title: PQQ glucose dehydrogenase with novel electron transfer ability.
abstract: PQQ glucose dehydrogenase from Acinetobacter calcoaceticus (GDH-B) is one of the most industrially attractive enzymes, as a sensor constituent for glucose sensing, because of its high catalytic activity and insensitivity to oxygen. We attempted to engineer GDH-B to enable electron transfer to the electrode in the absence of artificial electron mediator by mimicking the domain structure of the quinohemoprotein ethanol dehydrogenase (QH-EDH) from Comamonas testosteroni, which is composed of a PQQ-containing catalytic domain and a cytochrome c domain. We genetically fused the cytochrome c domain of QH-EDH to the C-terminal of GDH-B. The constructed fusion protein showed not only intra-molecular electron transfer, between PQQ and heme of the cytochrome c domain, but also electron transfer from heme to the electrode, thereby allowing the construction of a direct electron transfer-type glucose sensor.
authors.affiliation: Department of Biotechnology, Faculty of Technology, Tokyo University of Agriculture and Technology, 2-24-16 Nakamachi, Koganei, 184-8588, Tokyo, Japan.
authors.forename: Junko
authors.initials: J
authors.lastname: Okuda
authors.forename: Koji
authors.initials: K
authors.lastname: Sode
mesh_terms: D016954:Acinetobacter calcoaceticus / Q000201:enzymology; D000429:Alcohol Oxidoreductases / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D015374:Biosensing Techniques / Q000379:methods; D020604:Comamonas testosteroni / Q000201:enzymology; D045304:Cytochromes c / Q000235:genetics; D004566:Electrodes; D004579:Electron Transport; D005947:Glucose / Q000032:analysis; D005948:Glucose Dehydrogenases / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D007700:Kinetics; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000235:genetics; D013053:Spectrophotometry / Q000379:methods
chemical_list: D011993:Recombinant Fusion Proteins; D045304:Cytochromes c; D000429:Alcohol Oxidoreductases; D005948:Glucose Dehydrogenases; C019641:alcohol dehydrogenase (acceptor); C033955:glucose dehydrogenase (pyrroloquinoline-quinone); D005947:Glucose