Circular Permutated PQQ-Glucose Dehydrogenase as an Ultrasensitive Electrochemical Biosensor
Dr. Zhong Guo
CSIRO-QUT Synthetic Biology Alliance, ARC Centre of Excellence in Synthetic Biology, Centre for Agriculture and the Bioeconomy, Centre for Genomics and Personalised Health, School of Biology and Environmental Science, Queensland University of Technology, Brisbane, QLD, 4001 Australia
Search for more papers by this authorDr. Oleh Smutok
Department of Chemistry and Biomolecular Science, Clarkson University, 8 Clarkson Ave., Potsdam, NY, 13699 USA
Search for more papers by this authorDr. Wayne A. Johnston
CSIRO-QUT Synthetic Biology Alliance, ARC Centre of Excellence in Synthetic Biology, Centre for Agriculture and the Bioeconomy, Centre for Genomics and Personalised Health, School of Biology and Environmental Science, Queensland University of Technology, Brisbane, QLD, 4001 Australia
Search for more papers by this authorCagla Ergun Ayva
CSIRO-QUT Synthetic Biology Alliance, ARC Centre of Excellence in Synthetic Biology, Centre for Agriculture and the Bioeconomy, Centre for Genomics and Personalised Health, School of Biology and Environmental Science, Queensland University of Technology, Brisbane, QLD, 4001 Australia
Search for more papers by this authorDr. Patricia Walden
CSIRO-QUT Synthetic Biology Alliance, ARC Centre of Excellence in Synthetic Biology, Centre for Agriculture and the Bioeconomy, Centre for Genomics and Personalised Health, School of Biology and Environmental Science, Queensland University of Technology, Brisbane, QLD, 4001 Australia
Search for more papers by this authorBrett McWhinney
Department of Chemical Pathology, Pathology Queensland, Brisbane, QLD, 4001 Australia
Search for more papers by this authorProf. Jacobus P. J. Ungerer
Department of Chemical Pathology, Pathology Queensland, Brisbane, QLD, 4001 Australia
Faculty of Health and Behavioural Sciences, University of Queensland, Brisbane, QLD, 4072 Australia
Search for more papers by this authorProf. Artem Melman
Department of Chemistry and Biomolecular Science, Clarkson University, 8 Clarkson Ave., Potsdam, NY, 13699 USA
Search for more papers by this authorProf. Evgeny Katz
Department of Chemistry and Biomolecular Science, Clarkson University, 8 Clarkson Ave., Potsdam, NY, 13699 USA
Search for more papers by this authorCorresponding Author
Prof. Kirill Alexandrov
CSIRO-QUT Synthetic Biology Alliance, ARC Centre of Excellence in Synthetic Biology, Centre for Agriculture and the Bioeconomy, Centre for Genomics and Personalised Health, School of Biology and Environmental Science, Queensland University of Technology, Brisbane, QLD, 4001 Australia
Search for more papers by this authorDr. Zhong Guo
CSIRO-QUT Synthetic Biology Alliance, ARC Centre of Excellence in Synthetic Biology, Centre for Agriculture and the Bioeconomy, Centre for Genomics and Personalised Health, School of Biology and Environmental Science, Queensland University of Technology, Brisbane, QLD, 4001 Australia
Search for more papers by this authorDr. Oleh Smutok
Department of Chemistry and Biomolecular Science, Clarkson University, 8 Clarkson Ave., Potsdam, NY, 13699 USA
Search for more papers by this authorDr. Wayne A. Johnston
CSIRO-QUT Synthetic Biology Alliance, ARC Centre of Excellence in Synthetic Biology, Centre for Agriculture and the Bioeconomy, Centre for Genomics and Personalised Health, School of Biology and Environmental Science, Queensland University of Technology, Brisbane, QLD, 4001 Australia
Search for more papers by this authorCagla Ergun Ayva
CSIRO-QUT Synthetic Biology Alliance, ARC Centre of Excellence in Synthetic Biology, Centre for Agriculture and the Bioeconomy, Centre for Genomics and Personalised Health, School of Biology and Environmental Science, Queensland University of Technology, Brisbane, QLD, 4001 Australia
Search for more papers by this authorDr. Patricia Walden
CSIRO-QUT Synthetic Biology Alliance, ARC Centre of Excellence in Synthetic Biology, Centre for Agriculture and the Bioeconomy, Centre for Genomics and Personalised Health, School of Biology and Environmental Science, Queensland University of Technology, Brisbane, QLD, 4001 Australia
Search for more papers by this authorBrett McWhinney
Department of Chemical Pathology, Pathology Queensland, Brisbane, QLD, 4001 Australia
Search for more papers by this authorProf. Jacobus P. J. Ungerer
Department of Chemical Pathology, Pathology Queensland, Brisbane, QLD, 4001 Australia
Faculty of Health and Behavioural Sciences, University of Queensland, Brisbane, QLD, 4072 Australia
Search for more papers by this authorProf. Artem Melman
Department of Chemistry and Biomolecular Science, Clarkson University, 8 Clarkson Ave., Potsdam, NY, 13699 USA
Search for more papers by this authorProf. Evgeny Katz
Department of Chemistry and Biomolecular Science, Clarkson University, 8 Clarkson Ave., Potsdam, NY, 13699 USA
Search for more papers by this authorCorresponding Author
Prof. Kirill Alexandrov
CSIRO-QUT Synthetic Biology Alliance, ARC Centre of Excellence in Synthetic Biology, Centre for Agriculture and the Bioeconomy, Centre for Genomics and Personalised Health, School of Biology and Environmental Science, Queensland University of Technology, Brisbane, QLD, 4001 Australia
Search for more papers by this authorAbstract
Protein biosensors play an increasingly important role as reporters for research and clinical applications. Here we present an approach for the construction of fully integrated but modular electrochemical biosensors based on the principal component of glucose monitors PQQ-glucose dehydrogenase (PQQ-GDH). We designed allosterically regulated circular permutated variants of PQQ-GDH that show large (>10-fold) changes in enzymatic activity following intramolecular scaffolding of the newly generated N- and C termini by ligand binding domain/ligand complexes. The developed biosensors demonstrated sub-nanomolar affinities for small molecules and proteins in colorimetric and electrochemical assays. For instance, the concentration of Cyclosporine A could be measured in 1 μL of undiluted blood with the same accuracy as the leading diagnostic technique that uses 50 times more sample. We further used this biosensor to construct highly porous gold bioelectrodes capable of robustly detecting concentrations of Cyclosporine A as low as 20 pM and retained functionality in samples containing at least 60 % human serum.
Conflict of interest
KA and ZG are inventors on the patent covering the presented designs.
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