Reconstitution of apo-glucose oxidase with a nitrospiropyran-modified FAD cofactor yields a photoswitchable biocatalyst for amperometric transduction of recorded optical signals

Here we wish to report on a novel method to tailor photoswitchable redox biocatalysts by reconstitution of apo-flavoenzymes with a nitrospiropyran modified flavin adenine dinucleotide, FAD, cofactor. We demonstrate a novel method a organize photoswitchable redox proteins for amperometric transduction of optical signals. The method consists of reconstitution of a flavo-apoenzyme, apo-GOD, with a synthetic FAD analog modified by a nitrospiropyran photosiomerizable unit. The protein implanted photoisomerizable group delicately controls the protein structure in the FAD cofactor surrounding. The site-specific modification of the enzyme by the photoisomerizable units through the application of the reconstitution methodology reconstitution methodology represents a major advance in designing photoswitchable enzymes. It enables further structural characterization of the biocatalyst and elucidation of the steric perturbations of the protein stimulated by photoisomerizable components.