Enantioselective Trifluoromethylazidation of Styrenyl Olefins Catalyzed by an Engineered Nonheme Iron Enzyme

Angew Chem Int Ed Engl. 2025 Feb 24;64(9):e202423507. doi: 10.1002/anie.202423507. Epub 2025 Feb 5.

Abstract

Organofluorines, particularly those containing trifluoromethyl (CF3) groups, play a critical role in medicinal chemistry. While trifluoromethylation of alkenes provides a powerful synthetic route to construct CF3-containing compounds with broad structural and functional diversity, achieving enantioselective control in these reactions remains a formidable challenge. In this study, we engineered a nonheme iron enzyme, quercetin 2,3-dioxygenase from Bacillus subtilis (BsQueD), for the enantioselective trifluoromethylazidation of alkenes. Through directed evolution, the final variant BsQueD-CF3 exhibited excellent enantioselectivity, with an enantiomeric ratio (e.r.) of up to 98 : 2. Preliminary mechanistic studies suggest the involvement of radical intermediates. This work expands biocatalytic organofluorine chemistry by reprogramming metalloenzymes for innovative trifluoromethylation reactions.

Keywords: Alkenes; Azidation; Biocatalysis; Nonheme iron enzymes; Trifluoromethylation.

MeSH terms

  • Alkenes* / chemistry
  • Alkenes* / metabolism
  • Bacillus subtilis / enzymology
  • Biocatalysis
  • Dioxygenases* / chemistry
  • Dioxygenases* / genetics
  • Dioxygenases* / metabolism
  • Hydrocarbons, Fluorinated* / chemistry
  • Hydrocarbons, Fluorinated* / metabolism
  • Molecular Structure
  • Nonheme Iron Proteins* / chemistry
  • Nonheme Iron Proteins* / metabolism
  • Protein Engineering
  • Stereoisomerism

Substances

  • Alkenes
  • Dioxygenases
  • Hydrocarbons, Fluorinated
  • Nonheme Iron Proteins