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Third Generation ATP Sensor with Enzymatic Analyte Recycling |
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| Authors: | Aysu Yarman Christopher Schulz Cristoph Sygmund Roland Ludwig Lo Gorton Ulla Wollenberger Frieder W Scheller |
| Affiliation: | 1. Fraunhofer Institute for Cell Therapy and Immunology (IZI), Branch Bioanalytics and Bioprocesses (IZI‐BB), Am Mühlenberg 13, 14476 Potsdam, Germany;2. Institute for Biochemistry and Biology, University of Potsdam, Karl‐Liebknecht Strasse 24–25, 14476 Potsdam, Germany;3. Department of Analytical Chemistry/Biochemistry and Structural Biology, Lund University, P.O. Box 124, 226 46, Lund, Sweden;4. Department of Food Science and Technology, BOKU – University of Natural Resources and Life Sciences, Vienna, Muthgasse 18, 1190, Vienna, Austria |
| Abstract: | For the first time the direct electron transfer of an enzyme ‐ cellobiose dehydrogenase, CDH ‐ has been coupled with the hexokinase catalyzed competition for glucose in a sensor for ATP. To enhance the signal output for ATP, pyruvate kinase was coimmobilized to recycle ADP by the phosphoenolpyruvate driven reaction. The new sensor overcomes the limit of 1 : 1 stoichiometry of the sequential or competitive conversion of ATP by effective enzymatic recycling of the analyte. The anodic oxidation of the glucose converting CDH proceeds at electrode potentials below 0 mV vs. Ag|AgCl thus potentially interfering substances like ascorbic acid or catecholamines do not influence the measuring signal. The combination of direct electron transfer of CDH with the enzymatic recycling results in an interference‐free and oxygen‐independent measurement of ATP in the lower µmolar concentration range with a lower limit of detection of 63.3 nM (S/N=3). |
| Keywords: | ATP Third generation sensor Enzymatic recycling Cellobiose dehydrogenase Hexokinase Pyruvate kinase |
