Genetically Tunable Enzymatic C‒H Amidation for Lactam Synthesis
- Creators
-
Cho, Inha
-
Jia, Zhi-Jun
-
Arnold, Frances H.
Abstract
A major challenge in carbon‒hydrogen (C‒H) bond functionalization is to have the catalyst control precisely where a reaction takes place. Here we report engineered cytochrome P450 enzymes that perform unprecedented enantioselective C‒H amidation reactions and control the site selectivity to divergently construct β-, γ- and δ-lactams, completely overruling the inherent reactivities of the C‒H bonds. The enzymes, expressed in Escherichia coli cells, accomplish this abiological carbon‒nitrogen (C‒N) bond formation via reactive iron-bound carbonyl nitrenes generated from nature-inspired acyl-protected hydroxamate precursors. This transformation is exceptionally efficient (up to 1,020,000 total turnovers) and selective (up to 25:1 regioselectivity and 96% enantiomeric excess), and can be performed easily on preparative scale.
Additional Information
Preprint submitted on 12.02.2019, 21:02 and posted on 13.02.2019, 09:02. Declaration of Conflict of Interest: Caltech has filed a provisional patent application (CIT-8187-P). We thank K. Chen, S. Brinkmann-Chen, D. C. Miller and D. K. Romney for comments on the manuscript; X. Huang for helpful discussions; K. Chen for the parent variant E10FA; The Caltech Center for Catalysis and Chemical Synthesis and the Caltech Mass Spectrometry Laboratory for analytical support. Funding: Supported by NSF Division of Molecular and Cellular Biosciences grant MCB-1513007; Joseph J. Jacobs Institute for Molecular Engineering for Medicine (I.C.); and Deutsche Forschungsgemeinschaft (JI 289/1-1 to Z.-J.J.). Author contributions: I.C. and Z.-J.J. designed the research and performed the experiments with guidance from F.H.A.. I.C., Z.-J.J. and F.H.A. wrote the manuscript. Competing interests: Caltech has filed a provisional patent application (CIT-8187-P); and Data and materials availability: All data are available in the main text or the supplementary materials.Attached Files
Submitted - Cho_Genetically_Tunable_Enzymatic.pdf
Files
| Name | Size | Download all |
|---|---|---|
|
md5:a1770af62b07777a68baa54c6d614807
|
917.4 kB | Preview Download |
Additional details
- Eprint ID
- 93736
- Resolver ID
- CaltechAUTHORS:20190312-131401661
- MCB-1513007
- NSF
- Jacobs Institute for Molecular Engineering for Medicine
- JI 289/1-1
- Deutsche Forschungsgemeinschaft (DFG)
- Created
-
2019-03-12Created from EPrint's datestamp field
- Updated
-
2019-10-14Created from EPrint's last_modified field
- Caltech groups
- Jacobs Institute for Molecular Engineering for Medicine