Highly Regioselective Trifluoromethylation of Unprotected Phenols and Anilines Enabled by an Engineered P450 Peroxizyme System

The trifluoromethyl (CF 3 ) group is a pivotal motif in pharmaceuticals, agrochemicals, and functional materials, yet achieving direct and regioselective aromatic C─H trifluoromethylation—especially for phenols and anilines—remains a formidable challenge. Here, we transform a dual‐functional small molecule (DFSM)‐facilitated P450BM3 peroxizyme into a versatile biocatalytic platform that enables highly ortho ‐selective trifluoromethylation of diverse phenols and anilines. Through protein engineering, we achieve up to 98% ortho ‐selectivity for phenol—the first highly selective enzymatic trifluoromethylation reported. The system exhibits broad substrate scope and functional‐group tolerance, delivering >99% regioselectivity and over 6700 turnovers for leading substrates. Mechanistic studies confirm a radical pathway and reveal that selectivity is engineered via synergistic mutations‐DFSM interactions that pre‐organize substrates in a confined active site. This work establishes a programmable strategy to bridge biocatalysis and organofluorine chemistry, offering a sustainable route to valuable fluorinated building blocks.