Stereoselective Enzymatic Synthesis of Heteroatom-Substituted Cyclopropanes
Abstract
The repurposing of hemoproteins for non-natural carbene transfer activities has generated enzymes for functions previously accessible only to chemical catalysts. With activities constrained to specific substrate classes, however, the synthetic utility of these new biocatalysts has been limited. To expand the capabilities of non-natural carbene transfer biocatalysis, we engineered variants of Cytochrome P450_(BM3) that catalyze the cyclopropanation of heteroatom-bearing alkenes, providing valuable nitrogen-, oxygen-, and sulfur-substituted cyclopropanes. Four or five active-site mutations converted a single parent enzyme into selective catalysts for the synthesis of both cis and trans heteroatom-substituted cyclopropanes, with high diastereoselectivities and enantioselectivities and up to 40 000 total turnovers. This work highlights the ease of tuning hemoproteins by directed evolution for efficient cyclopropanation of new substrate classes and expands the catalytic functions of iron heme proteins.
Additional Information
© 2018 American Chemical Society. This is an open access article published under an ACS AuthorChoice License, which permits copying and redistribution of the article or any adaptations for non-commercial purposes. Received: December 23, 2017; Revised: February 7, 2018; Publication Date (Web): February 24, 2018. We thank Dr. Stephan Hammer for help with chiral separations of cyclopropylamines and all members of the Arnold Laboratory for stimulating comments and discussions. We thank Dr. David Rozzell for suggestions on cyclopropanation substrate scope. Author Contributions: The manuscript was written with contributions of all authors. All authors have given approval to the final version of the manuscript. This work was supported in part by the National Science Foundation, Division of Molecular and Cellular Biosciences (Grant No. MCB-1513007) and the Defense Advanced Research Projects Agency Biological Robustness in Complex Settings Contract HR0011-15-C-0093. O.F.B. acknowledges support from the Deutsche Forschungsgemeinschaft (DFG Grant No. BR 5238/1-1) and the Swiss National Science Foundation (SNF Grant No. P300PA-171225). C.K.P. thanks the Resnick Sustainability Institute for a postdoctoral fellowship. A.M.K. and Z.W. acknowledge support from the NSF Graduate Research Fellowship (Grant No. 1745301), and A.M.K. acknowledges support from Caltech's Center for Environmental Microbial Interactions. The authors declare no competing financial interest.Attached Files
Published - acscatal.7b04423.pdf
Supplemental Material - cs7b04423_si_001.cif
Supplemental Material - cs7b04423_si_002.pdf
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Additional details
- Eprint ID
- 85240
- Resolver ID
- CaltechAUTHORS:20180312-085606768
- NSF
- MCB-1513007
- Defense Advanced Research Projects Agency (DARPA)
- HR0011-15-C-0093
- Deutsche Forschungsgemeinschaft (DFG)
- BR 5238/1-1
- Swiss National Science Foundation (SNSF)
- P300PA-171225
- Resnick Sustainability Institute
- NSF Graduate Research Fellowship
- DGE-1745301
- Caltech Center for Environmental Microbial Interactions (CEMI)
- Created
-
2018-03-12Created from EPrint's datestamp field
- Updated
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2021-11-15Created from EPrint's last_modified field
- Caltech groups
- Resnick Sustainability Institute, Caltech Center for Environmental Microbial Interactions (CEMI), Rosen Bioengineering Center