Electrochemically and Catalytically Active Reconstituted Horseradish Peroxidase with Ferrocene-Modified Hemin and an Artificial Binding Site

The cofactor hemin modified with aminomethylferrocene was loaded into the apo form of horseradish peroxidase (HRP) to give Fc-HRP (scheme); this reconstituted enzyme is electrochemically and catalytically active with respect to ABTS and water-soluble ferrocenes. The ferrocenes display kinetically meaningful affinity to the reconstituted enzyme owing to creation of a novel artificial hydrophobic binding site at the protein surface.