Engineering cytochrome P450s for enantioselective cyclopropenation of internal alkynes
- Creators
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Chen, Kai
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Arnold, Frances H.
Abstract
We report a biocatalytic platform of engineered cytochrome P450 enzymes to carry out efficient cyclopropene synthesis via carbene transfer to internal alkynes. Directed evolution of a serine-ligated P450 variant, P411-C10, yielded a lineage of engineered P411 enzymes that together accommodate a variety of internal aromatic alkynes as substrates for cyclopropenation with unprecedented efficiencies and stereoselectivities (up to 5760 TTN, and all with >99.9% ee). Using an internal aliphatic alkyne bearing a propargylic ether group, different P411 variants can selectively catalyze cyclopropene formation, carbene insertion into a propargylic C–H bond or [3 + 2]-cycloaddition. This tunable reaction selectivity further highlights the benefit of using genetically encoded catalysts to address chemoselectivity challenges.
Additional Information
© 2020 American Chemical Society. Received: February 3, 2020; Published: March 29, 2020. This work was supported by NSF Division of Molecular and Cellular Biosciences grant MCB-1513007, US Army Research Office Institute for Collaborative Biotechnologies cooperative agreement W911NF-19-2-0026, and US Army Research Office Institute for Collaborative Biotechnologies contract W911NF-19-D-0001. K.C. thanks the Resnick Sustainability Institute at Caltech for fellowship support. We thank R. K. Zhang, N. P. Dunham, D. J. Wackelin, Y. Yang, and M. Garcia-Borràs for helpful discussions and comments. The authors declare no competing financial interest.Attached Files
Supplemental Material - ja0c01313_si_001.pdf
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Additional details
- Eprint ID
- 102170
- DOI
- 10.1021/jacs.0c01313
- Resolver ID
- CaltechAUTHORS:20200330-105819372
- NSF
- MCB-1513007
- Army Research Office (ARO)
- W911NF-19-2-0026
- Army Research Office (ARO)
- W911NF-19-D-0001
- Resnick Sustainability Institute
- Created
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2020-03-30Created from EPrint's datestamp field
- Updated
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2021-11-16Created from EPrint's last_modified field
- Caltech groups
- Resnick Sustainability Institute, Division of Biology and Biological Engineering