Computational Modeling of the Enzymatic Achmatowicz Rearrangement by Heme-Dependent Chloroperoxidase: Reaction Mechanism, Enantiopreference, Regioselectivity, and Substrate Specificity

The chloroperoxidase from Caldariomyces fumago (CfCPO) catalyzes the oxidative ring expansion of α-heterofunctionalized furans via the Achmatowicz rearrangement, providing an elegant tool to convert furan rings into complex-prefunctionalized scaffolds. However, the mechanism of this transformation remains unclear. Herein, the CfCPO-catalyzed reaction of rac-1-(2-furyl)ethanol (1a) is studied by quantum chemical calculations and molecular dynamics simulations. The calculations reveal that the conversion follows the general mechanism of the Achmatowicz reaction. Notably, the binding of 1a to the enzyme's active site influences the Compound I (Cpd I) formation, and the (R)-1a enantiomer binding results in a lower barrier compared to (S)-1a, explaining the observed (R)-enantiopreference toward a racemic substrate. Additionally, due to the weaker steric hindrance between the porphyrin ring and substrate, the nucleophilic attack of Cpd I on the furan core of 1a is preferred at the less-substituted C4=C5 bond, providing a rationale for the experimentally observed regioselectivity. Finally, the bottleneck residues in the substrate delivery channel and also the active site surroundings are proposed to be responsible for the substrate specificity of CfCPO. This study lays a theoretical foundation for the rational design of new CPOs that catalyze the Achmatowicz rearrangement with a broader substrate spectrum or specific stereopreference.