Biocatalytic Synthesis of α-Amino Esters via Nitrene C–H Insertion

Alfonzo, Edwin; Hanley, Deirdre; Li, Zi-Qi; Sicinski, Kathleen M.; Gao, Shilong; Arnold, Frances H.

doi:10.1021/jacs.4c09989

Title: Biocatalytic Synthesis of α-Amino Esters via Nitrene C–H Insertion

Journal Article · 27 September 2024 · Journal of the American Chemical Society

DOI: https://doi.org/10.1021/jacs.4c09989 · OSTI ID:2452751

^[1]; Hanley, Deirdre ^[2];

^[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)

α-Amino esters are precursors to noncanonical amino acids used in developing small-molecule therapeutics, biologics, and tools in chemical biology. α-C–H amination of abundant and inexpensive carboxylic acid esters through nitrene transfer presents a direct approach to α-amino esters. Methods for nitrene-mediated amination of the protic α-C–H bonds in carboxylic acid esters, however, are underdeveloped. This gap arises because hydrogen atom abstraction (HAA) of protic C–H bonds by electrophilic metal-nitrenoids is slow: metal-nitrenoids preferentially react with polarity-matched, hydridic C–H bonds, even when weaker protic C–H bonds are present. This study describes the discovery and evolution of highly stable protoglobin nitrene transferases that catalyze the enantioselective intermolecular amination of the α-C–H bonds in carboxylic acid esters. We developed a high-throughput assay to evaluate the activity and enantioselectivity of mutant enzymes together with their sequences using the Every Variant Sequencing (evSeq) method. Furthermore, the assay enabled the identification of enantiodivergent enzymes that function at ambient conditions in Escherichia coli whole cells and whose activities can be enhanced by directed evolution for the amination of a range of substrates.

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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:: 2452751

Journal Information:: Journal of the American Chemical Society, Journal Name: Journal of the American Chemical Society Journal Issue: 40 Vol. 146; ISSN 0002-7863

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: English

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