biostudies-literatureRen XNational Institutes of HealthNIH HHSNIGMS NIH HHSNational Science Foundation8550-8556https://www.ebi.ac.uk/biostudies/studies/S-EPMC9337741biostudies-literatureUnknown13(29)Organophosphonate compounds have represented a rich source of biologically active compounds, including enzyme inhibitors, antibiotics, and antimalarial agents. Here, we report the development of a highly stereoselective strategy for olefin cyclopropanation in the presence of a phosphonyl diazo reagent as carbene precursor. In combination with a 'substrate walking' protein engineering strategy, two sets of efficient and enantiodivergent myoglobin-based biocatalysts were developed for the synthesis of both (1<i>R</i>,2<i>S</i>) and (1<i>S</i>,2<i>R</i>) enantiomeric forms of the desired cyclopropylphosphonate ester products. This methodology enables the efficient transformation of a broad range of vinylarene substrates at a preparative scale (<i>i.e.</i> gram scale) with up to 99% de and ee. Mechanistic studies provide insights into factors that contribute to make this reaction inherently more challenging than hemoprotein-catalyzed olefin cyclopropanation with ethyl diazoacetate investigated previously. This work expands the range of synthetically useful, enzyme-catalyzed transformations and paves the way to the development of metalloprotein catalysts for abiological carbene transfer reactions involving non-canonical carbene donor reagents.Chemical scienceHighly stereoselective and enantiodivergent synthesis of cyclopropylphosphonates with engineered carbene transferases.PMC9337741CHE-0946653S10OD030302S10 OD025242GM098628CHE-1725028R01 GM098628S10 OD030302CBET-1929256CBET 1929237Ren XKhare SDShen ZChandgude ALFasan RCarminati DMfalseHighly stereoselective and enantiodivergent synthesis of cyclopropylphosphonates with engineered carbene transferases.Organophosphonate compounds have represented a rich source of biologically active compounds, including enzyme inhibitors, antibiotics, and antimalarial agents. Here, we report the development of a highly stereoselective strategy for olefin cyclopropanation in the presence of a phosphonyl diazo reagent as carbene precursor. In combination with a 'substrate walking' protein engineering strategy, two sets of efficient and enantiodivergent myoglobin-based biocatalysts were developed for the synthesis of both (1<i>R</i>,2<i>S</i>) and (1<i>S</i>,2<i>R</i>) enantiomeric forms of the desired cyclopropylphosphonate ester products. This methodology enables the efficient transformation of a broad range of vinylarene substrates at a preparative scale (<i>i.e.</i> gram scale) with up to 99% de and ee. Mechanistic studies provide insights into factors that contribute to make this reaction inherently more challenging than hemoprotein-catalyzed olefin cyclopropanation with ethyl diazoacetate investigated previously. This work expands the range of synthetically useful, enzyme-catalyzed transformations and paves the way to the development of metalloprotein catalysts for abiological carbene transfer reactions involving non-canonical carbene donor reagents.2022-01-01T00:00:00Z2022 Jul2025-06-01T01:57:39.355Z2024-11-14T15:34:13.672ZS-EPMC93377413597476410.1039/d2sc01965e