A Simple Fabrication Method for Three‐Dimensional Gold Nanoparticle Electrodes and Their Application to the Study of the Direct Electrochemistry of Cytochrome c

A simple method for constructing gold nanoparticle‐modified electrodes with three‐dimensional nanostructures is demonstrated. The electrodes were prepared by casting citrate‐reduced AuNPs onto polycrystalline gold electrodes. The resultant electrodes had a large surface area‐to‐volume ratio, adequate for high protein loading and conferring high stability. The gold nanoparticle electrodes were covered with a self‐assembled monolayer of 11‐mercaptoundecanoic acid for electrostatic immobilization of cytochrome c (cyt c). At the electrode, direct, reversible electron transfer from cyt c was observed with remarkable stability. Moreover, an extremely high surface coverage of electrochemically active cyt c, 167 fully packed monolayers, was obtained through use of the electrode.     

Direct and Cytochrome c Mediated Electrochemistry of Bilirubin Oxidase on Gold

Direct electron transfer (DET) of bilirubin oxidase from Myrothecium verrucaria (BOD) was established on promoter‐modified gold electrodes. The electrochemical behavior of the enzyme in solution was studied by means of cyclic voltammetry evaluating the biocatalytic reduction of dioxygen. The reaction of BOD at Au electrodes was shown to be efficient only at low pH. In addition, a novel interaction between BOD and cytochrome c (cyt.c) was found. It was shown that BOD efficiently accepts cyt.c as an electron donor in both cases when cyt.c is in solution and electrostatically adsorbed. The results suggest that cyt.c can play the role of a mediator facilitating electron transfer in a pH range where no DET could be observed between the enzyme and the electrode. For the interaction between cyt.c and BOD in solution the reaction kinetics has been studied electrochemically and spectrophotometrically.     

Highly Active Bidirectional Electron Transfer by a Self‐Assembled Electroactive Reduced‐Graphene‐Oxide‐Hybridized Biofilm

Low extracellular electron transfer performance is often a bottleneck in developing high‐performance bioelectrochemical systems. Herein, we show that the self‐assembly of graphene oxide and Shewanella oneidensis MR‐1 formed an electroactive, reduced‐graphene‐oxide‐hybridized, three‐dimensional macroporous biofilm, which enabled highly efficient bidirectional electron transfers between Shewanella and electrodes owing to high biomass incorporation and enhanced direct contact‐based extracellular electron transfer. This 3D electroactive biofilm delivered a 25‐fold increase in the outward current (oxidation current, electron flux from bacteria to electrodes) and 74‐fold increase in the inward current (reduction current, electron flux from electrodes to bacteria) over that of the naturally occurring biofilms.     

Direct Electrochemistry of Heme Proteins on Electrodes Modified with Didodecyldimethyl Ammonium Bromide and Carbon Black

A novel matrix based on commercially available carbon black (CB) N220 and didodecyldimethyl ammonium bromide (DDAB) was shown to be a reliable support for direct electron transfer reactions between screen printed electrode (SPE) and Fe(III)‐heme proteins. Cytochrome c (cyt c), myoglobin (Mb), horseradish peroxidase (HRP) and cytochromes P450 (CYP 51A1, CYP 3A4, CYP 2B4) generated well‐shaped cyclic voltammograms on SPE/CB/DDAB electrodes (both in solution and in immobilized state). The attractive performance characteristics of CB modified electrodes are advantageous over single‐walled carbon nanotubes (SW CNT) based ones. The achieved direct electrochemistry of heme proteins on CB/DDAB‐modified electrodes provided successful elaboration of the immunosensor for cardiac Mb. The immunosensor showed applicability for diagnostics of myocardial infarction displaying significant difference in cardiac Mb content of human blood plasma samples taken from the corresponding patients.     

Macroscopic Hexagonal Tubes of 3 d–4 f Metallocycles

The self‐assembly of three giant hexagonal 3d–4f metallocycles with inner diameters of 16.4, 16.5, and 16.4 Å, is described. Hexagonal metallocycles were stacked along the crystallographic c axis, producing unique hexagonal macroscopic tubular single crystals. The assembly mechanism of the tubular crystals was investigated. Remarkably, all three hexagonal metallocycles show typical single‐molecule magnet behavior, benefiting from the ferromagnetic couplings between the 3d and 4f ions.     

Functionalization of gold electrode surface with heterobifunctional poly(ethylene oxide)s having both mercapto and aldehyde groups

We synthesized heterobifunctional poly(ethylene oxide) (PEO) (α‐formyl‐ω‐mercapto‐PEO; CHO‐PEO₄₀₀‐SH, average molecular weight of PEO part being 400), which had both an aldehyde group as a binding site with amino group of protein and a mercapto group for gold electrode surface. The CHO‐PEO₄₀₀‐SH was adsorbed on a gold electrode surface and cytochrome c (cyt.c) was fixed on this modified electrode. The redox response of covalently immobilized cyt.c was observed on the cyclic voltammetry measurement, showing that CHO‐PEO₄₀₀‐SH can be used as a linker to fix cyt.c on an electrode. Another type of heterobifunctional PEO (α‐formyl‐ω‐(2‐pyridyldithio)‐PEO; CHO‐PEO₃₀₀‐SS‐Py), which had an aldehyde group and a 2‐pyridinethiol (2‐Py) through disulfide bond, was synthesized to form co‐adsorbed monolayer of PEO chain and 2‐Py on an electrode surface. It was expected, due to the spacer with shorter PEO chain and lower surface density, that better redox response of the fixed cyt.c was obtained. However, the redox response of fixed cyt.c was not detected on the CHO‐PEO₃₀₀‐SS‐Py modified gold electrode. Instead, this heterobifunctional PEO was found to function as a good promoter for cyt.c dissolved in phosphate buffer solution. Copyright © 2003 John Wiley & Sons, Ltd.     

Electorchemical Studies of Cytochrome c on Electodes Modified by Single—Wall Carbon Naotubes

Single-wall carbon nanotubes(SWNTs) modified gold electrodes were prepared by using two different methods.The electrochemical behavior of cytochrome c on the modified gold electrodes was investigated.The first kind of SWNT-modified electrode (noted as SWNT/Au electrode)was prepared by the adsorption of carboxylterminated SWNTs from DMF dispersion on the gold electrode.The oxidatively processed SWNT tips were covalently modified by coupling with amines (AET) to form amide linkage.Via Au-S chemical bonding,the self-assembled monolayer of thiol-unctionalized nanotubes on gold surface was fabricated so as to prepare the others SWNT-modified electrode (noted as SWNT/AET/Au electrode).It was shown from cyclic voltammetry cxperiments that cytochrome c exhibited direct electrochemical responses on the both electrodes, but only the current of controlled diffusion existed on the SWNT/Au electrode while both the currents of controlled diffusion and adsorption of cytochrome c occurred on the SWNT/AET/Au electrode.Photoelastic Modulation Infared Reflection Absorpthion Spectroscopy (PEM-IRRAS) and Quartz Crystal Microbalance (QCM) were employed to verify the adsorption of SWNTs on the gold electrodes.The results proved that SWNTs could enhance the direct electron transfer proecss between the electrodes and redox proteins.     

Electrochemical Quartz Crystal Microbalance Studies on Cytochrome c/Polyelectrolyte Multilayer Assemblies on Gold Electrodes

Polyelectrolyte multilayer assemblies containing proteins are of interest for applications such as sensors, bioreactors, and bioelectronics. A multilayer electrode was built up by the layer‐by‐layer strategy consisting of alternating layers of cytochrome c and poly(aniline sulfonic acid). The electrode showed a linear increase of redox active protein with the number of deposited layers. The principle of electrode preparation was transferred from needle electrodes to planar surfaces in order to further the understanding of electron transfer through the layer assembly by means of electrochemical quartz crystal microbalance studies. The deposition process was followed on‐line by detection of the frequency shift of the crystals and was found to be rather fast (minutes). The total mass deposited was found to correlate well with the electrochemical response of the immobilized cyt.c. Furthermore, the influence of the polyelectrolyte was investigated by addition of PSS to the PASA solution. The strong interaction of the former polyelectrolyte seemed to hinder the electron transfer although a multilayer formation was proved. Dilution of the protein solution with redox inactive apo‐cyt.c led to a strong decrease of the voltammetric signal, well beyond the percentage of apo‐cyt.c inside the assembly. Thus, arguments for an electron transfer via protein–protein interaction were found.     

Electroanalytical properties of cytochrome c by direct electrochemistry on multi-walled carbon nanotubes incorporated with DNA biocomposite film

A novel conductive biocomposite film (MWCNTs–DNA–cyt c) which contains multi-walled carbon nanotubes (MWCNTs) along with the incorporation of DNA and cytochrome c (cyt c) has been synthesized on glassy carbon electrode (GCE), gold (Au), indium tin oxide (ITO) and screen printed carbon electrode (SPCE) by potentiostatic methods. The presence of both MWCNTs and DNA in the biocomposite film enhances the surface coverage concentration (Γ), increases the electron transfer rate constant (K_s) up to 21% and decreases the degradation of cyt c during the cycling. The biocomposite film also exhibits a promising enhanced electrocatalytic activity towards the reduction of halogen oxyanions and oxidation of biochemical compounds such as ascorbic acid and l-cysteine. The cyclic voltammetry has been used for the measurement of electroanalytical properties of analytes by means of biocomposite film modified GCEs. The sensitivity of MWCNTs–DNA–cyt c modified GCE possess higher values than the values obtained for DNA–cyt c film modified GCE. Further, the reduction potentials of halogen oxyanions E_pc, clearly shows that the activity of the biocomposite is dependent on the electronegativity of halogen oxyanions. Electrochemical quartz crystal microbalance studies revealed the enhancements in the functional properties of MWCNTs, DNA and cyt c. We have studied the surface morphology of the biocomposite films using scanning electron microscopy and atomic force microscopy, which revealed that DNA and cyt c have been incorporated on MWCNTs. Finally, the flow injection analysis has been used for the amperometric detection of analytes at MWCNTs–DNA–cyt c film modified SPCE.     

Characterization of the Cytochrome c Membrane‐Binding Site Using Cardiolipin‐Containing Bicelles with NMR

Cytochrome (cyt) c transports electrons from Complex III to Complex IV in mitochondria. Cyt c is ordinarily anchored to the mitochondrial membrane through interaction with cardiolipin (CL), however its release into the cytosol initiates apoptosis. The cyt c interaction site with CL‐containing bicelles was characterized by NMR spectroscopy. Chemical shift perturbations in cyt c signals upon interaction with bicelles revealed that a relatively wide region, which includes the A‐site, the CXXCH motif, and the N‐ and C‐terminal helices, and contains multiple Lys residues, interacts cooperatively with CL. The specific cyt c–CL interaction increased with increasing CL molecules in the bicelles. The location of the cyt c interaction site for CL was similar to those for Complex III and Complex IV, thus indicating that cyt c recognizes lipids and partner proteins in a similar way. In addition to elucidating the cyt c membrane‐binding site, these results provide insight into the dynamic aspect of cyt c interactions in mitochondria.     

A new route of manipulation of poly(L‐lactic acid) crystallization by self‐assembly of p‐tert‐butylcalix[8]arene and toluene

A novel nucleating agent (TBC8‐t), self‐assembled with p‐tert‐butylcalix[8]arene (TBC8) and toluene, was used to manipulate the crystallization behavior of poly(L ‐lactic acid) (PLLA). Toluene molecules were used to adjust the crystallization structure of TBC8. Differential scanning calorimetry results show that the crystallization peak temperature (T_c) and crystallization rate (ΔH_c/time) of PLLA nucleated with TBC8‐t are 132.3 °C and 0.24 J/gs, respectively, which are much higher than that with conventional nucleating agent‐talc (T_c = 119.3 °C, ΔH_c/time = 0.13 J/gs). The results of polarized optical microscopy demonstrate that TBC8‐t could greatly enhance the crystallization rate of PLLA by increasing the nucleation rate rather than crystal growth rate. Along with an improvement of the crystallization rate, the crystalline morphology of PLLA is also affected by TBC8‐t. The addition of TBC8‐t transforms most of the original spherulite crystals into sheaf‐like crystals. © 2010 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 48: 1235–1243, 2010     

Synthesis,Crystal Structures,and Optical/Electronic Properties of Sphere–Rod Shape Amphiphiles Based on a [60]Fullerene?Oligofluorene Conjugate

A series of sphere–rod shape amphiphiles, in which a [60]fullerene (C₆₀) sphere was connected to the center of an oligofluorene (OF) rod through a rigid linkage (OF‐C₆₀), were designed and synthesized. Alkyl chains of various lengths were attached onto the OFs on both sides of the C₆₀ spheres. These compounds, denoted as alkyl‐OF‐C₆₀, were fully characterized by ¹H NMR, ¹³C NMR, and FTIR spectroscopy and by MALDI‐TOF mass spectrometry. The morphologies and structures of their crystals were elucidated by wide‐angle X‐ray diffraction (WAXD) and by electron diffraction in transmission electron microscopy (TEM). Butyl‐OF‐C₆₀ forms a monoclinic unit cell (a=1.86, b=3.96, c=2.24 nm; α=γ=90°, β=68°; space group P2), octyl‐OF‐C₆₀ also forms a monoclinic unit cell (a=2.21, b=4.06, c=1.81 nm; α=γ=90°, β=75.5°; space group C2m), and dodecanyl‐OF‐C₆₀ forms a triclinic structure (a=1.82, b=4.35, c=2.26 nm; α=93.1°, β=94.5°, γ=92.7°; space group P1). The inequivalent spheres and rods were found to pack into an alternating layered structure of C₆₀ and OF in the crystals, thus resembling a “double‐cable” structure. UV/Vis absorption spectroscopy revealed an electron perturbation between the two individual chromophores (C₆₀ and OF) in their ground states. Fluorescence spectroscopy exhibited complete fluorescence quenching of their solutions in toluene, thus suggesting an effective energy transfer from OF to C₆₀. Cyclic voltammetry indicated that the energy‐level profiles of C₆₀ and OF remained essentially unchanged. This work has broad implications in terms of understanding the self‐assembly and molecular packing of conjugated materials in crystals and has potential applications in organic field‐effect transistors and bulk heterojunction solar cells.     

Voltammetry of immobilized cytochrome c on novel binary self-assembled monolayers of thioctic acid and thioctic amide modified gold electrodes

Horse heart cytochrome c (cyt c) was adsorbed on the binary self-assembled monolayers (SAMs) composed of thioctic acid (T-COOH) and thioctic amide (T-NH₂) at gold electrodes via electrostatic interaction. The cyt c adsorbed on the modified gold electrode exhibited well-defined reversible electrochemical behavior in 10 mM phosphate buffer solution (PBS, pH 7.0). The surface concentration (Γ) of electroactive species, cyt c, on the binary SAMs was higher than that in single-component SAMs of T-COOH, and reached a maximum value of 9.2 × 10⁻¹² mol cm⁻² when the ratio of T-COOH to T-NH₂ in adsorption solution was of 3:2, and the formal potential (E^0′=(E_pa+E_pc)/2) of cyt c was −0.032 V (vs. Ag|AgCl (3 M NaCl)) in a 10 mM PBS. The interaction between cyt c and the binary SAMs made the E^0′ shift negatively when compared with that of cyt c in solution (+0.258 V vs. NHE, i.e., +0.058 V vs. Ag|AgCl (3 M NaCl)). The fractional coverage of bound cyt c was a 0.64 theoretical monolayer. The standard electron transfer rate constant of cyt c immobilized on the binary SAMs was also higher than that on single-component SAMs of T-COOH, and the maximum value of 15.8 ± 0.6 s⁻¹ was obtained when the ratio of T-COOH to T-NH₂ in adsorption solution was at 3:2. The results suggest that the electrode modified with the binary SAMs functions better than the electrode modified with single-component SAMs of T-COOH.     

Superoxide sensor based on cytochrome c immobilized on mixed-thiol SAM with a new calibration method

Cytochrome c was immobilized on a mixed-thiol (mercaptoundecanoic acid/mercaptoundecanol) modified gold electrode (MUA:MU/cyt c electrode). Characterization of the cyt c electrode showed a quasi-reversible, electrochemical redox behavior with a formal potential of −13±5 mV (versus Ag/AgCl) for the surface adsorbed protein and 3±5 mV for covalently immobilized cyt c. The heterogeneous electron transfer rate constants were determined to be about 70 and 40 s⁻¹ for both states of the protein, respectively. They were found to be significantly higher than those of pure MUA-modified cyt c electrodes (MUA/cyt c electrodes). The interaction of superoxide radicals (O₂⁻) with the (MUA:MU)/cyt c electrode was characterized and used for an amperometric O₂⁻ detection. The influence of H₂O₂ and uric acid on the sensor signal was investigated. The sensitivity of the (MUA:MU)/cyt c electrode to O₂⁻ was significantly improved compared with that of the MUA/cyt c electrode. Based on a kinetic model for the superoxide detection system, a new calibration method was established. This simple and fast method used the spontaneous dismutation of KO₂ and was compared with the enzymatic superoxide generation system using xanthine oxidase.     

Vibronic polarons: Self‐trapping,local rotation,and band features

We revisit basic theoretical concepts of local and itinerant vibronic polarons in crystals. The following results may be regarded as novel: (1) The electron self‐trapping rate to a small polaron is calculated via the reaction rate method; subsequently, self‐trapped on‐center small polarons relax to an off‐center vibronic polaron state. (2) The general vibronic Hamiltonian is redefined so as to incorporate both local and itinerant behavior and pairing into bipolarons or Cooper pairs. (3) The planar rotation and diametral tunneling of an off‐center polaron around and across its centrosymmetrical site are dealt with to adiabatic approximation. (4) Variational calculations are made for vibronic polarons itinerant along 1‐D chains by means of a two‐band extension of Merrifield's ansatz. This investigation of vibronic polarons is undertaken in view of their presumed role in high‐temperature superconductivity and colossal magnetoresistance. © 2002 Wiley Periodicals, Inc. Int J Quantum Chem, 2002     

Spectroscopic,Thermal, and Structural Studies of Two New Co‐Crystals of [4,4′‐Bithiazole]‐2,2′‐diamine

Two new 2 : 1 co‐crystals based on [4,4′‐bithiazole]‐2,2′‐diamine (=2,2′‐diamino‐4,4′‐bithiazole (DABTZ)) with 2,2′‐bipyridine (bipy) and benzo‐18‐crown‐6 (bk) were synthesized by slow‐evaporation method in MeOH. These co‐crystals were characterized by means of elemental analysis, and IR, and ¹H‐ and ¹³C‐NMR spectroscopy. Also, thermal analyses under air atmosphere and X‐ray crystallography have been performed on these structures. X‐Ray single‐crystal analyses revealed that these networks contain large vacant voids. These structures, [(DABTZ)₂(bipy)] and [(DABTZ)₂(bk)(MeOH)], crystallized in monoclinic and triclinic forms with space groups of P2₁/c and P , respectively. The self‐assembly of these compounds in the solid state is likely caused by both H‐bonding and π? π stacking.     

Photoinduced Triplet–Triplet Energy Transfer in a 2‐Ureido‐4(1H)‐Pyrimidinone‐Bridged,Quadruply Hydrogen‐Bonded Ferrocene–Fullerene Assembly

2‐Ureido‐4(1H)‐pyrimidinone‐bridged ferrocene–fullerene assembly I is designed and synthesized for elaborating the photoinduced electron‐transfer processes in self‐complementary quadruply hydrogen‐bonded modules. Unexpectedly, steady‐state and time‐resolved spectroscopy reveal an inefficient electron‐transfer process from the ferrocene to the singlet or triplet excited state of the fullerene, although the electron‐transfer reactions are thermodynamically feasible. Instead, an effective intra‐assembly triplet–triplet energy‐transfer process is found to be operative in assembly I with a rate constant of 9.2×10⁵ s^?1 and an efficiency of 73 % in CH₂Cl₂ at room temperature.     

Growth of Large Polymer Extended‐Chain Single Crystals in a Poly(ethylene terephthalate)/Bisphenol A Polycarbonate Blend under High Pressure

Summary: High‐quality synthetic polymer extended‐chain single crystals with c‐axis thickness exceeding 100 µm are obtained in a polyethylene terephthalate/polycarbonate (PET/PC) blend rapidly. Some crystals comprise of PET, while others are made up by the copolymers with PET and PC blocks. The fast growth of large crystals is attributed to the mechanisms including transesterification, segment nucleation, and chain‐sliding diffusion, which is a self‐assembling process induced by chemical reactions in multiphase polymer system at high pressure. Similar to the role of enzyme in biosystem, the PC plays as a macromolecular catalyst. This process provides a new direction to grow large polymer single crystals and suggests that self‐assembly under high pressure is a promising method to create materials with new structures and properties.

SEI for a wedge‐shaped extended‐chain single crystal grown in a PET/PC blend at 200 MPa, 623 K for 6 h.     

Electron‐Driven Self‐Assembly of Salt Nanocrystals in Liquid Helium

The self‐assembly of salt nanocrystals from chemical reactions inside liquid helium is reported for the first time. Reaction is initiated by an electron impacting a helium nanodroplet containing sodium atoms and SF₆ molecules, leading to preferential production of energetically favorable structures based on the unit cell of crystalline NaF. These favorable structures are observed as magic number ions (anomalously intense peaks) in mass spectra and are seen in both cationic and anionic channels in mass spectra, for example, (NaF)_nNa⁺ and (NaF)_nF^?. In the case of anions the self‐assembly is not directly initiated by electrons: the dominant process involves resonant electron‐induced production of metastable electronically excited He^? anions, which then initiate anionic chemistry by electron transfer.     

Template‐Free Supracolloidal Self‐Assembly of Atomically Precise Gold Nanoclusters: From 2D Colloidal Crystals to Spherical Capsids

We report supracolloidal self‐assembly of atomically precise and strictly monodisperse gold nanoclusters involving p‐mercaptobenzoic acid ligands (Au₁₀₂‐pMBA₄₄) under aqueous conditions into hexagonally packed monolayer‐thick two‐dimensional facetted colloidal crystals (thickness 2.7 nm) and their bending to closed shells leading to spherical capsids (d ca. 200 nm), as controlled by solvent conditions. The 2D colloidal assembly is driven in template‐free manner by the spontaneous patchiness of the pMBA ligands around the Au₁₀₂‐pMBA₄₄ nanoclusters preferably towards equatorial plane, thus promoting inter‐nanocluster hydrogen bonds and high packing to planar sheets. More generally, the findings encourage to explore atomically precise nanoclusters towards highly controlled colloidal self‐assemblies.