Electrochemical performance of Paenibacillus profundus YoMME encapsulated in alginate polymer.

Gram-positive bacterium Paenibacillus profundus YoMME, entrapped in an alginate polymer onto graphite paper, preserves its extracellular electron transfer capabilities. A current density of up to 30 mA m-2 was generated at an applied potential of -200 mV (vs. SHE). Fivefold higher initial current density values were recorded at applied potentials of +220 mV and +600 mV. The electrochemical behavior of the encapsulated bioelectrodes has been also explored by cyclic voltammetry, differential pulse voltammetry, and electrochemical impedance spectroscopy and evaluated by the parameters of the best-fitted equivalent electric circuit model. Over time the bacteria grow and divide within the alginate matrix, which affects considerably the capacitance and the charge transfer resistance of the coating. The impedance spectra follow the dynamics of the bacterial culture development within the alginate polymer and are useful for the prediction of the bioelectrode performance. A current density of 150 mA m-2 was achieved when the alginate was functionalized by a mixture of the redox dyes thiazolyl blue (MTT) formazan and phenazine methosulfate (PMS). It is supposed that the added artificial mediators facilitate the electron transfer from the bacteria to the electrode surface by forming conductive cascade conduits through the alginate matrix.

Efficient gold recovery by microbial electrochemical technologies.

The applicability of microbial electrochemical technologies for the recovery of gold was investigated. Two-chamber microbial fuel cells (MFC) with bioanodes buried in sediment were used in two operating modes. The cathodes (gold foil or graphitized paper), submerged in HAuCl4, solutions, were short-circuited with the bioanodes, and thus for the first time, the microbial electrochemical snorkel (MES) was applied for gold recovery. Operation in MFC mode, where the cathode and the anode were connected through an external resistor equal to the internal resistance of the system was also implemented. The electrochemical results along with the microscopic analyses, XPS data, and the estimated rate constants show the better performance of the MES over the MFC and predict the putative mechanism of the cathodic gold deposition. The gold removal and recovery reached ca. 95% within a day and the cathodic efficiency approached almost 100%. 7% higher gold recovery and 5% higher gold removal were achieved in the MES mode, which reveals the advantage of the operation under short-circuit conditions. The deposited on the cathode gold is in its elemental state. The similar results obtained with the two types of cathodes justify replacing the gold electrodes with much cheaper graphitized paper to reduce the cost. In addition, it has been demonstrated that gold can be also recovered by MES from the aqueous solutions of its complex Na3[Au(S2O3)2], simulating leachates from printed circuit board waste, which expands the limits of its practical application.

Draft Genome Sequence of Paenibacillus profundus YoMME, a New Exoelectrogenic Gram-Positive Bacterium.

Paenibacillus profundus YoMME was isolated from the anodic biofilm of a sediment microbial fuel cell and recognized as one of the few exoelectrogenic Gram-positive bacteria, capable of transferring electrons extracellularly. Here, we report its draft genome sequence. The genome project is deposited at DDBJ/ENA/GenBank under the accession number JAJNBZ000000000.

Gram-positive bacteria covered bioanode in a membrane-electrode assembly for use in bioelectrochemical systems.

A novel strain of Gram-positive bacteria Paenibacillus profundus YoMME was recognized by sequencing of 16S rRNA gene and after that tested for exoelectrogenicity for the first time. It was found that at an applied potential of -0.195 V (vs. SHE) the bacteria are capable of generating electricity and forming electroactive biofilms for 3-4 days. A tendency for the decrease in double-layer capacitance and the increase in the charge transfer resistance during the maturation of the biofilm was established. The formed bioanodes were used as a part of a membrane-electrode assembly (MEA) together with a selected cathode (E-Tek) and a separator (Zirfon). The applicability of MEA with the bioanode was tested by operating a newly designed bioelectrochemical system in a microbial fuel cell (MFC) or microbial electrolysis cell (MEC) mode. A current density of 200 mA m-2 was generated by the MFC after the improvement of the cathodic reaction through facilitated air access. The Coulombic efficiency in different MFC runs ranged from 5.2 to 7.4%. It was also determined that 0.65 V applied cell voltage is appropriate for the operation of the cell in the electrolysis mode, during which a current density of 2-3 Am-2 was reached. This, along with the evolved gas on the cathode, shows that as an anodic biocatalyst P. profundus YoMME assists the electrolysis processes at a significantly lower voltage than the theoretical one (1.23 V) for water decomposition. The hydrogen production rate varied between 0.5 and 0.7 m3/m3d and the cathodic hydrogen recovery ranged from 49.5 to 61.5 %. The estimated energy efficiency based on the electricity input exceeds 100 %, which indicates that additional energy is being gained from the biotic oxidation of the available organics.

Electroactivity of the Gram-positive bacterium Paenibacillus dendritiformis MA-72.

Whilst most of the microorganisms recognized as exoelectrogens are Gram-negative bacteria, the electrogenicity of Gram-positive bacteria has not been sufficiently explored. In this study, the putative electroactivity of the Gram-positive Paenibacillus dendritiformis MA-72 strain, isolated from the anodic biofilm of long-term operated Sediment Microbial Fuel Cell (SMFC), has been investigated. SEM observations show that under polarization conditions P. dendritiformis forms a dense biofilm on carbon felt electrodes. A current density, reaching 5 mA m-2, has been obtained at a prolonged applied potential of -0.195 V (vs. SHE), which represents 35% of the value achieved with the SMFC. The voltammetric studies confirm that the observed Faradaic current is associated with the electrochemical activity of the bacterial biofilm and not with a soluble redox mediator. The results suggest that a direct electron transfer takes place through the conductive extracellular polymer matrix via pili/nanowires and multiple cytochromes. All these findings demonstrate for the first time that the Gram-positive Paenibacillus dendritiformis MA-72 is a new exoelectrogenic bacterial strain.

Electrodeposited styrylquinolinium dye as molecular electrocatalyst for coupled redox reactions.

Modification of carbonaceous materials with different conductive coatings is a successful approach to enhance their electrocatalytic activity and thus to increase the electrical outputs when used as electrodes in biofuel cells. In this study, a methodology for electrodeposition of styrylquinolinium dye on carbon felt was developed. The produced dye electrodeposits were characterized by means of AFM, ESI-MS/MS and NMR spectroscopy. The obtained data reveal that the dye forms overlaid layers consisting of monomer molecules most likely with an antiparallel orientation. The UV-VIS spectroscopy, CV and EIS analyses show that the dye molecules preserve their redox activity within the coating and a charge transfer between NADH/NAD+ and electrodeposit is possible as a coupled redox reaction. The fabricated nano-modified electrodes were also tested as anodes in batch-mode operating yeast-based biofuel cell. The results indicate that the electrodeposited dye acts as an immobilized exogenous mediator, contributing to enhanced extracellular electron transfer.

Light-dependent processes on the cathode enhance the electrical outputs of sediment microbial fuel cells.

In this study, we explored in details the influence of the light irradiation on the SMFCs electrical outputs. The experiments at both natural and artificial illumination firmly show that during the photoperiods the current grows up. The intensity of the current increase depends on the duration of the photoperiod as well as on the wavelength of the monochromatic light source applied. The highest influence of the light irradiation has been obtained at wavelengths, corresponding to the absorption peaks of essential pigments in the light-harvesting system of oxygenic photosynthesizing microorganisms. The decreased values as well as the discontinued fluctuations of the current as a result of suppressed illumination or substitution of the biocathode with a new one suggest that photosynthesizing microorganisms, co-existing in the cathodic biofilm consortium, contribute to the overall SMFC performance. The microscopic observations confirm the existence of chlorophyll-containing microorganisms on the cathode surface. Though the performed metagenomics DNA analysis has not certified a dominance of photosynthesizing microorganisms, all other results support the hypothesis that the current enhance during the photoperiods is due to the in situ bio-oxygen production on the cathode surface, thus lowering the mass transport limitations for the oxygen reduction reaction.

The glyoxylate pathway contributes to enhanced extracellular electron transfer in yeast-based biofuel cell.

This study provides a new insight into our understanding of yeast response to starvation conditions (sole acetate as carbon source) and applied polarization and offers important information about the role of the glyoxylate cycle in the carbohydrate synthesis and extracellular charge transfer processes in biofuel cells. The biosynthetic capabilities of yeast C. melibiosica 2491 and the up/down-regulation of the glyoxylate cycle are evaluated by modifying the cellular metabolism by feedback inhibition or carbohydrate presence and establishing the malate dehydrogenase activity and carbohydrate content together with the electric charge passed through bioelectrochemical system. 10mM malate leads to a decrease of the produced quantity of electricity with ca. 55%. At the same time, 24-times lower intracellular malate dehydrogenase activity is established. At polarization conditions the glyoxylate pathway is up-regulated and huge amount of malate is intra-converted into oxaloacetate. The yeasts are able to synthesize carbohydrates from acetate and a part of them is used for the electricity generation. It is recognized that the enhanced charge transfer in acetate fed yeast-based biofuel cell is implemented by secreted endogenous mediator and changes in the cellular surface redox activity depending on the addition of carbohydrate in the medium.

Mechanisms of electron transfer between a styrylquinolinium dye and yeast in biofuel cell.

In the present study, the influence of the recently synthesized styrylquinolinium dye 4-{(E)-2-[4-(dimethylamino)naphthalen-1-yl]ethenyl}-1-methylquinolinium iodide (DANSQI) on the intracellular processes as well as the electrical outputs of Candida melibiosica 2491 yeast-based biofuel cell was investigated. The addition of nanomolar quantities of DANSQI to the yeast suspension results in an increase of the current outputs right after the startup of the biofuel cells, associated with an electrooxidation of the dye on the anode. After that, the formed cation radical of the dye penetrates the yeast cells, provoking a set of intracellular changes. Studies of the subcellular anolyte fractions show that 1µM dye increased the peroxisomal catalase activity 30-times (1.15±0.06Unit/mg protein) and over twice the mitochondrial cytochrome c oxidase activity (92±5Unit/mg protein). The results obtained by electrochemical and spectrophotometric analyses let to the supposition that the dye acts as subcellular shuttle, on account of its specific intramolecular charge transfer properties. The transition between its benzoid, quinolyl radical and ion forms and their putative role for the extracellular and intracellular charge transfer mechanisms are discussed.

Chemometrical assessment of the electrical parameters obtained by long-term operating freshwater sediment microbial fuel cells.

The electrical parameters of nine freshwater sediment microbial fuel cells (SMFCs) were monitored for a period of over 20 months. The developed SMFCs, divided into three groups, were started up and continuously operated under different constant loads (100, 510 and 1100 Ω) for 2.5 months. At this stage of the experiment, the highest power density values, reaching 1.2 ± 0.2 mW/m(2), were achieved by the SMFCs loaded with 510 Ω. The maximum power obtained at periodical polarization during the rest period, however, ranged between 26.2 ± 2.8 and 35.3 ± 2.8 mW/m(2), strongly depending on the internal cell resistance. The statistical evaluation of data derived from the polarization curves shows that after 300 days of operation all examined SMFCs reached a steady-state and the system might be assumed as homoscedastic. The estimated values of standard and expanded uncertainties of the electric parameters indicate a high repeatability and reproducibility of the SMFCs' performance. Results obtained in subsequent discharge-recovery cycles reveal the opportunity for practical application of studied SMFCs as autonomous power sources.

Extracellular electron transfer in yeast-based biofuel cells: A review.

This paper reviews the state-of-the art of the yeast-based biofuel cell research and development. The established extracellular electron transfer (EET) mechanisms in the presence and absence of exogenous mediators are summarized and discussed. The approaches applied for improvement of mediator-less yeast-based biofuel cells performance are also presented. The overview of the literature shows that biofuel cells utilizing yeasts as biocatalysts generate power density in the range of 20 to 2440 mW/m(2), which values are comparable with the power achieved when bacteria are used instead. The electrons' origin and the contribution of the glycolysis, fermentation, aerobic respiration, and phosphorylation to the EET are commented. The reported enhanced current generation in aerobic conditions presumes reconsideration of some basic MFC principles. The challenges towards the practical application of the yeast-based biofuel cells are outlined.

Enhanced metabolic and redox activity of vascular aquatic plant Lemna valdiviana under polarization in Direct Photosynthetic Plant Fuel Cell.

In this study, duckweed species Lemna valdiviana was investigated as a photoautotrophycally grown biocatalyst in recently developed Direct Photosynthetic Plant Fuel Cell. Stable current outputs, reaching maximum of 226±11 mА/m(2), were achieved during the operating period. The electricity production is associated with electrons generated through the light-dependent reactions in the chloroplasts as well as the respiratory processes in the mitochondria and transferred to the anode via endogenous electron shuttle, synthesized by the plants as a specific response to the polarization. In parallel, a considerable increase in the content of proteins (47%) and reserve carbohydrates (44%) of duckweeds grown under polarization conditions was established by means of biochemical analyses. This, combined with the electricity generation, makes the technology a feasible approach for the duckweed farming.

Mitochondrial origin of extracelullar transferred electrons in yeast-based biofuel cells.

The influence of mitochondrial electron transport chain inhibitors on the electricity outputs of Candida melibiosica yeast-based biofuel cell was investigated. The addition of 30 µM rotenone or antimycin A to the yeast suspension results in a decrease in the current generation, corresponding to 25.7±1.3%, respectively 38.8±1.9% reduction in the electric charge passed through the bioelectrochemical system. The latter percentage coincides with the share of aerobic respiration in the yeast catabolic processes, determined by the decrease of the ethanol production during cultivation in the presence of oxygen compared with that obtained under strict anaerobic conditions. It was established that the presence of both inhibitors leads to almost complete mitochondrial dysfunction, expressed by inactivation of cytochrome c oxidase and NADH:ubiquinone oxidoreductase as well as reduced electrochemical activity of isolated yeast mitochondria. It was also found that methylene blue partially neutralized the rotenone poisoning, probably serving as alternative intracellular electron shuttle for by-passing the complex I blockage. Based on the obtained results, we suppose that electrons generated through the aerobic respiration processes in the mitochondria participate in the extracellular electron transfer from the yeast cells to the biofuel cell anode, which contributes to higher current outputs at aerobic conditions.

Application of solid-phase extraction for the concentration of chromophores, fluorophores, and photosensitizers from lens protein digests.

Solid-phase extraction was applied for the separation of protein digests obtained from aged human lenses, cataractous human lenses, calf lens proteins in vitro glycated with dehydroascorbic acid and native calf lens proteins. Four fractions were collected after stepwise elution with different solvents. The first fraction contained about 80% of the digested material possessing free amino groups. At the same time, the third and the fourth fractions were enriched in chromophores, fluorophores, and photosensitizing structures that originate mainly from advanced protein glycation. The comparison between the total digest and the fourth fraction based on their UV absorption at 330 nm, intensity of fluorescence (excitation/emission 350/450 nm), and production of singlet oxygen upon UVA irradiation argues that the solid-phase extraction was capable of concentrating the advanced glycation end-products about a hundredfold. Thus, this technique is a useful step for separation and concentration of fluorophores, chromophores, and photosensitizers from aged and glycated lens protein digests.

Stable current outputs and phytate degradation by yeast-based biofuel cell.

In this paper, we report for the first time that Candida melibiosica 2491 yeast strain expresses enhanced phytase activity when used as a biocatalyst in biofuel cells. The polarization also results in an increase of the yeast biomass. Higher steady-state electrical outputs, assigned to earlier production of an endogenous mediator, were achieved at continuous polarization under constant load. The obtained results prove that the C. melibiosica yeast-based biofuel cell could be used for simultaneous electricity generation and phytate bioremediation. In addition, the higher phytase activity obtained by interruptive polarization suggests a new method for increasing the phytase yield from microorganisms.

Enhanced phytate dephosphorylation by using Candida melibiosica yeast-based biofuel cell.

We report for the first time that Candida melibiosica expresses enhanced phytase activity when grown under biofuel cell polarization in a nutrient-poor medium, containing only fructose as a carbohydrate source. Phytase activity during the cultivation under polarization reached up to 25 U per g dry biomass, exceeding with 20 ± 3 % those of the control. A participation of the enzyme in the adaptation processes to the stress conditions is proposed. In addition, steady-state electrical outputs were achieved during biofuel cell operation at continuous polarization under constant load. The obtained results show that C. melibiosica yeast-based biofuel cell could be used for simultaneous electricity generation and phytate bioremediation.

Conversion of solar energy into electricity by using duckweed in Direct Photosynthetic Plant Fuel Cell.

In the present study we demonstrate for the first time the possibility for conversion of solar energy into electricity on the principles of Direct Photosynthetic Plant Fuel Cell (DPPFC) technology by using aquatic higher plants. Lemna minuta duckweed was grown autotrophically in specially constructed fuel cells under sunlight irradiation and laboratory lighting. Current and power density up to 1.62±0.10 A.m(-2) and 380±19 mW.m(-2), respectively, were achieved under sunlight conditions. The influence of the temperature, light intensity and day/night sequencing on the current generation was investigated. The importance of the light intensity was demonstrated by the higher values of generated current (at permanently connected resistance) during daytime than those through the nights, indicating the participation of light-dependent photosynthetic processes. The obtained DPPFC outputs in the night show the contribution of light-independent reactions (respiration). The electron transfer in the examined DPPFCs is associated with a production of endogenous mediator, secreted by the duckweed. The plants' adaptive response to the applied polarization is also connected with an enhanced metabolism resulting in an increase of the protein and carbohydrate intracellular content. Further investigations aiming at improvement of the DPPFC outputs and elucidation of the electron transfer mechanism are required for practical application.

Influence of artificial mediators on yeast-based fuel cell performance.

Soluble artificial mediators are often applied to enhance the electron transfer from living cells to an anode in microbial fuel cells. Recently, we have demonstrated that the Candida melibiosica 2491 yeast strain possesses electrogenic properties and can be used as a biocatalyst in yeast-based fuel cells even in the absence of artificial mediators. To enhance the generated electrical power, the potential application of several organic compounds as mediators in a C. melibiosica-based fuel cell was examined in this study. The choice of compounds was based upon observed cyclic voltammetry reversible electrochemical behavior at potentials appropriate for mediated electron transfer. Among the studied mediators, methylene blue, methyl orange, methyl red and neutral red significantly increased the current and power outputs in comparison with those obtained with a mediatorless yeast-based fuel cell.

Potential application of Candida melibiosica in biofuel cells.

Various prokaryote species have been widely studied for microbial fuel cell (MFC) application. However, the information about yeast utilization into biofuel cells is still scanty. The aim of this investigation is to verify if Candida melibiosica 2491, a yeast strain, possessing high phytase activity, could be applied as a biocatalyst in a yeast biofuel cell. The microbiological requirements were coupled with the electrochemical ones tracing main biochemical pathway metabolites such as different carbohydrate and inorganic phosphates and their assimilation with time. The obtained results show that from the three carbohydrates investigated - glucose, fructose and sucrose, fructose is the most suitable for the yeast cultivation. The presence of yeast extract and peptone improves the performance into the biofuel cell. The relationship between the yeast cell amount and the biofuel cell characteristics was determined. Analyses showed that electricity was generated by the yeast culture even in the absence of an artificial mediator. The addition of methylene blue at concentrations higher than 0.1 mM improves the current and power density output. The obtained experimental results proved that C. melibiosica 2491 belongs to the electrogenic strains.