title: Interaction of Flavin-Dependent Fructose Dehydrogenase with Cytochrome c as Basis for the Construction of Biomacromolecular Architectures on Electrodes.
abstract: The creation of electron transfer (ET) chains based on the defined arrangement of enzymes and redox proteins on electrode surfaces represents an interesting approach within the field of bioelectrocatalysis. In this study, we investigated the ET reaction of the flavin-dependent enzyme fructose dehydrogenase (FDH) with the redox protein cytochrome c (cyt c). Two different pH optima were found for the reaction in acidic and neutral solutions. When cyt c was adsorbed on an electrode surface while the enzyme remained in solution, ET proceeded efficiently in media of neutral pH. Interprotein ET was also observed in acidic media; however, it appeared to be less efficient. These findings suggest that two different ET pathways between the enzyme and cyt c may occur. Moreover, cyt c and FDH were immobilized in multiple layers on an electrode surface by means of another biomacromolecule: DNA (double stranded) using the layer-by-layer technique. The biprotein multilayer architecture showed a catalytic response in dependence on the fructose concentration, indicating that the ET reaction between both proteins is feasible even in the immobilized state. The electrode showed a defined response to fructose and a good storage stability. Our results contribute to the better understanding of the ET reaction between FDH and cyt c and provide the basis for the creation of all-biomolecule based fructose sensors the sensitivity of which can be controlled by the layer preparation.
authors.affiliation: Biosystems Technology, Institute of Applied Life Sciences, Technical University of Applied Sciences Wildau , Hochschulring 1, 15745 Wildau, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Wettstein
authors.affiliation: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University , Sakyo, Kyoto 606-8502, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Kano
authors.affiliation: Biosystems Technology, Institute of Applied Life Sciences, Technical University of Applied Sciences Wildau , Hochschulring 1, 15745 Wildau, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Schäfer
authors.affiliation: Institute of Biochemistry and Biology, University Potsdam , Karl-Liebknecht-Strasse 24-25, 14476 Potsdam/Golm, Germany.
authors.forename: Ulla
authors.initials: U
authors.lastname: Wollenberger
authors.affiliation: Biosystems Technology, Institute of Applied Life Sciences, Technical University of Applied Sciences Wildau , Hochschulring 1, 15745 Wildau, Germany.
authors.forename: Fred
authors.initials: F
authors.lastname: Lisdat
mesh_terms: D000327:Adsorption; D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D002237:Carbohydrate Dehydrogenases / Q000737:chemistry* / Q000378:metabolism; D045304:Cytochromes c / Q000737:chemistry* / Q000378:metabolism; D004058:Diffusion; D004566:Electrodes; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism; D020576:Gluconobacter / Q000737:chemistry / Q000201:enzymology* / Q000378:metabolism; D006736:Horses; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular
chemical_list: D004800:Enzymes, Immobilized; D045304:Cytochromes c; D002237:Carbohydrate Dehydrogenases; C028761:D-fructose 5-dehydrogenase