site variants that function as C−H aminases. Furthermore, this enzyme could be applied to execute this non‐native transformation on a gram scale in a bioreactor, thus demonstrating its potential for synthetic applications. These studies highlight the functional versatility of non‐heme iron‐dependent enzymes and pave the way to their further investigation and development as promising biocatalysts for non‐native metal‐catalyzed transformations.