RESUMEN
We have compared the properties and resistance to DA fouling of a carbon nanotube fiber (CNTF) microelectrode to a traditional carbon fiber (CF) microelectrode. These two materials show comparable electrochemical activities for outer-sphere and inner-sphere redox reactions. Although the CNTF might have a higher intrinsic RC constant, thus limiting its high-frequency behavior, the CNTF shows a significantly higher durability than the CF in terms of electrode stability. During constant oxidation of 100 µM DA, the signal measured by the CNTF microelectrode shows a 2-h window over which no decrease in current is observed. Under the same conditions, the current obtained at the CF microelectrode decreases by almost 50%. A model of the fouling process, assuming the formation of growing patches of insulator on the surface, has been compared to the data. This model is found to be in good agreement with our results and indicates a growth rate of the patches in the 0.1-2 nm s(-1) range.
Asunto(s)
Dopamina/química , Electroquímica/instrumentación , Nanotubos de Carbono/química , Carbono/química , Fibra de Carbono , Dopamina/análisis , Cinética , MicroelectrodosRESUMEN
The flavocytochrome cellobiose dehydrogenase (CDH) is a versatile biorecognition element capable of detecting carbohydrates as well as quinones and catecholamines. In addition, it can be used as an anode biocatalyst for enzymatic biofuel cells to power miniaturised sensor-transmitter systems. Various electrode materials and designs have been tested in the past decade to utilize and enhance the direct electron transfer (DET) from the enzyme to the electrode. Additionally, mediated electron transfer (MET) approaches via soluble redox mediators and redox polymers have been pursued. Biosensors for cellobiose, lactose and glucose determination are based on CDH from different fungal producers, which show differences with respect to substrate specificity, pH optima, DET efficiency and surface binding affinity. Biosensors for the detection of quinones and catecholamines can use carbohydrates for analyte regeneration and signal amplification. This review discusses different approaches to enhance the sensitivity and selectivity of CDH-based biosensors, which focus on (1) more efficient DET on chemically modified or nanostructured electrodes, (2) the synthesis of custom-made redox polymers for higher MET currents and (3) the engineering of enzymes and reaction pathways. Combination of these strategies will enable the design of sensitive and selective CDH-based biosensors with reduced electrode size for the detection of analytes in continuous on-site and point-of-care applications.
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Técnicas Biosensibles/instrumentación , Deshidrogenasas de Carbohidratos/metabolismo , Enzimas Inmovilizadas/metabolismo , Hongos/enzimología , Animales , Fuentes de Energía Bioeléctrica , Técnicas Biosensibles/métodos , Deshidrogenasas de Carbohidratos/química , Electrodos , Enzimas Inmovilizadas/química , Hongos/química , Humanos , Modelos Moleculares , Nanoestructuras/química , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismoRESUMEN
Cellobiose dehydrogenase (CDH) is a bioelectrocatalyst that enables direct electron transfer (DET) in biosensors and biofuel cells. The application of this bidomain hemoflavoenzyme for physiological glucose measurements is limited by its acidic pH optimum and slow interdomain electron transfer (IET) at pH 7.5. The reason for this rate-limiting electron transfer step is electrostatic repulsion at the interface between the catalytic dehydrogenase domain and the electron mediating cytochrome domain (CYT). We applied rational interface engineering to accelerate the IET for the pH prevailing in blood or interstitial fluid. Phylogenetic and structural analyses guided the design of 17 variants in which acidic amino acids were mutated at the CYT domain. Five mutations (G71K, D160K, Q174K, D177K, M180K) increased the pH optimum and IET rate. Structure-based analysis of the variants suggested two mechanisms explaining the improvements: electrostatic steering and stabilization of the closed state by hydrogen bonding. Combining the mutations into six combinatorial variants with up to five mutations shifted the pH optimum from 4.5 to 7.0 and increased the IET at pH 7.5 over 12-fold from 0.1 to 1.24 s-1 . While the mutants sustained a high enzymatic activity and even surpassed the IET of the wild-type enzyme, the accumulated positive charges on the CYT domain decreased DET, highlighting the importance of CYT for IET and DET. This study shows that interface engineering is an effective strategy to shift the pH optimum and improve the IET of CDH, but future work needs to maintain the DET of the CYT domain for bioelectronic applications.
Asunto(s)
Deshidrogenasas de Carbohidratos , Electrones , Filogenia , Deshidrogenasas de Carbohidratos/genética , Deshidrogenasas de Carbohidratos/química , Citocromos/metabolismo , Transporte de Electrón/fisiologíaRESUMEN
The genome of Neurospora crassa encodes two different cellobiose dehydrogenases (CDHs) with a sequence identity of only 53%. So far, only CDH IIA, which is induced during growth on cellulose and features a C-terminal carbohydrate binding module (CBM), was detected in the secretome of N. crassa and preliminarily characterized. CDH IIB is not significantly upregulated during growth on cellulosic material and lacks a CBM. Since CDH IIB could not be identified in the secretome, both CDHs were recombinantly produced in Pichia pastoris. With the cytochrome domain-dependent one-electron acceptor cytochrome c, CDH IIA has a narrower and more acidic pH optimum than CDH IIB. Interestingly, the catalytic efficiencies of both CDHs for carbohydrates are rather similar, but CDH IIA exhibits 4- to 5-times-higher apparent catalytic constants (k(cat) and K(m) values) than CDH IIB for most tested carbohydrates. A third major difference is the 65-mV-lower redox potential of the heme b cofactor in the cytochrome domain of CDH IIA than CDH IIB. To study the interaction with a member of the glycoside hydrolase 61 family, the copper-dependent polysaccharide monooxygenase GH61-3 (NCU02916) from N. crassa was expressed in P. pastoris. A pH-dependent electron transfer from both CDHs via their cytochrome domains to GH61-3 was observed. The different properties of CDH IIA and CDH IIB and their effect on interactions with GH61-3 are discussed in regard to the proposed in vivo function of the CDH/GH61 enzyme system in oxidative cellulose hydrolysis.
Asunto(s)
Deshidrogenasas de Carbohidratos/metabolismo , Celulosa/metabolismo , Neurospora crassa/enzimología , Neurospora crassa/metabolismo , Deshidrogenasas de Carbohidratos/química , Deshidrogenasas de Carbohidratos/genética , Estabilidad de Enzimas , Concentración de Iones de Hidrógeno , Cinética , Neurospora crassa/genética , Oxidación-Reducción , Pichia/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismoRESUMEN
Cellobiose dehydrogenase (CDH) is capable of direct electron transfer (DET) on various carbon and thiol-modified gold electrodes. As a result, these systems have been utilized as biocatalyst in biosensors and biofuel cell anodes. Class I CDHs, from basidiomycetous fungi, are highly specific to cellulose or lactose, and DET is only observed at pH values below 5.5. To extend the applicability of CDH-based electrodes, the catalytic properties and the behavior on electrode surfaces of ascomycetous class II CDHs from Chaetomium attrobrunneum, Corynascus thermophilus, Dichomera saubinetii, Hypoxylon haematostroma, Neurospora crassa, and Stachybotrys bisbyi were investigated. We found that class II CDHs have diverse properties but generally show a lower substrate specificity than class I CDHs by converting also glucose and maltose. Intramolecular electron transfer (IET) and DET at neutral and alkaline pH were observed and elucidated by steady-state kinetics, pre-steady-state kinetics, and electrochemical measurements. The CDHs ability to interact with the electron acceptor cytochrome c and to communicate with electrode surfaces through DET at various pH conditions was used to classify the investigated enzymes. In combination with stopped-flow measurements, a model for the kinetics of the pH-dependent IET is developed. The efficient glucose turnover at neutral/alkaline pH makes some of these new CDHs potential candidates for glucose biosensors and biofuel cell anodes.
Asunto(s)
Deshidrogenasas de Carbohidratos/química , Ascomicetos/enzimología , Deshidrogenasas de Carbohidratos/metabolismo , Chaetomium/enzimología , Electrodos , Transporte de Electrón , Concentración de Iones de Hidrógeno , Modelos Moleculares , Neurospora crassa/enzimología , Stachybotrys/enzimología , Xylariales/enzimologíaRESUMEN
One of the challenges in the field of biosensors and biofuel cells is to establish a highly efficient electron transfer rate between the active site of redox enzymes and electrodes to fully access the catalytic potential of the biocatalyst and achieve high current densities. We report on very efficient direct electron transfer (DET) between cellobiose dehydrogenase (CDH) from Phanerochaete sordida (PsCDH) and surface modified single walled carbon nanotubes (SWCNT). Sonicated SWCNTs were adsorbed on the top of glassy carbon electrodes and modified with aryl diazonium salts generated in situ from p-aminobenzoic acid and p-phenylenediamine, thus featuring at acidic pH (3.5 and 4.5) negative or positive surface charges. After adsorption of PsCDH, both electrode types showed excellent long-term stability and very efficient DET. The modified electrode presenting p-aminophenyl groups produced a DET current density of 500 µA cm(-2) at 200 mV vs normal hydrogen reference electrode (NHE) in a 5 mM lactose solution buffered at pH 3.5. This is the highest reported DET value so far using a CDH modified electrode and comes close to electrodes using mediated electron transfer. Moreover, the onset of the electrocatalytic current for lactose oxidation started at 70 mV vs NHE, a potential which is 50 mV lower compared to when unmodified SWCNTs were used. This effect potentially reduces the interference by oxidizable matrix components in biosensors and increases the open circuit potential in biofuel cells. The stability of the electrode was greatly increased compared with unmodified but cross-linked SWCNTs electrodes and lost only 15% of the initial current after 50 h of constant potential scanning.
Asunto(s)
Deshidrogenasas de Carbohidratos/metabolismo , Compuestos de Diazonio/metabolismo , Nanotubos de Carbono/química , Adsorción , Biocatálisis , Deshidrogenasas de Carbohidratos/química , Carbono/química , Compuestos de Diazonio/química , Electrodos , Transporte de Electrón , Modelos Moleculares , Estructura Molecular , Phanerochaete/enzimología , Propiedades de SuperficieRESUMEN
Putative cellobiose dehydrogenase (CDH) genes are frequently discovered in various fungi by genome sequencing projects. The expression of CDH, an extracellular flavocytochrome, is well studied in white rot basidiomycetes and is attributed to extracellular lignocellulose degradation. CDH has also been reported for plant-pathogenic or saprotrophic ascomycetes, but the molecular and catalytic properties of these enzymes are currently less investigated. This study links various ascomycetous cdh genes with the molecular and catalytic characteristics of the mature proteins and suggests a differentiation of ascomycete class II CDHs into two subclasses, namely, class IIA and class IIB, in addition to the recently introduced class III of hypothetical ascomycete CDHs. This new classification is based on sequence and biochemical data obtained from sequenced fungal genomes and a screening of 40 ascomycetes. Thirteen strains showed CDH activity when they were grown on cellulose-based media, and Chaetomium atrobrunneum, Corynascus thermophilus, Dichomera saubinetii, Hypoxylon haematostroma, Neurospora crassa, and Stachybotrys bisbyi were selected for detailed studies. In these strains, one or two cdh-encoding genes were found that stem either from class IIA and contain a C-terminal carbohydrate-binding module or from class IIB without such a module. In several strains, both genes were found. Regarding substrate specificity, class IIB CDHs show a less pronounced substrate specificity for cellobiose than class IIA enzymes. A pH-dependent pattern of the intramolecular electron transfer was also observed, and the CDHs were classified into three groups featuring acidic, intermediate, or alkaline pH optima. The pH optimum, however, does not correlate with the CDH subclasses and is most likely a species-dependent adaptation to different habitats.
Asunto(s)
Ascomicetos/enzimología , Deshidrogenasas de Carbohidratos/clasificación , Deshidrogenasas de Carbohidratos/metabolismo , Secuencia de Aminoácidos , Secuencia de Bases , Deshidrogenasas de Carbohidratos/genética , Deshidrogenasas de Carbohidratos/aislamiento & purificación , Celulosa/metabolismo , Medios de Cultivo/química , ADN de Hongos , Cinética , Datos de Secuencia Molecular , Análisis de Secuencia de ADN , Especificidad por SustratoRESUMEN
A third generation glucose biosensor working under physiological conditions with a linear range of 0.1-30 mM, a detection limit of 0.05 mM, and a sensitivity of 222 nA µM(-1) cm(-2) has been developed by co-adsorption of cellobiose dehydrogenase (CDH) from the ascomycete Corynascus thermophilus (CtCDH) and oxidatively shortened single-walled carbon nanotubes (SWCNTs).
Asunto(s)
Ascomicetos/enzimología , Técnicas Biosensibles/métodos , Deshidrogenasas de Carbohidratos/metabolismo , Glucosa/análisis , Nanotubos de Carbono/química , Deshidrogenasas de Carbohidratos/química , Técnicas Electroquímicas/métodos , Electrodos , Enzimas Inmovilizadas/química , Enzimas Inmovilizadas/metabolismo , Grafito/químicaRESUMEN
Cellobiose dehydrogenase catalyses the oxidation of aldoses--a simple reaction, a boring enzyme? No, neither for the envisaged bioelectrochemical applications nor mechanistically. The catalytic cycle of this flavocytochrome is complex and modulated by its flexible cytochrome domain, which acts as a built-in redox mediator. This intramolecular electron transfer is modulated by the pH, an adaptation to the environmental conditions encountered or created by the enzyme-producing fungi. The cytochrome domain forms the base from which electrons can jump to large terminal electron acceptors, such as redox proteins, and also enables by that path direct electron transfer from the catalytically active flavodehydrogenase domain to electrode surfaces. The application of electrochemical techniques to the elucidation of the molecular and catalytic properties of cellobiose dehydrogenase is discussed and compared to biochemical methods. The results lead to valuable insights into the function of this cellulose-bound enzyme, but also form the basis of exciting applications in biosensors, biofuel cells and bioelectrocatalysis.
Asunto(s)
Deshidrogenasas de Carbohidratos/metabolismo , Electroquímica/métodos , Biocatálisis , Fuentes de Energía Bioeléctrica , Técnicas Biosensibles , Deshidrogenasas de Carbohidratos/química , Celulosa/química , Celulosa/metabolismo , Oxidación-ReducciónRESUMEN
Direct and mediated electron transfer (DET and MET) between the enzyme and electrodes were compared for cellobiose dehydrogenase (CDH) from the basidiomycete Phanerochaete sordida (PsCDH). For DET, PsCDH was adsorbed at pyrolytic graphite (PG) electrodes while for MET the enzyme was covalently linked to a low potential Os redox polymer. Both types of electrodes were prepared in the presence of single walled carbon nanotubes (SWCNTs). DET requires the oxidation of the heme domain, while MET occurs partially via the heme and the flavin domain at pH 3.5. At pH 6 MET occurs solely via the flavin domain. Most probably, the interaction of the domains decreases from pH 3.5 to 6.0 due to electrostatic repulsion of deprotonated amino acid residues, covering the surfaces of both domains. MET starts at a lower potential than DET. The midpoint potentials at pH 3.5 for the flavin (40 mV) and the heme domain (170 mV) were determined with spectroelectrochemistry. The electrochemical and spectroelectrochemical measurements presented in this work are in conformity. The pH dependency of DET and MET was investigated for PsCDH. The optimum was observed between pH 4 and 4.5 pH for DET and in the range of pH 5-6 for MET. The current densities obtained by MET are 1 order of magnitude higher than by DET. During multicycle cyclic voltammetry experiments carried out at different pHs, the PsCDH modified electrode working by MET turned out to be very stable. In order to characterize a PsCDH modified anode working by MET with respect to biofuel cell applications, this electrode was combined with a Pt-black cathode as model for a membraneless biofuel cell. In comparison to DET, a 10 times higher maximum current and maximum power density in a biofuel cell application could be achieved by MET. While CDH modified electrodes working by DET are highly qualified for applications in amperometric biosensors, a much better performance as biofuel cell anodes can be obtained by MET. The use of CDH modified electrodes working by MET for biofuel cell applications results in a less positive onset of the electrocatalytic current (which may lead to an increased cell voltage), higher current and power density, and much better long-term stability over a broad range of pH.
Asunto(s)
Deshidrogenasas de Carbohidratos/química , Phanerochaete/enzimología , Fuentes de Energía Bioeléctrica , Deshidrogenasas de Carbohidratos/metabolismo , Electroquímica , Electrodos , Transporte de Electrón , Estabilidad de Enzimas , Flavinas/química , Hemo/química , Concentración de Iones de Hidrógeno , Estructura Terciaria de Proteína , Análisis EspectralRESUMEN
Cellobiose dehydrogenase (CDH), an extracellular flavocytochrome produced by several wood-degrading fungi, was detected in cultures of the selective delignifier Ceriporiopsis subvermispora when grown on a cellulose- and yeast extract-based liquid medium. CDH amounted to up to 2.5% of total extracellular protein during latter phases of the cultivation and thus suggested an important function for the fungus under the given conditions. The enzyme was purified 44-fold to apparent homogeneity. It was found to be present in two glycoforms of 98 kDa and 87 kDa with carbohydrate contents of 16 and 4%, respectively. The isoelectric point of both glycoforms is around 3.0, differing by 0.1 units, which is the most acidic value so far reported for a CDH. By using degenerated primers of known CDH sequences, one cdh gene was found in the genomic DNA, cloned, and sequenced. Alignment of the 774-amino-acid protein sequence revealed a high similarity to CDH from other white rot fungi. One notable difference was found in the longer interdomain peptide linker, which might affect the interdomain electron transfer at higher temperatures. The preferred substrate of C. subvermispora CDH is cellobiose, while glucose conversion is strongly discriminated by a 155,000-fold-lower catalytic efficiency. This is a typical feature of a basidiomycete CDH, as are the acidic pH optima for all tested electron acceptors in the range from 2.5 to 4.5.
Asunto(s)
Deshidrogenasas de Carbohidratos/genética , Deshidrogenasas de Carbohidratos/metabolismo , Coriolaceae/enzimología , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Deshidrogenasas de Carbohidratos/química , Deshidrogenasas de Carbohidratos/aislamiento & purificación , Celobiosa/metabolismo , Coriolaceae/genética , ADN de Hongos/química , ADN de Hongos/genética , Estabilidad de Enzimas , Proteínas Fúngicas/química , Proteínas Fúngicas/aislamiento & purificación , Glucosa/metabolismo , Concentración de Iones de Hidrógeno , Punto Isoeléctrico , Isoenzimas/química , Cinética , Datos de Secuencia Molecular , Peso Molecular , Filogenia , Análisis de Secuencia de ADN , Homología de Secuencia de Aminoácido , Especificidad por Sustrato , TemperaturaRESUMEN
OBJECTIVE: Electrochemical sensors for glucose monitoring employ different signal transduction strategies for electron transfer from the biorecognition element to the electrode surface. We present a biosensor that employs direct electron transfer and evaluate its response to various interfering substances known to affect glucose biosensors. METHODS: The enzyme cellobiose dehydrogenase (CDH) was adsorbed on the surface of a carbon working electrode and covalently bound by cross linking. The response of CDH-modified electrodes to glucose and possible interfering compounds was measured by flow-injection analysis, linear sweep, and chronoamperometry. RESULTS: Chronoamperometry showed initial swelling/wetting of the electrode. After stabilization, the signal was stable and a sensitivity of 0.21 µA mM-1 cm-2 was obtained. To investigate the influence of the interfering substances on the biorecognition element, the simplest possible sensor architecture was used. The biosensor showed little (<5% signal deviation) or no response to various reported electroactive or otherwise interfering substances. CONCLUSIONS: Direct electron transfer from the biorecognition element to the electrode is a new principle applied to glucose biosensors, which can be operated at a low polarization potential of -100 mV versus silver/silver chloride. The reduction of interferences by electrochemically active substances is an attractive feature of this promising technology for the development of continuous glucose biosensors.
Asunto(s)
Técnicas Biosensibles/instrumentación , Glucosa/análisis , Deshidrogenasas de Carbohidratos , Electroquímica , Electrodos , Electrones , Enzimas Inmovilizadas , Especificidad por SustratoRESUMEN
Cellobiose dehydrogenase (CDH) is an emerging enzyme in the field of bioelectrocatalysis. Due to its flexible cytochrome domain, which acts as a built-in redox mediator, CDH is capable of direct electron transfer (DET) to electrode surfaces. This rare property is employed in mediatorless "third generation" biosensors. The ability of Corynascus thermophilus CDH to oxidize glucose under physiological conditions makes it a promising candidate for miniaturized glucose biosensors or glucose powered biofuel cell anodes. We report for the first time the electrochemical application and characterization of a recombinantly produced CDH in a glucose biosensor. Recombinant CDH from C. thermophilus (rCtCDH) was expressed by the methylotrophic yeast Pichia pastoris (376 U L(-1) , 132 mg L(-1) ). A comparative characterization of rCtCDH and CtCDH shows identical pH optima, K(M) values and heme b midpoint potentials. In contrast, the specific activity of rCtCDH (2.84 U mg(-1) ) and consequently the turnover numbers were ~five-times lower than for CtCDH, which was caused by a sub-stoichiometric occupation of catalytic sites with flavin-adenin-dinukleotid (FAD). The performance of rCtCDH-modified electrodes demonstrates the suitability for electrochemical studies. This opens the possibility to engineer the substrate specificity of C. thermophilus CDH for specific carbohydrates by rational engineering or directed evolution.
Asunto(s)
Fuentes de Energía Bioeléctrica , Técnicas Biosensibles/métodos , Deshidrogenasas de Carbohidratos/biosíntesis , Glucosa/metabolismo , Proteínas Recombinantes/biosíntesis , Sordariales/enzimología , Deshidrogenasas de Carbohidratos/química , Deshidrogenasas de Carbohidratos/genética , Celobiosa/metabolismo , Electrodos , Fermentación , Glucosa/química , Cinética , Peso Molecular , Pichia/genética , Pichia/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Sordariales/genéticaRESUMEN
The electrochemical characterization of a class II cellobiose dehydrogenase (CDH), from the ascomycete fungus Neurospora crassa, adsorbed on graphite (G), was performed in regard to direct (DET) and mediated electron transfer (MET). The effects of the applied potential, mediator (1,4 benzoquinone) concentration and flow carrier pH on the amperometric response of the G/CDH modified electrodes were investigated under flow conditions. From the calibration curves, recorded at two pH values (5.2 and 7.0) for nine different sugars, the kinetic and the analytical parameters were evaluated under DET and MET operation modes. These results together with those obtained from long term operational stability measurements showed that: (i) for all nine investigated sugars the sensitivity was higher for MET than for DET and for pH 5.2 compared to pH 7.0; (ii) irrespective of DET or MET operation mode, the sensitivity of the new enzyme towards the investigated sugars decreased in the following sequence: cellobiose>lactose>(cellotriose≈cellopentaose) >>(maltotriose≈maltotetraose≈maltopentaose)>(maltose≈glucose); (iii) for all tested substrates, the apparent CDH affinity was roughly higher in DET than in MET operation mode.
Asunto(s)
Deshidrogenasas de Carbohidratos/química , Deshidrogenasas de Carbohidratos/metabolismo , Electroquímica/métodos , Grafito/química , Neurospora crassa/enzimología , Adsorción , Biocatálisis , Técnicas Biosensibles , Carbohidratos/análisis , Carbohidratos/química , Electroquímica/instrumentación , Electrodos , Transporte de Electrón , Estabilidad de Enzimas , Concentración de Iones de Hidrógeno , Propiedades de SuperficieRESUMEN
A highly efficient anode for glucose biofuel cells has been developed by a combination of pyranose dehydrogenase from Agaricus meleagris (AmPDH) and cellobiose dehydrogenase from Myriococcum thermophilum (MtCDH). These two enzymes differ in how they oxidize glucose. AmPDH oxidizes glucose at the C(2) and C(3) carbon, whereas MtCDH at the C(1) carbon. Both enzymes oxidize efficiently a number of other mono- and disaccharides. They do not react directly with oxygen and produce no H(2)O(2). Electrodes were prepared by embedding (i) only AmPDH (in order to study this enzyme separately) and (ii) a mixture of AmPDH and MtCDH in an Os redox polymer hydrogel. Single-walled carbon nanotubes (SWCNTs) were added in order to enhance the current density. The electrodes were investigated with linear sweep and cyclic voltammetry in the presence of different substrates at physiological conditions. The electrochemical measurements revealed that the product of one enzyme can serve as a substrate for the other. In addition, a kinetic pathway analysis was performed by spectrophotometric measurements leading to the conclusion that up to six electrons can be gained from one glucose molecule through a combination of AmPDH and MtCDH. Hence, the combination of redox enzymes can lead to an enzymatic biofuel cell anode with an increased coulombic efficiency far beyond the usual yields of two electrons per substrate molecule.
Asunto(s)
Fuentes de Energía Bioeléctrica , Deshidrogenasas de Carbohidratos/química , Electrodos , Glucosa/química , Oxidorreductasas/química , Diseño de Equipo , Análisis de Falla de Equipo , Oxidación-Reducción , Electricidad EstáticaRESUMEN
We report the fabrication and characterisation of a non-compartmentalised, mediator and cofactor free glucose-oxygen biofuel cell based on adsorbed enzymes exhibiting direct bioelectrocatalysis, viz. cellobiose dehydrogenase from Dichomera saubinetii and laccase from Trametes hirsuta as the anodic and cathodic bioelements, respectively, with the following characteristics: an open-circuit voltage of 0.73 V; a maximum power density of 5 microW cm(-2) at 0.5 V of the cell voltage and an estimated half-life of > 38 h in air-saturated 0.1 M citrate-phosphate buffer, pH 4.5 containing 5 mM glucose.