Third Generation ATP Sensor with Enzymatic Analyte Recycling

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).

Transition metal oxide based non‐enzymatic electrochemical sensors: An arising approach for the meticulous detection of neurotransmitter biomarkers

“…. [54][55][56][57] Here, the judicious selection of the immobilized enzyme is imperative as its stability and selectivity influence the efficiency of the sensor. The electroactivity of the analyte is an inevitable requirement for the electrochemical analysis.…”

“…. [ 54–57 ] Here, the judicious selection of the immobilized enzyme is imperative as its stability and selectivity influence the efficiency of the sensor.…”

A precursor-approach in constructing 3D ITO electrodes for the improved performance of photosystem I-cyt c photobioelectrodes

“…One possibility for communication between multiple enzymes has been exploited by using intermediates of the reaction and establishing enzyme cascades, enzyme competition or recycling schemes on electrodes. [340][341][342] As a further advancement the concept of metabolic channeling can be considered by constructing multi-enzyme complexes in an artificial way with the aim to reduce the diffusion pathways. 343,344 Another step in the development of multi-protein biohybrid systems represents the establishment of direct electron exchange between immobilized proteins on electrode surfaces.…”

Studying Direct Electron Transfer by Site‐Directed Immobilization of Cellobiose Dehydrogenase

“…The growing interest in this enzyme derives from its ability to participate in DET at electrode surfaces, thanks to its haem cofactor that acts as an “internal mediator” shuttling electrons from the catalytic‐site to the electrode. Thus CDH has been used to construct biosensors for the detection of different carbohydrates such as glucose and lactose, as well as for ATP, catecholamines and phenolic pollutants . In biofuel cells it has been used as an anodic biocatalyst in glucose, lactose or cellobiose biofuel cells .…”