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Enantioselective Enzyme-Catalyzed Aziridination Enabled by Active-Site Evolution of a Cytochrome P450

Farwell, Christopher C. and Zhang, Ruijie K. and McIntosh, John A. and Hyster, Todd K. and Arnold, Frances H. (2015) Enantioselective Enzyme-Catalyzed Aziridination Enabled by Active-Site Evolution of a Cytochrome P450. ACS Central Science, 1 (2). pp. 89-93. ISSN 2374-7951. PMCID PMC4571169. https://resolver.caltech.edu/CaltechAUTHORS:20150504-091714076

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Abstract

One of the greatest challenges in protein design is creating new enzymes, something evolution does all the time, starting from existing ones. Borrowing from nature’s evolutionary strategy, we have engineered a bacterial cytochrome P450 to catalyze highly enantioselective intermolecular aziridination, a synthetically useful reaction that has no natural biological counterpart. The new enzyme is fully genetically encoded, functions in vitro or in whole cells, and can be optimized rapidly to exhibit high enantioselectivity (up to 99% ee) and productivity (up to 1,000 catalytic turnovers) for intermolecular aziridination, demonstrated here with tosyl azide and substituted styrenes. This new aziridination activity highlights the remarkable ability of a natural enzyme to adapt and take on new functions. Once discovered in an evolvable enzyme, this non-natural activity was improved and its selectivity tuned through an evolutionary process of accumulating beneficial mutations.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1021/acscentsci.5b00056DOIArticle
http://pubs.acs.org/doi/abs/10.1021/acscentsci.5b00056PublisherArticle
http://pubs.acs.org/doi/suppl/10.1021/acscentsci.5b00056PublisherSupporting Information
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4571169/PubMed CentralArticle
ORCID:
AuthorORCID
Zhang, Ruijie K.0000-0002-7251-5527
McIntosh, John A.0000-0002-9487-490X
Hyster, Todd K.0000-0003-3560-355X
Arnold, Frances H.0000-0002-4027-364X
Additional Information:© 2015 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: February 23, 2015. Publication Date (Web): April 22, 2015. We thank Dr. S. Virgil and the 3CS Catalysis Center at Caltech for assistance with HPLC, chiral HPLC, and LC−MS analysis. We thank Yufan Liang for assistance with chiral HPLC, and Hans Renata, Christopher Prier, and Sheel Dodani for helpful discussions. This work was supported by the Department of the Navy, Office of Naval Research (Grant N00014-11-0205), and the Jacobs Institute for Molecular Engineering for Medicine at Caltech. C.C.F and R.K.Z. are supported by NSF graduate research fellowships. T.K.H and J.A.M. are supported by Ruth L. Kirschstein National Research Service Awards (F32GM108143) (F32GM101792).
Group:Jacobs Institute for Molecular Engineering for Medicine
Funders:
Funding AgencyGrant Number
Office of Naval Research (ONR)N00014-11-0205
Jacobs Institute for Molecular Engineering for MedicineUNSPECIFIED
NSF Graduate Research FellowshipUNSPECIFIED
NIH Predoctoral FellowshipF32GM108143
NIH Predoctoral FellowshipF32GM101792
Issue or Number:2
PubMed Central ID:PMC4571169
Record Number:CaltechAUTHORS:20150504-091714076
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20150504-091714076
Official Citation:Enantioselective Enzyme-Catalyzed Aziridination Enabled by Active-Site Evolution of a Cytochrome P450 Christopher C. Farwell, Ruijie K. Zhang, John A. McIntosh, Todd K. Hyster, and Frances H. Arnold ACS Central Science 2015 1 (2), 89-93 DOI: 10.1021/acscentsci.5b00056
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:57182
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:04 May 2015 17:07
Last Modified:14 Oct 2019 17:10

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