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Enzymatic Metal-Hydrogen Atom Transfer with a Cobalt Protoporphyrin Cofactor
Carly L Masonheimer1, Michael J Rourke1, Reece S Gardner1
1Department of Chemistry, University of Wisconsin─Madison, Madison, Wisconsin 53706, United States.
Researchers engineered a P450 enzyme using an unnatural cobalt cofactor (CoPPIX) to perform metal-hydrogen atom transfer (MHAT) reactions. This biocatalytic approach enabled new deallylation and reductive dearomatization reactions, expanding enzymatic capabilities.
Area of Science:
- Biocatalysis and enzyme engineering
- Bioinorganic chemistry
- Synthetic biology
Background:
- Unnatural cofactors can create novel enzymatic reactivity.
- Metal-hydrogen atom transfer (MHAT) is a key synthetic reaction not observed in nature.
- P450 enzymes are versatile biocatalysts.
Purpose of the Study:
- To introduce unnatural cofactors for new biocatalytic reactions.
- To engineer a P450 enzyme (CYP119) for metal-hydrogen atom transfer (MHAT) reactivity.
- To develop enzymes for deallylation and reductive dearomatization.
Main Methods:
- De novo biosynthesis of cobalt protoporphyrin IX (CoPPIX) cofactor.
- Directed evolution using a colorimetric screen for MHAT activity.
- Mechanistic studies of enzyme variants.
Main Results:
- Generated a mononuclear cobalt hydride intermediate in CYP119.
- Engineered CYP119 variants for MHAT-mediated deallylation of nitrophenols.
- Discovered MHAT-mediated reductive dearomatization of aromatic rings by enzyme variants.
Conclusions:
- Biocatalytic MHAT is achievable using unnatural cofactors.
- Enzyme engineering can create novel catalytic functions like reductive dearomatization.
- Metal substitution in enzymes offers a route to tune reactivity.
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