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Third generation ATP sensor with enzymatic analyte recycling

: Yarman, Aysu; Schulz, Christopher; Sygmund, Christoph; Ludwig, Roland; Gorton, L.; Wollenberger, Ulla; Scheller, Frieder W.


Electroanalysis 26 (2014), Nr.9, S.2043-2048
ISSN: 1040-0397
ISSN: 1521-4109
Fraunhofer IZI ()
ATP; cellobiose dehydrogenase; enzymatic recycling; hexokinase; third generation sensor

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 vertical bar 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 mu molar concentration range with a lower limit of detection of 63.3 nM (S/N=3).