Evolved Aliphatic Halogenases Enable Regiocomplementary C-H Functionalization of a Pharmaceutically Relevant Compound.
- Hayashi T Competence Center for Biocatalysis, Institute of Chemistry and Biotechnology, Zürich University of Applied Sciences, Einsiedlerstrasse 31, 8820, Wädenswil, Switzerland.
- Ligibel M Novartis Institutes for BioMedical Research, Global Discovery Chemistry, 4056, Basel, Switzerland.
- Sager E Novartis Institutes for BioMedical Research, Global Discovery Chemistry, 4056, Basel, Switzerland.
- Voss M Competence Center for Biocatalysis, Institute of Chemistry and Biotechnology, Zürich University of Applied Sciences, Einsiedlerstrasse 31, 8820, Wädenswil, Switzerland.
- Hunziker J Novartis Institutes for BioMedical Research, Global Discovery Chemistry, 4056, Basel, Switzerland.
- Schroer K Novartis Institutes for BioMedical Research, Global Discovery Chemistry, 4056, Basel, Switzerland.
- Snajdrova R Novartis Institutes for BioMedical Research, Global Discovery Chemistry, 4056, Basel, Switzerland.
- Buller R Competence Center for Biocatalysis, Institute of Chemistry and Biotechnology, Zürich University of Applied Sciences, Einsiedlerstrasse 31, 8820, Wädenswil, Switzerland.
- 2019-10-08
Published in:
- Angewandte Chemie (International ed. in English). - 2019
C−H activation
biocatalysis
directed evolution
halogenases
non-heme iron enzymes
Bacterial Proteins
Halogenation
Humans
Molecular Structure
Substrate Specificity
English
Non-heme iron halogenases are synthetically valuable biocatalysts that are capable of halogenating unactivated sp3 -hybridized carbon centers with high stereo- and regioselectivity. The reported substrate scope of these enzymes, however, is limited primarily to the natural substrates and their analogues. We engineered the halogenase WelO5* for chlorination of a martinelline-derived fragment. Using structure-guided evolution, a halogenase variant with a more than 290-fold higher total turnover number and a 400-fold higher apparent kcat compared to the wildtype enzyme was generated. Moreover, we identified key positions in the active site that allow direction of the halogen to different positions in the target substrate. This is the first example of enzyme engineering to expand the substrate scope of a non-heme iron halogenase beyond the native indole-alkaloid-type substrates. The highly evolvable nature of WelO5* underscores the usefulness of this enzyme family for late-stage halogenation.
- Language
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- English
- Open access status
- green
- Identifiers
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- DOI 10.1002/anie.201907245
- PMID 31589798
- Persistent URL
- https://folia.unifr.ch/global/documents/279184
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