A Chemoselective and Stereodivergent Platform of Heme-Nitrene Transferases to Access Chiral Aryl-β-Amino Esters and An Investigation of the Sequence-Activity Landscape.

Engineered biocatalysts can utilize nitrene precursors to access enantioenriched amination products, yet they have not been applied to produce valuable, enantiomerically enriched noncanonical β-amino esters. Current approaches to synthesizing β-amino acids rely on pre-oxidized precursors and multistep synthetic approaches involving various protecting groups. We engineered a platform of heme enzymes for stereoselective C-H bond amination of readily available carboxylic ester derivatives to install primary amines. A directed evolution campaign coupled with sequencing of over 1000 variants enabled us to develop engineered variants that use either O-pivaloylhydroxylamine triflic acid (PONT) or hydroxylamine hydrochloride (H2NOH∙HCl) as aminating reagents. An analysis of the resulting sequence-activity dataset revealed additional improvements that could be made to the final variant, highlighting the utility of sequencing data to guide future steps in directed evolution campaigns. The evolved nitrene transferases expand the scope of accessible chiral β-amino acid building blocks for peptidomimetic applications and provide new starting points for the design and synthesis of enantioenriched β-amino acid motifs.

• Publication year

  2026

• Venue

  Angewandte Chemie

• Publication date

  2026-02-17

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1002/anie.202525505PMID 41703710
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar

• No claims are published for this paper.

• No concepts are published for this paper.

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