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Third Generation ATP Sensor with Enzymatic Analyte Recycling
Author(s) -
Yarman Aysu,
Schulz Christopher,
Sygmund Cristoph,
Ludwig Roland,
Gorton Lo,
Wollenberger Ulla,
Scheller Frieder W.
Publication year - 2014
Publication title -
electroanalysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.574
H-Index - 128
eISSN - 1521-4109
pISSN - 1040-0397
DOI - 10.1002/elan.201400231
Subject(s) - analyte , chemistry , detection limit , gluconic acid , pyruvate kinase , electron transfer , ascorbic acid , cellobiose dehydrogenase , inorganic chemistry , enzyme , biochemistry , cellobiose , chromatography , photochemistry , glycolysis , cellulase , food science
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).