Direct Electrochemistry and Electrocatalysis Behaviors of Glucose Oxidase Based on Hyaluronic Acid-Carbon Nanotubes-Ionic Liquid Composite Film

Multi‐walled carbon nanotubes (MWNTs) were dispersed in the ionic liquid [BMIM][BF₄] to form a uniform black suspension. Based on it, a novel glucose oxidase (GOx)‐hyaluronic (HA)‐[BMIM][BF₄]‐MWNTs/GCE modified electrode was fabricated. UV‐vis spectroscopy confirmed that GOx immobilized in the composite film retained its native structure. The experimental results of EIS indicated MWNTs, [BMIM][BF₄] and HA were successfully immobilized on the surface of GCE and [BMIM][BF₄]‐MWNTs could obviously improve the diffusion of ferricyanide toward the electrode surface. The experimental results of CV showed that a pair of well‐defined and quasi‐reversible peaks of GOx at the modified electrode was exhibited, and the redox reaction of GOx at the modified electrode was surface‐confined and quasi‐reversible electrochemical process. The average surface coverage of GOx and the apparent Michaelis‐Menten constant were 8.5×10⁻⁹ mol/cm² and 9.8 mmol/L, respectively. The cathodic peak current of GOx and the glucose concentration showed linear relationship in the range from 0.1 to 2.0 mmol/L with a detection limit of 0.03 mmol/L (S/N=3). As a result, the method presented here could be easily extended to immobilize and obtain the direct electrochemistry of other redox enzymes or proteins.     

Direct electrochemical sensing of glucose using glucose oxidase immobilized on functionalized carbon nanotubes via a novel metal chelate-based affinity method

We report on a novel amperometric glassy carbon biosensing electrode for glucose. It is based on the immobilization of a highly sensitive glucose oxidase (GOx) by affinity interaction on carbon nanotubes (CNTs) functionalized with iminodiacetic acid and metal chelates. The new technique for immobilization is exploiting the affinity of Co(II) ions to the histidine and cysteine moieties on the surface of GOx. The direct electrochemistry of immobilized GOx revealed that the functionalized CNTs greatly improve the direct electron transfer between GOx and the surface of the electrode to give a pair of well-defined and almost reversible redox peaks and undergoes fast heterogeneous electron transfer with a rate constant (k _s) of 0.59?s^?1. The GOx immobilized in this way fully retained its activity for the oxidation of glucose. The resulting biosensor is capable of detecting glucose at levels as low as 0.01?mM, and has excellent operational stability (with no decrease in the activity of enzyme over a 10?days period). The method of immobilizing GOx is easy and also provides a model technique for potential use with other redox enzymes and proteins.

Transparent and flexible glucose biosensor via layer-by-layer assembly of multi-wall carbon nanotubes and glucose oxidase

This paper reports a transparent and flexible glucose biosensor of which multi-wall carbon nanotubes (MWNTs) and glucose oxidase (GOx) is layer-by-layer (LBL) self-assembled on a polymer substrate. A thin Ti and Au layers is firstly deposited on the polymer substrate through plasma immersion ion implantation (PIII) and sputtering, respectively. An organic monolayer then forms on the gold surface using thiol chemistry. Subsequently, negatively charged MWNTs and GOx are stably LBL assembled on the modified Au surface, respectively, via alternative electrostatic interaction of the positively charged polyelectrolyte with the oppositely charged MWNTs and GOx. Electrochemical studies show that the multilayer membrane exhibits remarkable electrocatalytic activity to detect glucose molecule. The biosensor displays a linear response range of 0.02–2.2 mM (a correlation coefficient of 0.998) with a low detection limit of 10 μM. This remarkable performance, combined with the large area preparation process, demonstrates this CNT-based multilayer biosensor is well suited for commercial applications.     

Electrochemical glucose biosensor based on silver nanoparticles/multiwalled carbon nanotubes modified electrode

A novel glucose biosensor was fabricated by immobilizing glucose oxidase (GOx) on Ag nanoparticles-decorated multiwalled carbon nanotube (AgNP-MWNT) modified glass carbon electrode (GCE). The AgNP-MWNT composite membrane showed an improving biocompatibility for GOx immobilization and an enhancing electrocatalytic activity toward reduction of oxygen due to decoration of AgNPs on MWNT surfaces. The AgNPs also accelerated the direct electron transfer between redox-active site of GOx and GCE surface because of their excellent conductivity and large capacity for protein loading, leading to direct electrochemistry of GOx. The glucose biosensor of this work showed a lower limit of detection of 0.01 mM (S/N?=?3) and a wide linear range from 0.025 to 1.0 mM, indicating an excellent analytical performance of the obtained biosensor to glucose detection. The resulting biosensor exhibits good stability and excellent reproducibility. Such bionanocomposite provides us good candidate material for fabrication of biosensors based on direct electrochemistry of immobilized enzymes.     

Direct Electrochemistry and Electrocatalysis of Myoglobin Immobilized on Gold Nanoparticles/Carbon Nanotubes Nanohybrid Film

A novel nanohybrid material, constructed by gold nanoparticles (GNPs) and multiwalled carbon nanotubes (MWNTs), was designed for immobilization and biosensing of myoglobin (Mb). Morphology of the nanohybrid film was characterized by SEM. UV‐vis spectroscopy demonstrated that Mb on the composite film could retain its native structure. Direct electrochemistry of Mb immobilized on the GNPs/MWNTs film was investigated. The immobilized Mb showed a couple of quasireversible and well‐defined cyclic voltammetry peaks with a formal potential of about ?0.35 V (vs. Ag/AgCl) in pH 6.0 phosphate buffer solution (PBS) solution. Furthermore, the modified electrode also displayed good sensitivity, wide linear range and long‐term stability to the detection of hydrogen peroxide. The experiment results demonstrated that the hybrid matrix provided a biocompatible microenvironment for protein and supplied a necessary pathway for its direct electron transfer.     

Antibacterial effects of carbon nanotubes: size does matter!

We provide the first evidence that the size (diameter) of carbon nanotubes (CNTs) is a key factor governing their antibacterial effects and that the likely main CNT-cytotoxicity mechanism is cell membrane damage by direct contact with CNTs. Experiments with well-characterized single-walled carbon nanotubes (SWNTs) and multiwalled carbon nanotubes (MWNTs) demonstrate that SWNTs are much more toxic to bacteria than MWNTs. Gene expression data show that in the presence of both MWNTs and SWNTs, Escherichia coli expresses high levels of stress-related gene products, with the quantity and magnitude of expression being much higher in the presence of SWNTs.     

Direct electrochemistry of glucose oxidase immobilized on NdPO4 nanoparticles/chitosan composite film on glassy carbon electrodes and its biosensing application

The direct electrochemistry of glucose oxidase (GOx) immobilized on a composite matrix based on chitosan (CHIT) and NdPO(4) nanoparticles (NPs) underlying on glassy carbon electrode (GCE) was achieved. The cyclic voltammetry and electrochemical impedance spectroscopy were used to characterize the modified electrode. In deaerated buffer solutions, the cyclic voltammetry of the composite films of GOx/NdPO(4) NPs/CHIT showed a pair of well-behaved redox peaks that are assigned to the redox reaction of GOx, confirming the effective immobilization of GOx on the composite film. The electron transfer rate constant was estimated to be 5.0 s(-1). The linear dynamic range for the detection of glucose was 0.15-10 mM with a correlation coefficient of 0.999 and the detection limit was estimated at about 0.08 mM (S/N=3). The calculated apparent Michaelis-Menten constant was 2.5 mM, which suggested a high affinity of the enzyme-substrate. The immobilized GOx in the NdPO(4) NPs/CHIT composite film retained its bioactivity. Furthermore, the method presented here can be easily extended to immobilize and obtain the direct electrochemistry of other redox enzymes or proteins.     

In Situ Fabrication of Noble Metal Nanoparticles Modified Multiwalled Carbon Nanotubes and Related Electrocatalysis

Using multiwalled carbon nanotubes (MWNTs) as templates, noble metal (Au, Ag, Pt or Pd) nanoparticles (NPs) were fabricated in situ by electrochemistry with a diameter of 40–60 nm. Further, catalytic behaviors of these composite materials were investigated. Experiments showed that such carbon nanotubes decorated with Pd NPs modified glassy carbon electrodes exhibited higher electrocatalytic ability to some molecules such as evolution of hydrogen, reduction of oxygen and oxidation of ascorbic acid. Atomic force microscopy, X‐ray photoelectron spectroscopy and cyclic voltammetry were used to characterize the film formation and their properties.     

Sensitive voltammetric determination of xanthinol nicotinate at a carbon nanotubes-ionic liquid gel modified electrode

The electrochemistry of xanthinol nicotinate(Xan) was studied by cyclic voltammetry at a glassy carbon electrode modified by a gel containing multi-walled carbon nanotubes(MWNTs) and room-temperature ionic liquid of 1-butyl-3-methylimidazolium hexafluorophosphate(BMTMPF_6).The modified electrode exhibited good promotion to the electrochemical oxidation of Xan and an ultrasensitive electrochemical method was proposed for the determination of Xan.This method was successfully applied to the determination of...     

酶法合成的葡萄糖氧化酶-纳米金复合物的直接电化学与生物传感

将NaAuCl₄、葡萄糖氧化酶（GOx）和葡萄糖混合,借一步酶促反应制得吸附GOx的金纳米颗粒（AuNPs）,再通过滴干修饰法研制了Nafion/GOx-AuNPs修饰的玻碳（GC）电极,并考察了该酶电极上GOx的直接电化学和生物传感性能. 这种酶法合成的GOx-AuNPs复合物有良好的酶直接电化学活性,也保持了GOx的生物活性,似可归因于酶法合成的纳米金更接近酶氧化还原活性中心的缘故. 该酶电极在-0.4 V（vs. SCE）电位下,其稳态电流下降与葡萄糖浓度（0.5 4 mmol·L^-1）成正比,检测下限0.2 mol·L^-1.     

A biosensor prepared by co-entrapment of a glucose oxidase and a carbon nanotube within an electrochemically deposited redox polymer multilayer

A glucose biosensor based on a nanocomposite made by layer-by-layer electrodeposition of the redox polymer into a multilayer containing glucose oxidase (GOx) and single-walled carbon nanotubes (SWCNT) on a screen-printed carbon electrode (SPCE) surface was developed. The objectives of the electrodeposition of redox polymer are to stabilize further the multilayer using a coordinative cross-linked redox polymer and to wire the GOx. The electrochemistry of the layer-by-layer assembly of the GOx/SWCNT/redox polymer nanocomposite was followed by cyclic voltammetry. The resultant biosensor provided stable and reproducible electrocatalytic responses to glucose, and the electrocatalytic current for glucose oxidation was enhanced with an increase in the number of layers. The biosensor displayed a linear range from 0.5 to 6.0mM, a sensitivity of 16.4μA/(mMcm(2)), and a response time of about 5s. It shows no response to 0.05mM of ascorbic acid, 0.32mM of uric acid and 0.20mM of acetaminophen using a Nafion membrane covering the nanocomposite-modified electrode surface.     

Electrochemical characteristics of the immobilization of calf thymus DNA molecules on multi-walled carbon nanotubes

Immobilization of DNA on carbon nanotubes plays an important role in the development of new types of miniature DNA biosensors. Electrochemical characteristics of the immobilization of calf thymus DNA molecules on the surfaces of multi-walled carbon nanotubes (MWNTs) have been investigated by cyclic voltammetry and electrochemical impedance analysis. The peak currents for Fe(CN)(6)(3-)/Fe(CN)(6)(4-) redox couple observed in the cyclic voltammograms decrease and the electron-transfer resistance (R(et)) obtained from the Nyquist plots increase due to the immobilization of DNA molecules (dsDNA or ssDNA) on the surfaces of MWNTs. Most of calf thymus DNA are covalently immobilized on MWNTs via diimide-activated amidation between the carboxylic acid groups on the carbon nanotubes and the amino groups on DNA bases, though the direct adsorption of the DNA molecules on MWNTs can be observed. Additionally, the interaction between DNA molecules immobilized on MWNTs and small biomolecules (ethidium bromide) can be observed obviously by cyclic voltammetry and electrochemical impedance analysis. This implies that the DNA molecules immobilized at the surface of MWNTs, with little structure change, still has the ability to interact with small biomolecules.     

Direct electron transfer of glucose oxidase on the carbon nanotube electrode

The direct electrochemistry of redox enzymes (or proteins) has received more and more attention[1—9]. These studies developed an electrochemical basis for the investigation of enzyme structure, mechanisms of redox transformations of enzyme molecules and metabolic processes involving redox transformations. From these studies, one can also find potential appli-cations of enzymes in biotechnology. For example, if an enzyme immobilized on electrode surface is ca-pable of the direct electron tra…     

Direct electron transfer of glucose oxidase promoted by carbon nanotubes is without value in certain mediator-free applications

We have investigated the direct electron transfer (DET) promoted by carbon nanotubes (CNTs) on an electrode containing immobilized glucose oxidase (GOx) with the aim to develop a third-generation glucose biosensor and a mediator-free glucose biofuel cell anode. GOx was immobilized via chitosan (CS) on a glassy carbon electrode (GCE) modified with multi-walled carbon nanotubes (MWCNTs). Cyclic voltammetric revealed that the GOx on the surface of such an electrode is unable to simultaneously demonstrate DET with the electrode and to retain its catalytic activity towards glucose, although the MWCNTs alone can promote electron transfer between GOx and electrode. This is interpreted in terms of two types of GOx on the surface, the distribution and properties of which are quite different. The first type exhibits DET capability that results from the collaboration of MWCNTs and metal impurities, but is unable to catalyze the oxidation of glucose. The second type maintains its glucose-specific catalytic capability in the presence of a mediator, which can be enhanced by MWCNTs, but cannot undergo DET with the electrode. As a result, the MWCNTs are capable of promoting the electron transfer, but this is without value in some mediator-free applications such as in third-generation glucose biosensors and in mediator-free anodes for glucose biofuel cells.

Mediatorless amperometric bienzyme glucose biosensor based on horseradish peroxidase and glucose oxidase cross-linked to multiwall carbon nanotubes

We report on a bienzyme-channeling sensor for sensing glucose without the aid of mediator. It was fabricated by cross-linking horseradish peroxidase (HRP) and glucose oxidase (GOx) on a glassy carbon electrode modified with multiwalled carbon nanotubes (MWNTs). The bienzyme was cross-linked with the MWNTs by glutaraldehyde and bovine serum albumin. The MWNTs were employed to accelerate the electron transfer between immobilized HRP and electrode. Glucose was sensed by amperometric reduction of enzymatically generated H₂O₂ at an applied voltage of ?50 mV (vs. Ag/AgCl). Factors influencing the preparation and performance of the bienzyme electrode were investigated in detail. The biosensor exhibited a fast and linear response to glucose in the concentration range from 0.4 to 15 mM, with a detection limit of 0.4 mM. The sensor exhibited good selectivity and durability, with a long-term relative standard deviation of <5 %. Analysis of glucose-spiked human serum samples yielded recoveries between 96 and 101 %.

Synergistic contributions of multiwall carbon nanotubes and gold nanoparticles in a chitosan–ionic liquid matrix towards improved performance for a glucose sensor

We report an ingenious approach for the fabrication of a promising glucose sensor, GOx/Au/CS–IL–MWNT(SH), that exploits the synergistic beneficial characteristics of multiwalled-carbon nanotubes (MWNTs), gold nanoparticles (AuNPs), chitosan (CS) and room temperature ionic liquid (RTIL). Direct electron transfer between glucose oxidase (GOx) and electrode was achieved. Scanning electron microscopy and atomic force microscopy images of GOx/Au/CS–IL–MWNT(SH) reveal that MWNTs and AuNPs are dispersed in CS–IL matrix. Cyclic voltammetry, impedance spectroscopy and chronoamperometry were used to evaluate the performance of biosensor. The GOx/Au/CS–IL–MWNT(SH) biosensor exhibits a linear current response to glucose concentration (1–10 mM) at a low potential of 0.10 V and precludes interferences from uric acid and ascorbic acid. The GOx/Au/CS–IL–MWNT(SH) biosensor has superior performances over GOx/CS–IL–MWNT(SH).     

New Nanocomposite Based on Prussian Blue Nanoparticles/Carbon Nanotubes/Chitosan and Its Application for Assembling of Amperometric Glucose Biosensor

Abstract

A new nanocomposite was developed by combination of prussian blue (PB) nanoparticles and multiwalled carbon nanotubes (MWNTs) in the matrix of biopolymer chitosan (CHIT). The PB and MWNTs had a synergistic electrocatalytic effect toward the reduction of hydrogen peroxide. The CHIT/MWNTs/PB nanocomposite‐modified glassy carbon (GC) electrode could amplify the reduction current of hydrogen peroxide by ～35 times compared with that of CHIT/MWNTs/GC electrode and reduce the response time from ～60 s for CHIT/PB/GC to 3 s. Besides, the CHIT/MWNTs/PB nanocomposite‐modified GC electrode could reduce hydrogen peroxide at a much lower applied potential and inhibit the responses of interferents such as ascorbic acid (AA) uric acid (UA) and acetaminophen (AC). With glucose oxidase (GOx) as an enzyme model, a new glucose biosensor was fabricated. The biosensor exhibited excellent sensitivity (the detection limit is down to 2.5 µM), fast response time (less than 5 s), wide linear range (from 4 µM to 2 mM), and good selection.     

Direct electrochemistry and electrochemical biosensing of glucose oxidase based on CdSe@CdS quantum dots and MWNT-modified electrode

The direct electron transfer of glucose oxidase (GOx) was achieved based on the immobilization of CdSe@CdS quantum dots on glassy carbon electrode by multi-wall carbon nanotubes (MWNTs)-chitosan (Chit) film. The immobilized GOx displayed a pair of well-defined and reversible redox peaks with a formal potential (E ^θ’) of ?0.459 V (versus Ag/AgCl) in 0.1 M pH 7.0 phosphate buffer solution. The apparent heterogeneous electron transfer rate constants (k _s) of GOx confined in MWNTs-Chit/CdSe@CdS membrane were evaluated as 1.56 s^?1 according to Laviron's equation. The surface concentration (Γ*) of the electroactive GOx in the MWNTs-Chit film was estimated to be (6.52?±?0.01)?×?10^?11?mol?cm^?2. Meanwhile, the catalytic ability of GOx toward the oxidation of glucose was studied. Its apparent Michaelis–Menten constant for glucose was 0.46?±?0.01 mM, showing a good affinity. The linear range for glucose determination was from 1.6?×?10^?4 to 5.6?×?10^?3?M with a relatively high sensitivity of 31.13?±?0.02 μA?mM^?1?cm^?2 and a detection limit of 2.5?×?10^?5?M (S/N=3).     

Adsorption and Electrooxidation of DNA at Glassy Carbon Electrodes Modified with Multiwall Carbon Nanotubes Dispersed in Glucose Oxidase

We are proposing for the first time the successful immobilization of DNA at glassy carbon electrodes (GCE) modified with carbon nanotubes (CNT) dispersed in glucose oxidase (GOx) (GCE/CNT‐GOx) either by direct adsorption or by layer‐by‐layer self‐assembling using polydiallyldimethylamine (PDDA). The presence of GOx allows an efficient dispersion of CNT and gives a most favorable environment that promotes the adsorption and makes possible a more sensitive electrooxidation of DNA. The PDDA incorporated in the self‐assembled architecture largely facilitates the adsorption and electrooxidation of dsDNA and the adsorbed layer can be successfully used for evaluating the interaction of DNA with methylene blue.     

Enzymatic degradation of multiwalled carbon nanotubes

Because of their unique properties, carbon nanotubes and, in particular, multiwalled carbon nanotubes (MWNTs) have been used for the development of advanced composite and catalyst materials. Despite their growing commercial applications and increased production, the potential environmental and toxicological impacts of MWNTs are not fully understood; however, many reports suggest that they may be toxic. Therefore, a need exists to develop protocols for effective and safe degradation of MWNTs. In this article, we investigated the effect of chemical functionalization of MWNTs on their enzymatic degradation with horseradish peroxidase (HRP) and hydrogen peroxide (H(2)O(2)). We investigated HRP/H(2)O(2) degradation of purified, oxidized, and nitrogen-doped MWNTs and proposed a layer-by-layer degradation mechanism of nanotubes facilitated by side wall defects. These results provide a better understanding of the interaction between HRP and carbon nanotubes and suggest an eco-friendly way of mitigating the environmental impact of nanotubes.