Hydrazines probe of interaction of horseradish peroxidase with cryo-hydrogel

The interaction between horseradish peroxidase (HRP) and the cryo-hydrogel was probed by using hydrazines which show high specificity of the reaction of the edge in the prosthetic heme of horseradish peroxidase. For comparison, the interaction of hydrazine with the horseradish peroxidase adsorbed on graphite electrode was also carried out by using steady-state response of the enzyme electrode and cyclic voltammetry. In order to obtain a proper explanation of the kinetic parameters for the enzymatic reaction, the theoretical expressions of I-max and K-M' in the Michaelis-Menten equation for the experimental system were provided. (C) 1997 Elsevier Science B.V.

The electrochemical study of oxidation-reduction properties of horseradish peroxidase

Oxidation-reduction properties of horseradish peroxidase (HRP) have been investigated by using direct electrochemical methods. Two successive separated distinct one-electron processes of HRP were obtained and the related physiological processes were described. The monolayer coverage of HRP at the electrode surface is about 50 pmol/cm(2). UV-Vis spectrophotometry and stable amperometry prove that the enzyme electrode possesses catalytic activity for H2O2 in the absence of a mediator and it might offer an opportunity to build the third generation of biosensors for analytes, such as H2O2, glucose and cholesterol etc. (C) 1997 Elsevier Science S.A.

Electrochemical Study of the Self-exchange between Viologen-thiol Self assembled Monolayers and Alkanethiol Gold Electrodes

The preparation and cyclic voltammetric behaviors of self assembled monolayers (SAMs) containing electroactive viologen group have been investigated. Treatment of this viologen SAM with solutions of alkanethiols remits in replacing the electroactive third, shifting negatively its formal potentials and decreasing its heterogeneous elixtron transfer constants along with the immersion time. The aim of the work is to understand the exchange regularity of the mixed SANK on gold electrode surface.

Self-assembled monolayers of thiols on gold electrodes for bioelectrochemistry and biosensors

Monolayers of biological compounds including redox proteins and enzymes, and phospholipids have been immobilized on a gold electrode surface through self-assembling. These proteins and enzymes, such as cytochrome c, cytochrome c oxidase and horseradish peroxidase (HRP), immobilized covalently to the self-assembled monolayers (SAMs) of 3-mercaptopropionic acid on a gold electrode, communicate directly electrons with the electrode surface without mediators and keep their physiological activities. The electron transfer of HRP with the gold electrode can also be mediated by the alkanethiol SAMs with electroactive group viologens on the gold electrode surface. All these direct electrochemistries of proteins and enzymes might offer an opportunity to build a third generation of biosensors without mediators for analytes, such as H2O2, glucose and cholesterol. Monensin and valinomycin have been incorporated into the bilayers on the gold electrode consisting of the SAMs of alkanethiol and a lipid monolayer, which have high selectivity for monovalent ions, and the resulting Na+ or K+ sensor has a wide linear range and high stability. These self-assembly systems provide a good mimetic model for studying the physiological function of a membrane and its associated enzyme. (C) 1997 Elsevier Science S.A.

Electrochemical study of the interfacial characteristics of redox-active viologen thiol self-assembled monolayers

The electrochemical behavior of the electroactive self-assembled monolayers (SAMs) of thiol-functionalized viologen, CH3(CH2)(9)V2+(CH2)(8)SH, where V2+ is a viologen group, on the gold electrodes is examined by cyclic voltammetry and electrochemical a.c. impedance. A monolayer of viologen is immobilized on the gold electrode surface via the Au-S bond and the normal potentials corresponding to the two successive one-electron transfer processes of the viologen active centers are -310 mV and -652 mV (vs. Ag/AgCl) in 0.1 mol l(-1) phosphate buffer solution (pH 6.96) respectively. These results suggest that the viologen SAMs are stable and well-behaved monolayers. The experimental impedance data corresponding to different forms of viologen group have been fitted to equivalent electrical circuits, and the surface capacitances and resistances have been given. The heterogenous electron transfer rates of the first and the second redox processes are 7.57 s(-1) and 1.49 s(-1) respectively through a.c. impedance.

Electrochemical studies on vitamin B-12 and its derivatives

The electrochemical studies on vitamin B-12 and its derivatives were reviewed in this paper. The importance of electrochemical studies for explaining the mechanism of B-12 coenzyme in body was discussed. The latest results of electrochemical studies on vitamin B-12 and its derivatives was reviewed. A prospect for the electrochemical studies in vitamin B-12 and its derivatives was suggested.

Studies on the electron transfer centers of amino oxidase immobilized on self-assembly monolayer using electrochemical methods

The direct electron transfer of amino oxidase on electrode surface based on self-assembly technique occurs at 505 mW(vs. Ag/AgCl), indicating that copper atoms are the electron transfer centers and catalytic centers of amino oxidase.

Direct electron transfer to cytochrome c oxidase in self-assembled monolayers on gold electrodes

The monolayer of cytochrome c oxidase maintaining physiological activity and attached covalently to the self-assembled monolayers of 3-mercaptopropionic acid (MPA) on a gold electrode was obtained. The results of cyclic voltammetry show that direct electron transfer between cytochrome c oxidase and the electrode surface is a fast and diffusionless process. MPA has a dual role as both electrode modifier and the bridging molecule which: keeps cytochrome c oxidase at an appropriate orientation without denaturation and enables direct electron transfer between the protein and the modified electrode. Immobilized cytochrome c oxidase exhibits biphasic phenomena between the concentration of the electrolyte and the normal potentials; meanwhile its electrochemical behavior is also influenced by the buffer components. The quasi-reversible electron transfer process of cytochrome c oxidase with formal potential 385 mV vs. SHE in 5mM phosphate buffer solution (pH 6.4) corresponds to the redox reaction of cyt a(3) in cytochrome c oxidase, and the heterogeneous electron transfer rate constant obtained is 1.56 s(-1). By cyclic voltammetry measurements, it was observed that oxidation and reduction of cytochrome c in solution were catalyzed by the immobilized cytochrome c oxidase. This cytochrome c oxidase/MPA/Au system provides a good mimetic model to study the physiological functions of membrane-associated enzymes and hopefully to build a third-generation biosensor without using a mediator.

Electrochemical identification of intermediate forms of urea denaturation of horse heart cytochrome c

The electrochemical identification of the urea denaturation of horse heart cytochrome c in bulk solution at the 4,4'-dithiodipyridine-modified gold electrode is reported. The results are similar to the three-step transitions of equilibrium studies (Myer et al., Biochemistry, 19 (1980) 199) of urea denaturation of cytochrome c in bulk solution. This method permits a clear resolution of which of the three steps of urea denaturation is electrochemically related. In addition, by analysing the effects of urea on the structural forms of cytochrome c and on the solution properties, as well as the cyclic voltammetric responses of the protein, the individual forms of the urea denaturation of cytochrome c can be understood. The results reflect the superposition of protein denaturation on the electrode surface and in solution.

The ion selectivity of monensin incorporated phospholipid/alkanethiol bilayers

Monensin was incorporated into phospholipid/alkanethiol bilayers on the gold electrode surface by a new, paint-freeze method to deposit a lipid monolayer on the self-assembled monolayers (SAMs) of alkanethiol. The advantages of this assembly system with a suitable function for investigating the ion selective transfer across the mimetic biomembrane are based on the characteristics of SAMs of alkanethiols and monensin. On the one hand, the SAMs of alkanethiols bring out their efficiency of packing and coverage of the metal substrate and relatively long-term stability; on the other hand, monensin improves the ion selectivity noticeably. The selectivity coefficients K-Na+,K-K+, K-Na+,K-Rb+ and K-Na+,K-Ag+ are 6 x 10(-2), 7.2 x 10(-3) and 30 respectively. However, the selectivity coefficient K-Na+,K-Li+ could not be obtained by a potentiometric method due to the specific interaction between Li+ and phospholipid and the lower degree of complexion between Li+ and monensin. The potential response of this bilayer system to monovalent ions is fairly good. For example, the slope of the response to Na+ is close to 60 mV per decade and its linearity range is from 10(-1) to 10(-5) M with a detection limit of 2 x 10(-6) M, The bilayer is stable for at least two months without changing its properties. This monensin incorporated lipid/alkanethiol bilayer is a good mimetic biomembrane system, which provides great promise for investigating the ion transfer mechanism across the biomembrane and developing a practical biosensor.

Effect of structures of modifiers on electrochemistry of di-mu-oxo dimanganese 2,2'-bipyridine complex

The gold electrodes modified with 2-picolinic acid , nicotinic acid, iso-nicotinic or thiophene were prepared using membrane transfer method, The electrochemistry of di-mu-oxodimanganese 2,2'-bipyridine complex was studied in the acetic acid buffer solution at different modified gold electrodes, It was found that the modifiers which can promote the electrochemical reaction of the complex should be of at least two functional groups, One group can be bound to the electrode surface and the other can form electron transfer pathway between the modifier and the complex through sal; bridge or hydrogen bond, In addition, the mechanism of the electrochemical reaction was discussed.

Electrochemical study of isopoly- and heteropoly-oxometallates film modified microelectrodes .6. Preparation and redox properties of 12-molybdophosphoric acid and 12-molybdosilicic acid modified carbon fiber microelectrodes

The redox behaviours of 12-molybdophosphoric acid (12-MPA) and 12-molybdosilicic acid (12-MSA) in aqueous acid media are characterized at the carbon fiber (CF) microelectrode. The preparation of CF microelectrode modified with 12-MPA or 12-MSA monolayer and the oxidation-reduction properties of the modified electrode in aqueous acid media or 50% (v/v) water-organic media containing some inorganic acids are studied by cyclic voltammetry. 12-MPA or 12-MSA monolayer modified CF microelectrode with high stability and redox reversibility in aqueous acidic media can be prepared by simple dip coating. The cyclic voltammograms of 12-MPA and 12-MSA and their modified CF microelectrodes in aqueous acid solution exhibit three two-electron reversible waves with the same half-wave potentials, which defines that the species adsorbed on the CF electrode surface are 12-MPA and 12-MSA themselves. The acidity of electrolyte solution, the organic solvents in the electrolyte solution, and the scanning potential range strongly influence on the redox behaviours and stability of 12-MPA or 12-MSA monolayer modified electrodes. On the other hand, the catalytic effects of the 12-MPA and 12-MSA and chlorate anions in aqueous acidic solution on the electrode reaction processes of 12-MPA or 12-MSA are described.

Electrocatalytic Oxidation of Dihydric Phenol on Polypyrrole Film Modified Electrodes

The electrochemical behavior of catechol, hydroquinone and resorcinol on GC and PPy/GC electrode surface were studied by CV and RDE method. The results indicated that these three substance could be oxidized electrocatalytically on PPy film electrode. The possibility of fabrication of amperometric electrochemical sensor for catechol was also studied.

INVESTIGATION OF ELECTRIC DICHROISM OF CETYLPYRIDINIUM BROMIDE BY SPECTROELECTROCHEMISTRY WITH A LONG OPTICAL-PATH LENGTH THIN-LAYER CELL

In this paper, the electric dichroism of cetylpyridinium bromide (CPB) has been found and studied by spectroelectrochemistry with a long optical path length thin-layer cell (LOPTLC) for the first time. The CPB molecule with a long carbon chain and a polar pyridinium ring is anisotropic in molecular configuration or in polarizability. In the electric field of a thin-layer cell, the CPB molecule reorientates along the direction of the electric field and exhibits electric dichroism, which results in the increase of absorbance of CPB in the UV-vis range. By use of in situ measurement of spectroelectrochemistry, the order parameters of long molecular axis (S = 0.845) and short molecular axis (D = 0.155) and the angle between the long axis direction of the CPB molecule and the direction normal to the electrode surface (theta = 18-degrees 44') have been determined. These data were used to describe the state of arrangement of the molecules in the solution. The reorientation of CPB molecules is the result of the interaction between the anisotropic molecules and electric field. The effects of the concentration of CPB and of the applied electric field on the electric dichroism have been investigated.

SPECTROELECTROCHEMISTRY WITH CYLINDRICAL CARBON-FIBER MICROELECTRODES

Transmittance spectroelectrochemistry can be performed using a group of cylindrical microelectrodes. A dependence of absorbance on electrolytic charge during the potential step was derived. The rate constant of catalytic reaction of the ferrocyanide-ascorbic acid system was determined using single potential step-open circuit relaxation chronoabsorptometry. This is the first report that the reaction can still be considered as a pseudo-first-order reaction when the concentration of ascorbic acid is close to and even slightly lower than the concentration of ferrocyanide. The determined rate constant is in agreement with the reported value. The reason is that the diffusion of ascorbic acid toward electrode surface is contractive and the diffusion of the electrogenerated ferricyanide from the electrode surface to the bulk of solution is expansive.