Graphene Paste Electrode: Analytical Applications for the Quantification of Dopamine,Phenolic Compounds and Ethanol

This work reports the analytical applications of a graphene paste electrode (GrPE) for the quantification of dopamine, ethanol and phenolic compounds. Dopamine was detected by differential pulse voltammetry‐adsorptive stripping with medium exchange at submicromolar levels even in the presence of high excess of ascorbic acid and serotonin. The electrocatalytic activity of graphene towards the oxidation of NADH and the reduction of quinones allowed the sensitive amperometric determination of ethanol and phenols using GrPE modified with alcohol dehydrogenase/NAD⁺ or polyphenol oxidase, respectively, with successful applications in real samples like alcoholic beverages and tea.     

具胆酸单元氢化烟酰胺腺嘌呤二核苷酸模型物的合成及其仿酶不对称还原性

设计并合成了5种具有胆酸单元的氢化烟酰胺腺嘌呤二核苷酸(NADH)模型化合物,并以苯甲酰甲酸甲酯为底物,镁离子为催化剂,研究了模型化合物的不对称还原能力.结果表明这些模型化合物能够将α-酮酸酯还原为α-羟基酸酯,有一定的负氢转移能力,但它们的手性还原能力不佳,对映体过量值最高为26%.     

Bridging the Two Worlds: A Universal Interface between Enzymatic and DNA Computing Systems

Molecular computing based on enzymes or nucleic acids has attracted a great deal of attention due to the perspectives of controlling living systems in the way we control electronic computers. Enzyme‐based computational systems can respond to a great variety of small molecule inputs. They have the advantage of signal amplification and highly specific recognition. DNA computing systems are most often controlled by oligonucleotide inputs/outputs and are capable of sophisticated computing as well as controlling gene expressions. Here, we developed an interface that enables communication of otherwise incompatible nucleic‐acid and enzyme‐computational systems. The enzymatic system processes small molecules as inputs and produces NADH as an output. The NADH output triggers electrochemical release of an oligonucleotide, which is accepted by a DNA computational system as an input. This interface is universal because the enzymatic and DNA computing systems are independent of each other in composition and complexity.     

Standardized Characterization of Electrocatalytic Electrodes

This paper investigates the utility of ‘cross–lab’ comparative analysis of electrocatalytic electrode performance using standardized modular stack cells and test protocols. Using poly(methylene green)‐modified glassy carbon electrodes as the model system, we characterized electrode fabrication and performance with respect to the catalytic oxidation of NADH at neutral pH and low overpotential. Three sets of experiments were duplicated across four independent laboratories and the experimental results from each set were analyzed and compared in terms of key electroanalytical parameters. Statistical analyses were performed at three distinct levels: 1) the standard deviation among repetitive cycles within an experiment; 2) the standard deviation among repetitive experiments in the same laboratory, and 3) the standard deviation among experiments performed across all four laboratories. Using predefined criteria of ‘reproducibility’ for each level, most parameters were found to be statistically reproducible at most levels. When a particular parameter was found to be irreproducible in a given level, commentary is given on how that information can be used diagnose what chemical/physical aspects of the process were uncontrolled or poorly understood and therefore candidates for future research. This exercise, which is presented as a ‘proof–of‐principle’ step towards the concept of standardizing electrocatalytic evaluation, illustrates the importance of executing electrochemical characterization protocols across several labs and using fixed geometry and dimensions, system configuration, and applied electrochemical conditions. Future work is under way to extend these principles to systems with fluid flow.     

肌红蛋白作催化剂荧光分析法高灵敏检测NADH

基于肌红蛋白对NADH与H2O2之间的反应具有催化作用,提出了一种催化荧光光谱测定NADH的新方法.在实验选定的最佳反应条件下,该反应体系的荧光强度与NADH的浓度在1.6×10-7～2.0×10-5mol/L范围内呈良好的线性关系,检出限为4.1×10-8mol/L.对1.0×10-6mol/LNADH的标准样品作了9次重复测定,其相对标准偏差RSD=4.6%.     

Free‐standing Graphene/Poly(methylene blue)/AgNPs Composite Paper for Electrochemical Sensing of NADH

Highly flexible graphene/poly(methylene blue)/AgNPs composite paper was successfully prepared for amperometric biosensing of NADH. For this purpose, a dispersion including graphene oxide (GO), methylene blue (MB) and silver nanoparticles (AgNPs) was prepared and GO/MB/AgNPs paper was acquired by vacuum‐filtration of this dispersion through a suitable membrane. After peeling it off from membrane, it was transformed to rGO/MB/AgNPs paper by performing reduction with hydriodic acid. In a three‐electrode cell, which is containing 0.1 M phosphate buffer solution (pH: 9.0), rGO/MB/AgNPs paper was used as working electrode and rGO/poly(MB)/AgNPs composite paper was generated by surface‐confined electropolymerization of MB using successive cyclic voltammetry approach in a suitable potential window. Characterization of this composite paper was carried out by using scanning electron microscopy, scanning tunneling microscopy, X‐ray photoelectron spectroscopy, powder X‐ray diffraction spectroscopy, Raman spectroscopy, four‐point probe conductivity measurement and cyclic voltammetry techniques. Flexible rGO/poly(MB)/AgNPs composite paper has demonstrated high sensitivity, wide linear range and low detection limit for amperometric quantification of NADH.     

Directional Regulation of Enzyme Pathways through the Control of Substrate Channeling on a DNA Origami Scaffold

Artificial multi‐enzyme systems with precise and dynamic control over the enzyme pathway activity are of great significance in bionanotechnology and synthetic biology. Herein, we exploit a spatially addressable DNA nanoplatform for the directional regulation of two enzyme pathways (G6pDH–MDH and G6pDH–LDH) through the control of NAD⁺ substrate channeling by specifically shifting NAD⁺ between the two enzyme pairs. We believe that this concept will be useful for the design of regulatory biological circuits for synthetic biology and biomedicine.     

Optimized carbon nanotube fiber microelectrodes as potential analytical tools

The preparation and interesting electrochemical properties of carbon nanotube (CNT) fiber microelectrodes are reported. By combining the advantages of CNT with those of fiber electrodes, this type of microelectrode differs from CNT-modified or CNT-containing composite electrodes, because they are made solely of CNT without other components, for example additives or binders. The performance of these electrodes has been characterized with regard to, among others, the electrocatalytic oxidation of analytes via dehydrogenase-mediated reactions. In this context the reversible regeneration of the coenzyme NAD(+) using a mediator is a key step in the development of new amperometric sensor devices and we have successfully immobilized mediator molecules that are very efficient for this purpose on the surface of the CNT fiber electrode. The microelectrodes thus obtained have been compared with classic carbon microelectrodes and have promising behavior in biosensing applications, especially after specific pretreatments such as CNT alignment inside the fiber or expansion of the specific surface by chemically induced swelling.     

Evaluation of Meldola Blue‐Carbon Nanotube‐Sol‐Gel Composite for Electrochemical NADH Sensors and Their Application for Lactate Dehydrogenase‐Based Biosensors

A novel MB‐SWNT‐sol‐gel nanocomposite material was prepared by the sol‐gel process incorporating a redox mediator and carbon nanotubes. The electrocatalytic properties of the nanomaterial based sensor toward NADH oxidation were studied by electrochemical measurements. Significant enhancement of oxidation current is obtained at electrodes modified by MB‐SWNT‐sol‐gel in comparison with the analogous carbon black and/or graphite composite modified electrode. The usefulness of the nanocomposite material as a matrix for immobilizing enzymes is also demonstrated. Analytical parameters of D ‐lactate biosensors with and without SWNT in the hybrid film were compared demonstrating that performance of the biosensor was significantly improved when introducing SWNT.     

Electrocatalytic detection of NADH and ethanol at glassy carbon electrode modified with electropolymerized films from methylene green

The oxidation of NADH on electropolymerizing methylene green (MG)-modified glassy carbon electrode (GCE) is described. The modified electrode shows an excellent electrocatalytic activity toward NADH oxidation, reducing its overpotential by about 650 mV and exhibits a wide linear range of 5.6–420 μM NADH with the detection limit of 3.8 μM. The electrode displays a good reproducibility and stability and the coexisting species does not affect the determination of NADH. The application in the amperometric biosensing of ethanol using alcohol dehydrogenase enzyme (ADH) also has been demonstrated with this electrode. MG-modified GCE can not only be used to detect NADH in biochemical reaction, but also can be used as the potential matrix of the construction of dehydrogenases biosensor.