Enzymatic Nitrogen Incorporation Using Hydroxylamine

Gao, Shilong; Das, Anuvab; Alfonzo, Edwin; Sicinski, Kathleen M.; Rieger, Dominic; Arnold, Frances H.

doi:10.1021/jacs.3c08053

Enzymatic Nitrogen Incorporation Using Hydroxylamine

Journal Article · Wed Sep 06 00:00:00 EDT 2023 · Journal of the American Chemical Society

DOI:https://doi.org/10.1021/jacs.3c08053· OSTI ID:1999523

^[1]; ^[2]; Alfonzo, Edwin ^[2]; Sicinski, Kathleen M. ^[2]; ^[2]; ^[2]

California Institute of Technology (CalTech), Pasadena, CA (United States); California Institute of Technology
California Institute of Technology (CalTech), Pasadena, CA (United States)

Hydroxylamine-derived reagents have enabled versatile nitrene transfer reactions for introducing nitrogen-containing functionalities in small-molecule catalysis as well as biocatalysis. These reagents, however, result in poor atom economy and stoichiometric organic waste. Activating hydroxylamine (NH₂OH) for nitrene transfer offers a low-cost and sustainable route to amine synthesis, since water is the sole byproduct. Despite its presence in nature, however, hydroxylamine is not known to be used for enzymatic nitrogen incorporation in biosynthesis. Here, we report an engineered heme enzyme that can utilize hydroxylammonium chloride, an inexpensive commodity chemical, for nitrene transfer. Directed evolution of Pyrobaculum arsenaticum protoglobin generated efficient enzymes for benzylic C–H primary amination and styrene aminohydroxylation. Mechanistic studies supported a stepwise radical pathway involving rate-limiting hydrogen atom transfer. Furthermore, this unprecedented activity is a useful addition to the ‘nitrene transferase’ repertoire and hints at possible future discovery of natural enzymes that use hydroxylamine for amination chemistry.

View Accepted Manuscript (DOE)

Research Organization:: California Institute of Technology (CalTech), Pasadena, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES)

Grant/Contract Number:: SC0021141

OSTI ID:: 1999523

Journal Information:: Journal of the American Chemical Society, Journal Name: Journal of the American Chemical Society Journal Issue: 37 Vol. 145; ISSN 0002-7863

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: English

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