RESUMO
In contrast to their molybdenum dependent relatives, tungsten enzymes operate at significantly lower redox potentials, and in some cases they can carry out reversible redox transformations of their substrates and products. Still, the electrochemical properties of W enzymes have received much less attention than their Mo relatives. Herein we analyse the tungsten enzyme aldehyde oxidoreductase (AOR) from the mesophilic bacterium Aromatoleum aromaticum which has been immobilised on a glassy carbon working electrode. This generates a functional system that electrochemically oxidises a wide variety of aromatic and aliphatic aldehydes in the presence of the electron transfer mediators benzyl viologen, methylene blue or dichlorophenol indophenol. Simulation of the cyclic voltammetry has enabled a thorough kinetic analysis of the system, which reveals that methylene blue acts as a two-electron acceptor. In contrast, the other two mediators act as single electron oxidants. The different electrochemical driving forces imparted by these mediators also lead to significantly different outer sphere electron transfer rates with AOR. This work shows that electrocatalytic aldehyde oxidation can be achieved at a low applied electrochemical potential leading to an extremely energy efficient catalytic process.
Assuntos
Aldeído Oxirredutases , Aldeídos , Aldeído Oxirredutases/química , Aldeído Oxirredutases/metabolismo , Tungstênio , Azul de Metileno , Cinética , Oxirredução , Aldeído DesidrogenaseRESUMO
MtsZ is a molybdenum-containing methionine sulfoxide reductase that supports virulence in the human respiratory pathogen Haemophilus influenzae (Hi). HiMtsZ belongs to a group of structurally and spectroscopically uncharacterized S-/N-oxide reductases, all of which are found in bacterial pathogens. Here, we have solved the crystal structure of HiMtsZ, which reveals that the HiMtsZ substrate-binding site encompasses a previously unrecognized part that accommodates the methionine sulfoxide side chain via interaction with His182 and Arg166. Charge and amino acid composition of this side chain-binding region vary and, as indicated by electrochemical, kinetic, and docking studies, could explain the diverse substrate specificity seen in closely related enzymes of this type. The HiMtsZ Mo active site has an underlying structural flexibility, where dissociation of the central Ser187 ligand affected catalysis at low pH. Unexpectedly, the two main HiMtsZ electron paramagnetic resonance (EPR) species resembled not only a related dimethyl sulfoxide reductase but also a structurally unrelated nitrate reductase that possesses an Asp-Mo ligand. This suggests that contrary to current views, the geometry of the Mo center and its primary ligands, rather than the specific amino acid environment, is the main determinant of the EPR properties of mononuclear Mo enzymes. The flexibility in the electronic structure of the Mo centers is also apparent in two of three HiMtsZ EPR-active Mo(V) species being catalytically incompetent off-pathway forms that could not be fully oxidized.
Assuntos
Proteínas de Bactérias/química , Haemophilus influenzae/enzimologia , Metaloproteínas/química , Molibdênio/metabolismo , Oxirredutases/química , Sequência de Aminoácidos , Proteínas de Bactérias/metabolismo , Catálise , Domínio Catalítico , Cinética , Ligantes , Metaloproteínas/metabolismo , Molibdênio/química , Oxirredução , Oxirredutases/metabolismo , Conformação Proteica , Homologia de Sequência de Aminoácidos , Especificidade por SubstratoRESUMO
Carbon-based nanocomposites have developed as the most promising and emerging materials in nanoscience and technology during the last several years. They are microscopic materials that range in size from 1 to 100 nanometers. They may be distinguished from bulk materials by their size, shape, increased surface-to-volume ratio, and unique physical and chemical characteristics. Carbon nanocomposite matrixes are often created by combining more than two distinct solid phase types. The nanocomposites that were constructed exhibit unique properties, such as significantly enhanced toughness, mechanical strength, and thermal/electrochemical conductivity. As a result of these advantages, nanocomposites have been used in a variety of applications, including catalysts, electrochemical sensors, biosensors, and energy storage devices, among others. This study focuses on the usage of several forms of carbon nanomaterials, such as carbon aerogels, carbon nanofibers, graphene, carbon nanotubes, and fullerenes, in the development of hydrogen fuel cells. These fuel cells have been successfully employed in numerous commercial sectors in recent years, notably in the car industry, due to their cost-effectiveness, eco-friendliness, and long-cyclic durability. Further; we discuss the principles, reaction mechanisms, and cyclic stability of the fuel cells and also new strategies and future challenges related to the development of viable fuel cells.
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Proteases are often used as biomarkers of many pathologies as well as of microbial contamination and infection. Therefore, extensive efforts are devoted to the development of protease sensors. Some applications would benefit from wireless monitoring of proteolytic activity at minimal cost, e.g., sensors embedded in care products like wound dressings and diapers to track wound and urinary infections. Passive (batteryless) and chipless transponders stand out among wireless sensing technologies when low cost is a requirement. Here, we developed and extensively characterized a composite material that is biodegradable but still highly stable in aqueous media, whose proteolytic degradation could be used in these wireless transponders as a transduction mechanism of proteolytic activity. This composite material consisted of a cross-linked gelatin network with incorporated caprylic acid. The digestion of the composite when exposed to proteases results in a change of its resistivity, a quantity that can be wirelessly monitored by coupling the composite to an inductor-capacitor resonator, i.e., an antenna. We experimentally proved this wireless sensor concept by monitoring the presence of a variety of proteases in aqueous media. Moreover, we also showed that detection time follows a relationship with protease concentration, which enables quantification possibilities for practical applications.
Assuntos
Ácidos Graxos/química , Gelatina/química , Peptídeo Hidrolases/análise , Máquina de Vetores de Suporte , Tecnologia sem Fio , Aspergillus/enzimologia , Ácidos Graxos/metabolismo , Gelatina/metabolismo , Peptídeo Hidrolases/metabolismo , Técnicas de Microbalança de Cristal de QuartzoRESUMO
We report the first electrochemical study of a lanthanoid-dependent methanol dehydrogenase (Eu-MDH) from the acidophilic verrucomicrobial methanotroph Methylacidiphilum fumariolicum SolV with its own physiological cytochromeâ cGJ electron acceptor. Eu-MDH harbours a redox active 2,7,9-tricarboxypyrroloquinoline quinone (PQQ) cofactor which is non-covalently bound but coordinates trivalent lanthanoid elements including Eu3+ . Eu-MDH and the cytochrome were co-adsorbed with the biopolymer chitosan and cast onto a mercaptoundecanol (MU) monolayer modified Au working electrode. Cyclic voltammetry of cytochrome cGJ reveals a well-defined quasi-reversible FeIII/II redox couple at +255â mV vs. NHE at pHâ 7.5 and this response is pH independent. The reversible one-electron response of the cytochrome cGJ transforms into a sigmoidal catalytic wave in the presence of Eu-MDH and its substrates (methanol or formaldehyde). The catalytic current was pH-dependent and pHâ 7.3 was found to be optimal. Kinetic parameters (pH dependence, activation energy) obtained by electrochemistry show the same trends as those obtained from an artificial phenazine ethosulfate/dichlorophenol indophenol assay.
Assuntos
Oxirredutases do Álcool/metabolismo , Citocromos c/química , Európio/química , Oxirredutases do Álcool/química , Biocatálise , Domínio Catalítico , Citocromos c/metabolismo , Técnicas Eletroquímicas , Eletrodos , Cinética , Metanol/química , Metanol/metabolismo , Oxirredução , Cofator PQQ/química , Cofator PQQ/metabolismo , Espectrofotometria , Especificidade por Substrato , Temperatura , Verrucomicrobia/enzimologiaRESUMO
A central conserved arginine, first identified as a clinical mutation leading to sulfite oxidase deficiency, is essential for catalytic competency of sulfite oxidizing molybdoenzymes, but the molecular basis for its effects on turnover and substrate affinity have not been fully elucidated. We have used a bacterial sulfite dehydrogenase, SorT, which lacks an internal heme group, but transfers electrons to an external, electron accepting cytochrome, SorU, to investigate the molecular functions of this arginine residue (Arg78). Assay of the SorT Mo centre catalytic competency in the absence of SorU showed that substitutions in the central arginine (R78Q, R78K and R78M mutations) only moderately altered SorT catalytic properties, except for R78M which caused significant reduction in SorT activity. The substitutions also altered the Mo-centre redox potentials (MoVI/V potential lowered by ca. 60-80mV). However, all Arg78 mutations significantly impaired the ability of SorT to transfer electrons to SorU, where activities were reduced 17 to 46-fold compared to SorTWT, precluding determination of kinetic parameters. This was accompanied by the observation of conformational changes in both the introduced Gln and Lys residues in the crystal structure of the enzymes. Taking into account data collected by others on related SOE mutations we propose that the formation and maintenance of an electron transfer complex between the Mo centre and electron accepting heme groups is the main function of the central arginine, and that the reduced turnover and increases in KMsulfite are caused by the inefficient operation of the oxidative half reaction of the catalytic cycle in enzymes carrying these mutations.
Assuntos
Arginina/química , Proteínas de Bactérias/química , Sinorhizobium meliloti/enzimologia , Sulfito Desidrogenase/química , Substituição de Aminoácidos , Arginina/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Domínio Catalítico , Transporte de Elétrons , Cinética , Molibdênio , Mutação de Sentido Incorreto , Oxirredução , Sinorhizobium meliloti/genética , Sulfito Desidrogenase/genética , Sulfito Desidrogenase/metabolismoRESUMO
The electrochemically driven catalysis of the complex molybdoenzyme steroid C25 dehydrogenase (S25DH) from the ß-Proteobacterium Sterolibacterium denitrificans is reported. S25DH catalyses the oxygen-independent regioselective hydroxylation of the tertiary C25 atom of sterols and also their derivatives. Cholest-4-en-3-one is a native substrate for S25DH, which produces 25-hydroxycholest-4-en-3-one as a product of catalytic turnover. Cholecalciferol (vitD3 ) is also a substrate. S25DH was immobilised on a modified gold working electrode with the co-adsorbent chitosan. The complexes ferricyanide ([Fe(CN)6 ]3- ) and ferrocenium methanol (FM+ ) are effective artificial electron acceptors from S25DH and act as mediators of electron transfer between the electrode and the enzyme. 2-Hydroxypropyl-ß-cyclodextrin (HPCD) was employed as a sterol solubiliser, in addition to 2-methoxyethanol. The catalytic activity varied, depending upon the concentration of solubiliser in the reaction mixture. Parallel studies with [Fe(CN)6 ]3- as a chemical (as opposed to electrochemical) oxidant coupled to HPLC analysis show that S25DH is capable of oxidising both vitD3 and its less stable isomer, pre-vitD3 , and that the former substrate is stabilised by HPCD.
Assuntos
Colestenonas/química , Bactérias Gram-Negativas/química , Oxirredutases/química , Oxirredutases/metabolismo , Rhodocyclaceae/química , Esteroides/química , Esteróis/química , Catálise , Hidroxilação , Oxirredução , Esteroides/metabolismo , Esteróis/metabolismoRESUMO
We report the first direct (unmediated) catalytic electrochemistry of a eukaryotic nitrate reductase (NR). NR from the filamentous fungus Neurospora crassa, is a member of the mononuclear molybdenum enzyme family and contains a Mo, heme and FAD cofactor which are involved in electron transfer from NAD(P)H to the (Mo) active site where reduction of nitrate to nitrite takes place. NR was adsorbed on an edge plane pyrolytic graphite (EPG) working electrode. Non-turnover redox responses were observed in the absence of nitrate from holo NR and three variants lacking the FAD, heme or Mo cofactor. The FAD response is due to dissociated cofactor in all cases. In the presence of nitrate, NR shows a pronounced cathodic catalytic wave with an apparent Michaelis constant (KM) of 39µM (pH7). The catalytic cathodic current increases with temperature from 5 to 35°C and an activation enthalpy of 26kJmol(-1) was determined. In spite of dissociation of the FAD cofactor, catalytically activity is maintained.
Assuntos
Neurospora crassa/enzimologia , Nitrato Redutase/química , Catálise , Eletroquímica , Concentração de Íons de Hidrogênio , Oxirredução , Oxigênio/farmacologia , TemperaturaRESUMO
We describe the catalytic voltammograms of the periplasmic arsenite oxidase (Aio) from the chemolithoautotrophic bacterium Rhizobium sp. str. NT-26 that oxidizes arsenite to arsenate. Electrochemistry of the enzyme was accomplished using its native electron transfer partner, cytochrome c552 (cyt c552), as a mediator. The protein cyt c552 adsorbed on a mercaptoundecanoic acid (MUA) modified Au electrode exhibited a stable, reversible one-electron voltammetric response at +275mV vs NHE (pH6). In the presence of arsenite and Aio the voltammetry of cyt c552 is transformed from a transient response to an amplified sigmoidal (steady state) wave consistent with an electro-catalytic system. Digital simulation was performed using a single set of parameters for all catalytic voltammetries obtained at different sweep rates and various substrate concentrations. The obtained kinetic constants from digital simulation provide new insight into the kinetics of the NT-26 Aio catalytic mechanism.
Assuntos
Catálise , Grupo dos Citocromos c/química , Transporte de Elétrons , Oxirredutases/química , Adsorção , Arsenitos/química , Arsenitos/metabolismo , Grupo dos Citocromos c/metabolismo , Eletroquímica , Cinética , Oxidantes/química , Oxirredução , Oxirredutases/metabolismo , Rhizobium/enzimologiaRESUMO
Nitrate reductase (NR) from the plant Arabidopsis thaliana has been employed in the development of an amperometric nitrate biosensor that functions at physiological pH. The anion anthraquinone-2-sulfonate (AQ) is used as an effective artificial electron transfer partner for NR at a glassy carbon (GC) electrode. Nitrate is enzymatically reduced to nitrite and the oxidized form of NR is electrochemically reduced by the hydroquinone form of the mediator (AQH2). The GC/NR electrode shows a pronounced cathodic wave for nitrate reduction and the catalytic current increases linearly in the nitrate concentration range of 10-400 µM with a correlation coefficient of 0.989. Using an amperometric method, a low detection limit of 0.76 nM (S/N = 3) was achieved. The practical application of the present electrochemical biosensor was demonstrated by the determination of nitrate concentration in natural water samples and the results agreed well with a standard spectroscopic method.
Assuntos
Técnicas Biossensoriais , Nitrato Redutase/química , Nitratos/isolamento & purificação , Arabidopsis/química , Arabidopsis/enzimologia , Hidroquinonas/química , Nitratos/químicaRESUMO
Photoluminescent-carbon nanoparticles (PL-CNPs) are a new class of materials that received immense interest among researchers due to their distinct characteristics, including photoluminescence, high surface-to-volume ratio, low cost, ease of synthesis, high quantum yield, and biocompatibility. By exploiting these outstanding properties, many studies have been reported on its utility as sensors, photocatalysts, probes for bio-imaging, and optoelectronics applications. From clinical applications to point-of-care test devices, drug loading to tracking of drug delivery, and other research innovations demonstrated PL-CNPs as an emerging material that could substitute conventional approaches. However, some of the PL-CNPs have poor PL properties and selectivity due to the presence of impurities (e.g., molecular fluorophores) and unfavourable surface charges by the passivation molecules, which impede their applications in many fields. To address these issues, many researchers have been paying great attention to developing new PL-CNPs with different composite combinations to achieve high PL properties and selectivity. Herein, we thoroughly discussed the recent development of various synthetic strategies employed to prepare PL-CNPs, doping effects, photostability, biocompatibility, and applications in sensing, bioimaging, and drug delivery fields. Moreover, the review discussed the limitations, future direction, and perspectives of PL-CNPs in possible potential applications.
Assuntos
Pontos Quânticos , CarbonoRESUMO
The bacterial molybdenum (Mo)-containing formate dehydrogenase (FdsDABG) from Cupriavidus necator is a soluble NAD+-dependent enzyme belonging to the DMSO reductase family. The holoenzyme is complex and possesses nine redox-active cofactors including a bis(molybdopterin guanine dinucleotide) (bis-MGD) active site, seven iron-sulfur clusters, and 1 equiv of flavin mononucleotide (FMN). FdsDABG catalyzes the two-electron oxidation of HCOO- (formate) to CO2 and reversibly reduces CO2 to HCOO- under physiological conditions close to its thermodynamic redox potential. Here we develop an electrocatalytically active formate oxidation/CO2 reduction system by immobilizing FdsDABG on a glassy carbon electrode in the presence of coadsorbents such as chitosan and glutaraldehyde. The reversible enzymatic interconversion between HCOO- and CO2 by FdsDABG has been realized with cyclic voltammetry using a range of artificial electron transfer mediators, with methylene blue (MB) and phenazine methosulfate (PMS) being particularly effective as electron acceptors for FdsDABG in formate oxidation. Methyl viologen (MV) acts as both an electron acceptor (MV2+) in formate oxidation and an electron donor (MV+â¢) for CO2 reduction. The catalytic voltammetry was reproduced by electrochemical simulation across a range of sweep rates and concentrations of formate and mediators to provide new insights into the kinetics of the FdsDABG catalytic mechanism.
Assuntos
Cupriavidus necator , Formiato Desidrogenases , Formiato Desidrogenases/química , Dióxido de Carbono/química , Oxirredução , FormiatosRESUMO
The bacterial xanthine dehydrogenase (XDH) from Rhodobacter capsulatus was immobilized on an edge-plane pyrolytic graphite (EPG) electrode to construct a hypoxanthine/xanthine biosensor that functions at physiological pH. Phenazine methosulfate (PMS) was used as a mediator which acts as an artificial electron-transfer partner for XDH. The enzyme catalyzes the oxidation of hypoxanthine to xanthine and also xanthine to uric acid by an oxidative hydroxylation mechanism. The present electrochemical biosensor was optimized in terms of applied potential and pH. The electrocatalytic oxidation response showed a linear dependence on the xanthine concentration ranging from 1.0 × 10(-5) to 1.8 × 10(-3) M with a correlation coefficient of 0.994. The modified electrode shows a very low detection limit for xanthine of 0.25 nM (signal-to-noise ratio = 3) using controlled potential amperometry.
Assuntos
Técnicas Biossensoriais , Hipoxantina/metabolismo , Rhodobacter capsulatus/enzimologia , Xantina Desidrogenase/metabolismo , Xantina/metabolismo , Catálise , Eletroquímica , Eletrodos , Grafite/química , Concentração de Íons de Hidrogênio , Oxirredução , Razão Sinal-RuídoRESUMO
Metal-air batteries and fuel cells are considered the most promising highly efficient energy storage systems because they possess long life cycles, high carbon monoxide (CO) tolerance, and low fuel crossover ability. The use of energy storage technology in the transport segment holds great promise for producing green and clean energy with lesser greenhouse gas (GHG) emissions. In recent years, nanoscale based electrocatalysts have shown remarkable electrocatalytic performance towards the construction of sustainable energy-related devices/applications, including fuel cells, metal-air battery and water-splitting processes. This review summarises the recent advancement in the development of nanoscale-based electrocatalysts and their energy-related electrocatalytic applications. Further, we focus on different synthetic approaches employed to fabricate the nanomaterial catalysts and also their size, shape and morphological related electrocatalytic performances. Following this, we discuss the catalytic reaction mechanism of the electrochemical energy generation process, which provides close insight to develop a more efficient catalyst. Moreover, we outline the future perspectives and challenges pertaining to the development of highly efficient nanoscale-based electrocatalysts for green energy storage technology.
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The Mo-dependent enzyme YiiM enzyme from Escherichia coli is a member of the sulfite oxidase family and shares many similarities with the well-studied human mitochondrial amidoxime reducing component (mARC). We have investigated YiiM catalysis using electrochemical and spectroscopic methods. EPR monitored redox potentiometry found the active site redox potentials to be MoVI/V -0.02 V and MoV/IV -0.12 V vs NHE at pH 7.2. In the presence of methyl viologen as an electrochemically reduced electron donor, YiiM catalysis was studied with a range of potential substrates. YiiM preferentially reduces N-hydroxylated compounds such as hydroxylamines, amidoximes, N-hydroxypurines and N-hydroxyureas but shows little or no activity against amine-oxides or sulfoxides. The pH optimum for catalysis was 7.1 and a bell-shaped pH profile was found with pKa values of 6.2 and 8.1 either side of this optimum that are associated with protonation/deprotonations that modulate activity. Simulation of the experimental voltammetry elucidated kinetic parameters associated with YiiM catalysis with the substrates 6-hydroxyaminopurine and benzamidoxime.
Assuntos
Escherichia coli , Molibdênio , Catálise , Domínio Catalítico , Humanos , Cinética , Molibdênio/química , OxirreduçãoRESUMO
Molybdenum-dependent enzymes that can reduce N-hydroxylated substrates (e.g. N-hydroxyl-purines, amidoximes) are found in bacteria, plants and vertebrates. They are involved in the conversion of a wide range of N-hydroxylated organic compounds into their corresponding amines, and utilize various redox proteins (cytochrome b5, cyt b5 reductase, flavin reductase) to deliver reducing equivalents to the catalytic centre. Here we present catalytic electrochemistry of the bacterial enzyme YcbX from Escherichia coli utilizing the synthetic electron transfer mediator methyl viologen (MV2+). The electrochemically reduced form (MV+.) acts as an effective electron donor for YcbX. To immobilize YcbX on a glassy carbon electrode, a facile protein crosslinking approach was used with the crosslinker glutaraldehyde (GTA). The YcbX-modified electrode showed a catalytic response for the reduction of a broad range of N-hydroxylated substrates. The catalytic activity of YcbX was examined at different pH values exhibiting an optimum at pHâ¯7.5 and a bell-shaped pH profile with deactivation through deprotonation (pKa1 9.1) or protonation (pKa2 6.1). Electrochemical simulation was employed to obtain new biochemical data for YcbX, in its reaction with methyl viologen and the organic substrates 6-N-hydroxylaminopurine (6-HAP) and benzamidoxime (BA).
Assuntos
Oxirredutases , Paraquat , Animais , Catálise , Eletroquímica , Escherichia coli/metabolismo , Oxirredutases/metabolismo , Paraquat/químicaRESUMO
Nitrate reductase (NR) from the fungus Neurospora crassa is a complex homodimeric metallo-flavoenzyme, where each protomer contains three distinct domains; the catalytically active terminal molybdopterin cofactor, a central heme-containing domain, and an FAD domain which binds with the natural electron donor NADPH. Here, we demonstrate the catalytic voltammetry of variants of N. crassa NRs on a modified Au electrode with the electrochemically reduced forms of benzyl viologen (BV2+) and anthraquinone sulfonate (AQS-) acting as artificial electron donors. The biopolymer chitosan used to entrap NR on the electrode non-covalently and the enzyme film was both stable and highly active. Electrochemistry was conducted on two distinct forms; one lacking the FAD cofactor and the other lacking both the FAD and heme cofactors. While both enzymes showed catalytic nitrate reductase activity, removal of the heme cofactor resulted in a more significant effect on the rate of nitrate reduction. Electrochemical simulation was carried out to enable kinetic characterisation of both the NR:nitrate and NR:mediator reactions.
Assuntos
Complexo de Proteínas da Cadeia de Transporte de Elétrons/química , Proteínas Fúngicas/química , Neurospora crassa/enzimologia , Nitrato Redutase/química , Benzil Viologênio/química , OxirreduçãoRESUMO
Presently, the global energy demand for increasing clean and green energy consumption lies in the development of low-cost, sustainable, economically viable and eco-friendly natured electrochemical conversion process, which is a significant advancement in different morphological types of advanced electrocatalysts to promote their electrocatalytic properties. Herein, we overviewed the recent advancements in oxygen evolution reactions (OERs), including easy electrode fabrication and significant action in water-splitting devices. To date, various synthetic approaches and modern characterization techniques have effectively been anticipated for upgraded OER activity. Moreover, the discussed electrode catalysts have emerged as the most hopeful constituents and received massive appreciation in OER with low overpotential and long-term cyclic stability. This review article broadly confers the recent progress research in OER, the general mechanistic approaches, challenges to enhance the catalytic performances and future directions for the scientific community.
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High efficient, low-cost and environmentally friendly-natured bi-functional-based perovskite electrode catalysts (BFPEC) are receiving increasing attention for oxygen reduction/oxygen evolution reaction (ORR/OER), playing an important role in the electrochemical energy conversion process using fuel cells and rechargeable batteries. Herein, we highlighted the different kinds of synthesis routes, morphological studies and electrode catalysts with A-site and B-site substitution co-substitution, generating oxygen vacancies studies for boosting ORR and OER activities. However, perovskite is a novel type of oxide family, which shows the state-of-art electrocatalytic performances in energy storage device applications. In this review article, we go through different types of BFPECs that have received massive appreciation and various strategies to promote their electrocatalytic activities (ORR/OER). Based on these various properties and their applications of BFPEC for ORR/OER, the general mechanism, catalytic performance and future outlook of these electrode catalysts have also been discussed.
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This paper describes a highly sensitive electrochemical (voltammetric) determination of sulfite using a combination of Starkeya novella sulfite dehydrogenase (SDH), horse heart cytochrome c (cyt c), and a self-assembled monolayer of 11-mercaptoundecanol (MU) cast on a gold electrode. The biosensor was optimized in terms of pH and the ratio of cyt c/SDH. The electrocatalytic oxidation current of sulfite increased linearly from 1 to 6 microM at the enzyme-modified electrode with a correlation coefficient of 0.9995 and an apparent Michaelis constant (K(M,app)) of 43 microM. Using an amperometric method, the low detection limit for sulfite at the enzyme-modified electrode was 44 pM (signal-to-noise ratio = 3). The modified electrode retained a stable response for 3 days while losing only ca. 4% of its initial sensitivity during a 2 week storage period in 50 mM Tris buffer solution at 4 degrees C. The enzyme electrode was successfully used for the determination of sulfite in beer and white wine samples. The results of these electrochemical analyses agreed well with an independent spectrophotometric method using Ellman's reagent, but the detection limit was far superior using the electrochemical method.