In nature, pyruvate oxidase employs two cofactors thiamine pyrophosphate (ThDP) and Flavin adenine dinucleotide (FAD) to catalyze a conversion of pyruvate to acetyl phosphate. First, ThDP mediates a decarboxylation of pyruvate and generates an active aldehyde as a product. The aldehyde is then oxidized by FAD and is subsequently attacked by phosphate to yield acetyl phosphate.
Diederich and coworkers mimicked this system with a supramolecular catalyst based on cyclophane. The catalyst has thiazolium ion, a reactive part of ThDP and flavin, a bare-bones core of FAD, in close proximity and near the substrate binding site. The catalytic cycle is almost the same as that in nature, except the substrate is an aromatic aldehyde rather than pyruvate. First, the catalyst binds the substrate within its cyclophane ring. Then, it uses thiazolium ion to condense with the substrate generating an active aldehyde. This aldehyde is oxidized by flavin and then attacked by methanol to yield a methyl ester.