title: Scanning electrochemical microscopy of quinoprotein glucose dehydrogenase.
abstract: The activity of immobilized glucose dehydrogenase (GDH), a typical PQQ-dependent quinoprotein, was studied qualitatively and quantitatively by scanning electrochemical microscopy (SECM). PQQ-dependent GDH is of interest because of its high activity and independence of dissolved oxygen in catalyzing the transfer of electrons from glucose to an electron mediator. Biotinylated glucose dehydrogenase was bound to streptavidin-coated paramagnetic beads (surface concentration > or = 1.8 x 10(-11) mol cm(-2)) which were deposited as microscopic microspots on a hydrophobic surface. The catalytic activity of immobilized GDH was mapped in SECM feedback mode and generation-collection mode using ferrocenemethanol, ferrocenecarboxylic acid, p-aminophenol, and ferricyanide as electron mediators, respectively. The apparent steady-state kinetics of catalysis were measured under conditions of high d-glucose concentration using the theory developed for the SECM feedback and generation collection (GC) modes. In feedback mode, curves of the kinetically controlled substrate current against normalized distance were plotted, and it was found that GDH catalysis follows pseudo-first-order kinetics. In GC mode detection, the catalysis follows zero-order kinetics in the presence of high concentration of both substrates for GDH. The turnover rate obtained for immobilized GDH is lower than that of native GDH but much higher than that generally observed for glucose oxidase.
authors.affiliation: Department of Chemistry and Institute of Chemistry and Biology of the Marine Environment, Carl Von Ossietzky University of Oldenburg, D-26111 Oldenburg, Germany.
authors.forename: Chuan
authors.initials: C
authors.lastname: Zhao
authors.forename: Gunther
authors.initials: G
authors.lastname: Wittstock
mesh_terms: D004563:Electrochemistry; D004579:Electron Transport; D005948:Glucose Dehydrogenases / Q000737:chemistry* / Q000378:metabolism; D008853:Microscopy / Q000379:methods*
chemical_list: D005948:Glucose Dehydrogenases; C033955:glucose dehydrogenase (pyrroloquinoline-quinone)