TY - JOUR
T1 - Development of a simple bioelectrode for the electrochemical detection of hydrogen peroxide using Pichia pastoris catalase immobilized on gold nanoparticle nanotubes and polythiophene hybrid
AU - Nandini, Seetharamaiah
AU - Nalini, Seetharamaiah
AU - Sanetuntikul, Jakkid
AU - Shanmugam, Sangaraju
AU - Niranjana, Pathappa
AU - Melo, Jose Savio
AU - Suresh, Gurukar Shivappa
PY - 2014/10/15
Y1 - 2014/10/15
N2 - In this paper, a simple and innovative electrochemical hydrogen peroxide biosensor has been proposed using catalase (CATpp) derived from Pichia pastoris as bioelectrocatalyst. The model biocomponent was immobilized on gold nanoparticle nanotubes (AuNPNTs) and polythiophene composite using 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide and N-hydroxysuccinimide (EDC–NHS) coupling reagent. In this present work, we have successfully synthesized gold nanoparticles (AuNPs) by ultrasonic irradiation. The tubular gold nanostructures containing coalesced AuNPs were obtained by sacrificial template synthesis. The assembly of AuNPNTs onto the graphite (Gr) electrode was achieved via S–Au chemisorption. The latter was pre-coated with electropolymerized thiophene (PTh) to enable S groups to bind AuNPNTs. The combination of AuNPNTs–PTh, i.e., an inorganic–organic hybrid, provides a stable enzyme immobilization platform. The physical morphology of the fabricated biosensor Gr/PTh/AuNPNTs/EDC–NHS/CATpp was investigated using scanning electron microscopy and energy-dispersive microscopy. The analytical performance of the bioelectrode was examined using cyclic voltammetry, differential pulse voltammetry and chronoamperometry. Operational parameters such as working potential, pH, and thermal stability of the modified electrode were examined. The beneficial analytical characteristics of the proposed electrode were demonstrated. Our results indicate that the Gr/PTh/AuNPNTs/EDC–NHS/CATpp bioelectrode exhibits a wide linear range from 0.05 mM to 18.5 mM of H2O2, fast response time of 7 s, excellent sensitivity of 26.2 mA mM−1 cm−2, good detection limit of 0.12 μM and good Michaelis–Menten constant of 1.4 mM. In addition, the bioelectrode displayed good repeatability, high stability and acceptable reproducibility, which can be attributed to the AuNPNTs–PTh composite that provides a biocompatible micro-environment.
AB - In this paper, a simple and innovative electrochemical hydrogen peroxide biosensor has been proposed using catalase (CATpp) derived from Pichia pastoris as bioelectrocatalyst. The model biocomponent was immobilized on gold nanoparticle nanotubes (AuNPNTs) and polythiophene composite using 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide and N-hydroxysuccinimide (EDC–NHS) coupling reagent. In this present work, we have successfully synthesized gold nanoparticles (AuNPs) by ultrasonic irradiation. The tubular gold nanostructures containing coalesced AuNPs were obtained by sacrificial template synthesis. The assembly of AuNPNTs onto the graphite (Gr) electrode was achieved via S–Au chemisorption. The latter was pre-coated with electropolymerized thiophene (PTh) to enable S groups to bind AuNPNTs. The combination of AuNPNTs–PTh, i.e., an inorganic–organic hybrid, provides a stable enzyme immobilization platform. The physical morphology of the fabricated biosensor Gr/PTh/AuNPNTs/EDC–NHS/CATpp was investigated using scanning electron microscopy and energy-dispersive microscopy. The analytical performance of the bioelectrode was examined using cyclic voltammetry, differential pulse voltammetry and chronoamperometry. Operational parameters such as working potential, pH, and thermal stability of the modified electrode were examined. The beneficial analytical characteristics of the proposed electrode were demonstrated. Our results indicate that the Gr/PTh/AuNPNTs/EDC–NHS/CATpp bioelectrode exhibits a wide linear range from 0.05 mM to 18.5 mM of H2O2, fast response time of 7 s, excellent sensitivity of 26.2 mA mM−1 cm−2, good detection limit of 0.12 μM and good Michaelis–Menten constant of 1.4 mM. In addition, the bioelectrode displayed good repeatability, high stability and acceptable reproducibility, which can be attributed to the AuNPNTs–PTh composite that provides a biocompatible micro-environment.
UR - https://www.scopus.com/pages/publications/84907963263
U2 - 10.1039/c4an01262c
DO - 10.1039/c4an01262c
M3 - Article
C2 - 25208248
AN - SCOPUS:84907963263
SN - 0003-2654
VL - 139
SP - 5800
EP - 5812
JO - Analyst
JF - Analyst
IS - 22
ER -