Biosensor Based on Cellobiose Dehydrogenase for Detection of Catecholamines

Abstract

A cellobiose dehydrogenase (CDH)-modified graphite electrode was designed for amperometric detection of catecholamines in the flow injection mode, by their recycling between the graphite electrode (+300 mV vs Ag|AgCl) and the reduced FAD cofactor of adsorbed CDH, resulting in an amplified response signal. The high efficiency of the enzyme-catecholamine reaction leads to a detection limit below 1 nM and a sensitivity of 15.8 A.M(-1) x cm(-2) (approximately 1150 nA/microM) for noradrenaline, with a coverage of less than 2.5 microg of CDH adsorbed on the electrode surface (0.073 cm(2)). Working parameters such as pH, cellobiose concentration, carrier buffer, and applied potential were optimized, using hydroquinone as a model analyte. The sensitivity, linear range, and amplification factor can be modulated by the steady-state concentration of cellobiose in the flow buffer. The response of the sensor decreases only 2% when run continuously for 4 h in the flow injection mode. The response peak maximum is obtained within 6 s at a flow rate of 0.5 mL/min, representing the time of the entire sample segment to pass the electrode. CDH enzymes from Phanerochaete chrysosporium and Sclerotium rolfsii were investigated, providing different characteristics of the sensor, with sensors made with CDH from P. chrysosporium being the better ones.

Cellobiose Dehydrogenase: A Versatile Catalyst for Electrochemical Applications

“…In the first case, a variety of quinones (Q) and related compounds, for example DCIP, have been reported to have high reactions rates with CDH(F r H o ) [reaction (2)]. [6,10,22] This reaction corresponds to the first step in the MET reaction [reaction (7)]:…”

“…This fungal enzyme has been applied in biosensors for the detection of cellobiose, [1] cellodextrins, [1a, 2] maltose, [3] lactose, [4] diphenolic compounds [5] and catecholamines, such as dopamine, adrenaline and noradrenaline, [6] as well as in biofuel cell (BFC) anodes fuelled by glucose, lactose or cellobiose. [7] Electric contact between its catalytic site and the electrode has been provided by direct electron transfer (DET) [3, 4, 6, 7a,c,d, 8] by dissolved redox mediators [3] and by redox polymers.…”

Comparison of Enzymatic Recycling Electrodes for Measuring Aminophenol: Development of a Highly Sensitive Natriuretic Peptide Assay System

“…In enzymatic recycling, an electrochemically active small molecule is used as a redox mediator, and the electrochemically oxidized (or reduced) mediator is then reduced (or oxidized) back by the redox enzyme(s), which leads to the redox cycling of the mediator to amplify the electrical current. [18][19][20] Quinone derivatives such as p-aminophenol (PAP) are the most frequently studied mediators. We compared the abilities of several redox enzymes to function in an enzymatic recycling electrode system for PAP, and coupled the recycling system to an enzyme immunoassay to measure ANP.…”

Highly Efficient Membraneless Glucose Bioanode Based on Corynascus thermophilus Cellobiose Dehydrogenase on Aryl Diazonium‐Activated Single‐Walled Carbon Nanotubes

“…[20b] Various CDHs have been applied for making different biosensors, for example, for lactose, [21] glucose, [22] catecholamines, [23] and phenolic compounds, [24] and for use in EFCs in combination with laccase or bilirubin-oxidase-based cathodes. [2a-c, 25] In this work, CtCDH-based bioanodes were characterized for future construction of EFCs.…”