Diversity-Oriented Enzymatic Synthesis of Cyclopropane Building Blocks
Abstract
While biocatalysis is increasingly incorporated into drug development pipelines, it is less commonly used in the early stages of drug discovery. By engineering a protein to produce a chiral motif with a derivatizable functional handle, biocatalysts can be used to help generate diverse building blocks for drug discovery. Here we show the engineering of two variants of Rhodothermus marinus nitric oxide dioxygenase (RmaNOD) to catalyze the formation of cis- and trans-diastereomers of a pinacolboronate-substituted cyclopropane which can be readily derivatized to generate diverse stereopure cyclopropane building blocks.
Additional Information
© 2020 American Chemical Society. Received: April 28, 2020; Revised: June 4, 2020; Published: June 4, 2020. This work was supported by NSF MCB (grant 1513007 to F.H.A.), NSF STTR (grant 1738308 to F.H.A.), and the NIH (grant R35GM118191-01 to S.E.R.). Graduate student support from NIH T32 training grants GM112592 (A.M.K.) and GM07616 (B.J.W.), and the NSF Graduate Research Fellowship Program (A.M.K. and J.L.H., DGE-1144469), is acknowledged. The authors thank Mr. Lawrence M. Henling for assistance with small-molecule X-ray crystallography, as well as Dr. Mona Shahgholi and Naseem Torian for assistance with mass spectrometry measurements. Crystallography experiments were supported by Jens Kaiser and the Caltech Molecular Observatory. The authors thank Jingzhou Wang for technical assistance, David Rozzell, Nathaniel Goldberg, Ferdinand Huber, Nicholas Porter, and Kari Hernandez for valuable discussions, and Sabine Brinkmann-Chen and Zhen Liu for critical reading of the manuscript. Author Contributions: B.J.W., A.M.K.: These authors contributed equally. The manuscript was written through contributions of all authors. All authors have given approval to the final version of the manuscript. The authors declare the following competing financial interest(s): A provisional patent, on which A.M.K., B.J.W., and S.B.J.K. are inventors, has been filed through the California Institute of Technology based on the results presented here.Attached Files
Accepted Version - nihms-1604406.pdf
Supplemental Material - cs0c01888_si_001.pdf
Supplemental Material - cs0c01888_si_002.xlsx
Supplemental Material - cs0c01888_si_003.cif
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Additional details
- PMCID
- PMC7709968
- Eprint ID
- 103768
- Resolver ID
- CaltechAUTHORS:20200608-115355402
- NSF
- MCB-1513007
- NSF
- IIP-1738308
- NIH
- R35GM118191-01
- NIH Predoctoral Fellowship
- T32 GM112592
- NIH Predoctoral Fellowship
- T32 GM07616
- NSF Graduate Research Fellowship
- DGE-1144469
- Created
-
2020-06-08Created from EPrint's datestamp field
- Updated
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2022-02-09Created from EPrint's last_modified field
- Caltech groups
- Division of Biology and Biological Engineering (BBE)