Amperometric ATP Biosensors Based on Coimmobilizations of p-Hydroxybenzoate Hydroxylase, Glucose-6-Phosphate Dehydrogenase, and Hexokinase on Clark-Type and Screen-Printed Electrodes

Abstract

Two types of amperometric trienzyme ATP biosensors were developed based on new combinations of enzymes and electrodes by using the coimmobilizations of p-hydroxybenzoate hydroxylase (HBH), glucose-6-phosphate dehydrogenase (G6PDH), and hexokinase (HEX) on a Clark-type oxygen electrode and on a screen-printed electrode. The principles of the determinations are as follows: HEX transfers the phosphate group from ATP to glucose to form glucose-6-phosphate. G6PDH catalyzes the specific dehydrogenation of glucose-6-phosphate by consuming NADP + . The product, NADPH initiates the irreversible hydroxylation of p-hydroxybenzoate by HBH to consume dissolved oxygen which results in a detectable signal on a Clark-type electrode and generate 3, 4-dihydroxybenzoate which results in a detectable signal on a screen-printed electrode. Both sensors show high-performance characteristics with broad detection ranges, short measuring times and good specificities.Index Terms-Adenosine-5'-triphosphate, biosensor, Clark-type electrode, glucose-6-phosphate dehydrogenase, hexokinase, p-hydroxybenzoate hydroxylase, screen-printed electrode.

Forizymes – functionalised artificial forisomes as a platform for the production and immobilisation of single enzymes and multi-enzyme complexes

“…S3 ). For example, our model HXK2 and G6PDH forizymes could be developed into lab-on-a-chip devices for the detection of glucose or ATP, because such devices require micromolecular pump and valve systems as well as enzymatic cascades for the measurement of glucose/ATP concentrations 48 49 . The construction of such devices remains a challenging task that will need to be addressed in the future, but our novel forizymes combine biosensor and catalytic properties that would be highly advantageous when integrated into such devices.…”

A review on amperometric-type immunosensors based on screen-printed electrodes

“…104,105 Because of its simplicity, electrochemical transduction has gained more popularity over other mechanisms and has been widely employed in various low-cost devices. 106 The main electrochemical transducers are amperometric 107 (measuring the current), potentiometric 108 (measuring the electrode potential or voltage differences) and conductometric (measuring the conductivity or resistance); 109 however, amperometric transducers based on signal generation aer an enzymatic reaction are mostly developed 106 owing to their high sensitivity and wide applicability. In the case of screen-printed electrodes based on the amperometric principle, the WE serves as the transduction element in biochemical reactions.…”