Enhanced stability of enzyme organophosphate hydrolase interfaced on the carbon nanotubes.
In this paper we demonstrate that SWNTs and a covalent immobilization strategy enable very sensitive sensors with excellent long term stability. Organophosphorus hydrolase (OPH) functionalized single and multi-walled carbon nanotube (CNT) conjugates were exploited for direct amperometric detection of paraoxon, a model organophosphate. The catalytic hydrolysis of paraoxon produces equimoles of p-nitrophenol; oxidation was monitored amperometrically in real time under flow-injection (FIA) mode. OPH covalently immobilized on single-walled carbon nanotubes (SWNTs) demonstrated much higher activity than OPH conjugated to multi-walled carbon nanotubes (MWNTs). The dynamic concentration range for SWNT-OPH was 0.5-8.5 micromolL(-1) with a detection limit of 0.01 micromolL(-1) (S/N=3). In addition to this high sensitivity, the immobilized OPH retained a significant degree of enzymatic activity, and displayed remarkable stability with only 25% signal loss over 7 months. These results suggest that covalent immobilization of OPH on CNTs can be used for specific immobilization with advantages of long term stability, high sensitivity, and simplicity.
Pedrosa
Valber A
VA
Department of Materials Engineering, Auburn University, AL 36849, USA.
Paliwal
Sheetal
S
Balasubramanian
Shankar
S
Nepal
Dhriti
D
Davis
Virginia
V
Wild
James
J
Ramanculov
Erlan
E
Simonian
Aleksandr
A
D055162:Biocatalysis; D015374:Biosensing Techniques; D004563:Electrochemistry; D004795:Enzyme Stability; D006868:Hydrolysis; D057230:Limit of Detection; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D037742:Nanotubes, Carbon; D010261:Paraoxon / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*
D037742:Nanotubes, Carbon; C045290:phosphorylphosphatase; D010744:Phosphoric Monoester Hydrolases; D010261:Paraoxon