title: Engineered PQQ-Glucose Dehydrogenase as a Universal Biosensor Platform.
abstract: Biosensors with direct electron output hold promise for nearly seamless integration with portable electronic devices. However, so far, they have been based on naturally occurring enzymes that significantly limit the spectrum of detectable analytes. Here, we present a novel biosensor architecture based on analyte-driven intermolecular recombination and activity reconstitution of a re-engineered component of glucometers: PQQ-glucose dehydrogenase. We demonstrate that this sensor architecture can be rapidly adopted for the detection of immunosuppressant drugs, α-amylase protein, or protease activity of thrombin and Factor Xa. The biosensors could be stored in dried form without appreciable loss of activity. We further show that ligand-induced activity of the developed biosensors could be directly monitored by chronoamperometry, enabling construction of disposable sensory electrodes. We expect that this architecture could be expanded to the detection of other biochemical activities, post-translational modifications, nucleic acids, and inorganic molecules.
authors.affiliation: Institute for Molecular Bioscience and Australian Institute for Bioengineering and Nanotechnology, The University of Queensland , Brisbane, QLD 4072, Australia.
authors.forename: Zhong
authors.initials: Z
authors.lastname: Guo
authors.affiliation: Molecular Warehouse Ltd. , 40 Occam Road, Guildford GU2 7YG, U.K.
authors.forename: Lindy
authors.initials: L
authors.lastname: Murphy
authors.affiliation: Institute for Molecular Bioscience and Australian Institute for Bioengineering and Nanotechnology, The University of Queensland , Brisbane, QLD 4072, Australia.
authors.forename: Viktor
authors.initials: V
authors.lastname: Stein
authors.affiliation: Institute for Molecular Bioscience and Australian Institute for Bioengineering and Nanotechnology, The University of Queensland , Brisbane, QLD 4072, Australia.
authors.forename: Wayne A
authors.initials: WA
authors.lastname: Johnston
authors.affiliation: Molecular Warehouse Ltd. , 40 Occam Road, Guildford GU2 7YG, U.K.
authors.forename: Siro
authors.initials: S
authors.lastname: Alcala-Perez
authors.affiliation: Institute for Molecular Bioscience and Australian Institute for Bioengineering and Nanotechnology, The University of Queensland , Brisbane, QLD 4072, Australia.
authors.forename: Kirill
authors.initials: K
authors.lastname: Alexandrov
mesh_terms: D000409:Alanine / Q000737:chemistry; D015415:Biomarkers / Q000378:metabolism; D015374:Biosensing Techniques; D004566:Electrodes; D004583:Electrons; D015951:Factor Xa / Q000737:chemistry; D005947:Glucose / Q000737:chemistry; D042843:Glucose 1-Dehydrogenase / Q000737:chemistry*; D006801:Humans; D007166:Immunosuppressive Agents; D007700:Kinetics; D000072417:Protein Domains; D015202:Protein Engineering / Q000379:methods*; D011499:Protein Processing, Post-Translational; D011995:Recombination, Genetic; D012680:Sensitivity and Specificity; D020123:Sirolimus / Q000737:chemistry; D013917:Thrombin / Q000737:chemistry; D000516:alpha-Amylases / Q000378:metabolism
chemical_list: D015415:Biomarkers; D007166:Immunosuppressive Agents; D042843:Glucose 1-Dehydrogenase; D000516:alpha-Amylases; D013917:Thrombin; D015951:Factor Xa; D005947:Glucose; D000409:Alanine; D020123:Sirolimus