Acetylcholinesterase biosensor based on 3-carboxyphenylboronic acid/reduced graphene oxide–gold nanocomposites modified electrode for amperometric detection of organophosphorus and carbamate pesticides

Abstract A novel amperometric biosensor based on immobilizing acetylcholinesterase on 3-carboxyphenylboronic/reduced graphene oxide–gold nanocomposites modified electrode was developed for the detection of organophosphorus and carbamate pesticides. Acetylcholinesterase was successfully immobilized on the electrode with relatively high activity by the specific binding between the boronic acid group of 3-carboxyphenylboronic and the glycosyl of acetylcholinesterase. The biosensor enjoyed good sensitivity owing to the excellent properties of gold nanoparticles and reduced graphene oxide, which promoted electron transfer reaction and enhanced the electrochemical response. Parameters affecting the biosensor response such as pH, applied potential, enzyme loading and inhibition time were optimized. Based on the inhibition of an organophosphorus insecticide on the enzymatic activity of AChE, organophosphorus and carbamate pesticides were determined with satisfactory results in the presence of acetylthiocholine chloride as a substrate. Moreover, the fabricated biosensor had good repeatability and high stability.

[1]  K. Müllen,et al.  Transparent, conductive graphene electrodes for dye-sensitized solar cells. , 2008, Nano letters.

[2]  Jun Liu,et al.  Glucose biosensor based on immobilization of glucose oxidase in platinum nanoparticles/graphene/chitosan nanocomposite film. , 2009, Talanta.

[3]  B. Pletschke,et al.  Review on the use of enzymes for the detection of organochlorine, organophosphate and carbamate pesticides in the environment. , 2011, Chemosphere.

[4]  Dan Du,et al.  Immobilization of acetylcholinesterase based on the controllable adsorption of carbon nanotubes onto an alkanethiol monolayer for carbaryl sensing. , 2008, The Analyst.

[5]  Richard D. Smith,et al.  Enrichment and analysis of nonenzymatically glycated peptides: boronate affinity chromatography coupled with electron-transfer dissociation mass spectrometry. , 2007, Journal of proteome research.

[6]  S. Mohan,et al.  Development of acetylcholinesterase silica sol-gel immobilized biosensor--an application towards oxydemeton methyl detection. , 2004, Biosensors & bioelectronics.

[7]  H. Dai,et al.  Highly conducting graphene sheets and Langmuir-Blodgett films. , 2008, Nature nanotechnology.

[8]  Dan Du,et al.  Acetylcholinesterase biosensor design based on carbon nanotube-encapsulated polypyrrole and polyaniline copolymer for amperometric detection of organophosphates. , 2010, Biosensors & bioelectronics.

[9]  Juewen Liu,et al.  Preparation of aptamer-linked gold nanoparticle purple aggregates for colorimetric sensing of analytes , 2006, Nature Protocols.

[10]  Arben Merkoçi,et al.  Determination of Toxic Substances Based on Enzyme Inhibition. Part II. Electrochemical Biosensors for the Determination of Pesticides Using Flow Systems , 2003 .

[11]  Giuseppe Palleschi,et al.  Determination of Organophosphorus and Carbamic Pesticides with a Choline and Acetylcholine Electrochemical Biosensor , 1991 .

[12]  Prashant V Kamat,et al.  Anchoring semiconductor and metal nanoparticles on a two-dimensional catalyst mat. Storing and shuttling electrons with reduced graphene oxide. , 2010, Nano letters.

[13]  Gajendra S Shekhawat,et al.  Immunoanalytical techniques for analyzing pesticides in the environment , 2009 .

[14]  S. Shinkai,et al.  Saccharide Sensing with Molecular Receptors Based on Boronic Acid , 1996 .

[15]  Jean-Louis Marty,et al.  Enzyme immobilization procedures on screen-printed electrodes used for the detection of anticholinesterase pesticides Comparative study , 2004 .

[16]  Shengyang Tao,et al.  Ultra-sensitive biosensor based on mesocellular silica foam for organophosphorous pesticide detection. , 2011, Biosensors & bioelectronics.

[17]  Human erythrocyte bisphosphoglycerate mutase: inactivation by glycation in vivo and in vitro. , 1998, Journal of biochemistry.

[18]  Hua Zhang,et al.  Organic photovoltaic devices using highly flexible reduced graphene oxide films as transparent electrodes. , 2010, ACS nano.

[19]  C. Geula,et al.  Neurobiology of butyrylcholinesterase , 2003, Nature Reviews Neuroscience.

[20]  Yee Ying Tan,et al.  Using detonation nanodiamond for the specific capture of glycoproteins. , 2008, Analytical chemistry.

[21]  Prashant V. Kamat,et al.  Decorating Graphene Sheets with Gold Nanoparticles , 2008 .

[22]  Danila Moscone,et al.  Detection of carbamic and organophosphorous pesticides in water samples using a cholinesterase biosensor based on Prussian Blue-modified screen-printed electrode. , 2006, Analytica chimica acta.

[23]  Akira Matsumoto,et al.  Noninvasive sialic acid detection at cell membrane by using phenylboronic acid modified self-assembled monolayer gold electrode. , 2009, Journal of the American Chemical Society.

[24]  Yuehua Qin,et al.  One-step synthesis of multiwalled carbon nanotubes-gold nanocomposites for fabricating amperometric acetylcholinesterase biosensor , 2010 .

[25]  S. Khondaker,et al.  High yield fabrication of chemically reduced graphene oxide field effect transistors by dielectrophoresis , 2010, Nanotechnology.

[26]  H. Lin,et al.  Development of an amperometric micro-biodetector for pesticide monitoring and detection , 2009 .

[27]  J. Klein-Seetharaman,et al.  The enzymatic oxidation of graphene oxide. , 2011, ACS nano.

[28]  X. Xia,et al.  Two-step pyrolysis process to synthesize highly dispersed Pt–Ru/carbon nanotube catalysts for methanol electrooxidation , 2006 .

[29]  H. Kuivila,et al.  ARENEBORONATES FROM DIOLS AND POLYOLS1 , 1954 .

[30]  Jean-Louis Marty,et al.  Multidisciplinary collaboration for environmental protection using biosensors: detection of organophosphate insecticides in aqueous medium , 2006 .

[31]  S. Khondaker,et al.  Schottky diode via dielectrophoretic assembly of reduced graphene oxide sheets between dissimilar metal contacts , 2011 .

[32]  Xi Chen,et al.  Application of chemisorption/desorption process of thiocholine for pesticide detection based on acetylcholinesterase biosensor , 2008 .

[33]  G. S. Wilson,et al.  Rotating ring-disk enzyme electrode for biocatalysis kinetic studies and characterization of the immobilized enzyme layer , 1980 .

[34]  B. H. Weiller,et al.  Practical chemical sensors from chemically derived graphene. , 2009, ACS nano.

[35]  Félix Pariente,et al.  Determination of Organophosphorus and Carbamate Pesticides Using a Piezoelectric Biosensor , 1998 .

[36]  Dongxue Han,et al.  Electrochemical determination of NADH and ethanol based on ionic liquid-functionalized graphene. , 2010, Biosensors & bioelectronics.

[37]  Giuseppe Palleschi,et al.  Determination of organophosphorus insecticides with a choline electrochemical biosensor , 1992 .

[38]  S. Dong,et al.  Electrochemical sensing and biosensing platform based on chemically reduced graphene oxide. , 2009, Analytical chemistry.

[39]  Fei Xiao,et al.  Layer-by-Layer self-assembled acetylcholinesterase/PAMAM-Au on CNTs modified electrode for sensing pesticides. , 2010, Bioelectrochemistry.

[40]  Tse-Chuan Chou,et al.  Amperometric protein sensor - fabricated as a polypyrrole, poly-aminophenylboronic acid bilayer. , 2006, Biosensors & bioelectronics.

[41]  Jing‐Juan Xu,et al.  A Reusable Interface Constructed by 3‐Aminophenylboronic Acid‐Functionalized Multiwalled Carbon Nanotubes for Cell Capture, Release, and Cytosensing , 2010 .

[42]  Huangxian Ju,et al.  Binding of acetylcholinesterase to multiwall carbon nanotube-cross-linked chitosan composite for flow-injection amperometric detection of an organophosphorous insecticide. , 2006, Chemistry.

[43]  S. Khondaker,et al.  Position dependent photodetector from large area reduced graphene oxide thin films , 2010, 1002.3191.

[44]  S. Khondaker,et al.  Graphene based materials: Past, present and future , 2011 .

[45]  P. Millner,et al.  Acetylecholinesterase-based biosensor electrodes for organophosphate pesticide detection. II. Immobilization and stabilization of acetylecholinesterase. , 2005, Biosensors & bioelectronics.

[46]  Masahiro Fujiwara,et al.  Thin-film particles of graphite oxide 1:: High-yield synthesis and flexibility of the particles , 2004 .

[47]  S. Stankovich,et al.  Graphene-based composite materials , 2006, Nature.

[48]  Michael S Freund,et al.  Potentiometric saccharide detection based on the pK(a) changes of poly(aniline boronic acid). , 2002, Journal of the American Chemical Society.

[49]  Brad Miller,et al.  Phenylboronic acid self-assembled layer on glassy carbon electrode for recognition of glycoprotein peroxidase , 2005 .