Teaching natural enzymes new radical tricks

Area of Science:

• Biocatalysis and enzyme engineering
• Organic chemistry
• Green chemistry

Background:

• Cytochromes P450 are versatile heme-containing enzymes involved in diverse oxidative reactions.
• Traditional P450 applications often require biological systems or cofactors.
• Abiotic radical reactions are crucial in organic synthesis but challenging to control stereoselectively.

Purpose of the Study:

• To engineer cytochromes P450 for catalyzing abiotic radical reactions.
• To achieve high stereoselectivity in these engineered enzymatic reactions.
• To develop novel biocatalytic tools for synthetic chemistry.

Main Methods:

• Directed evolution and protein engineering of cytochrome P450 variants.
• Development of abiotic reaction conditions compatible with enzyme stability.
• Stereochemical analysis of reaction products using chiral chromatography and spectroscopy.

Main Results:

• Engineered P450 variants demonstrated significant catalytic activity in abiotic radical reactions.
• High levels of stereoselectivity (enantiomeric and diastereomeric excess) were achieved.
• The engineered enzymes enabled the formation of complex chiral molecules via radical pathways.

Conclusions:

• Cytochromes P450 can be successfully engineered for abiotic stereoselective radical catalysis.
• This work expands the synthetic utility of P450 enzymes beyond their natural biological roles.
• The developed biocatalysts offer a sustainable and precise alternative for synthesizing chiral compounds.