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Diverse engineered heme proteins enable stereodivergent cyclopropanation of unactivated alkenes

Knight, Anders M. and Kan, S. B. Jennifer and Lewis, Russell D. and Brandenberg, Oliver F. and Chen, Kai and Arnold, Frances H. (2018) Diverse engineered heme proteins enable stereodivergent cyclopropanation of unactivated alkenes. ACS Central Science, 4 (3). pp. 372-377. ISSN 2374-7943. PMCID PMC5879470. http://resolver.caltech.edu/CaltechAUTHORS:20171220-092532762

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Abstract

Developing catalysts that produce each stereoisomer of a desired product selectively is a longstanding synthetic challenge. Biochemists have addressed this challenge by screening nature’s diversity to discover enzymes that catalyze the formation of complementary stereoisomers. We show here that the same approach can be applied to a new-to-nature enzymatic reaction, alkene cyclopropanation via carbene transfer. By screening diverse native and engineered heme proteins, we identified globins and serine-ligated “P411” variants of cytochromes P450 with promiscuous activity for cyclopropanation of unactivated alkene substrates. We then enhanced their activities and stereoselectivities by directed evolution: just 1–3 rounds of site-saturation mutagenesis and screening generated enzymes that transform unactivated alkenes and electron-deficient alkenes into each of the four stereoisomeric cyclopropanes with up to 5,400 total turnovers and 98% enantiomeric excess. These fully genetically encoded biocatalysts function in whole Escherichia coli cells in mild, aqueous conditions and provide the first example of enantioselective, intermolecular iron-catalyzed cyclopropanation of unactivated alkenes.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1021/acscentsci.7b00548DOIArticle
https://pubs.acs.org/doi/suppl/10.1021/acscentsci.7b00548PublisherSupporting Information
https://doi.org/10.26434/chemrxiv.5718076.v1DOIDiscussion Paper
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5879470PubMed CentralArticle
ORCID:
AuthorORCID
Knight, Anders M.0000-0001-9665-8197
Kan, S. B. Jennifer0000-0001-6371-8042
Lewis, Russell D.0000-0002-5776-7347
Brandenberg, Oliver F.0000-0001-5662-1234
Chen, Kai0000-0002-3325-3536
Arnold, Frances H.0000-0002-4027-364X
Additional Information:© 2018 American Chemical Society. ACS AuthorChoice - 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: November 10, 2017; Publication Date (Web): February 21, 2018. This work was supported by the National Science Foundation Division of Molecular and Cellular Biosciences (Grant MCB-1513007) and the Office of Chemical, Bioengineering, Environmental and Transport Systems SusChEM Initiative (Grant CBET-1403077). The authors thank Dr. Nathan Dalleska, Aurapat Ngamnithiporn, and Dr. Scott C. Virgil for analytical chiral GC support, and Dr. Stephan C. Hammer and Dr. Xiongyi Huang for helpful discussions and critical reading of the manuscript. A.M.K. gratefully acknowledges support from Caltech’s Center for Environmental Microbial Interactions and the NSF Graduate Research Fellowship (Grant No. DGE-1745301). R.D.L. is supported by an NIH–National Research Service Award training grant (5 T32 GM07616). O.F.B. acknowledges support from the Deutsche Forschungsgemeinschaft (Grant No. BR 5238/1-1) and the Swiss National Science Foundation (Grant No. P300PA-171225). A provisional patent application has been filed through the California Institute of Technology based on the results presented here. The authors declare no competing financial interest.
Group:Caltech Center for Environmental Microbial Interactions (CEMI), Rosen Bioengineering Center
Funders:
Funding AgencyGrant Number
NSFMCB-1513007
NSFCBET-1403077
Caltech Center for Environmental Microbial Interactions (CEMI)UNSPECIFIED
NSF Graduate Research FellowshipDGE-1745301
NIH Predoctoral Fellowship5 T32 GM07616
Deutsche Forschungsgemeinschaft (DFG)BR 5238/1-1
Swiss National Science Foundation (SNSF)P300PA-171225
Subject Keywords:Stereodivergence, Biocatalysis, Carbene Transfer, Heme Protein, Cyclopropanation, Directed Evolution
PubMed Central ID:PMC5879470
Record Number:CaltechAUTHORS:20171220-092532762
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20171220-092532762
Official Citation:Diverse Engineered Heme Proteins Enable Stereodivergent Cyclopropanation of Unactivated Alkenes Anders M. Knight, S. B. Jennifer Kan, Russell D. Lewis, Oliver F. Brandenberg, Kai Chen, and Frances H. Arnold ACS Central Science 2018 4 (3), 372-377 DOI: 10.1021/acscentsci.7b00548
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:83972
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:20 Dec 2017 17:53
Last Modified:14 May 2019 19:56

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