Study on the Relationship between Particulate Methane Monooxygenase and Methanobactin on Gold-Nanoparticles-Modified Electrodes.

(1) Background: Particulate methane monooxygenase (pMMO) has a strong dependence on the natural electron transfer path and is prone to denaturation, which results in its redox activity centers being unable to transfer electrons with bare electrodes directly and making it challenging to observe an electrochemical response; (2) Methods: Using methanobactin (Mb) as the electron transporter between gold electrodes and pMMO, a bionic interface with high biocompatibility and stability was created. The Mb-AuNPs-modified functionalized gold net electrode as a working electrode, the kinetic behaviors of pMMO bioelectrocatalysis, and the effect of Mb on pMMO were analyzed. The CV tests were performed at different scanning rates to obtain electrochemical kinetics parameters. (3) Results: The values of the electron transfer coefficient (α) and electron transfer rate constant (ks) are relatively large in test environments containing only CH4 or O2. In contrast, in the test environment containing both CH4 and O2, the bioelectrocatalysis of pMMO is a two-electron transfer process with a relatively small α and ks; (4) Conclusions: It was inferred that Mb formed the complex with pMMO. More importantly, Mb not only played a role in electron transfer but also in stabilizing the enzyme structure of pMMO and maintaining a specific redox state. Furthermore, the continuous catalytic oxidation of natural substrate methane was realized.

Electrochemical Biosensor Based on Horseradish Peroxidase and Black Phosphorene Quantum Dot Modified Electrode.

Black phosphorene quantum dots (BPQDs) were prepared by ultrasonic-assisted liquid-phase exfoliation and centrifugation with morphologies proved by TEM results. Furthermore, an electrochemical enzyme sensor was prepared by co-modification of BPQDs with horseradish peroxidase (HRP) on the surface of a carbon ionic liquid electrode (CILE) for the first time. The direct electrochemical behavior of HRP was studied with a pair of well-shaped voltammetric peaks that appeared, indicating that the existence of BPQDs was beneficial to accelerate the electron transfer rate between HRP and the electrode surface. This was due to the excellent properties of BPQDs, such as small particle size, high interfacial reaction activity, fast conductivity, and good biocompatibility. The presence of BPQDs on the electrode surface provided a fast channel for direct electron transfer of HRP. Therefore, the constructed electrochemical HRP biosensor was firstly used to investigate the electrocatalytic behavior of trichloroacetic acid (TCA) and potassium bromate (KBrO3), and the wide linear detection ranges of TCA and KBrO3 were 4.0-600.0 mmol/L and 2.0-57.0 mmol/L, respectively. The modified electrode was applied to the actual samples detection with satisfactory results.

Achievements of Graphene and Its Derivatives Materials on Electrochemical Drug Assays and Drug-DNA Interactions.

Graphene, emerging as a true two-dimensional (2D) material, has attracted increasing attention due to its unique physical and electrochemical properties such as high surface area, excellent conductivity, high mechanical strength, and ease of functionalization and mass production. The entire scientific community recognizes the significance and potential impact of graphene. Electrochemical detection strategies have advantages such as being simple, fast, and low-cost. The use of graphene as an excellent interface for electrode modification provides a promising way to construct more sensitive and stable electrochemical (bio)sensors. The review presents sensors based on graphene and its derivatives for electrochemical drug assays from pharmaceutical dosage forms and biological samples. Future perspectives in this rapidly developing field are also discussed. In addition, the interaction of several important anticancer drug molecules with deoxyribonucleic acid (DNA) that was immobilized onto graphene-modified electrodes has been detailed in terms of dosage regulation and utility purposes.

In vitro electrochemical detection of the degradation of amyloid-ß oligomers.

The clearance of overloaded amyloid ß (Aß) oligomers is thought to be an attractive and potential strategy for the therapy of Alzheimer's disease (AD). A variety of strategies have already been utilized to study Aß degradation in vitro. Here, the electrochemical detection based on direct electrooxidation of specific Tyr residues within Aß peptide has been developed as a simple and robust approach for monitoring the oligomers' degradation. C60 was employed for photodegrading Aß oligomers due to the generated ROS under light irradiation. The oxidation current of Tyr residues by square wave voltammetry (SWV) increased upon the Aß degradation, confirming that the structure variation of Aß peptide indeed influenced the exposure of those redox species to the electrode surface and final signal output. Chronoamperometric assay also found the electrooxidation of Tyr undergone an irreversible process. Additionally, the direct electrochemistry was capable of detecting the aggregation with rapid test and better sensitivity in compared with dynamic light scattering (DLS), atomic force microscopy (AFM) and thioflavin T (ThT) based fluorescence assay. Thus, this work indicated the potential application of direct electrochemistry in the in vitro measurement of Aß degradation and clearance, providing new insights and a complementary means into the AD theranostics.

Electrochemical and DFT studies of andrographolide on electrochemically reduced graphene oxide for anti-viral herbaceutical sensor.

Herbal extracts are re-emerging as potential remedies for various vector-borne diseases. Amongst several phytochemicals, active ingredients of Andrographis paniculata extract is regarded as promising for dengue fever, caused by Aedes species. However, fingerprinting the active phytochemicals from herbal extracts are often relies on sophisticated analytical techniques which are not universally accessible. Herein, an electrochemically reduced graphene oxide on glassy carbon electrode (ErGO/GCE) has been devised as user-friendly and cost-effective sensor platform for fingerprinting of andrographolide (AG) in anti-dengue polyherbal formulation, i.e., Nilavembu kudineer powder. Confocal laser Raman and X-ray photoelectron spectral analyses revealed that the ErGO surfaces exert structural defects augmenting the conductivity at the electrode interface. DFT investigations enabled that C-3 and C-18 OH groups in AG is involved in the electrooxidation and adsorption-diffusion at the ErGO interface, respectively. Complementary electrochemical studies revealed that the diffusion-controlled process follows 1e-/1H+ transfer. Under optimal experimental conditions, ErGO sensor platform exhibit an amplified current sensitivity of 13.3 µA µM-1. cm-2 in the studied analyte concentration range of 10-400 µM. From the polyherbal extract and clinical sample analysis, the proposed sensor system offers selective, and sensitive detection of target AG regardless of common interferents.

Expression, purification, characterization and direct electrochemistry of two HiPIPs from Acidithiobacillus caldus SM-1.

High-potential iron-sulfur proteins (HiPIPs) from extremely acidophilic chemolithotrophic non-photosynthetic Acidithiobacillus commonly play a crucial role in ferrous or sulfurous biooxidation. Acidithiobacillus exhibit important industrial applications for bioleaching valuable metals from sulfide ores. In this study, two HiPIP genes from thermophilic Acidithiobacillus caldus SM-1 were cloned and successfully expressed, and their proteins were purified. The proteins displayed a brownish color with an optical absorbance peak at approximately 385 nm and an electronic paramagnetic resonance (EPR) g value of approximately 2.01, which confirmed that the iron-sulfur cluster was correctly inserted into the active site when the proteins were generated in E. coli. The proteins were more thermostable than HiPIPs from mesophilic Acidithiobacillus. The direct electron transfer (DET) between HiPIPs and electrode was achieved by the 2-mercaptopyrimidine (MP) surface-modified gold electrodes; the redox potentials of the HiPIP1 and HiPIP2 measured by cyclic voltammetry were approximately 304.5 mV and 400.5 mV, respectively. The electron transfer rate constant was estimated to be 0.75 s-1 and 0.66 s-1, respectively. The MP/Au electrode and Au electrode showed consistent differences in heterogeneous electron transfer rates and electron transfer resistances. Bioinformatics and molecular simulations further explained the direct electron transfer between the proteins and surface-modified electrode.

Direct electrochemistry of covalently immobilized hemoglobin on a naphthylimidazolium butyric acid ionic liquid/MWCNT matrix.

Monitoring the concentration levels of hydrogen peroxide (H2O2) is significant in both clinical and industrial applications. Herein, we develop a facile biosensor for the detection of H2O2 based on direct electron transfer of hemoglobin (Hb), which was covalently immobilized on a hydrophobic naphthylimidazolium butyric acid ionic liquid (NIBA-IL) over a multiwalled carbon nanotube (MWCNT) modified glassy carbon electrode (GCE) to obtain an Hb/NIBA-IL/MWCNT/GCE. Highly water-soluble Hb protein was firmly immobilized on NIBA-IL via stable amide bonding between the free NH2 groups of Hb and COOH groups of NIBA-IL via EDC/NHS coupling. Thus fabricated biosensor showed a well resolved redox peak with a cathodic peak potential (Epc) at -0.35 V and anodic peak potential (Epa) at -0.29 V with a formal potential (E°') of -0.32 V, which corresponds to the deeply buried FeIII/FeII redox centre of Hb, thereby direct electrochemistry of Hb was established. Further, the modified electrode demonstrated very good electrocatalytic activity towards H2O2 reduction and showed a wide linear range of detection from 0.01 to 6.3 mM with a limit of detection and sensitivity of 3.2 µM and 111 µA mM-1 cm-2, respectively. Moreover, the developed biosensor displayed high operational stability under dynamic conditions as well as during continuous potential cycles and showed reliable reproducibility. The superior performance of the fabricated biosensor is attributed to the effective covalent immobilization of Hb on the newly developed highly conducting and biocompatible NIBA-IL/MWCNT/GCE platform.

Facile strategy for immobilizing horseradish peroxidase on a novel acetate functionalized ionic liquid/MWCNT matrix for electrochemical biosensing.

Facile yet simple platforms for the immobilization of biomolecules have always been a substantial requirement for the fabrication of proficient biosensors. In this study, we report a naphthyl substituted acetate functionalized ionic liquid (NpAc-IL) for the covalent anchoring of horseradish peroxidase (HRP), using which the direct electrochemistry of HRP was successfully accomplished and a H2O2 biosensor was developed. The naphthyl substitution on the NpAc-IL was utilized for the π-π stacking with the MWCNT modified GCE and the terminal -OCH3 group of NpAc-IL was used for the covalent attachment with the free -NH2 group of HRP via amide bond formation. High conducting nature of the newly designed ionic liquid (NpAc-IL), facilitated an improved communication with the deeply buried redox centre of the HRP, while the covalent bonding provided enhanced stability to the fabricated biosensor by stably holding the water soluble HRP enzyme on the electrode surface. Furthermore, incorporation of MWCNT on the sensor setup synergistically enhanced the sensitivity of the developed biosensor. Under optimized conditions, the fabricated biosensor showed an enhanced electrocatalytic reduction of H2O2 in the range of 0.01 to 2.07 mM with a limit of detection and sensitivity of 2.7 µM and 55.98 µA mM-1 cm-2 respectively. Further, the proposed biosensor was utilized for the sensing of H2O2 spiked in real samples. Moreover, the newly fabricated biosensor demonstrated excellent stability with improved sensitivity and selectivity towards H2O2 reduction. The superior analytical characteristics are attributed to the facile fabrication strategy using this newly developed acetate functionalized ionic liquid platform.

Proton-coupled electron transfer mechanisms of the copper centres of nitrous oxide reductase from Marinobacter hydrocarbonoclasticus - An electrochemical study.

Reduction of N2O to N2 is catalysed by nitrous oxide reductase in the last step of the denitrification pathway. This multicopper enzyme has an electron transferring centre, CuA, and a tetranuclear copper-sulfide catalytic centre, "CuZ", which exists as CuZ*(4Cu1S) or CuZ(4Cu2S). The redox behaviour of these metal centres in Marinobacter hydrocarbonoclasticus nitrous oxide reductase was investigated by potentiometry and for the first time by direct electrochemistry. The reduction potential of CuA and CuZ(4Cu2S) was estimated by potentiometry to be +275 ± 5 mV and +65 ± 5 mV vs SHE, respectively, at pH 7.6. A proton-coupled electron transfer mechanism governs CuZ(4Cu2S) reduction potential, due to the protonation/deprotonation of Lys397 with a pKox of 6.0 ± 0.1 and a pKred of 9.2 ± 0.1. The reduction potential of CuA, in enzyme samples with CuZ*(4Cu1S), is controlled by protonation of the coordinating histidine residues in a two-proton coupled electron transfer process. In the cyclic voltammograms, two redox pairs were identified corresponding to CuA and CuZ(4Cu2S), with no additional signals being detected that could be attributed to CuZ*(4Cu1S). However, an enhanced cathodic signal for the activated enzyme was observed under turnover conditions, which is explained by the binding of nitrous oxide to CuZ0(4Cu1S), an intermediate species in the catalytic cycle.

Multiwalled carbon nanotubes coated with cobalt(II) sulfide nanoparticles for electrochemical sensing of glucose via direct electron transfer to glucose oxidase.

Multiwalled carbon nanotubes coated with cobalt(II) sulfide nanoparticles were prepared and used for immobilization of glucose oxidase (GOx) to obtain an electrochemical glucose biosensor. The nanocomposite was synthesized through an in-situ hydrothermal method and characterized by X-ray diffraction, scanning electron microscopy, transmission electron microscopy, and electrochemical impedance spectroscopy. The results show that the nanocomposite possesses a large specific surface area and apparently enhances the direct electron transfer between GOx and the surface of the electrode, best at a potential near -0.43 V (vs. SCE). The immobilized GOx retains its good bioactivity even at a high surface coverage of 30 pmol cm-2. Under the optimum conditions. The biosensor exhibits a wide linear range (from 8 µM to 1.5 mM), a high sensitivity (15 mA M -1 cm-2), and a 5 µM detection limit (at S/N = 3). The sensor is selective, acceptably repeatable, specific and stable. Graphical abstractMultiwalled carbon nanotubes coated with cobalt(II) sulfide nanoparticles (CoS-MWCNTs) were synthesized through in situ hydrothermal method for the construction of a sensitive electrochemical glucose biosensor.

Synthesis and utilization of Co3O4 doped carbon nanofiber for fabrication of hemoglobin-based electrochemical sensor.

In this paper cobalt oxide (Co3O4) nanoparticles were mixed with polyacrylonitrile to prepare Co3O4 doped carbon nanofiber (CNF) composite by electrospinning and carbonization, which was further used to modify on carbon ionic liquid electrode (CILE). Hemoglobin (Hb) was immobilized on Co3O4-CNF/CILE surface with Nafion acted as the protective film to fabricate an electrochemical biosensor (Nafion/Hb/Co3O4-CNF/CILE). Electrochemical behavior of Hb on the electrode was investigated with a pair of quasi-reversible redox peak appeared on cyclic voltammogram and electrochemical parameters were calculated. Moreover, this biosensor had good analytical capabilities for electrocatalytic reduction of different substrates including trichloroacetic acid, potassium bromate and sodium nitrite with wider detection range from 40.0 to 260.0 mmol L-1, 0.1 to 48.0 mmol L-1 and 1.0 to 12.0 mmol L-1 by cyclic voltammetry, respectively. The proposed method showed excellent anti-interferences ability with good selectivity and was successful used for quantitative detection of real samples, which displayed the potential applications to develop into a new analytical device.

Electrochemical Characterization of a Complex FeFe Hydrogenase, the Electron-Bifurcating Hnd From Desulfovibrio fructosovorans.

Hnd, an FeFe hydrogenase from Desulfovibrio fructosovorans, is a tetrameric enzyme that can perform flavin-based electron bifurcation. It couples the oxidation of H2 to both the exergonic reduction of NAD+ and the endergonic reduction of a ferredoxin. We previously showed that Hnd retains activity even when purified aerobically unlike other electron-bifurcating hydrogenases. In this study, we describe the purification of the enzyme under O2-free atmosphere and its biochemical and electrochemical characterization. Despite its complexity due to its multimeric composition, Hnd can catalytically and directly exchange electrons with an electrode. We characterized the catalytic and inhibition properties of this electron-bifurcating hydrogenase using protein film electrochemistry of Hnd by purifying Hnd aerobically or anaerobically, then comparing the electrochemical properties of the enzyme purified under the two conditions via protein film electrochemistry. Hydrogenases are usually inactivated under oxidizing conditions in the absence of dioxygen and can then be reactivated, to some extent, under reducing conditions. We demonstrate that the kinetics of this high potential inactivation/reactivation for Hnd show original properties: it depends on the enzyme purification conditions and varies with time, suggesting the coexistence and the interconversion of two forms of the enzyme. We also show that Hnd catalytic properties (Km for H2, diffusion and reaction at the active site of CO and O2) are comparable to those of standard hydrogenases (those which cannot catalyze electron bifurcation). These results suggest that the presence of the additional subunits, needed for electron bifurcation, changes neither the catalytic behavior at the active site, nor the gas diffusion kinetics but induces unusual rates of high potential inactivation/reactivation.

Direct electrochemistry of bacterial surface displayed cytokinin oxidase and its application in the sensitive electrochemical detection of cytokinins.

Cytokinin oxidase from Nipponbare (OsCKX4) was successfully displayed on the surface of E. coli cells by an ice nucleation protein from Pseudomonas borealis DL7 as an anchoring motif and a maltodextrin-binding protein(MBP) from E. coli as a solubility enhancer. The OsCKX4-displayed bacteria can be directly immobilized onto an electrode to selectively detect cytokinins, thus eliminating the need for enzyme extraction and purification. Direct electrochemistry of the cofactor FADH2 in OsCKX4 has been achieved on an edge-plane pyrolytic graphite electrode (PGE) with a formal potential (E0') of -0.45 V at pH 7.0 in phosphate buffer. With the addition of isopentenyladenine, the reduction peak current for FADH2 decreased, and the oxidative peak current increased gradually. Therefore, a bacteria-OsCKX4-modified PGE has been developed for the detection of isopentenyladenine with a linear range of 1.0-11.0 µM and a lower limit of detection of 0.7 µM (S/N = 3). Slight interference was observed in the presence of other phytohormones, including brassinosteroid, abscisic acid, methylene jasminate and gibberellin. The proposed bacterial biosensor is stable, specific and simple and has great potential for applications that require the detection of cytokinins.

Glucose Oxidase Immobilized on a Functional Polymer Modified Glassy Carbon Electrode and Its Molecule Recognition of Glucose.

In the present study, a glucose oxidase (GluOx) direct electron transfer was realized on an aminated polyethylene glycol (mPEG), carboxylic acid functionalized multi-walled carbon nanotubes (fMWCNTs), and ionic liquid (IL) composite functional polymer modified glassy carbon electrode (GCE). The amino groups in PEG, carboxyl groups in multi-walled carbon nanotubes, and IL may have a better synergistic effect, thus more effectively adjust the hydrophobicity, stability, conductivity, and biocompatibility of the composite functional polymer film. The composite polymer membranes were characterized by cyclic voltammetry (CV), ultraviolet-visible (UV-Vis) spectrophotometer, fluorescence spectroscopy, electrochemical impedance spectroscopy (EIS), and transmission electron microscopy (TEM), respectively. In 50 mM, pH 7.0 phosphate buffer solution, the formal potential and heterogeneous electron transfer constant (ks) of GluOx on the composite functional polymer modified GCE were -0.27 V and 6.5 s-1, respectively. The modified electrode could recognize and detect glucose linearly in the range of 20 to 950 µM with a detection limit of 0.2 µM. The apparent Michaelis-Menten constant (Kmapp) of the modified electrode was 143 µM. The IL/mPEG-fMWCNTs functional polymer could preserve the conformational structure and catalytic activity of GluOx and lead to high sensitivity, stability, and selectivity of the biosensors for glucose recognition and detection.

Direct Electron Transfer of Enzymes Facilitated by Cytochromes.

The direct electron transfer (DET) of enzymes has been utilized to develop biosensors and enzymatic biofuel cells on micro- and nanostructured electrodes. Whereas some enzymes exhibit direct electron transfer between their active-site cofactor and an electrode, other oxidoreductases depend on acquired cytochrome domains or cytochrome subunits as built-in redox mediators. The physiological function of these cytochromes is to transfer electrons between the active-site cofactor and a redox partner protein. The exchange of the natural electron acceptor/donor by an electrode has been demonstrated for several cytochrome carrying oxidoreductases. These multi-cofactor enzymes have been applied in third generation biosensors to detect glucose, lactate, and other analytes. This review investigates and classifies oxidoreductases with a cytochrome domain, enzyme complexes with a cytochrome subunit, and covers designed cytochrome fusion enzymes. The structurally and electrochemically best characterized proponents from each enzyme class carrying a cytochrome, that is, flavoenzymes, quinoenzymes, molybdenum-cofactor enzymes, iron-sulfur cluster enzymes, and multi-haem enzymes, are featured, and their biochemical, kinetic, and electrochemical properties are compared. The cytochromes molecular and functional properties as well as their contribution to the interdomain electron transfer (IET, between active-site and cytochrome) and DET (between cytochrome and electrode) with regard to the achieved current density is discussed. Protein design strategies for cytochrome-fused enzymes are reviewed and the limiting factors as well as strategies to overcome them are outlined.

Voltammetric sensing performances of a carbon ionic liquid electrode modified with black phosphorene and hemin.

A black phosphorene (BPE) and poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate) (PEDOT:PSS) hybrid was used for the immobilization of hemin on a carbon ionic liquid electrode (CILE). BPE inside the PEDOT:PSS film was stable without adverse effects of water and oxygen. The hemin-modified electrode facilitates electrochemical communication with a couple of well-shaped and enhanced redox waves. Therefore BPE exhibits an accelerating function to the electron movement. This sensor exhibits excellent electrocatalytic effects on the reduction of various substrates including trichloroacetic acid (TCA), nitrite and H2O2. As for TCA, the reduction current at -0.36 V (vs. Ag/AgCl) increases linearly in the concentration range from 2.0 to 180 mmol·L-1 with a detection limit of 0.67 mmol·L-1 (at 3σ). As for nitrite, the reduction current at -0.59 Vis linear in the 1.0 to 10.5 mmol·L-1 concentration range, and the detection limit is 0.33 mmol·L-1 (at 3σ). As for H2O2, the reduction current at -0.033 V (vs. Ag/AgCl) is linear in the concentration range from 4.0 to 35.0 mmol·L-1 and the detection limit is 1.3 mmol·L-1 (at 3σ). The real sample was analyzed with satisfactory results, which indicated that BPE had potential applications in the field of electrochemical biosensor. Graphical abstract Photos of (a) black phosphorene (BPE) solution, (b) poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate) (PEDOT:PSS), (c) BPE-PEDOT:PSS (1:5) dispersion, and the fabrication procedure of this electrochemical sensor. It was applied to the determination of trichloroacetic acid, nitrite and hydrogen peroxide.

Direct electrochemical reduction of carbon dioxide by a molybdenum-containing formate dehydrogenase.

Formate dehydrogenase enzymes catalyse the reversible two-electron oxidation of formate to carbon dioxide. The class of metal-dependent formate dehydrogenases comprises prokaryotic enzymes holding redox-active centres and a catalytic site, containing either molybdenum or tungsten ion, that mediates the formate/carbon dioxide interconversion. The carbon dioxide reduction is of a particular interest, since it may be a route for its atmospheric mitigation with the simultaneous production of added-value products, as formate-derived compounds. Recently, the periplasmic formate dehydrogenase from Desulfovibrio desulfuricans, a molybdenum-containing enzyme, was proven to be an efficient enzyme for the CO2 reduction to formate. In this work, the immobilized formate dehydrogenase isolated from Desulfovibrio desulfuricans direct electrochemical behaviour was attained in the presence and absence of substrates and the formal potentials associated with the catalytic centre transitions were determined in non-turnover conditions. The enzyme catalytic activity towards carbon dioxide reduction was observed using direct electrochemical methods.

Construction of a Biosensor Based on a Combination of Cytochrome c, Graphene, and Gold Nanoparticles.

A biosensor based on a combination of cytochrome c (Cyt c), electrochemical reduced graphene oxides (ERGO), and gold nanoparticles (AuNPs) on a glassy carbon electrode (GCE) was fabricated. The proposed biosensor electrode was denoted as GCE/ERGO-Nafion/AuNPs/Cyt c/Nafion, where ERGO-Nafion was deposited by dropping graphene oxides-Nafion mixed droplet first and following electrochemical reduction, AuNPs were directly deposited on the surface of the ERGO-Nafion modified electrode by electrochemical reduction, and other components were deposited by the dropping-dry method. The effect of the deposition amount of AuNPs on direct electrochemistry of Cyt c in the proposed electrode was investigated. The hydrogen peroxide was taken to evaluate the performance of the proposed biosensor. The results showed that the biosensor has great analytical performance, including a high sensitivity, a wide linear range, a low detection limit, and good stability, reproducibility, and reliability.

Nitrogen doped chiral carbonaceous nanotube for ultrasensitive DNA direct electrochemistry, DNA hybridization and damage study.

In the interest of developing novel electrocatalyst for high performance DNA biosensing, with distinctive chiral double helix nanostructure, nitrogen doped chiral carbonaceous nanotube (Chiral-CNT) was employed for ultrasensitive label-free DNA biosensing research. Chiral-CNT can quantitative detection of four DNA bases with high sensitivity and selectivity. Without any prehydrolysis and labeling process, direct electrochemistry of single-stranded DNA and double-stranded DNA, qualitative and quantitative detection of DNA hybridization (low detection limit: 0.0268 g L-1) were realized. Moreover, sensitive detection of DNA damage induced by fenton reagent was also realized with low detection limit of 0.0350 mg mL-1 and high sensitivity of 7.42 µA mg-1 mL. The high biosensing performance attributes to the unique chiral structure of Chiral-CNT, leads to efficient interreaction between Chiral-CNT and DNA molecule.

Enhanced direct electron transfer of redox protein based on multiporous SnO2 nanofiber-carbon nanotube nanocomposite and its application in biosensing.

A novel third generation H2O2 biosensor is fabricated using multiporous SnO2 nanofiber/carbon nanotubes (CNTs) composite as a matrix for the immobilization of redox protein onto glassy carbon electrode. The multiporous nanofiber (MPNFs) of SnO2 is synthesized by electrospinning technique from the tin precursor. This nanofiber shows high surface area and good electrical conductivity. The SnO2 nanofiber/CNT composite increases the efficiency of biomolecule loading due to its high surface area. The morphology of the nanofiber has been evaluated by scanning electron microscopy (SEM). Cyclic Voltammetry and amperometry technique are employed to study and optimize the performance of the fabricated electrode. A direct electron transfer between the protein's redox centre and the glassy carbon electrode is established after fabrication of the electrode. The fabricated electrode shows excellent electrocatalytic reduction to H2O2. The catalysis currents increases linearly to the H2O2 concentration in a wide range of 1.0 10-6-1.4×10-4M and the lowest detection limit was 30nM (S/N=3). Moreover, the biosensor showed a rapid response to H2O2, a good stability and reproducibility.