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Sample records for nife hydrogenase genes

  1. Deletion of a gene cluster for [Ni-Fe] hydrogenase maturation in the anaerobic hyperthermophilic bacterium Caldicellulosiruptor bescii identifies its role in hydrogen metabolism.

    Science.gov (United States)

    Cha, Minseok; Chung, Daehwan; Westpheling, Janet

    2016-02-01

    The anaerobic, hyperthermophlic, cellulolytic bacterium Caldicellulosiruptor bescii grows optimally at ∼80 °C and effectively degrades plant biomass without conventional pretreatment. It utilizes a variety of carbohydrate carbon sources, including both C5 and C6 sugars, released from plant biomass and produces lactate, acetate, CO2, and H2 as primary fermentation products. The C. bescii genome encodes two hydrogenases, a bifurcating [Fe-Fe] hydrogenase and a [Ni-Fe] hydrogenase. The [Ni-Fe] hydrogenase is the most widely distributed in nature and is predicted to catalyze hydrogen production and to pump protons across the cellular membrane creating proton motive force. Hydrogenases are the key enzymes in hydrogen metabolism and their crystal structure reveals complexity in the organization of their prosthetic groups suggesting extensive maturation of the primary protein. Here, we report the deletion of a cluster of genes, hypABFCDE, required for maturation of the [Ni-Fe] hydrogenase. These proteins are specific for the hydrogenases they modify and are required for hydrogenase activity. The deletion strain grew more slowly than the wild type or the parent strain and produced slightly less hydrogen overall, but more hydrogen per mole of cellobiose. Acetate yield per mole of cellobiose was increased ∼67 % and ethanol yield per mole of cellobiose was decreased ∼39 %. These data suggest that the primary role of the [Ni-Fe] hydrogenase is to generate a proton gradient in the membrane driving ATP synthesis and is not the primary enzyme for hydrogen catalysis. In its absence, ATP is generated from increased acetate production resulting in more hydrogen produced per mole of cellobiose.

  2. Structure and function of [NiFe] hydrogenases.

    Science.gov (United States)

    Ogata, Hideaki; Lubitz, Wolfgang; Higuchi, Yoshiki

    2016-11-01

    Hydrogenases catalyze the reversible conversion of molecular hydrogen to protons and electrons via a heterolytic splitting mechanism. The active sites of [NiFe] hydrogenases comprise a dinuclear Ni-Fe center carrying CO and CN(-) ligands. The catalytic activity of the standard (O2-sensitive) [NiFe] hydrogenases vanishes under aerobic conditions. The O2-tolerant [NiFe] hydrogenases can sustain H2 oxidation activity under atmospheric conditions. These hydrogenases have very similar active site structures that change the ligand sphere during the activation/catalytic process. An important structural difference between these hydrogenases has been found for the proximal iron-sulphur cluster located in the vicinity of the active site. This unprecedented [4Fe-3S]-6Cys cluster can supply two electrons, which lead to rapid recovery of the O2 inactivation, to the [NiFe] active site. © The Authors 2016. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.

  3. Structural and functional models for [NiFe] hydrogenase

    NARCIS (Netherlands)

    Angamuthu, Raja

    2009-01-01

    The main goal of the research presented in this thesis is the synthesis of suitable structural and functional models for the enzyme [NiFe] hydrogenase, which can reduce protons into dihydrogen. A brief survey of the roles of all the known nickel containing enzymes in biological systems with a focus

  4. Three different [NiFe] hydrogenases confer metabolic flexibility in the obligate aerobe Mycobacterium smegmatis.

    Science.gov (United States)

    Berney, Michael; Greening, Chris; Hards, Kiel; Collins, Desmond; Cook, Gregory M

    2014-01-01

    Mycobacterium smegmatis is an obligate aerobe that harbours three predicted [NiFe] hydrogenases, Hyd1 (MSMEG_2262–2263), Hyd2 (MSMEG_2720-2719) and Hyd3 (MSMEG_3931-3928). We show here that these three enzymes differ in their phylogeny, regulation and catalytic activity. Phylogenetic analysis revealed that Hyd1 groups with hydrogenases that oxidize H2 produced by metabolic processes, and Hyd2 is homologous to a novel group of putative high-affinity hydrogenases. Hyd1 and Hyd2 respond to carbon and oxygen limitation, and, in the case of Hyd1, hydrogen supplementation. Hydrogen consumption measurements confirmed that both enzymes can oxidize hydrogen. In contrast, the phylogenetic analysis and activity measurements of Hyd3 are consistent with the enzyme evolving hydrogen. Hyd3 is controlled by DosR, a regulator that responds to hypoxic conditions. The strict dependence of hydrogen oxidation of Hyd1 and Hyd2 on oxygen suggests that the enzymes are oxygen tolerant and linked to the respiratory chain. This unique combination of hydrogenases allows M. smegmatis to oxidize hydrogen at high (Hyd1) and potentially tropospheric (Hyd2) concentrations, as well as recycle reduced equivalents by evolving hydrogen (Hyd3). The distribution of these hydrogenases throughout numerous soil and marine species of actinomycetes suggests that oxic hydrogen metabolism provides metabolic flexibility in environments with changing nutrient fluxes.

  5. Purification, crystallization and preliminary X-ray analysis of the membrane-bound [NiFe] hydrogenase from Allochromatium vinosum

    Energy Technology Data Exchange (ETDEWEB)

    Kellers, Petra; Ogata, Hideaki; Lubitz, Wolfgang, E-mail: lubitz@mpi-muelheim.mpg.de [Max-Planck-Institut für Bioanorganische Chemie, Stiftstrasse 34-36, D-45470 Mülheim an der Ruhr (Germany)

    2008-08-01

    This article describes the first successful crystallization of a membrane-bound [NiFe] hydrogenase isolated from a photosynthetic organism (A. vinosum). The crystals obtained produced diffraction patterns up to 2.5 Å resolution. The membrane-bound [NiFe] hydrogenase is a unique metalloprotein that is able to catalyze the reversible oxidation of hydrogen to protons and electrons during a complex reaction cycle. The [NiFe] hydrogenase was isolated from the photosynthetic purple sulfur bacterium Allochromatium vinosum and its crystallization and preliminary X-ray analysis are reported. It was crystallized by the hanging-drop vapour-diffusion method using sodium citrate and imidazole as crystallization agents. The crystals belong to space group P2{sub 1}2{sub 1}2, with unit-cell parameters a = 205.00, b = 217.42, c = 120.44 Å. X-ray diffraction data have been collected to 2.5 Å resolution.

  6. Modelling NiFe hydrogenases: nickel-based electrocatalysts for hydrogen production

    Energy Technology Data Exchange (ETDEWEB)

    Canaguier, S.; Artero, V.; Fontecave, M. [CEA, DSV, iRTSV, Lab Chim Biol Metaux, CEA-CNRS-Univ Grenoble 1, UMR 5249, F-38054 Grenoble 9 (France)

    2008-07-01

    NiFe hydrogenases are unique enzymes that catalyze the H{sup +}/H{sub 2} interconversion with remarkable efficiency. The determination of the tridimensional structure of their active site (a sulfur-rich dinuclear nickel-iron cluster with diatomic cyanide and carbonyl ligands) has stimulated the synthesis of a variety of nickel-based complexes as potential electrocatalysts for hydrogen production. These catalysts may provide an adequate alternative to platinum. This paper gives an historical perspective of this biomimetic structural approach and then focusses on recently reported bio-inspired functional mimics displaying electrocatalytic activity for hydrogen production. (authors)

  7. Isotopic fractionation associated with [NiFe]- and [FeFe]-hydrogenases

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    Yang, Hui; Gandhi, Hasand; Cornish, Adam J.; Moran, James J.; Kreuzer, Helen W.; Ostrom, Nathaniel; Hegg, Eric L.

    2016-01-30

    Hydrogenases catalyze the reversible formation of H2 from electrons and protons with high efficiency. Understanding the relationships between H2 production, H2 uptake, and H2-H2O exchange can provide insight into the metabolism of microbial communities in which H2 is an essential component in energy cycling. In this manuscript, we used stable H isotopes (1H and 2H) to probe the isotope effects associated with three [FeFe]-hydrogenases and three [NiFe]-hydrogenases. All six hydrogenases displayed fractionation factors for H2 formation that were significantly less than 1, producing H2 that was severely depleted in 2H relative to the substrate, water. Consistent with differences in their active site structure, the fractionation factors for each class appear to cluster, with the three [NiFe]-hydrogenases (α = 0.27-0.40) generally having smaller values than the three [FeFe]-hydrogenases (α = 0.41-0.55). We also obtained isotopic fractionation factors associated with H2 uptake and H2-H2O exchange under conditions similar to those utilized for H2 production, providing us with a more complete picture of the three reactions catalyzed by hydrogenases. The fractionation factors determined in our studies can be used as signatures for different hydrogenases to probe their activity under different growth conditions and to ascertain which hydrogenases are most responsible for H2 production and/or uptake in complex microbial communities.

  8. Modulation of Active Site Electronic Structure by the Protein Matrix to Control [NiFe] Hydrogenase Reactivity

    Energy Technology Data Exchange (ETDEWEB)

    Smith, Dayle MA; Raugei, Simone; Squier, Thomas C.

    2014-09-30

    Control of the reactivity of the nickel center of the [NiFe] hydrogenase and other metalloproteins commonly involves outer coordination sphere ligands that act to modify the geometry and physical properties of the active site metal centers. We carried out a combined set of classical molecular dynamics and quantum/classical mechanics calculations to provide quantitative estimates of how dynamic fluctuations of the active site within the protein matrix modulate the electronic structure at the catalytic center. Specifically we focused on the dynamics of the inner and outer coordination spheres of the cysteinate-bound Ni–Fe cluster in the catalytically active Ni-C state. There are correlated movements of the cysteinate ligands and the surrounding hydrogen-bonding network, which modulate the electron affinity at the active site and the proton affinity of a terminal cysteinate. On the basis of these findings, we hypothesize a coupling between protein dynamics and electron and proton transfer reactions critical to dihydrogen production.

  9. In search of metal hydrides: an X-ray absorption and emission study of [NiFe] hydrogenase model complexes.

    Science.gov (United States)

    Hugenbruch, Stefan; Shafaat, Hannah S; Krämer, Tobias; Delgado-Jaime, Mario Ulises; Weber, Katharina; Neese, Frank; Lubitz, Wolfgang; DeBeer, Serena

    2016-04-28

    Metal hydrides are invoked as important intermediates in both chemical and biological H2 production. In the [NiFe] hydrogenase enzymes, pulsed EPR and high-resolution crystallography have argued that the hydride interacts primarily at the Ni site. In contrast, in [NiFe] hydrogenase model complexes, it is observed that the bridging hydride interacts primarily with the Fe. Herein, we utilize a combination of Ni and Fe X-ray absorption (XAS) and emission (XES) spectroscopies to examine the contribution of the bridging hydride to the observed spectral features in [(dppe)Ni(μ-pdt)(μ-H)Fe(CO)3](+). The corresponding data on (dppe)Ni(μ-pdt)Fe(CO)3 are used as a reference for the changes that occur in the absence of a hydride bridge. For further interpretation of the observed spectral features, all experimental spectra were calculated using a density functional theory (DFT) approach, with excellent agreement between theory and experiment. It is found that the iron valence-to-core (VtC) XES spectra reveal clear signatures for the presence of a Fe-H interaction in the hydride bridged model complex. In contrast, the Ni VtC XES spectrum largely reflects changes in the local Ni geometry and shows little contribution from a Ni-H interaction. A stepwise theoretical analysis of the hydride contribution and the Ni site symmetry provides insights into the factors, which govern the different metal-hydride interactions in both the model complexes and the enzyme. Furthermore, these results establish the utility of two-color XES to reveal important insights into the electronic structure of various metal-hydride species.

  10. Proton Inventory and Dynamics in the Nia-S to Nia-C Transition of a [NiFe] Hydrogenase.

    Science.gov (United States)

    Greene, Brandon L; Wu, Chang-Hao; Vansuch, Gregory E; Adams, Michael W W; Dyer, R Brian

    2016-03-29

    Hydrogenases (H2ases) represent one of the most striking examples of biological proton-coupled electron transfer (PCET) chemistry, functioning in facile proton reduction and H2 oxidation involving long-range proton and electron transport. Spectroscopic and electrochemical studies of the [NiFe] H2ases have identified several catalytic intermediates, but the details of their interconversion are still a matter of debate. Here we use steady state and time-resolved infrared spectroscopy, sensitive to the CO ligand of the active site iron, as a probe of the proton inventory as well as electron and proton transfer dynamics in the soluble hydrogenase I from Pyrococcus furiosus. Subtle shifts in infrared signatures associated with the Nia-C and Nia-S states as a function of pH revealed an acid-base equilibrium associated with an ionizable amino acid near the active site. Protonation of this residue was found to correlate with the photoproduct distribution that results from hydride photolysis of the Nia-C state, in which one of the two photoproduct states becomes inaccessible at low pH. Additionally, the ability to generate Nia-S via PCET from Nia-C was weakened at low pH, suggesting prior protonation of the proton acceptor. Kinetic and thermodynamic analysis of electron and proton transfer with respect to the various proton inventories was utilized to develop a chemical model for reversible hydride oxidation involving two intermediates differing in their hydrogen bonding character.

  11. Reduction of unusual iron-sulfur clusters in the H2-sensing regulatory Ni-Fe hydrogenase from Ralstonia eutropha H16.

    Science.gov (United States)

    Buhrke, Thorsten; Löscher, Simone; Lenz, Oliver; Schlodder, Eberhard; Zebger, Ingo; Andersen, Lars K; Hildebrandt, Peter; Meyer-Klaucke, Wolfram; Dau, Holger; Friedrich, Bärbel; Haumann, Michael

    2005-05-20

    The regulatory Ni-Fe hydrogenase (RH) from Ralstonia eutropha functions as a hydrogen sensor. The RH consists of the large subunit HoxC housing the Ni-Fe active site and the small subunit HoxB containing Fe-S clusters. The heterolytic cleavage of H(2) at the Ni-Fe active site leads to the EPR-detectable Ni-C state of the protein. For the first time, the simultaneous but EPR-invisible reduction of Fe-S clusters during Ni-C state formation was demonstrated by changes in the UV-visible absorption spectrum as well as by shifts of the iron K-edge from x-ray absorption spectroscopy in the wild-type double dimeric RH(WT) [HoxBC](2) and in a monodimeric derivative designated RH(stop) lacking the C-terminal 55 amino acids of HoxB. According to the analysis of iron EXAFS spectra, the Fe-S clusters of HoxB pronouncedly differ from the three Fe-S clusters in the small subunits of crystallized standard Ni-Fe hydrogenases. Each HoxBC unit of RH(WT) seems to harbor two [2Fe-2S] clusters in addition to a 4Fe species, which may be a [4Fe-3S-3O] cluster. The additional 4Fe-cluster was absent in RH(stop). Reduction of Fe-S clusters in the hydrogen sensor RH may be a first step in the signal transduction chain, which involves complex formation between [HoxBC](2) and tetrameric HoxJ protein, leading to the expression of the energy converting Ni-Fe hydrogenases in R. eutropha.

  12. H₂-dependent azoreduction by Shewanella oneidensis MR-1: involvement of secreted flavins and both [Ni-Fe] and [Fe-Fe] hydrogenases.

    Science.gov (United States)

    Le Laz, Sébastien; Kpebe, Arlette; Lorquin, Jean; Brugna, Myriam; Rousset, Marc

    2014-03-01

    In this paper, the hydrogen (H2)-dependent discoloration of azo dye amaranth by Shewanella oneidensis MR-1 was investigated. Experiments with hydrogenase-deficient strains demonstrated that periplasmic [Ni-Fe] hydrogenase (HyaB) and periplasmic [Fe-Fe] hydrogenase (HydA) are both respiratory hydrogenases of dissimilatory azoreduction in S. oneidensis MR-1. These findings suggest that HyaB and HydA can function as uptake hydrogenases that couple the oxidation of H2 to the reduction of amaranth to sustain cellular growth. This constitutes to our knowledge the first report of the involvement of [Fe-Fe] hydrogenase in a bacterial azoreduction process. Assays with respiratory inhibitors indicated that a menaquinone pool and different cytochromes were involved in the azoreduction process. High-performance liquid chromatography analysis revealed that flavin mononucleotide and riboflavin were secreted in culture supernatant by S. oneidensis MR-1 under H2-dependent conditions with concentration of 1.4 and 2.4 μmol g protein(-1), respectively. These endogenous flavins were shown to significantly accelerate the reduction of amaranth at micromolar concentrations acting as electron shuttles between the cell surface and the extracellular azo dye. This work may facilitate a better understanding of the mechanisms of azoreduction by S. oneidensis MR-1 and may have practical applications for microbiological treatments of dye-polluted industrial effluents.

  13. [NiFe] hydrogenase structural and functional models: new bio-inspired catalysts for hydrogen evolution; Modeles structuraux et fonctionnels du site actif des hydrogenases [NiFe]: de nouveaux catalyseurs bio-inspires pour la production d'hydrogene

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    Oudart, Y

    2006-09-15

    Hydrogenase enzymes reversibly catalyze the oxidation and production of hydrogen in a range close to the thermodynamic potential. The [NiFe] hydrogenase active site contains an iron-cyano-carbonyl moiety linked to a nickel atom which is in an all sulphur environment. Both the active site originality and the potential development of an hydrogen economy make the synthesis of functional and structural models worthy. To take up this challenge, we have synthesised mononuclear ruthenium models and more importantly, nickel-ruthenium complexes, mimicking some structural features of the [NiFe] hydrogenase active site. Ruthenium is indeed isoelectronic to iron and some of its complexes are well-known to bear hydrides. The compounds described in this study have been well characterised and their activity in proton reduction has been successfully tested. Most of them are able to catalyze this reaction though their electrocatalytic potentials remain much more negative compared to which of platinum. The studied parameters point out the importance of the complexes electron richness, especially of the nickel environment. Furthermore, the proton reduction activity is stable for several hours at good rates. The ruthenium environment seems important for this stability. Altogether, these compounds represent the very first catalytically active [NiFe] hydrogenase models. Important additional results of this study are the synergetic behaviour of the two metals in protons reduction and the evidence of a protonation step as the limiting step of the catalytic cycle. We have also shown that a basic site close to ruthenium improves the electrocatalytic potential of the complexes. (author)

  14. Photoactivation of the Ni-SIr state to the Ni-SIa state in [NiFe] hydrogenase: FT-IR study on the light reactivity of the ready Ni-SIr state and as-isolated enzyme revisited.

    Science.gov (United States)

    Tai, Hulin; Xu, Liyang; Inoue, Seiya; Nishikawa, Koji; Higuchi, Yoshiki; Hirota, Shun

    2016-08-10

    The Ni-SIr state of [NiFe] hydrogenase from Desulfovibrio vulgaris Miyazaki F was photoactivated to its Ni-SIa state by Ar(+) laser irradiation at 514.5 nm, whereas the Ni-SL state was light induced from a newly identified state, which was less active than any other identified state and existed in the "as-isolated" enzyme.

  15. Structural and gene expression analyses of uptake hydrogenases and other proteins involved in nitrogenase protection in Frankia

    Indian Academy of Sciences (India)

    K H Richau; R L Kudahettige; P Pujic; N P Kudahettige; A Sellstedt

    2013-11-01

    The actinorhizal bacterium Frankia expresses nitrogenase and can therefore convert molecular nitrogen into ammonia and the by-product hydrogen. However, nitrogenase is inhibited by oxygen. Consequently, Frankia and its actinorhizal hosts have developed various mechanisms for excluding oxygen from their nitrogen-containing compartments. These include the expression of oxygen-scavenging uptake hydrogenases, the formation of hopanoid-rich vesicles, enclosed by multi-layered hopanoid structures, the lignification of hyphal cell walls, and the production of haemoglobins in the symbiotic nodule. In this work, we analysed the expression and structure of the so-called uptake hydrogenase (Hup), which catalyses the in vivo dissociation of hydrogen to recycle the energy locked up in this ‘waste’ product. Two uptake hydrogenase syntons have been identified in Frankia: synton 1 is expressed under free-living conditions while synton 2 is expressed during symbiosis. We used qPCR to determine synton 1 hup gene expression in two Frankia strains under aerobic and anaerobic conditions. We also predicted the 3D structures of the Hup protein subunits based on multiple sequence alignments and remote homology modelling. Finally, we performed BLAST searches of genome and protein databases to identify genes that may contribute to the protection of nitrogenase against oxygen in the two Frankia strains. Our results show that in Frankia strain ACN14a, the expression patterns of the large (HupL1) and small (HupS1) uptake hydrogenase subunits depend on the abundance of oxygen in the external environment. Structural models of the membrane-bound hydrogenase subunits of ACN14a showed that both subunits resemble the structures of known [NiFe] hydrogenases (Volbeda et al. 1995), but contain fewer cysteine residues than the uptake hydrogenase of the Frankia DC12 and Eu1c strains. Moreover, we show that all of the investigated Frankia strains have two squalene hopane cyclase genes (shc1 and shc2

  16. O₂migration rates in [NiFe] hydrogenases. A joint approach combining free-energy calculations and kinetic modeling.

    Science.gov (United States)

    Topin, Jérémie; Diharce, Julien; Fiorucci, Sébastien; Antonczak, Serge; Golebiowski, Jérôme

    2014-01-23

    Hydrogenases are promising candidates for the catalytic production of green energy by means of biological ways. The major impediment to such a production is rooted in their inhibition under aerobic conditions. In this work, we model dioxygen migration rates in mutants of a hydrogenase of Desulfovibrio fructusovorans. The approach relies on the calculation of the whole potential of mean force for O2 migration within the wild-type as well as in V74M, V74F, and V74Q mutant channels. The three free-energy barriers along the entire migration pathway are converted into chemical rates through modeling based on Transition State Theory. The use of such a model recovers the trend of O2 migration rates among the series.

  17. Process and genes for expression and overexpression of active [FeFe] hydrogenases

    Science.gov (United States)

    Seibert, Michael; King, Paul W; Ghirardi, Maria Lucia; Posewitz, Matthew C; Smolinski, Sharon L

    2014-09-16

    A process for expression of active [FeFe]-hydrogenase in a host organism that does not contain either the structural gene(s) for [FeFe]-hydrogenases and/or homologues for the maturation genes HydE, HydF and HyG, comprising: cloning the structural hydrogenase gene(s) and/or the maturation genes HydE, HydF and HydG from an organisms that contains these genes into expression plasmids; transferring the plasmids into an organism that lacks a native [FeFe]-hydrogenase or that has a disrupted [FeFe]-hydrogenase and culturing it aerobically; and inducing anaerobiosis to provide [FeFe] hydrogenase biosynthesis and H?2#191 production.

  18. Process and genes for expression and overexpression of active [FeFe] hydrogenases

    Energy Technology Data Exchange (ETDEWEB)

    Seibert, Michael; King, Paul W; Ghirardi, Maria Lucia; Posewitz, Matthew C; Smolinski, Sharon L

    2014-09-16

    A process for expression of active [FeFe]-hydrogenase in a host organism that does not contain either the structural gene(s) for [FeFe]-hydrogenases and/or homologues for the maturation genes HydE, HydF and HyG, comprising: cloning the structural hydrogenase gene(s) and/or the maturation genes HydE, HydF and HydG from an organisms that contains these genes into expression plasmids; transferring the plasmids into an organism that lacks a native [FeFe]-hydrogenase or that has a disrupted [FeFe]-hydrogenase and culturing it aerobically; and inducing anaerobiosis to provide [FeFe] hydrogenase biosynthesis and H?2#191 production.

  19. Computational Design of Iron Diphosphine Complexes with Pendant Amines for Hydrogenation of CO2 to Methanol: A Mimic of [NiFe] Hydrogenase.

    Science.gov (United States)

    Chen, Xiangyang; Jing, Yuanyuan; Yang, Xinzheng

    2016-06-20

    Inspired by the active-site structure of the [NiFe] hydrogenase, we have computationally designed the iron complex [P(tBu) 2 N(tBu) 2 )Fe(CN)2 CO] by using an experimentally ready-made diphosphine ligand with pendant amines for the hydrogenation of CO2 to methanol. Density functional theory calculations indicate that the rate-determining step in the whole catalytic reaction is the direct hydride transfer from the Fe center to the carbon atom in the formic acid with a total free energy barrier of 28.4 kcal mol(-1) in aqueous solution. Such a barrier indicates that the designed iron complex is a promising low-cost catalyst for the formation of methanol from CO2 and H2 under mild conditions. The key role of the diphosphine ligand with pendent amine groups in the reaction is the assistance of the cleavage of H2 by forming a Fe-H(δ-) ⋅⋅⋅H(δ+) -N dihydrogen bond in a fashion of frustrated Lewis pairs.

  20. Atomic model of the F420-reducing [NiFe] hydrogenase by electron cryo-microscopy using a direct electron detector.

    Science.gov (United States)

    Allegretti, Matteo; Mills, Deryck J; McMullan, Greg; Kühlbrandt, Werner; Vonck, Janet

    2014-02-25

    The introduction of direct electron detectors with higher detective quantum efficiency and fast read-out marks the beginning of a new era in electron cryo-microscopy. Using the FEI Falcon II direct electron detector in video mode, we have reconstructed a map at 3.36 Å resolution of the 1.2 MDa F420-reducing hydrogenase (Frh) from methanogenic archaea from only 320,000 asymmetric units. Videos frames were aligned by a combination of image and particle alignment procedures to overcome the effects of beam-induced motion. The reconstructed density map shows all secondary structure as well as clear side chain densities for most residues. The full coordination of all cofactors in the electron transfer chain (a [NiFe] center, four [4Fe4S] clusters and an FAD) is clearly visible along with a well-defined substrate access channel. From the rigidity of the complex we conclude that catalysis is diffusion-limited and does not depend on protein flexibility or conformational changes. DOI: http://dx.doi.org/10.7554/eLife.01963.001.

  1. Minimal Influence of [NiFe] Hydrogenase on Hydrogen Isotope Fractionation in H2-Oxidizing Cupriavidus necator

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    Brian J. Campbell

    2017-10-01

    Full Text Available Fatty acids produced by H2-metabolizing bacteria are sometimes observed to be more D-depleted than those of photoautotrophic organisms, a trait that has been suggested as diagnostic for chemoautotrophic bacteria. The biochemical reasons for such a depletion are not known, but are often assumed to involve the strong D-depletion of H2. Here, we cultivated the bacterium Cupriavidus necator H16 (formerly Ralstonia eutropha H16 under aerobic, H2-consuming, chemoautotrophic conditions and measured the isotopic compositions of its fatty acids. In parallel with the wild type, two mutants of this strain, each lacking one of two key hydrogenase enzymes, were also grown and measured. In all three strains, fractionations between fatty acids and water ranged from -173‰ to -235‰, and averaged -217‰, -196‰, and -226‰, respectively, for the wild type, SH- mutant, and MBH- mutant. There was a modest increase in δD as a result of loss of the soluble hydrogenase enzyme. Fractionation curves for all three strains were constructed by growing parallel cultures in waters with δDwater values of approximately -25‰, 520‰, and 1100‰. These curves indicate that at least 90% of the hydrogen in fatty acids is derived from water, not H2. Published details of the biochemistry of the soluble and membrane-bound hydrogenases confirm that these enzymes transfer electrons rather than intact hydride (H- ions, providing no direct mechanism to connect the isotopic composition of H2 to that of lipids. Multiple lines of evidence thus agree that in this organism, and presumably others like it, environmental H2 plays little or no direct role in controlling lipid δD values. The observed fractionations must instead result from isotope effects in the reduction of NAD(PH by reductases with flavin prosthetic groups, which transfer two electrons and acquire H+ (or D+ from solution. Parallels to NADPH reduction in photosynthesis may explain why D/H fractionations in C. necator

  2. Ni l-edge soft x-ray spectroscopy of ni-fe hydrogenases and modelcompounds--evidence for high-spin ni(ii) in the active enzyme

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Hongxin; Ralston, C.Y.; Patil, D.S.; Jones, R.M.; Gu, M.; Verhagen, M.; Adams, M.; Ge, P.; Riordan, C.; Marganian, C.A.; Mascharak,P.; Kovacs, J.; Miller, C.G.; Collins, T.J.; Brooker, S.; Croucher, P.D.; Wang, Kun; Stiefel, E.I.; Cramer, S.P.

    2000-03-15

    L-edge X-ray absorption spectroscopy has been used to study, under a variety of conditions, the electronic structure of Ni in the Ni-Fe hydrogenases from Desulfovibrio gigas, Desulfovibrio baculatus, and Pyrococcus furiosus. The status of the enzyme films used for these measurements was monitored by FT-IR spectroscopy. The L-edge spectra were interpreted by ligand field multiplet simulations and by comparison with data for Ni model complexes. The spectrum for Ni in D. gigas enzyme ''form A'' is consistent with a covalent Ni(III) species. In contrast, all of the reduced enzyme samples exhibited high spin Ni(II) spectra. The significance of the Ni(II) spin state for the structure of the hydrogenase active site is discussed.

  3. Contributions of the [NiFe]- and [FeFe]-hydrogenase to H2 production in Shewanella oneidensis MR-1 as revealed by isotope ratio analysis of evolved H2

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    Kreuzer, Helen W.; Hill, Eric A.; Moran, James J.; Bartholomew, Rachel A.; Hui, Yang; Hegg, Eric L.

    2014-03-01

    Shewanella oneidensis MR-1 encodes both a [NiFe]- and an [FeFe]-hydrogenase. While the output of these proteins has been characterized in mutant strains expressing only one of the enzymes, the contribution of each to H2 synthesis in the wild-type organism is not clear. Here we use stable isotope analysis of H2 in the culture headspace, along with transcription data and measurements of the concentrations of gases in the headspace, to characterize H2 production in the wild-type strain. After most of the O2 in the headspace had been consumed, H2 was produced and then consumed by the bidirectional [NiFe]-hydrogenase. Once the cultures were completely anaerobic, a new burst of H2 synthesis catalyzed by both enzymes took place. Our data is consistent with the hypothesis that at this point in the culture cycle, a pool of electrons is shunted toward both hydrogenases in the wild-type organism, but that in the absence of one of the hydrogenases, the flux is redirected to the available enzyme. To our knowledge, this is the first use of stable isotope analysis of a metabolic product to elucidate substrate flux through two alternative enzymes in the same cellular system.

  4. Fundamental Studies of Recombinant Hydrogenases

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    Adams, Michael W

    2014-01-25

    This research addressed the long term goals of understanding the assembly and organization of hydrogenase enzymes, of reducing them in size and complexity, of determining structure/function relationships, including energy conservation via charge separation across membranes, and in screening for novel H2 catalysts. A key overall goal of the proposed research was to define and characterize minimal hydrogenases that are produced in high yields and are oxygen-resistant. Remarkably, in spite of decades of research carried out on hydrogenases, it is not possible to readily manipulate or design the enzyme using molecular biology approaches since a recombinant form produced in a suitable host is not available. Such resources are essential if we are to understand what constitutes a “minimal” hydrogenase and design such catalysts with certain properties, such as resistance to oxygen, extreme stability and specificity for a given electron donor. The model system for our studies is Pyrococcus furiosus, a hyperthermophile that grows optimally at 100°C, which contains three different nickel-iron [NiFe-] containing hydrogenases. Hydrogenases I and II are cytoplasmic while the other, MBH, is an integral membrane protein that functions to both evolve H2 and pump protons. Three important breakthroughs were made during the funding period with P. furiosus soluble hydrogenase I (SHI). First, we produced an active recombinant form of SHI in E. coli by the co-expression of sixteen genes using anaerobically-induced promoters. Second, we genetically-engineered P. furiosus to overexpress SHI by an order of magnitude compared to the wild type strain. Third, we generated the first ‘minimal’ form of SHI, one that contained two rather than four subunits. This dimeric form was stable and active, and directly interacted with a pyruvate-oxidizing enzyme with any intermediate electron carrier. The research resulted in five peer-reviewed publications.

  5. [FeFe]-hydrogenase gene quantification and melting curve analysis from hydrogen-fermenting bioreactor samples

    Energy Technology Data Exchange (ETDEWEB)

    Tolvanen, Katariina E.S.; Santala, Ville P.; Karp, Matti T. [Department of Chemistry and Bioengineering, Tampere University of Technology, P.O. Box 541, FI-33101 Tampere (Finland)

    2010-04-15

    In this study, quantitative PCR (qPCR) was used to quantify [FeFe]-hydrogenases and subsequently melting curves were analyzed from hydrogen-fermenting, mixed-culture bioreactor samples. Denaturing gradient gel electrophoresis (DGGE) analysis was also performed to the reactor samples revealing a clostridial dominance in the reactor. Primers targeting [FeFe]-hydrogenases were designed based on known clostridial [FeFe]-hydrogenase gene sequences and tested with several clostridial strains. The results show that amplification efficiencies of four different clostridia are highly similar and melting curves of the clostridial strains were within 1 C of each other. We compared the melting curves to the hydrogen percentage and observed a correlation between the results. The closer the melting curves were to those of clostridia, the better the hydrogen production. Based on these results, the primers and melting curve analysis of [FeFe]-hydrogenase amplicons can be used for analysing hydrogenase genes from bioreactor samples. (author)

  6. The [FeFe] hydrogenase of Nyctotherus ovalis has a chimeric origin

    Directory of Open Access Journals (Sweden)

    Jouany Jean-Pierre

    2007-11-01

    Full Text Available Abstract Background The hydrogenosomes of the anaerobic ciliate Nyctotherus ovalis show how mitochondria can evolve into hydrogenosomes because they possess a mitochondrial genome and parts of an electron-transport chain on the one hand, and a hydrogenase on the other hand. The hydrogenase permits direct reoxidation of NADH because it consists of a [FeFe] hydrogenase module that is fused to two modules, which are homologous to the 24 kDa and the 51 kDa subunits of a mitochondrial complex I. Results The [FeFe] hydrogenase belongs to a clade of hydrogenases that are different from well-known eukaryotic hydrogenases. The 24 kDa and the 51 kDa modules are most closely related to homologous modules that function in bacterial [NiFe] hydrogenases. Paralogous, mitochondrial 24 kDa and 51 kDa modules function in the mitochondrial complex I in N. ovalis. The different hydrogenase modules have been fused to form a polyprotein that is targeted into the hydrogenosome. Conclusion The hydrogenase and their associated modules have most likely been acquired by independent lateral gene transfer from different sources. This scenario for a concerted lateral gene transfer is in agreement with the evolution of the hydrogenosome from a genuine ciliate mitochondrion by evolutionary tinkering.

  7. Iron-dependent hydrogenases of Entamoeba histolytica and Giardia lamblia: activity of the recombinant entamoebic enzyme and evidence for lateral gene transfer.

    Science.gov (United States)

    Nixon, Julie E J; Field, Jessica; McArthur, Andrew G; Sogin, Mitchell L; Yarlett, Nigel; Loftus, Brendan J; Samuelson, John

    2003-02-01

    Entamoeba histolytica and Spironucleus barkhanus have genes that encode short iron-dependent hydrogenases (Fe-hydrogenases), even though these protists lack hydrogenosomes. To understand better the biochemistry of the protist Fe-hydrogenases, we prepared a recombinant E. histolytica short Fe-hydrogenase and measured its activity in vitro. A Giardia lamblia gene encoding a short Fe-hydrogenase was identified from shotgun genomic sequences, and RT-PCR showed that cultured entamoebas and giardias transcribe short Fe-hydrogenase mRNAs. A second E. histolytica gene, which encoded a long Fe-hydrogenase, was identified from shotgun genomic sequences. Phylogenetic analyses suggested that the short Fe-hydrogenase genes of entamoeba and diplomonads share a common ancestor, while the long Fe-hydrogenase gene of entamoeba appears to have been laterally transferred from a bacterium. These results are discussed in the context of competing ideas for the origins of genes encoding fermentation enzymes of these protists.

  8. Hydrogenase Gene Distribution and H2 Consumption Ability within the Thiomicrospira Lineage.

    Science.gov (United States)

    Hansen, Moritz; Perner, Mirjam

    2016-01-01

    Thiomicrospira were originally characterized as sulfur-oxidizing chemolithoautotrophs. Attempts to grow them on hydrogen failed for many years. Only recently we demonstrated hydrogen consumption among two of three tested Thiomicrospira and posited that hydrogen consumption may be more widespread among Thiomicrospira than previously assumed. Here, we investigate and compare the hydrogen consumption ability and the presence of group 1 [NiFe]-hydrogenase genes (enzyme catalyzes H2↔2H(+) + 2e(-)) for sixteen different Thiomicrospira species. Seven of these Thiomicrospira species encoded group 1 [NiFe]-hydrogenase genes and five of these species could also consume hydrogen. All Thiomicrospira species exhibiting hydrogen consumption were from hydrothermal vents along the Mid-Atlantic ridge or Eastern Pacific ridges. The tested Thiomicrospira from Mediterranean and Western Pacific vents could not consume hydrogen. The [NiFe]-hydrogenase genes were categorized into two clusters: those resembling the hydrogenase from Hydrogenovibrio are in cluster I and are related to those from Alpha- and other Gammaproteobacteria. In cluster II, hydrogenases found exclusively in Thiomicrospira crunogena strains are combined and form a monophyletic group with those from Epsilonproteobacteria suggesting they were acquired through horizontal gene transfer. Hydrogen consumption appears to be common among some Thiomicrospira, given that five of the tested sixteen strains carried this trait. The hydrogen consumption ability expands their competitiveness within an environment.

  9. Hydrogenase gene distribution and H2 consumption ability within the Thiomicrospira lineage

    Directory of Open Access Journals (Sweden)

    Moritz eHansen

    2016-02-01

    Full Text Available Thiomicrospira were originally characterized as sulfur-oxidizing chemolithoautotrophs. Attempts to grow them on hydrogen failed for many years. Only recently we demonstrated hydrogen consumption among two of three tested Thiomicrospira and posited that hydrogen consumption may be more widespread among Thiomicrospira than previously assumed. Here, we investigate and compare the hydrogen consumption ability and the presence of group 1 [NiFe]-hydrogenase genes (enzyme catalyzes H22H+ + 2e- for sixteen different Thiomicrospira species. Seven of these Thiomicrospira species encoded group 1 [NiFe]-hydrogenase genes and five of these species could also consume hydrogen. All Thiomicrospira species exhibiting hydrogen consumption were from hydrothermal vents along the Mid-Atlantic ridge or Eastern Pacific ridges. The tested Thiomicrospira from Mediterranean and Western Pacific vents could not consume hydrogen. The [NiFe]-hydrogenase genes were categorized into two clusters: those resembling the hydrogenase from Hydrogenovibrio are in cluster I and are related to those from Alpha- and other Gammaproteobacteria. In cluster II, hydrogenases found exclusively in T. crunogena strains are combined and form a monophyletic group with those from Epsilonproteobacteria suggesting they were acquired through horizontal gene transfer. Hydrogen consumption appears to be common among some Thiomicrospira, given that five of the tested sixteen strains carried this trait. The hydrogen consumption ability expands their competitiveness within an environment.

  10. Nickel availability to pea (Pisum sativum L.) plants limits hydrogenase activity of Rhizobium leguminosarum bv. viciae bacteroids by affecting the processing of the hydrogenase structural subunits.

    Science.gov (United States)

    Brito, B; Palacios, J M; Hidalgo, E; Imperial, J; Ruiz-Argüeso, T

    1994-01-01

    Rhizobium leguminosarum bv. viciae UPM791 induces the synthesis of an [NiFe] hydrogenase in pea (Pisum sativum L.) bacteroids which oxidizes the H2 generated by the nitrogenase complex inside the root nodules. The synthesis of this hydrogenase requires the genes for the small and large hydrogenase subunits (hupS and hupL, respectively) and 15 accessory genes clustered in a complex locus in the symbiotic plasmid. We show here that the bacteroid hydrogenase activity is limited by the availability of nickel to pea plants. Addition of Ni2+ to plant nutrient solutions (up to 10 mg/liter) resulted in sharp increases (up to 15-fold) in hydrogenase activity. This effect was not detected when other divalent cations (Zn2+, Co2+, Fe2+, and Mn2+) were added at the same concentrations. Determinations of the steady-state levels of hupSL-specific mRNA indicated that this increase in hydrogenase activity was not due to stimulation of transcription of structural genes. Immunoblot analysis with antibodies raised against the large and small subunits of the hydrogenase enzyme demonstrated that in the low-nickel situation, both subunits are mainly present in slow-migrating, unprocessed forms. Supplementation of the plant nutrient solution with increasing nickel concentrations caused the conversion of the slow-migrating forms of both subunits into fast-moving, mature forms. This nickel-dependent maturation process of the hydrogenase subunits is mediated by accessory gene products, since bacteroids from H2 uptake-deficient mutants carrying Tn5 insertions in hupG and hupK and in hypB and hypE accumulated the immature forms of both hydrogenase subunits even in the presence of high nickel levels. Images PMID:8071205

  11. Flow-FISH analysis and isolation of clostridial strains in an anaerobic semi-solid bio-hydrogen producing system by hydrogenase gene target.

    Science.gov (United States)

    Jen, Chang Jui; Chou, Chia-Hung; Hsu, Ping-Chi; Yu, Sian-Jhong; Chen, Wei-En; Lay, Jiunn-Jyi; Huang, Chieh-Chen; Wen, Fu-Shyan

    2007-04-01

    By using hydrogenase gene-targeted polymerase chain reaction (PCR) and reverse transcriptase PCR (RT-PCR), the predominant clostridial hydrogenase that may have contributed to biohydrogen production in an anaerobic semi-solid fermentation system has been monitored. The results revealed that a Clostridium pasteurianum-like hydrogenase gene sequence can be detected by both PCR and RT-PCR and suggested that the bacterial strain possessing this specific hydrogenase gene was dominant in hydrogenase activity and population. Whereas another Clostridium saccharobutylicum-like hydrogenase gene can be detected only by RT-PCR and suggest that the bacterial strain possessing this specific hydrogenase gene may be less dominant in population. In this study, hydrogenase gene-targeted fluorescence in situ hybridization (FISH) and flow cytometry analysis confirmed that only 6.6% of the total eubacterial cells in a hydrogen-producing culture were detected to express the C. saccharobutylicum-like hydrogenase, whereas the eubacteria that expressed the C. pasteurianum-like hydrogenase was 25.6%. A clostridial strain M1 possessing the identical nucleotide sequences of the C. saccharobutylicum-like hydrogenase gene was then isolated and identified as Clostridium butyricum based on 16S rRNA sequence. Comparing to the original inoculum with mixed microflora, either using C. butyricum M1 as the only inoculum or co-culturing with a Bacillus thermoamylovorans isolate will guarantee an effective and even better production of hydrogen from brewery yeast waste.

  12. Enhancing hydrogen production of Enterobacter aerogenes by heterologous expression of hydrogenase genes originated from Synechocystis sp.

    Science.gov (United States)

    Song, Wenlu; Cheng, Jun; Zhao, Jinfang; Zhang, Chuanxi; Zhou, Junhu; Cen, Kefa

    2016-09-01

    The hydrogenase genes (hoxEFUYH) of Synechocystis sp. PCC 6803 were cloned and heterologously expressed in Enterobacter aerogenes ATCC13408 for the first time in this study, and the hydrogen yield was significantly enhanced using the recombinant strain. A recombinant plasmid containing the gene in-frame with Glutathione-S-Transferase (GST) gene was transformed into E. aerogenes ATCC13408 to produce a GST-fusion protein. SDS-PAGE and western blot analysis confirm the successful expression of the hox genes. The hydrogenase activity of the recombinant strain is 237.6±9.3ml/(g-DW·h), which is 152% higher than the wild strain. The hydrogen yield of the recombinant strain is 298.3ml/g-glucose, which is 88% higher than the wild strain. During hydrogen fermentation, the recombinant strain produces more acetate and butyrate, but less ethanol. This is corresponding to the NADH metabolism in the cell due to the higher hydrogenase activity with the heterologous expression of hox genes.

  13. Synthesis and vibrational spectroscopy of 57Fe-labeled models of [NiFe] hydrogenase: first direct observation of a nickel–iron interaction† †Electronic supplementary information (ESI) available: Experimental procedures, spectral data, computational chemistry details, animated vibrational modes as GIFs. See DOI: 10.1039/c4cc04572f Click here for additional data file. Click here for additional data file.

    Science.gov (United States)

    Pelmenschikov, Vladimir; Wang, Hongxin; Meier, Florian; Gee, Leland B.; Yoda, Yoshitaka; Kaupp, Martin; Rauchfuss, Thomas B.

    2014-01-01

    A new route to iron carbonyls has enabled synthesis of 57Fe-labeled [NiFe] hydrogenase mimic (OC)3 57Fe(pdt)Ni(dppe). Its study by nuclear resonance vibrational spectroscopy revealed Ni–57Fe vibrations, as confirmed by calculations. The modes are absent for [(OC)3 57Fe(pdt)Ni(dppe)]+, which lacks Ni–57Fe bonding, underscoring the utility of the analyses in identifying metal–metal interactions. PMID:25237680

  14. Isolation of genes (nif/hup cosmids) involved in hydrogenase and nitrogenase activities in Rhizobium japonicum.

    Science.gov (United States)

    Hom, S S; Graham, L A; Maier, R J

    1985-03-01

    Recombinant cosmids containing a Rhizobium japonicum gene involved in both hydrogenase (Hup) and nitrogenase (Nif) activities were isolated. An R. japonicum gene bank utilizing broad-host-range cosmid pLAFR1 was conjugated into Hup- Nif- R. japonicum strain SR139. Transconjugants containing the nif/hup cosmid were identified by their resistance to tetracycline (Tcr) and ability to grow chemoautotrophically (Aut+) with hydrogen. All Tcr Aut+ transconjugants possessed high levels of H2 uptake activity, as determined amperometrically. Moreover, all Hup+ transconjugants tested possessed the ability to reduce acetylene (Nif+) in soybean nodules. Cosmid DNAs from 19 Hup+ transconjugants were transferred to Escherichia coli by transformation. When the cosmids were restricted with EcoRI, 15 of the 19 cosmids had a restriction pattern with 13.2-, 4.0-, 3.0-, and 2.5-kilobase DNA fragments. Six E. coli transformants containing the nif/hup cosmids were conjugated with strain SR139. All strain SR139 transconjugants were Hup+ Nif+. Moreover, one nif/hup cosmid was transferred to 15 other R. japonicum Hup- mutants. Hup+ transconjugants of six of the Hup- mutants appeared at a frequency of 1.0, whereas the transconjugants of the other nine mutants remained Hup-. These results indicate that the nif/hup gene cosmids contain a gene involved in both nitrogenase and hydrogenase activities and at least one and perhaps other hup genes which are exclusively involved in H2 uptake activity.

  15. Heterologous expression and maturation of an NADP-dependent [NiFe]-hydrogenase: a key enzyme in biofuel production.

    Directory of Open Access Journals (Sweden)

    Junsong Sun

    Full Text Available Hydrogen gas is a major biofuel and is metabolized by a wide range of microorganisms. Microbial hydrogen production is catalyzed by hydrogenase, an extremely complex, air-sensitive enzyme that utilizes a binuclear nickel-iron [NiFe] catalytic site. Production and engineering of recombinant [NiFe]-hydrogenases in a genetically-tractable organism, as with metalloprotein complexes in general, has met with limited success due to the elaborate maturation process that is required, primarily in the absence of oxygen, to assemble the catalytic center and functional enzyme. We report here the successful production in Escherichia coli of the recombinant form of a cytoplasmic, NADP-dependent hydrogenase from Pyrococcus furiosus, an anaerobic hyperthermophile. This was achieved using novel expression vectors for the co-expression of thirteen P. furiosus genes (four structural genes encoding the hydrogenase and nine encoding maturation proteins. Remarkably, the native E. coli maturation machinery will also generate a functional hydrogenase when provided with only the genes encoding the hydrogenase subunits and a single protease from P. furiosus. Another novel feature is that their expression was induced by anaerobic conditions, whereby E. coli was grown aerobically and production of recombinant hydrogenase was achieved by simply changing the gas feed from air to an inert gas (N2. The recombinant enzyme was purified and shown to be functionally similar to the native enzyme purified from P. furiosus. The methodology to generate this key hydrogen-producing enzyme has dramatic implications for the production of hydrogen and NADPH as vehicles for energy storage and transport, for engineering hydrogenase to optimize production and catalysis, as well as for the general production of complex, oxygen-sensitive metalloproteins.

  16. The surprising diversity of clostridial hydrogenases: a comparative genomic perspective.

    Science.gov (United States)

    Calusinska, Magdalena; Happe, Thomas; Joris, Bernard; Wilmotte, Annick

    2010-06-01

    Among the large variety of micro-organisms capable of fermentative hydrogen production, strict anaerobes such as members of the genus Clostridium are the most widely studied. They can produce hydrogen by a reversible reduction of protons accumulated during fermentation to dihydrogen, a reaction which is catalysed by hydrogenases. Sequenced genomes provide completely new insights into the diversity of clostridial hydrogenases. Building on previous reports, we found that [FeFe] hydrogenases are not a homogeneous group of enzymes, but exist in multiple forms with different modular structures and are especially abundant in members of the genus Clostridium. This unusual diversity seems to support the central role of hydrogenases in cell metabolism. In particular, the presence of multiple putative operons encoding multisubunit [FeFe] hydrogenases highlights the fact that hydrogen metabolism is very complex in this genus. In contrast with [FeFe] hydrogenases, their [NiFe] hydrogenase counterparts, widely represented in other bacteria and archaea, are found in only a few clostridial species. Surprisingly, a heteromultimeric Ech hydrogenase, known to be an energy-converting [NiFe] hydrogenase and previously described only in methanogenic archaea and some sulfur-reducing bacteria, was found to be encoded by the genomes of four cellulolytic strains: Clostridum cellulolyticum, Clostridum papyrosolvens, Clostridum thermocellum and Clostridum phytofermentans.

  17. HupW Protease Specifically Required for Processing of the Catalytic Subunit of the Uptake Hydrogenase in the Cyanobacterium Nostoc sp. Strain PCC 7120

    Science.gov (United States)

    Lindberg, Pia; Devine, Ellenor; Stensjö, Karin

    2012-01-01

    The maturation process of [NiFe] hydrogenases includes a proteolytic cleavage of the large subunit. We constructed a mutant of Nostoc strain PCC 7120 in which hupW, encoding a putative hydrogenase-specific protease, is inactivated. Our results indicate that the protein product of hupW selectively cleaves the uptake hydrogenase in this cyanobacterium. PMID:22020512

  18. [NiFe]-hydrogenases: spectroscopic and electrochemical definition of reactions and intermediates.

    NARCIS (Netherlands)

    F.A. Armstrong; S.P.J. Albracht

    2005-01-01

    Production and usage of di-hydrogen, H2, in micro-organisms is catalysed by highly active, 'ancient' metalloenzymes known as hydrogenases. Based on the number and identity of metal atoms in their active sites, hydrogenases fall into three main classes, [NiFe]-, [FeFe]- and [Fe]-. All contain the unu

  19. An analysis of the changes in soluble hydrogenase and global gene expression in Cupriavidus necator (Ralstonia eutropha) H16 grown in heterotrophic diauxic batch culture.

    Science.gov (United States)

    Jugder, Bat-Erdene; Chen, Zhiliang; Ping, Darren Tan Tek; Lebhar, Helene; Welch, Jeffrey; Marquis, Christopher P

    2015-03-25

    Soluble hydrogenases (SH) are enzymes that catalyse the oxidation of molecular hydrogen. The SH enzyme from Cupriavidus necator H16 is relatively oxygen tolerant and makes an attractive target for potential application in biochemical hydrogen fuel cells. Expression of the enzyme can be mediated by derepression of the hox promoter system under heterotrophic conditions. However, the overall impact of hox derepression, from a transcriptomic perspective, has never been previously reported. Derepression of hydrogenase gene expression upon fructose depletion was confirmed in replicate experiments. Using qRT-PCR, hoxF was 4.6-fold up-regulated, hypF2 was up-regulated in the cells grown 2.2-fold and the regulatory gene hoxA was up-regulated by a mean factor of 4.5. A full transcriptomic evaluation revealed a substantial shift in the global pattern of gene expression. In addition to up-regulation of genes associated with hydrogenase expression, significant changes were observed in genes associated with energy transduction, amino acid metabolism, transcription and translation (and regulation thereof), genes associated with cell stress, lipid and cell wall biogenesis and other functions, including cell motility. We report the first full transcriptome analysis of C. necator H16 grown heterotrophically on fructose and glycerol in diauxic batch culture, which permits expression of soluble hydrogenase under heterotrophic conditions. The data presented deepens our understanding of the changes in global gene expression patterns that occur during the switch to growth on glycerol and suggests that energy deficit is a key driver for induction of hydrogenase expression in this organism.

  20. Molecular study of the chlorella algae and determining its functional features with the approach of hydrogenase gene expression

    Directory of Open Access Journals (Sweden)

    Seyedeh Tayebeh Mousavi

    2016-09-01

    Full Text Available Biohydrogen production by biological processes are known as a renewable energy source. The aim of the investigation was to find and optimize the most appropriate medium for the algae growth to produce the maximum amount of hydrogen. First of all, the bioinformatics and biosystematics studies were taken for identifying the collected microalgae which was detected as Chlorella with the following features: spherical appearance, spent protozoan, Cup-shaped chloroplast with no flagella. On the other hand, the molecular analysis by PCR and 18S sequence typing of interested microalgae demonstrated 100% similarity to that well known sequences for Chlorella vulgaris. Second, we assessed some culture media including BBM, Chu10, TAP, and Sorokin and Krauss for optimum growth conditions for Chlorella vulgaris. In general, our results showed that BBM medium had the highest efficiency for producing microalgae biomass under following conditions: pH=8, temperature of 30 ° with 16 to 8 h light to darkness periods ratio.Third, we designed a more efficient photo-bioreactor apparatus toward inducing more powerful bio-hydrogen production by hydrogenase enzyme activity of our given microalgae. Then, the performance of the apparatus as well as the gene expression was scrutinized under different conditions (light, pH, sulphorous, etc.. For this, after extracting RNA and constructing cDNA, hydrogenase gene was amplified with PCR and the product was evaluated by agarose gel. However, the relative expression of the gene measured by Real Time PCR showed the influence of light, pH and sulphourous on the expression as compared with control.

  1. Mononuclear Ni(III) complexes [NiIII(L)(P(C6H3-3-SiMe3-2-S)3)]0/1- (L = thiolate, selenolate, CH2CN, Cl, PPh3): relevance to the nickel site of [NiFe] hydrogenases.

    Science.gov (United States)

    Lee, Chien-Ming; Chuang, Ya-Lan; Chiang, Chao-Yi; Lee, Gene-Hsiang; Liaw, Wen-Feng

    2006-12-25

    The stable mononuclear Ni(III)-thiolate complexes [NiIII(L)(P(C6H3-3-SiMe3-2-S)3)]- (L = SePh (2), Cl (3), SEt (4), 2-S-C4H3S (5), CH2CN (7)) were isolated and characterized by UV-vis, EPR, IR, SQUID, CV, 1H NMR, and single-crystal X-ray diffraction. The increased basicity (electronic density) of the nickel center of complexes [NiIII(L)(P(C6H3-3-SiMe3-2-S)3)]- modulated by the monodentate ligand L and the substituted groups of the phenylthiolate rings promotes the stability and reactivity. In contrast to the irreversible reduction at -1.17 V (vs Cp2Fe/Cp2Fe+) for complex 3, the cyclic voltammograms of complexes [NiIII(SePh)(P(o-C6H4S)3)]-, 2, 4, and 7 display reversible NiIII/II redox processes with E(1/2) = -1.20, -1.26, -1.32, and -1.34 V (vs Cp2Fe/Cp2Fe+), respectively. Compared to complex 2 containing a phenylselenolate-coordinated ligand, complex 4 with a stronger electron-donating ethylthiolate coordinated to the Ni(III) promotes dechlorination of CH2Cl2 to yield complex 3 (kobs = (6.01 +/- 0.03) x 10-4 s-1 for conversion of complex 4 into 3 vs kobs = (4.78 +/- 0.02) x 10-5 s-1 for conversion of complex 2 into 3). Interestingly, addition of CH3CN into complex 3 in the presence of sodium hydride yielded the stable Ni(III)-cyanomethanide complex 7 with a NiIII-CH2CN bond distance of 2.037(3) A. The NiIII-SEt bond length of 2.273(1) A in complex 4 is at the upper end of the 2.12-2.28 A range for the NiIII-S bond lengths of the oxidized-form [NiFe] hydrogenases. In contrast to the inertness of complexes 3 and 7 under CO atmosphere, carbon monoxide triggers the reductive elimination of the monodentate chalcogenolate ligand of complexes 2, 4, and 5 to produce the trigonal bipyramidal complex [NiII(CO)(P(C6H3-3-SiMe3-2-S)3]- (6).

  2. [NiFe]Hydrogenase from Citrobacter sp. S-77 surpasses platinum as an electrode for H2 oxidation reaction.

    Science.gov (United States)

    Matsumoto, Takahiro; Eguchi, Shigenobu; Nakai, Hidetaka; Hibino, Takashi; Yoon, Ki-Seok; Ogo, Seiji

    2014-08-18

    Reported herein is an electrode for dihydrogen (H2) oxidation, and it is based on [NiFe]Hydrogenase from Citrobacter sp. S-77 ([NiFe]S77). It has a 637 times higher mass activity than Pt (calculated based on 1 mg of [NiFe]S77 or Pt) at 50 mV in a hydrogen half-cell. The [NiFe]S77 electrode is also stable in air and, unlike Pt, can be recovered 100 % after poisoning by carbon monoxide. Following characterization of the [NiFe]S77 electrode, a fuel cell comprising a [NiFe]S77 anode and Pt cathode was constructed and shown to have a a higher power density than that achievable by Pt.

  3. Is engineering O{sub 2}-tolerant hydrogenases just a matter of reproducing the active sites of the naturally occurring O{sub 2}-resistant enzymes?

    Energy Technology Data Exchange (ETDEWEB)

    Leroux, Fanny; Liebgott, Pierre-Pol; Kpebe, Arlette; Leger, Christophe; Rousset, Marc; Dementin, Sebastien [CNRS, Laboratoire de Bioenergetique et Ingenierie des Proteines, Institut de Microbiologie de la Mediterranee, 31 chemin Joseph Aiguier, 13402 Marseille Cedex 20 (France); Cournac, Laurent; Richaud, Pierre [CEA, DSV, IBEB, Laboratoire de Bioenergetique et Biotechnologie des Bacteries et Microalgues, 13108 Saint-Paul-lez-Durance (France); Aix-Marseille Universite, 3 place Victor-Hugo, 13331 Marseille (France); CNRS, UMR Biologie Vegetale et Microbiologie Environnementales, 13108 Saint-Paul-lez-Durance (France); Burlat, Benedicte; Guigliarelli, Bruno; Bertrand, Patrick [CNRS, Laboratoire de Bioenergetique et Ingenierie des Proteines, Institut de Microbiologie de la Mediterranee, 31 chemin Joseph Aiguier, 13402 Marseille Cedex 20 (France); Aix-Marseille Universite, 3 place Victor-Hugo, 13331 Marseille (France)

    2010-10-15

    Reproducing the naturally occurring O{sub 2}-tolerant hydrogenases is a potential strategy to make the oxygen sensitive enzymes, produced by organisms of biotechnological interest, more resistant. The search for resistance ''hotspots'' that could be transposed into sensitive hydrogenases is underway. Here, we replaced two residues (Y77 and V78) of the oxygen sensitive [NiFe] hydrogenase from Desulfovibrio fructosovorans with Gly and with Cys, respectively, to copy the active site pocket of the resistant membrane-bound [NiFe] enzyme from Ralstonia eutropha and we examined how this affected oxygen sensitivity. The results are discussed in the light of a short review of the recent results dealing with the reactivity of hydrogenases towards oxygen. (author)

  4. Construction and use of a Cupriavidus necator H16 soluble hydrogenase promoter (PSH) fusion to gfp (green fluorescent protein).

    Science.gov (United States)

    Jugder, Bat-Erdene; Welch, Jeffrey; Braidy, Nady; Marquis, Christopher P

    2016-01-01

    Hydrogenases are metalloenzymes that reversibly catalyse the oxidation or production of molecular hydrogen (H2). Amongst a number of promising candidates for application in the oxidation of H2 is a soluble [Ni-Fe] uptake hydrogenase (SH) produced by Cupriavidus necator H16. In the present study, molecular characterisation of the SH operon, responsible for functional SH synthesis, was investigated by developing a green fluorescent protein (GFP) reporter system to characterise PSH promoter activity using several gene cloning approaches. A PSH promoter-gfp fusion was successfully constructed and inducible GFP expression driven by the PSH promoter under de-repressing conditions in heterotrophic growth media was demonstrated in the recombinant C. necator H16 cells. Here we report the first successful fluorescent reporter system to study PSH promoter activity in C. necator H16. The fusion construct allowed for the design of a simple screening assay to evaluate PSH activity. Furthermore, the constructed reporter system can serve as a model to develop a rapid fluorescent based reporter for subsequent small-scale process optimisation experiments for SH expression.

  5. Hydrogenase Enzymes and Their Synthetic Models: The Role of Metal Hydrides.

    Science.gov (United States)

    Schilter, David; Camara, James M; Huynh, Mioy T; Hammes-Schiffer, Sharon; Rauchfuss, Thomas B

    2016-08-10

    Hydrogenase enzymes efficiently process H2 and protons at organometallic FeFe, NiFe, or Fe active sites. Synthetic modeling of the many H2ase states has provided insight into H2ase structure and mechanism, as well as afforded catalysts for the H2 energy vector. Particularly important are hydride-bearing states, with synthetic hydride analogues now known for each hydrogenase class. These hydrides are typically prepared by protonation of low-valent cores. Examples of FeFe and NiFe hydrides derived from H2 have also been prepared. Such chemistry is more developed than mimicry of the redox-inactive monoFe enzyme, although functional models of the latter are now emerging. Advances in physical and theoretical characterization of H2ase enzymes and synthetic models have proven key to the study of hydrides in particular, and will guide modeling efforts toward more robust and active species optimized for practical applications.

  6. Reactions of H2, CO, and O2 with active [NiFe]-hydrogenase from Allochromatium vinosum. A stopped-flow infrared study.

    NARCIS (Netherlands)

    George, S.J.; Kurkin, S.; Thorneley, R.N.; Albracht, S.P.J.

    2004-01-01

    The Ni-Fe site in the active membrane-bound [NiFe]-hydrogenase from Allochromatium Vinosum can exist in three different redox states. In the most oxidized state (Nia-S) the nickel is divalent. The most reduced state (Nia-SR) likewise has Ni2+, while the intermediate state (Nia-C*) has Ni3+. The tran

  7. H2-Producing Bacterial Community during Rice Straw Decomposition in Paddy Field Soil: Estimation by an Analysis of [FeFe]-Hydrogenase Gene Transcripts.

    Science.gov (United States)

    Baba, Ryuko; Asakawa, Susumu; Watanabe, Takeshi

    2016-09-29

    The transcription patterns of [FeFe]-hydrogenase genes (hydA), which encode the enzymes responsible for H2 production, were investigated during rice straw decomposition in paddy soil using molecular biological techniques. Paddy soil amended with and without rice straw was incubated under anoxic conditions. RNA was extracted from the soil, and three clone libraries of hydA were constructed using RNAs obtained from samples in the initial phase of rice straw decomposition (day 1 with rice straw), methanogenic phase of rice straw decomposition (day 14 with rice straw), and under a non-amended condition (day 14 without rice straw). hydA genes related to Proteobacteria, Firmicutes, Bacteroidetes, Chloroflexi, and Thermotogae were mainly transcribed in paddy soil samples; however, their proportions markedly differed among the libraries. Deltaproteobacteria-related hydA genes were predominantly transcribed on day 1 with rice straw, while various types of hydA genes related to several phyla were transcribed on day 14 with rice straw. Although the diversity of transcribed hydA was significantly higher in the library on day 14 with rice straw than the other two libraries, the composition of hydA transcripts in the library was similar to that in the library on day 14 without rice straw. These results indicate that the composition of active H2 producers and/or H2 metabolic patterns dynamically change during rice straw decomposition in paddy soil.

  8. Homologous Cloning of Clostridium Hydrogenase Gene and Construction of Gene Knockout Vector%梭菌氢酶基因部分片段的同源克隆及敲除载体的构建

    Institute of Scientific and Technical Information of China (English)

    闫倩; 闫苗章; 王保莉; 曲东

    2012-01-01

    Dissimilatory iron-reducing bacteria grown in soil play an important role in bioremediation of organics and heavy metal pollution. The aim of this study is to explore the internal relationship between the iron-reducing ability and hydrogen-evolution of Clostridium, which is a typical iron-reducing bacteria. In this paper, a Clostridium strain isolated from paddy soil as the research object. Using homologous clone, a fragment (761 bp) of hydrogenase gene was obtained. Bioinformatics analysis showed that this gene fragment contained the active centers of hydrogenase, and was its major functional domain. Moreover, knockout vector (pMD-19-HTH) included tetracycline resistance gene was constructed by overlap PCR technique with the purpose of knockout hydrogenase function and lay the foundation for uncovering relationship between the iron-reducing ability and hydrogen-evolution.%为从分子水平探索典型铁还原菌一梭菌的铁还原能力与其氢酶产氢之间的关系,以从水稻土中分离得到一株具有高铁还原能力和高产氢能力的梭菌为材料,通过同源克隆获得长度为761 bp氢酶基因的部分序列.生物信息分析发现,该基因片段覆盖氢酶的活性中心,是氢酶的主要功能结构域.采用Overlap PCR的方法构建含有四环素抗性基因的氢酶基因敲除载体(pMD-19-HTH),以期进一步构建氢酶基因缺失的梭菌突变体,为分析氢酶产氢与铁还原的关系奠定基础.

  9. Development and application of a functional CE-SSCP fingerprinting method based on [Fe-Fe]-hydrogenase genes for monitoring hydrogen-producing Clostridium in mixed cultures

    Energy Technology Data Exchange (ETDEWEB)

    Quemeneur, Marianne; Hamelin, Jerome; Latrille, Eric; Steyer, Jean-Philippe; Trably, Eric [INRA, UR050, Laboratoire de Biotechnologie de l' Environnement, avenue des Etangs, Narbonne F-11100 (France)

    2010-12-15

    A Capillary Electrophoresis Single Strand Conformation Polymorphism (CE-SSCP) method based on functional [Fe-Fe]-hydrogenase genes was developed for monitoring the hydrogen (H{sub 2})-producing clostridial population in mixed-culture bioprocesses. New non-degenerated primers were designed and then validated on their specific PCR detection of a broad range of clostridial hydA genes. The hydA-based CE-SSCP method gave a specific and discriminating profile for each of the Clostridium strains tested. This method was validated using H{sub 2}-producing mixed cultures incubated at temperatures ranging from 25 C to 45 C. The hydA CE-SSCP profiles clearly differed between temperatures tested. Hence, they varied according to variations of the H{sub 2} production performances. The HydA sequences amplified with the new primer set indicated that diverse Clostridium strains impacted the H{sub 2} production yields. The highest performances were related to the dominance of Clostridium sporogenes-like hydA sequences. This CE-SSCP tool offers highly reliable and throughput analysis of the functional diversity and structure of the hydA genes for better understanding of the H{sub 2}-producing clostridial population dynamics in H{sub 2} dark fermentation bioreactors. (author)

  10. Polarized potential and electrode materials implication on electro-fermentative di-hydrogen production: Microbial assemblages and hydrogenase gene copy variation.

    Science.gov (United States)

    Arunasri, Kotakonda; Annie Modestra, J; Yeruva, Dileep Kumar; Vamshi Krishna, K; Venkata Mohan, S

    2016-01-01

    This study examined the changes in microbial diversity in response to different electrode materials viz., stainless steel mesh (SS) and graphite plate as anodes in two microbial electrolysis cell (MEC) each poised at 0.2V, 0.4V, 0.6V and 0.8V. Changes in microbiota prior to and after pretreatment along with microbiota enriched in response to various poised potentials with SS and graphite are monitored by 16S rRNA gene based DGGE profiling. Significant shifts in microbial community were noticed at all these experimental conditions. Correspondingly, the level of hydrogenase belonging to genera Bacillus, Pseudomonas, Rhodopseudomonas and Clostridium was studied by quantitative real time PCR (RT-PCR) at various applied potentials. DGGE based 16S rRNA gene profiling revealed enriched members belonging to phylum Firmicutes predominantly present at 0.8V in both MECs contributing to high hydrogen production. This study first time explored the growth behavior of mixed consortia in response to poised potentials and electrode materials.

  11. Synthetic Active Site Model of the [NiFeSe] Hydrogenase.

    Science.gov (United States)

    Wombwell, Claire; Reisner, Erwin

    2015-05-26

    A dinuclear synthetic model of the [NiFeSe] hydrogenase active site and a structural, spectroscopic and electrochemical analysis of this complex is reported. [NiFe('S2Se2')(CO)3] (H2'S2Se2' = 1,2-bis(2-thiabutyl-3,3-dimethyl-4-selenol)benzene) has been synthesized by reacting the nickel selenolate complex [Ni('S2Se2')] with [Fe(CO)3bda] (bda = benzylideneacetone). X-ray crystal structure analysis confirms that [NiFe('S2Se2')(CO)3] mimics the key structural features of the enzyme active site, including a doubly bridged heterobimetallic nickel and iron center with a selenolate terminally coordinated to the nickel center. Comparison of [NiFe('S2Se2')(CO)3] with the previously reported thiolate analogue [NiFe('S4')(CO)3] (H2'S4' = H2xbsms = 1,2-bis(4-mercapto-3,3-dimethyl-2-thiabutyl)benzene) showed that the selenolate groups in [NiFe('S2Se2')(CO)3] give lower carbonyl stretching frequencies in the IR spectrum. Electrochemical studies of [NiFe('S2Se2')(CO)3] and [NiFe('S4')(CO)3] demonstrated that both complexes do not operate as homogenous H2 evolution catalysts, but are precursors to a solid deposit on an electrode surface for H2 evolution catalysis in organic and aqueous solution. © 2015 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

  12. Proton-coupled electron transfer dynamics in the catalytic mechanism of a [NiFe]-hydrogenase.

    Science.gov (United States)

    Greene, Brandon L; Wu, Chang-Hao; McTernan, Patrick M; Adams, Michael W W; Dyer, R Brian

    2015-04-08

    The movement of protons and electrons is common to the synthesis of all chemical fuels such as H2. Hydrogenases, which catalyze the reversible reduction of protons, necessitate transport and reactivity between protons and electrons, but a detailed mechanism has thus far been elusive. Here, we use a phototriggered chemical potential jump method to rapidly initiate the proton reduction activity of a [NiFe] hydrogenase. Coupling the photochemical initiation approach to nanosecond transient infrared and visible absorbance spectroscopy afforded direct observation of interfacial electron transfer and active site chemistry. Tuning of intramolecular proton transport by pH and isotopic substitution revealed distinct concerted and stepwise proton-coupled electron transfer mechanisms in catalysis. The observed heterogeneity in the two sequential proton-associated reduction processes suggests a highly engineered protein environment modulating catalysis and implicates three new reaction intermediates; Nia-I, Nia-D, and Nia-SR(-). The results establish an elementary mechanistic understanding of catalysis in a [NiFe] hydrogenase with implications in enzymatic proton-coupled electron transfer and biomimetic catalyst design.

  13. Light-driven hydrogen production by a hybrid complex of a [NiFe]-hydrogenase and the cyanobacterial photosystem I.

    Science.gov (United States)

    Ihara, Masaki; Nishihara, Hirofumi; Yoon, Ki-Seok; Lenz, Oliver; Friedrich, Bärbel; Nakamoto, Hitoshi; Kojima, Kouji; Honma, Daisuke; Kamachi, Toshiaki; Okura, Ichiro

    2006-01-01

    In order to generate renewable and clean fuels, increasing efforts are focused on the exploitation of photosynthetic microorganisms for the production of molecular hydrogen from water and light. In this study we engineered a 'hard-wired' protein complex consisting of a hydrogenase and photosystem I (hydrogenase-PSI complex) as a direct light-to-hydrogen conversion system. The key component was an artificial fusion protein composed of the membrane-bound [NiFe] hydrogenase from the beta-proteobacterium Ralstonia eutropha H16 and the peripheral PSI subunit PsaE of the cyanobacterium Thermosynechococcus elongatus. The resulting hydrogenase-PsaE fusion protein associated with PsaE-free PSI spontaneously, thereby forming a hydrogenase-PSI complex as confirmed by sucrose-gradient ultracentrifuge and immunoblot analysis. The hydrogenase-PSI complex displayed light-driven hydrogen production at a rate of 0.58 mumol H(2).mg chlorophyll(-1).h(-1). The complex maintained its accessibility to the native electron acceptor ferredoxin. This study provides the first example of a light-driven enzymatic reaction by an artificial complex between a redox enzyme and photosystem I and represents an important step on the way to design a photosynthetic organism that efficiently converts solar energy and water into hydrogen.

  14. Study of hydrogenases activity inhibition by O{sub 2} and direct electrochemistry; Etude de l'inhibition par O{sub 2} de l'activite d'hydrogenases par electrochimie directe

    Energy Technology Data Exchange (ETDEWEB)

    Baffert, C.; Leger, Ch.; Leroux, F.; Bertrand, P.; Guigliarelli, B. [Laboratoire de Bioenergetique et d' Ingenierie des proteines, BIP-CNRS, 13 - Marseille (France)

    2007-07-01

    At the present time, a great effort of research is made on the identification and the design of enzymes insensitive or low sensitive to O{sub 2}. In parallel, it seems important to understand the inhibition mechanisms in order to propose mutations able to limit this inhibition. The Protein Film Voltametry allows to obtain data which can not be observed or quantitatively obtained by other techniques. The enzyme is immobilized directly on the electrode and the electronic transfer is direct. The redox state of the enzyme depends on the potential of the electrode and the catalytic current is proportional to the enzyme activity. The data obtained for the Ni-Fe hydrogenase (Desulfovibrio fructosovorans) and for the Fe hydrogenase (Clostridium Acetobutylicum) will be compared to the data obtained for hydrogenases of other organisms, by Protein Film Voltametry as well as by other techniques. (O.M.)

  15. Structure of an Actinobacterial-Type [NiFe]-Hydrogenase Reveals Insight into O2-Tolerant H2 Oxidation.

    Science.gov (United States)

    Schäfer, Caspar; Bommer, Martin; Hennig, Sandra E; Jeoung, Jae-Hun; Dobbek, Holger; Lenz, Oliver

    2016-02-01

    A novel group of bacterial [NiFe]-hydrogenases is responsible for high-affinity H2 uptake from the troposphere, and is therefore thought to play an important role in the global H2 cycle. Here we present the first crystal structure at 2.85-Å resolution of such an actinobacterial-type hydrogenase (AH), which was isolated from the dihydrogen oxidizing bacterium, Ralstonia eutropha. The enzyme has a dimeric structure carrying two active [NiFe] sites that are interconnected by six [4Fe4S] clusters over a range of approximately 90 Å. Unlike most other [NiFe]-hydrogenases, the [4Fe4S] cluster proximal to the [NiFe] site is coordinated by three cysteines and one aspartate. Mutagenesis experiments revealed that this aspartate residue is related to the apparent O2 insensitivity of the AH. Our data provide first structural insight into specialized hydrogenases that are supposed to consume atmospheric H2 under challenging conditions, i.e. at high O2 concentration and wide temperature and pH ranges.

  16. Gene Expression by the Sulfate-Reducing Bacterium Desulfovibrio vulgaris Hildenborough Grown on an Iron Electrode under Cathodic Protection Conditions▿ †

    Science.gov (United States)

    Caffrey, Sean M.; Park, Hyung Soo; Been, Jenny; Gordon, Paul; Sensen, Christoph W.; Voordouw, Gerrit

    2008-01-01

    The genome sequence of the sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough was reanalyzed to design unique 70-mer oligonucleotide probes against 2,824 probable protein-coding regions. These included three genes not previously annotated, including one that encodes a c-type cytochrome. Using microarrays printed with these 70-mer probes, we analyzed the gene expression profile of wild-type D. vulgaris grown on cathodic hydrogen, generated at an iron electrode surface with an imposed negative potential of −1.1 V (cathodic protection conditions). The gene expression profile of cells grown on cathodic hydrogen was compared to that of cells grown with gaseous hydrogen bubbling through the culture. Relative to the latter, the electrode-grown cells overexpressed two hydrogenases, the hyn-1 genes for [NiFe] hydrogenase 1 and the hyd genes, encoding [Fe] hydrogenase. The hmc genes for the high-molecular-weight cytochrome complex, which allows electron flow from the hydrogenases across the cytoplasmic membrane, were also overexpressed. In contrast, cells grown on gaseous hydrogen overexpressed the hys genes for [NiFeSe] hydrogenase. Cells growing on the electrode also overexpressed genes encoding proteins which promote biofilm formation. Although the gene expression profiles for these two modes of growth were distinct, they were more closely related to each other than to that for cells grown in a lactate- and sulfate-containing medium. Electrochemically measured corrosion rates were lower for iron electrodes covered with hyn-1, hyd, and hmc mutant biofilms than for wild-type biofilms. This confirms the importance, suggested by the gene expression studies, of the corresponding gene products in D. vulgaris-mediated iron corrosion. PMID:18310429

  17. Enhancement of photoheterotrophic biohydrogen production at elevated temperatures by the expression of a thermophilic clostridial hydrogenase.

    Science.gov (United States)

    Lo, Shou-Chen; Shih, Shau-Hua; Chang, Jui-Jen; Wang, Chun-Ying; Huang, Chieh-Chen

    2012-08-01

    The working temperature of a photobioreactor under sunlight can be elevated above the optimal growth temperature of a microorganism. To improve the biohydrogen productivity of photosynthetic bacteria at higher temperatures, a [FeFe]-hydrogenase gene from the thermophile Clostridium thermocellum was expressed in the mesophile Rhodopseudomonas palustris CGA009 (strain CGA-CThydA) using a log-phase expression promoter P( pckA ) to drive the expression of heterogeneous hydrogenase gene. In contrast, a mesophilic Clostridium acetobutylicum [FeFe]-hydrogenase gene was also constructed and expressed in R. palustris (strain CGA-CAhydA). Both transgenic strains were tested for cell growth, in vivo hydrogen production rate, and in vitro hydrogenase activity at elevated temperatures. Although both CGA-CThydA and CGA-CAhydA strains demonstrated enhanced growth over the vector control at temperatures above 38 °C, CGA-CThydA produced more hydrogen than the other strains. The in vitro hydrogenase activity assay, measured at 40 °C, confirmed that the activity of the CGA-CThydA hydrogenase was higher than the CGA-CAhydA hydrogenase. These results showed that the expression of a thermophilic [FeFe]-hydrogenase in R. palustris increased the growth rate and biohydrogen production at elevated temperatures. This transgenic strategy can be applied to a broad range of purple photosynthetic bacteria used to produce biohydrogen under sunlight.

  18. Presence and expression of hydrogenase specific C-terminal endopeptidases in cyanobacteria

    Directory of Open Access Journals (Sweden)

    Lindblad Peter

    2003-05-01

    Full Text Available Abstract Background Hydrogenases catalyze the simplest of all chemical reactions: the reduction of protons to molecular hydrogen or vice versa. Cyanobacteria can express an uptake, a bidirectional or both NiFe-hydrogenases. Maturation of those depends on accessory proteins encoded by hyp-genes. The last maturation step involves the cleavage of a ca. 30 amino acid long peptide from the large subunit by a C-terminal endopeptidase. Until know, nothing is known about the maturation of cyanobacterial NiFe-hydrogenases. The availability of three complete cyanobacterial genome sequences from strains with either only the uptake (Nostoc punctiforme ATCC 29133/PCC 73102, only the bidirectional (Synechocystis PCC 6803 or both NiFe-hydrogenases (Anabaena PCC 7120 prompted us to mine these genomes for hydrogenase maturation related genes. In this communication we focus on the presence and the expression of the NiFe-hydrogenases and the corresponding C-terminal endopeptidases, in the three strains mentioned above. Results We identified genes encoding putative cyanobacterial hydrogenase specific C-terminal endopeptidases in all analyzed cyanobacterial genomes. The genes are not part of any known hydrogenase related gene cluster. The derived amino acid sequences show only low similarity (28–41% to the well-analyzed hydrogenase specific C-terminal endopeptidase HybD from Escherichia coli, the crystal structure of which is known. However, computational secondary and tertiary structure modeling revealed the presence of conserved structural patterns around the highly conserved active site. Gene expression analysis shows that the endopeptidase encoding genes are expressed under both nitrogen-fixing and non-nitrogen-fixing conditions. Conclusion Anabaena PCC 7120 possesses two NiFe-hydrogenases and two hydrogenase specific C-terminal endopeptidases but only one set of hyp-genes. Thus, in contrast to the Hyp-proteins, the C-terminal endopeptidases are the only known

  19. The AbrB2 autorepressor, expressed from an atypical promoter, represses the hydrogenase operon to regulate hydrogen production in Synechocystis strain PCC6803.

    Science.gov (United States)

    Dutheil, Jérémy; Saenkham, Panatda; Sakr, Samer; Leplat, Christophe; Ortega-Ramos, Marcia; Bottin, Hervé; Cournac, Laurent; Cassier-Chauvat, Corinne; Chauvat, Franck

    2012-10-01

    We have thoroughly investigated the abrB2 gene (sll0822) encoding an AbrB-like regulator in the wild-type strain of the model cyanobacterium Synechocystis strain PCC6803. We report that abrB2 is expressed from an active but atypical promoter that possesses an extended -10 element (TGTAATAT) that compensates for the absence of a -35 box. Strengthening the biological significance of these data, we found that the occurrence of an extended -10 promoter box and the absence of a -35 element are two well-conserved features in abrB2 genes from other cyanobacteria. We also show that AbrB2 is an autorepressor that is dispensable to cell growth under standard laboratory conditions. Furthermore, we demonstrate that AbrB2 also represses the hox operon, which encodes the Ni-Fe hydrogenase of biotechnological interest, and that the hox operon is weakly expressed even though it possesses the two sequences resembling canonical -10 and -35 promoter boxes. In both the AbrB2-repressed promoters of the abrB2 gene and the hox operon, we found a repeated DNA motif [TT-(N(5))-AAC], which could be involved in AbrB2 repression. Supporting this hypothesis, we found that a TT-to-GG mutation of one of these elements increased the activity of the abrB2 promoter. We think that our abrB2-deleted mutant with increased expression of the hox operon and hydrogenase activity, together with the reporter plasmids we constructed to analyze the abrB2 gene and the hox operon, will serve as useful tools to decipher the function and the regulation of hydrogen production in Synechocystis.

  20. Cloning and knockout of formate hydrogen lyase and H{sub 2}-uptake hydrogenase genes in Enterobacter aerogenes for enhanced hydrogen production

    Energy Technology Data Exchange (ETDEWEB)

    Zhao, Hongxin; Ma, Kun; Lu, Yuan; Zhang, Chong; Wang, Liyan; Xing, Xin-Hui [Institute of Biochemical Engineering, Department of Chemical Engineering, Tsinghua University, Tsinghua Yuan, Beijing 100084 (China)

    2009-01-15

    A 5431-bp DNA fragment partially encoding the formate hydrogen lyase (FHL) gene cluster hycABCDE was isolated and identified from Enterobacter aerogenes IAM1183 chromosomal DNA. All the five putative gene products showed a high degree of homology to the reported bacterial FHL proteins. The gene hycA, encoding the FHL repressor protein, and hybO, encoding the small subunit of the uptake hydrogenase, were targeted for genetic knockout for improving the hydrogen production. The pYM-Red recombination system was adopted to form insertional mutations in the E. aerogenes genome, thereby creating mutant strains of IAM1183-A ({delta} hycA), IAM1183-O ({delta} hybO), and IAM1183-AO ({delta} hycA/ {delta} hybO double knockout). The hydrogen production experiments with these mutants showed that the maximum specific hydrogen productivities of IAM1183-A, IAM1183-O, and IAM1183-AO were 2879.466 {+-} 38.59, 2747.203 {+-} 13.25 and 3372.019 {+-} 4.39 (ml h{sup -1} g{sup -1}dry cell weight), respectively, higher than that of the wild strain (2321.861 {+-} 15.34 ml h{sup -1} g{sup -1}dry cell weight). The total H{sub 2} yields by the three mutants IAM1183-A, IAM1183-O and IAM1183-AO were 0.73, 0.78, and 0.83 mol-H{sub 2}/mol glucose, respectively, while the wild-type IAM1183 was only 0.65 mol-H{sub 2}/mol glucose. The metabolites of the mutants including acetate, ethanol, 2,3-butanediol and succinate were all increased compared with that of the wild type, implying the changed metabolic flux by the mutation. In the fermentor cultivation with IAM1183 {delta} hycA/ {delta} hybO, the total hydrogen volume after 16 h cultivation reached 4.4 L, while that for the wild type was only 2.9 L. (author)

  1. Structural and functional synthetic model of mono-iron hydrogenase featuring an anthracene scaffold

    Science.gov (United States)

    Seo, Junhyeok; Manes, Taylor A.; Rose, Michael J.

    2017-06-01

    Mono-iron hydrogenase was the third type of hydrogenase discovered. Its Lewis acidic iron(II) centre promotes the heterolytic cleavage of the H-H bond and this non-redox H2 activation distinguishes it from the well-studied dinuclear [FeFe] and [NiFe] hydrogenases. Cleavage of the H-H bond is followed by hydride transfer to the enzyme's organic substrate, H4MPT+, which serves as a CO2 'carrier' in methanogenic pathways. Here we report a scaffold-based synthetic approach by which to model mono-iron hydrogenase using an anthracene framework, which supports a biomimetic fac-C,N,S coordination motif to an iron(II) centre. This arrangement includes the biomimetic and organometallic Fe-C σ bond, which enables bidirectional activity reminiscent of the native enzyme: the complex activates H2 under mild conditions, and catalyses C-H hydride abstraction plus H2 generation from a model substrate. Notably, neither H2 activation nor C-H hydride abstraction was observed in the analogous complex with a pincer-type mer-C,N,S ligation, emphasizing the importance of the fac-C,N,S-iron(II) motif in promoting enzyme-like reactivity.

  2. Properties of hydrogenase from Megasphaera elsdenii

    NARCIS (Netherlands)

    Dijk, van C.

    1980-01-01

    This thesis is concerned with the purification and properties of hydrogenase from the obligate anaerobic rumen bacterium Megasphaera elsdenii. In chapter 1 the motives underlying this thesis, the physiological role of hydrogenase in some heterotrophs, including

  3. Production of biohydrogen by recombinant expression of [NiFe]-hydrogenase 1 in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Kim Jaoon YH

    2010-07-01

    Full Text Available Abstract Background Hydrogenases catalyze reversible reaction between hydrogen (H2 and proton. Inactivation of hydrogenase by exposure to oxygen is a critical limitation in biohydrogen production since strict anaerobic conditions are required. While [FeFe]-hydrogenases are irreversibly inactivated by oxygen, it was known that [NiFe]-hydrogenases are generally more tolerant to oxygen. The physiological function of [NiFe]-hydrogenase 1 is still ambiguous. We herein investigated the H2 production potential of [NiFe]-hydrogenase 1 of Escherichia coli in vivo and in vitro. The hyaA and hyaB genes corresponding to the small and large subunits of [NiFe]-hydrogenase 1 core enzyme, respectively, were expressed in BL21, an E. coli strain without H2 producing ability. Results Recombinant BL21 expressing [NiFe]-hydrogenase 1 actively produced H2 (12.5 mL H2/(h·L in 400 mL glucose minimal medium under micro-aerobic condition, whereas the wild type BL21 did not produce H2 even when formate was added as substrate for formate hydrogenlyase (FHL pathway. The majority of recombinant protein was produced as an insoluble form, with translocation of a small fraction to the membrane. However, the membrane fraction displayed high activity (~65% of total cell fraction, based on unit protein mass. Supplement of nickel and iron to media showed these metals contribute essentially to the function of [NiFe]-hydrogenase 1 as components of catalytic site. In addition, purified E. coli [NiFe]-hydrogenase 1 using his6-tag displayed oxygen-tolerant activity of ~12 nmol H2/(min·mg protein under a normal aeration environment, compared to [FeFe]-hydrogenase, which remains inactive under this condition. Conclusions This is the first report on physiological function of E. coli [NiFe]-hydrogenase 1 for H2 production. We found that [NiFe]-hydrogenase 1 has H2 production ability even under the existence of oxygen. This oxygen-tolerant property is a significant advantage because it is

  4. Molecular characterization and homologous overexpression of [FeFe]-hydrogenase in Clostridium tyrobutyricum JM1

    Energy Technology Data Exchange (ETDEWEB)

    Jo, Ji Hye [Biosciences Center, National Renewable Energy Laboratory, 1617 Cole Blvd, Golden, CO 80401-3393 (United States); Jeon, Che Ok [Department of Life Science, Chung-Ang University, Seoul 156-756 (Korea); Lee, Seung Yoon [K-water Research Institute, Korea Water Resources Corporation, 462-1, Jeonmin-dong, Yuseong-gu, Daejon 305-703 (Korea); Lee, Dae Sung [Department of Environmental Engineering, Kyungpook National University, 1370 Sankyuk-dong, Buk-gu, Daegu 702-701 (Korea); Park, Jong Moon [Department of Chemical Engineering, Pohang University of Science and Technology, San 31, Hyoja-dong, Nam-gu, Pohang, Gyeongbuk 790-784 (Korea)

    2010-02-15

    The H{sub 2}-evoving [FeFe]-hydrogenase in Clostridium tyrobutyricum JM1 was isolated to elucidate molecular characterization and modular structure of the hydrogenase. Then, homologous overexpression of the hydrogenase gene was for the first time performed to enhance hydrogen production. The hydA open reading frame (ORF) was 1734-bp, encodes 577 amino acids with a predicted molecular mass of 63,970 Da, and presents 80% and 75% identity at the amino acid level with the [FeFe]-hydrogenase genes of Clostridium kluyveri DSM 555 and Clostridium acetobutylicum ATCC 824, respectively. One histidine residue and 19 cysteine residues, known to fasten one [2Fe-2S] cluster, three [4Fe-4S] clusters and one H-cluster, were conserved in hydA of C. tyrobutyricum. A 2327-bp DNA region containing the ORF and the putative promoter region was amplified and subcloned into a pJIR418 shuttle vector. The gene transfer of the recombinant plasmid into C. tyrobutyricum JM1 was performed by a modified electrotransformation method. Homologous overexpression of the [FeFe]-hydrogenase gene resulted in a 1.7-fold and 1.5-fold increase in hydrogenase activity and hydrogen yield concomitant with the shift of metabolic pathway. (author)

  5. Heterologous expression of an algal hydrogenase in a heterocystous cyanobacterium

    Energy Technology Data Exchange (ETDEWEB)

    Thorsten Heidorn; Peter Lindblad [Dept. of Physiological Botany, Uppsala University, Villavogen 6, SE-752 36 Uppsala, (Sweden)

    2006-07-01

    For the expression of an active algal [FeFe] hydrogenase in the heterocystous cyanobacterium Nostoc punctiforme A TCC 29133 the Chlamydomonas reinhardtii hydrogenase gene hydA1 and the accessory genes hydEF and hydG are to be introduced into the cyanobacteria cells. The genes were amplified by PCR from EST clones, cloned into the cloning vector pBluescript SK+ and sequenced. An expression vector for multi-cistronic cloning, based on pSCR202, was constructed and for a functional test GFP was inserted as a reporter gene. The GFP construct was transformed into Nostoc punctiforme A TCC 29133 by electroporation and expression of GFP was visualized by fluorescence microscopy. (authors)

  6. Cyanobacterial Hydrogenases and Hydrogen Metabolism Revisited: Recent Progress and Future Prospects

    Directory of Open Access Journals (Sweden)

    Namita Khanna

    2015-05-01

    Full Text Available Cyanobacteria have garnered interest as potential cell factories for hydrogen production. In conjunction with photosynthesis, these organisms can utilize inexpensive inorganic substrates and solar energy for simultaneous biosynthesis and hydrogen evolution. However, the hydrogen yield associated with these organisms remains far too low to compete with the existing chemical processes. Our limited understanding of the cellular hydrogen production pathway is a primary setback in the potential scale-up of this process. In this regard, the present review discusses the recent insight around ferredoxin/flavodoxin as the likely electron donor to the bidirectional Hox hydrogenase instead of the generally accepted NAD(PH. This may have far reaching implications in powering solar driven hydrogen production. However, it is evident that a successful hydrogen-producing candidate would likely integrate enzymatic traits from different species. Engineering the [NiFe] hydrogenases for optimal catalytic efficiency or expression of a high turnover [FeFe] hydrogenase in these photo-autotrophs may facilitate the development of strains to reach target levels of biohydrogen production in cyanobacteria. The fundamental advancements achieved in these fields are also summarized in this review.

  7. A hydrogen-producing, hydrogenase-free mutant strain of Nostoc punctiforme ATCC 29133

    Energy Technology Data Exchange (ETDEWEB)

    Lindberg, P.; Lindblad, P. [Uppsala Univ. (Sweden). Dept. of Physiological Botany; Schuetz, K.; Happe, T. [Universitaet Bonn (Germany). Botanisches Inst.

    2002-12-01

    The hupL gene, encoding the uptake hydrogenase large subunit, in Nostoc sp. strain ATCC 29133, a strain lacking a bidirectional hydrogenase, was inactivated by insertional mutagenesis. Recombinant strains were isolated and analysed, and one hupL{sup -} strain, NHM5, was selected for further study. Cultures of NHM5 were grown under nitrogen-fixing conditions and H{sub 2} evolution under air was observed using an H{sub 2} electrode. (Author)

  8. Shewanella oneidensis: a new and efficient System for Expression and Maturation of heterologous [Fe-Fe] Hydrogenase from Chlamydomonas reinhardtii

    Directory of Open Access Journals (Sweden)

    Sybirna Kateryna

    2008-09-01

    Full Text Available Abstract Background The eukaryotic green alga, Chlamydomonas reinhardtii, produces H2 under anaerobic conditions, in a reaction catalysed by a [Fe-Fe] hydrogenase HydA1. For further biochemical and biophysical studies a suitable expression system of this enzyme should be found to overcome its weak expression in the host organism. Two heterologous expression systems used up to now have several advantages. However they are not free from some drawbacks. In this work we use bacterium Shewanella oneidensis as a new and efficient system for expression and maturation of HydA1 from Chlamydomonas reinhardtii. Results Based on codon usage bias and hydrogenase maturation ability, the bacterium S. oneidensis, which possesses putative [Fe-Fe] and [Ni-Fe] hydrogenase operons, was selected as the best potential host for C. reinhardtii [Fe-Fe] hydrogenase expression. Hydrogen formation by S. oneidensis strain AS52 (ΔhydAΔhyaB transformed with a plasmid bearing CrHydA1 and grown in the presence of six different substrates for anaerobic respiration was determined. A significant increase in hydrogen evolution was observed for cells grown in the presence of trimethylamine oxide, dimethylsulfoxide and disodium thiosulfate, showing that the system of S. oneidensis is efficient for heterologous expression of algal [Fe-Fe] hydrogenase. Conclusion In the present work a new efficient system for heterologous expression and maturation of C. reinhardtii hydrogenase has been developed. HydA1 of C. reinhardtii was purified and shown to contain 6 Fe atoms/molecule of protein, as expected. Using DMSO, TMAO or thiosulfate as substrates for anaerobic respiration during the cell growth, 0.4 – 0.5 mg l-1(OD600 = 1 of catalytically active HydA1 was obtained with hydrogen evolution rate of ~700 μmol H2 mg-1 min-1.

  9. Mutational analysis of the hyc-operon determining the relationship between hydrogenase-3 and NADH pathway in Enterobacter aerogenes.

    Science.gov (United States)

    Pi, Jian; Jawed, Muhammad; Wang, Jun; Xu, Li; Yan, Yunjun

    2016-01-01

    In this study, the hydrogenase-3 gene cluster (hycDEFGH) was isolated and identified from Enterobacter aerogenes CCTCC AB91102. All gene products were highly homologous to the reported bacterial hydrogenase-3 (Hyd-3) proteins. The genes hycE, hycF, hycG encoding the subunits of hydrogenase-3 were targeted for genetic knockout to inhibit the FHL hydrogen production pathway via the Red recombination system, generating three mutant strains AB91102-E (ΔhycE), AB91102-F (ΔhycF) and AB91102-G (ΔhycG). Deletion of the three genes affected the integrity of hydrogenase-3. The hydrogen production experiments with the mutant strains showed that no hydrogen was detected compared with the wild type (0.886 mol/mol glucose), demonstrating that knocking out any of the three genes could inhibit NADH hydrogen production pathway. Meanwhile, the metabolites of the mutant strains were significantly changed in comparison with the wild type, indicating corresponding changes in metabolic flux by mutation. Additionally, the activity of NADH-mediated hydrogenase was found to be nil in the mutant strains. The chemostat experiments showed that the NADH/NAD(+) ratio of the mutant strains increased nearly 1.4-fold compared with the wild type. The NADH-mediated hydrogenase activity and NADH/NAD(+) ratio analysis both suggested that NADH pathway required the involvement of the electron transport chain of hydrogenase-3.

  10. Inhibition of hydrogen uptake in Escherichia coli by expressing the hydrogenase from the cyanobacterium Synechocystis sp. PCC 6803

    Directory of Open Access Journals (Sweden)

    Wood Thomas K

    2007-05-01

    Full Text Available Abstract Background Molecular hydrogen is an environmentally-clean fuel and the reversible (bi-directional hydrogenase of the cyanobacterium Synechocystis sp. PCC 6803 as well as the native Escherichia coli hydrogenase 3 hold great promise for hydrogen generation. These enzymes perform the simple reaction 2H+ + 2e- ↔ H2 (g. Results Hydrogen yields were enhanced up to 41-fold by cloning the bidirectional hydrogenase (encoded by hoxEFUYH from the cyanobacterium into E. coli. Using an optimized medium, E. coli cells expressing hoxEFUYH also produced twice as much hydrogen as the well-studied Enterobacter aerogenes HU-101, and hydrogen gas bubbles are clearly visible from the cultures. Overexpression of HoxU alone (small diaphorase subunit accounts for 43% of the additional hydrogen produced by HoxEFUYH. In addition, hydrogen production in E. coli mutants with defects in the native formate hydrogenlyase system show that the cyanobacterial hydrogenase depends on both the native E. coli hydrogenase 3 as well as on its maturation proteins. Hydrogen absorption by cells expressing hoxEFUYH was up to 10 times lower than cells which lack the cloned cyanobacterial hydrogenase; hence, the enhanced hydrogen production in the presence of hoxEFUYH is due to inhibition of hydrogen uptake activity in E. coli. Hydrogen uptake by cells expressing hoxEFUYH was suppressed in three wild-type strains and in two hycE mutants but not in a double mutant defective in hydrogenase 1 and hydrogenase 2; hence, the active cyanobacterial locus suppresses hydrogen uptake by hydrogenase 1 and hydrogenase 2 but not by hydrogenase 3. Differential gene expression indicated that overexpression of HoxEFUYH does not alter expression of the native E. coli hydrogenase system; instead, biofilm-related genes are differentially regulated by expression of the cyanobacterial enzymes which resulted in 2-fold elevated biofilm formation. This appears to be the first enhanced hydrogen production

  11. Development of a Rhodobacter capsulatus self-reporting model system for optimizing light-dependent, [FeFe]-hydrogenase-driven H 2 production: A Model System for Optimizing H 2 Production

    Energy Technology Data Exchange (ETDEWEB)

    Wecker, Matt S. A. [GeneBiologics, LLC, Boulder Colorado; Beaton, Stephen E. [United States Air Force Academy, Department of Chemistry, Colorado Springs Colorado; Chado, Robert A. [United States Air Force Academy, Department of Chemistry, Colorado Springs Colorado; Ghirardi, Maria L. [National Renewable Energy Laboratory, MS 3313, 15013 Denver West Parkway Golden Colorado 80401

    2016-08-23

    The photosynthetic bacterium Rhodobacter capsulatus normally photoproduces H2 as a by-product of its nitrogenase-catalyzed nitrogen-fixing activity. Such H2 production, however, is expensive from a metabolic perspective, requiring nearly four times as many photons as the equivalent algal hydrogenase-based system (Ghirardi et al. 2009). Here we report the insertion of a Clostridium acetobutylicum [FeFe]-hydrogenase and its three attendant hydrogenase assembly proteins into an R. capsulatus strain lacking its native uptake hydrogenase. Further, this strain is modified to fluoresce upon sensing H2. The resulting strain photoproduces H2 and self-reports its own H2 production through fluorescence. This model system represents a unique method of developing hydrogenase-based H2 production in R. capsulatus, may serve as a powerful system for in vivo directed evolution of hydrogenases and hydrogenase-associated genes, and provides a means of screening for increased metabolic production of H2.

  12. Using directed evolution to improve hydrogen production in chimeric hydrogenases from Clostridia species.

    Science.gov (United States)

    Plummer, Scott M; Plummer, Mark A; Merkel, Patricia A; Hagen, Moira; Biddle, Jennifer F; Waidner, Lisa A

    2016-11-01

    Hydrogenases are enzymes that play a key role in controlling excess reducing equivalents in both photosynthetic and anaerobic organisms. This enzyme is viewed as potentially important for the industrial generation of hydrogen gas; however, insufficient hydrogen production has impeded its use in a commercial process. Here, we explore the potential to circumvent this problem by directly evolving the Fe-Fe hydrogenase genes from two species of Clostridia bacteria. In addition, a computational model based on these mutant sequences was developed and used as a predictive aid for the isolation of enzymes with even greater efficiency in hydrogen production. Two of the improved mutants have a logarithmic increase in hydrogen production in our in vitro assay. Furthermore, the model predicts hydrogenase sequences with hydrogen productions as high as 540-fold over the positive control. Taken together, these results demonstrate the potential of directed evolution to improve the native bacterial hydrogenases as a first step for improvement of hydrogenase activity, further in silico prediction, and finally, construction and demonstration of an improved algal hydrogenase in an in vivo assay of C. reinhardtii hydrogen production. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. Disclosure of key stereoelectronic factors for efficient H2 binding and cleavage in the active site of [NiFe]-hydrogenases.

    Science.gov (United States)

    Bruschi, Maurizio; Tiberti, Matteo; Guerra, Alessandro; De Gioia, Luca

    2014-02-05

    A comparative analysis of a series of DFT models of [NiFe]-hydrogenases, ranging from minimal NiFe clusters to very large systems including both the first and second coordination sphere of the bimetallic cofactor, was carried out with the aim of unraveling which stereoelectronic properties of the active site of [NiFe]-hydrogenases are crucial for efficient H2 binding and cleavage. H2 binding to the Ni-SIa redox state is energetically favored (by 4.0 kcal mol(-1)) only when H2 binds to Ni, the NiFe metal cluster is in a low spin state, and the Ni cysteine ligands have a peculiar seesaw coordination geometry, which in the enzyme is stabilized by the protein environment. The influence of the Ni coordination geometry on the H2 binding affinity was then quantitatively evaluated and rationalized analyzing frontier molecular orbitals and populations. Several plausible reaction pathways leading to H2 cleavage were also studied. It turned out that a two-step pathway, where H2 cleavage takes place on the Ni-SIa redox state of the enzyme, is characterized by very low reaction barriers and favorable reaction energies. More importantly, the seesaw coordination geometry of Ni was found to be a key feature for facile H2 cleavage. The discovery of the crucial influence of the Ni coordination geometry on H2 binding and activation in the active site of [NiFe]-hydrogenases could be exploited in the design of novel biomimetic synthetic catalysts.

  14. A novel endo-hydrogenase activity recycles hydrogen produced by nitrogen fixation.

    Directory of Open Access Journals (Sweden)

    Gordon Ng

    Full Text Available BACKGROUND: Nitrogen (N(2 fixation also yields hydrogen (H(2 at 1:1 stoichiometric amounts. In aerobic diazotrophic (able to grow on N(2 as sole N-source bacteria, orthodox respiratory hupSL-encoded hydrogenase activity, associated with the cell membrane but facing the periplasm (exo-hydrogenase, has nevertheless been presumed responsible for recycling such endogenous hydrogen. METHODS AND FINDINGS: As shown here, for Azorhizobium caulinodans diazotrophic cultures open to the atmosphere, exo-hydrogenase activity is of no consequence to hydrogen recycling. In a bioinformatic analysis, a novel seven-gene A. caulinodans hyq cluster encoding an integral-membrane, group-4, Ni,Fe-hydrogenase with homology to respiratory complex I (NADH: quinone dehydrogenase was identified. By analogy, Hyq hydrogenase is also integral to the cell membrane, but its active site faces the cytoplasm (endo-hydrogenase. An A. caulinodans in-frame hyq operon deletion mutant, constructed by "crossover PCR", showed markedly decreased growth rates in diazotrophic cultures; normal growth was restored with added ammonium--as expected of an H(2-recycling mutant phenotype. Using A. caulinodans hyq merodiploid strains expressing beta-glucuronidase as promoter-reporter, the hyq operon proved strongly and specifically induced in diazotrophic culture; as well, hyq operon induction required the NIFA transcriptional activator. Therefore, the hyq operon is constituent of the nif regulon. CONCLUSIONS: Representative of aerobic N(2-fixing and H(2-recycling alpha-proteobacteria, A. caulinodans possesses two respiratory Ni,Fe-hydrogenases: HupSL exo-hydrogenase activity drives exogenous H(2 respiration, and Hyq endo-hydrogenase activity recycles endogenous H(2, specifically that produced by N(2 fixation. To benefit human civilization, H(2 has generated considerable interest as potential renewable energy source as its makings are ubiquitous and its combustion yields no greenhouse gases. As

  15. Hydrogenases from the Hyperthermophilic Archaeon Pyrococcus furiosus

    NARCIS (Netherlands)

    van Haaster, D.J.

    2007-01-01

    Hydrogenase is an electron-transfer protein and catalyses the simplest chemical redox reaction, the reversible two-electron oxidation of molecular hydrogen in aerobic and anaerobic microorganisms. A kinetic study of the hydrogen oxidation reaction by Fe-hydrogenase from Desulfovibrio vulgaris

  16. Properties of hydrogenase from Megasphaera elsdenii

    NARCIS (Netherlands)

    Dijk, van C.

    1980-01-01

    This thesis is concerned with the purification and properties of hydrogenase from the obligate anaerobic rumen bacterium Megasphaera elsdenii. In chapter 1 the motives underlying this thesis, the physiological role of hydrogenase in some heterotrophs, including Megasphaera elsdenii, as well as a com

  17. Hydrogenases from the Hyperthermophilic Archaeon Pyrococcus furiosus

    NARCIS (Netherlands)

    van Haaster, D.J.

    2007-01-01

    Hydrogenase is an electron-transfer protein and catalyses the simplest chemical redox reaction, the reversible two-electron oxidation of molecular hydrogen in aerobic and anaerobic microorganisms. A kinetic study of the hydrogen oxidation reaction by Fe-hydrogenase from Desulfovibrio vulgaris (Hilde

  18. Diversity and transcription of proteases involved in the maturation of hydrogenases in Nostoc punctiforme ATCC 29133 and Nostoc sp. strain PCC 7120

    Directory of Open Access Journals (Sweden)

    Lindblad Peter

    2009-03-01

    Full Text Available Abstract Background The last step in the maturation process of the large subunit of [NiFe]-hydrogenases is a proteolytic cleavage of the C-terminal by a hydrogenase specific protease. Contrary to other accessory proteins these hydrogenase proteases are believed to be specific whereby one type of hydrogenases specific protease only cleaves one type of hydrogenase. In cyanobacteria this is achieved by the gene product of either hupW or hoxW, specific for the uptake or the bidirectional hydrogenase respectively. The filamentous cyanobacteria Nostoc punctiforme ATCC 29133 and Nostoc sp strain PCC 7120 may contain a single uptake hydrogenase or both an uptake and a bidirectional hydrogenase respectively. Results In order to examine these proteases in cyanobacteria, transcriptional analyses were performed of hupW in Nostoc punctiforme ATCC 29133 and hupW and hoxW in Nostoc sp. strain PCC 7120. These studies revealed numerous transcriptional start points together with putative binding sites for NtcA (hupW and LexA (hoxW. In order to investigate the diversity and specificity among hydrogeanse specific proteases we constructed a phylogenetic tree which revealed several subgroups that showed a striking resemblance to the subgroups previously described for [NiFe]-hydrogenases. Additionally the proteases specificity was also addressed by amino acid sequence analysis and protein-protein docking experiments with 3D-models derived from bioinformatic studies. These studies revealed a so called "HOXBOX"; an amino acid sequence specific for protease of Hox-type which might be involved in docking with the large subunit of the hydrogenase. Conclusion Our findings suggest that the hydrogenase specific proteases are under similar regulatory control as the hydrogenases they cleave. The result from the phylogenetic study also indicates that the hydrogenase and the protease have co-evolved since ancient time and suggests that at least one major horizontal gene transfer

  19. Diversity and transcription of proteases involved in the maturation of hydrogenases in Nostoc punctiforme ATCC 29133 and Nostoc sp. strain PCC 7120

    Science.gov (United States)

    2009-01-01

    Background The last step in the maturation process of the large subunit of [NiFe]-hydrogenases is a proteolytic cleavage of the C-terminal by a hydrogenase specific protease. Contrary to other accessory proteins these hydrogenase proteases are believed to be specific whereby one type of hydrogenases specific protease only cleaves one type of hydrogenase. In cyanobacteria this is achieved by the gene product of either hupW or hoxW, specific for the uptake or the bidirectional hydrogenase respectively. The filamentous cyanobacteria Nostoc punctiforme ATCC 29133 and Nostoc sp strain PCC 7120 may contain a single uptake hydrogenase or both an uptake and a bidirectional hydrogenase respectively. Results In order to examine these proteases in cyanobacteria, transcriptional analyses were performed of hupW in Nostoc punctiforme ATCC 29133 and hupW and hoxW in Nostoc sp. strain PCC 7120. These studies revealed numerous transcriptional start points together with putative binding sites for NtcA (hupW) and LexA (hoxW). In order to investigate the diversity and specificity among hydrogeanse specific proteases we constructed a phylogenetic tree which revealed several subgroups that showed a striking resemblance to the subgroups previously described for [NiFe]-hydrogenases. Additionally the proteases specificity was also addressed by amino acid sequence analysis and protein-protein docking experiments with 3D-models derived from bioinformatic studies. These studies revealed a so called "HOXBOX"; an amino acid sequence specific for protease of Hox-type which might be involved in docking with the large subunit of the hydrogenase. Conclusion Our findings suggest that the hydrogenase specific proteases are under similar regulatory control as the hydrogenases they cleave. The result from the phylogenetic study also indicates that the hydrogenase and the protease have co-evolved since ancient time and suggests that at least one major horizontal gene transfer has occurred. This co

  20. Functional Analysis by Site-Directed Mutagenesis of the NAD+-Reducing Hydrogenase from Ralstonia eutropha

    Science.gov (United States)

    Burgdorf, Tanja; De Lacey, Antonio L.; Friedrich, Bärbel

    2002-01-01

    The tetrameric cytoplasmic [NiFe] hydrogenase (SH) of Ralstonia eutropha couples the oxidation of hydrogen to the reduction of NAD+ under aerobic conditions. In the catalytic subunit HoxH, all six conserved motifs surrounding the [NiFe] site are present. Five of these motifs were altered by site-directed mutagenesis in order to dissect the molecular mechanism of hydrogen activation. Based on phenotypic characterizations, 27 mutants were grouped into four different classes. Mutants of the major class, class I, failed to grow on hydrogen and were devoid of H2-oxidizing activity. In one of these isolates (HoxH I64A), H2 binding was impaired. Class II mutants revealed a high D2/H+ exchange rate relative to a low H2-oxidizing activity. A representative (HoxH H16L) displayed D2/H+ exchange but had lost electron acceptor-reducing activity. Both activities were equally affected in class III mutants. Mutants forming class IV showed a particularly interesting phenotype. They displayed O2-sensitive growth on hydrogen due to an O2-sensitive SH protein. PMID:12399498

  1. Genomic and metagenomic surveys of hydrogenase distribution indicate H2 is a widely utilised energy source for microbial growth and survival.

    Science.gov (United States)

    Greening, Chris; Biswas, Ambarish; Carere, Carlo R; Jackson, Colin J; Taylor, Matthew C; Stott, Matthew B; Cook, Gregory M; Morales, Sergio E

    2016-03-01

    Recent physiological and ecological studies have challenged the long-held belief that microbial metabolism of molecular hydrogen (H2) is a niche process. To gain a broader insight into the importance of microbial H2 metabolism, we comprehensively surveyed the genomic and metagenomic distribution of hydrogenases, the reversible enzymes that catalyse the oxidation and evolution of H2. The protein sequences of 3286 non-redundant putative hydrogenases were curated from publicly available databases. These metalloenzymes were classified into multiple groups based on (1) amino acid sequence phylogeny, (2) metal-binding motifs, (3) predicted genetic organisation and (4) reported biochemical characteristics. Four groups (22 subgroups) of [NiFe]-hydrogenase, three groups (6 subtypes) of [FeFe]-hydrogenases and a small group of [Fe]-hydrogenases were identified. We predict that this hydrogenase diversity supports H2-based respiration, fermentation and carbon fixation processes in both oxic and anoxic environments, in addition to various H2-sensing, electron-bifurcation and energy-conversion mechanisms. Hydrogenase-encoding genes were identified in 51 bacterial and archaeal phyla, suggesting strong pressure for both vertical and lateral acquisition. Furthermore, hydrogenase genes could be recovered from diverse terrestrial, aquatic and host-associated metagenomes in varying proportions, indicating a broad ecological distribution and utilisation. Oxygen content (pO2) appears to be a central factor driving the phylum- and ecosystem-level distribution of these genes. In addition to compounding evidence that H2 was the first electron donor for life, our analysis suggests that the great diversification of hydrogenases has enabled H2 metabolism to sustain the growth or survival of microorganisms in a wide range of ecosystems to the present day. This work also provides a comprehensive expanded system for classifying hydrogenases and identifies new prospects for investigating H2

  2. Hydrogenase polypeptide and methods of use

    Energy Technology Data Exchange (ETDEWEB)

    Adams, Michael W.W.; Hopkins, Robert C.; Jenney, JR, Francis E.; Sun, Junsong

    2016-02-02

    Provided herein are polypeptides having hydrogenase activity. The polypeptide may be multimeric, and may have hydrogenase activity of at least 0.05 micromoles H.sub.2 produced min.sup.-1 mg protein.sup.-1. Also provided herein are polynucleotides encoding the polypeptides, genetically modified microbes that include polynucleotides encoding one or more subunits of the multimeric polypeptide, and methods for making and using the polypeptides.

  3. Improving cyanobacterail O2-tolerance using CBS hydrogenase for hydrogen production

    Energy Technology Data Exchange (ETDEWEB)

    Maness, Pin-Ching [National Renewable Energy Lab. (NREL), Golden, CO (United States); Eckert, Carrie [National Renewable Energy Lab. (NREL), Golden, CO (United States); Wawrousek, Karen [National Renewable Energy Lab. (NREL), Golden, CO (United States); Noble, Scott [National Renewable Energy Lab. (NREL), Golden, CO (United States); Pennington, Grant [National Renewable Energy Lab. (NREL), Golden, CO (United States); Yu, Jianping [National Renewable Energy Lab. (NREL), Golden, CO (United States)

    2016-11-11

    Cyanobacterial H2 production is a viable path to renewable H2 with water serving as the electron donor and sunlight the energy source. A grand challenge is the sensitivity of the underlying hydrogenase to O2, the latter an inherent byproduct of oxygenic photosynthesis. This challenge has been identified as a technical barrier in the Fuel Cell Technologies Office (FCTO) Multi-year Research, Development and Deployment Plan. One solution is to express in cyanobacterium an O2-tolerant hydrogenase to circumvent this barrier. We have uncovered an O2-tolerant hydrogenase from a photosynthetic bacterium Rubrivivax gelatinosus CBS (Casa Bonita Strain; hereafter “CBS”) with a half-life near 21 h when exposed to ambient O2. We sequenced the CBS genome and identified two sets of maturation machineries hyp1 and hyp2. Transcripts expression analysis and mutagenesis revealed that hyp1 is responsible for the assembly of the O2-tolerant CO-oxidation (Coo) hydrogenase and hyp2 is involved in the maturation of a H2-uptake hydrogenase. The structural genes encoding the O2-tolerant hydrogenase (cooLXUH) and maturation genes hyp1FABCDE were therefore cloned and expressed in the model cyanobacterium Synechocystis sp. PCC 6803. We obtained several recombinants displaying hydrogenase activity in a Synechocystis host lacking background activity, suggesting that the CBS hydrogenase is active in Synechocystis. Yet the activity is extremely low. To ensure balanced protein expression, we systematically optimized heterologous expression of 10 CBS genes by using stronger promoters and better ribosome binding site. Moreover we attempted the expression of cooM and cooK genes, verified to be important in CBS to afford activity. CooM is a very large protein and both CooM and CooK are membrane-associated. These properties limited our success in expressing both genes in Synechocystis, although they

  4. Roles of HynAB and Ech, the only two hydrogenases found in the model sulfate reducer Desulfovibrio gigas.

    Science.gov (United States)

    Morais-Silva, Fabio O; Santos, Catia I; Rodrigues, Rute; Pereira, Inês A C; Rodrigues-Pousada, Claudina

    2013-10-01

    Sulfate-reducing bacteria are characterized by a high number of hydrogenases, which have been proposed to contribute to the overall energy metabolism of the cell, but exactly in what role is not clear. Desulfovibrio spp. can produce or consume H2 when growing on organic or inorganic substrates in the presence or absence of sulfate. Because of the presence of only two hydrogenases encoded in its genome, the periplasmic HynAB and cytoplasmic Ech hydrogenases, Desulfovibrio gigas is an excellent model organism for investigation of the specific function of each of these enzymes during growth. In this study, we analyzed the physiological response to the deletion of the genes that encode the two hydrogenases in D. gigas, through the generation of ΔechBC and ΔhynAB single mutant strains. These strains were analyzed for the ability to grow on different substrates, such as lactate, pyruvate, and hydrogen, under respiratory and fermentative conditions. Furthermore, the expression of both hydrogenase genes in the three strains studied was assessed through quantitative reverse transcription-PCR. The results demonstrate that neither hydrogenase is essential for growth on lactate-sulfate, indicating that hydrogen cycling is not indispensable. In addition, the periplasmic HynAB enzyme has a bifunctional activity and is required for growth on H2 or by fermentation of pyruvate. Therefore, this enzyme seems to play a dominant role in D. gigas hydrogen metabolism.

  5. Molecular detection of the clostridia in an anaerobic biohydrogen fermentation system by hydrogenase mRNA-targeted reverse transcription-PCR.

    Science.gov (United States)

    Chang, Jui-Jen; Chen, Wei-En; Shih, Shiou-Yun; Yu, Sian-Jhong; Lay, Jiunn-Jyi; Wen, Fu-Shyan; Huang, Chieh-Chen

    2006-05-01

    Molecular biological approaches were developed to monitor the potential biohydrogen-producing clostridia in an anaerobic semisolid fermentation system that used brewery yeast waste as the fermentation substrate. The denaturing gradient gel electrophoresis with 16S rDNA gene-targeted polymerase chain reaction (PCR) analysis was employed to confirm the existence of clostridia in the system. Remarkably, reproducible nucleotide sequences of clostridia were obtained from different hydrogen production stages by using hydrogenase gene-targeted reverse transcription (RT)-PCR. These RNA-based information suggested that the predominant hydrogen-producing strains possess either a specific Clostridium pasteurianum-like or a specific Clostridium saccharobutylicum-like hydrogenase sequence. Comparison of the hydrogenase gene-targeted sequence profiles between PCR and RT-PCR revealed that the specific C. pasteurianum-like hydrogenase harboring bacterial strains were dominant in both mRNA and bacterial population level. On the other hand, the specific C. saccharobutylicum-like hydrogenase harboring strains expressed high level of hydrogenase mRNA but may not be dominant in population. Furthermore, quantitative real-time RT-PCR analysis showed the expression pattern of the clostridial hydrogenase mRNA and may serve as an activity index for the system.

  6. Expression of a clostridial [FeFe]-hydrogenase in Chlamydomonas reinhardtii prolongs photo-production of hydrogen from water splitting

    Energy Technology Data Exchange (ETDEWEB)

    Noone, Seth; Ratcliff, Kathleen; Davis, ReAnna; Subramanian, Venkataramanan; Meuser, Jonathan; Posewitz, Matthew C.; King, Paul W.; Ghirardi, Maria L.

    2017-03-01

    The high oxygen (O2) sensitivity of green algal [FeFe]-hydrogenases is a significant limitation for the sustained production of hydrogen gas (H2) from photosynthetic water splitting. To address this limitation we replaced the native [FeFe]-hydrogenases with a more O2-tolerant clostridial [FeFe]-hydrogenase CaI in Chlamydomonas reinhardtii strain D66..delta..HYD (hydA1-hydA2-) that contains insertionally inactivated [FeFe]-hydrogenases genes. Expression and translocation of CaI in D66..delta..HYD led to the recovery of H2 photoproduction at ~ 20% of the rates of the wild-type parent strain D66. We show for the first time that a bacterial [FeFe]-hydrogenase can be expressed, localized and matured to a catalytically active form that couples to photosynthetic electron transport in the green alga C. reinhardtii. The lower rates of O2 inactivation of CaI led to more sustained H2 photoproduction when cultures were challenged with O2 or kept under prolonged illumination at solar intensities. These results provide new insights into the requisites for attaining photobiological H2 production from water splitting using a more O2-tolerant hydrogenase.

  7. Cobaloxime-based artificial hydrogenases.

    Science.gov (United States)

    Bacchi, Marine; Berggren, Gustav; Niklas, Jens; Veinberg, Elias; Mara, Michael W; Shelby, Megan L; Poluektov, Oleg G; Chen, Lin X; Tiede, David M; Cavazza, Christine; Field, Martin J; Fontecave, Marc; Artero, Vincent

    2014-08-01

    Cobaloximes are popular H2 evolution molecular catalysts but have so far mainly been studied in nonaqueous conditions. We show here that they are also valuable for the design of artificial hydrogenases for application in neutral aqueous solutions and report on the preparation of two well-defined biohybrid species via the binding of two cobaloxime moieties, {Co(dmgH)2} and {Co(dmgBF2)2} (dmgH2 = dimethylglyoxime), to apo Sperm-whale myoglobin (SwMb). All spectroscopic data confirm that the cobaloxime moieties are inserted within the binding pocket of the SwMb protein and are coordinated to a histidine residue in the axial position of the cobalt complex, resulting in thermodynamically stable complexes. Quantum chemical/molecular mechanical docking calculations indicated a coordination preference for His93 over the other histidine residue (His64) present in the vicinity. Interestingly, the redox activity of the cobalt centers is retained in both biohybrids, which provides them with the catalytic activity for H2 evolution in near-neutral aqueous conditions.

  8. The weak, fluctuating, dipole moment of membrane-bound hydrogenase from Aquifex aeolicus accounts for its adaptability to charged electrodes.

    Science.gov (United States)

    Oteri, Francesco; Ciaccafava, Alexandre; de Poulpiquet, Anne; Baaden, Marc; Lojou, Elisabeth; Sacquin-Mora, Sophie

    2014-06-21

    [NiFe] hydrogenases from Aquifex aeolicus (AaHase) and Desulfovibrio fructosovorans (DfHase) have been mainly studied to characterize physiological electron transfer processes, or to develop biotechnological devices such as biofuel cells. In this context, it remains difficult to control the orientation of AaHases on electrodes to achieve a fast interfacial electron transfer. Here, we study the electrostatic properties of these two proteins based on microsecond-long molecular dynamics simulations that we compare to voltammetry experiments. Our calculations show weak values and large fluctuations of the dipole direction in AaHase compared to DfHase, enabling the AaHase to absorb on both negatively and positively charged electrodes, with an orientation distribution that induces a spread in electron transfer rates. Moreover, we discuss the role of the transmembrane helix of AaHase and show that it does not substantially impact the general features of the dipole moment.

  9. Photoinduced reduction of the medial FeS center in the hydrogenase small subunit HupS from Nostoc punctiforme.

    Science.gov (United States)

    Raleiras, Patrícia; Hammarström, Leif; Lindblad, Peter; Styring, Stenbjörn; Magnuson, Ann

    2015-07-01

    The small subunit from the NiFe uptake hydrogenase, HupSL, in the cyanobacterium Nostoc punctiforme ATCC 29133, has been isolated in the absence of the large subunit (P. Raleiras, P. Kellers, P. Lindblad, S. Styring, A. Magnuson, J. Biol. Chem. 288 (2013) 18,345-18,352). Here, we have used flash photolysis to reduce the iron-sulfur clusters in the isolated small subunit, HupS. We used ascorbate as electron donor to the photogenerated excited state of Ru(II)-trisbipyridine (Ru(bpy)3), to generate Ru(I)(bpy)3 as reducing agent. Our results show that the isolated small subunit can be reduced by the Ru(I)(bpy)3 generated through flash photolysis. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. Electrical resistivity of NiFe2O4 ceramic and NiFe2O4 based cermets

    Institute of Scientific and Technical Information of China (English)

    田忠良; 赖延清; 李劼; 张刚; 刘业翔

    2004-01-01

    NiFe2O4 ceramic and NiFe2O4 based cermets, expected to be used as the inert anodes in aluminum electrolysis, were prepared and their electrical resistivities were measured at different temperatures. The effects of temperature and composition on their electrical resistivities were investigated. The results indicate that the electrical resistivities of NiFe2O4 based cermets mainly depend on temperature, resistivity of ceramic matrix, composition and dispersion of the metal phase among ceramic matrix. The electrical resistivity of NiFe2O4 ceramic decreases from 10. 094 Ω · cm to 0. 475 Ω · em with increasing temperature from 573 K to 1 233 K. The electrical resistivities of NiFe2O4 based cermets are greatly lowered, but decrease with increasing the temperature with similar trend compared to that of NiFe2O4 ceramic. The resistivities of NiFe2O4 based cermets containing 5 % Ni, 5 % Cu and 5 % CuNi alloy are 0. 046 8, 0.066 8 and 0. 0532 Ω · cm at 1 233 K, respectively, which are all acceptable as inert anode materials compared to that of the current carbon anode used for aluminum electrolysis.

  11. Heterologous expression of an algal hydrogenase in a hetero-cystous cyanobacterium

    Energy Technology Data Exchange (ETDEWEB)

    Thorsten Heidorn; Peter Lindblad [Dept. of Physiological Botany, Uppsala University, V illavagen 6, SE-752 36 Uppsala, (Sweden)

    2006-07-01

    For the expression of an active algal [FeFe] hydrogenase in the hetero-cystous cyanobacterium Nostoc punctiforme A TCC 29133 the Chlamydomonas reinhardtii hydrogenase gene hydA1 and the accessory genes hydEF and hydG are to be introduced into the cyano-bacterial cells. The genes were amplified by PCR from EST clones, cloned into the cloning vector pBluescript SK+ and sequenced. An expression vector for multi-cistronic cloning, based on pSCR202, was constructed and for a functional test GFP was inserted as a reporter gene. The GFP construct was transformed into Nostoc punctiforme A TCC 29133 by electroporation and expression of GFP was visualized by fluorescence microscopy. (authors)

  12. Optimized over-expression of [FeFe] hydrogenases with high specific activity in Clostridium acetobutylicum

    Energy Technology Data Exchange (ETDEWEB)

    von Abendroth, Gregory; Stripp, Sven; Happe, Thomas [Ruhr-Universitaet Bochum, Lehrstuhl fuer Biochemie der Pflanzen, AG Photobiotechnologie, 44780 Bochum (Germany); Silakov, Alexey [Max-Planck-Institut fuer Bioanorganische Chemie, 45470 Muelheim an der Ruhr (Germany); Croux, Christian; Soucaille, Philippe; Girbal, Laurence [UMR5504, UMR792 Ingenierie des Systemes Biologiques et des Procedes, CNRS, INRA, INSA, 31400 Toulouse (France)

    2008-11-15

    It was previously shown that Clostridium acetobutylicum is capable to over-express various [FeFe] hydrogenases although the protein yield was low. In this study we report on doubling the yield of the clostridial hydrogenase by replacing the native gene hydA1{sub Ca} with a recombinant one via homologous recombination. The purified protein HydA1{sub Ca} shows an unexpected high specific activity (up to 2257 {mu}mol H{sub 2} min{sup -1} mg{sup -1}) for hydrogen evolution. Furthermore, the highly active green algal hydrogenase HydA1{sub Cr} from Chlamydomonas reinhardtii was heterologously expressed in C. acetobutylicum, and purified with increased yield (1 mg protein per liter of cells) and high activity (625 {mu}mol H{sub 2} min{sup -1} mg{sup -1}). EPR studies demonstrate intact H-clusters for homologously and heterologously expressed [FeFe] hydrogenases in the CO-inhibited oxidized redox state, and prove the high efficiency of the C. acetobutylicum expression system. (author)

  13. The surprising diversity of clostridial hydrogenases: a comparative genomic perspective

    OpenAIRE

    Calusinska, Magdalena; Happe, Thomas; Joris, Bernard; Wilmotte, Annick

    2010-01-01

    Among the large variety of micro-organisms capable of fermentative hydrogen production, strict anaerobes such as members of the genus Clostridium are the most widely studied. They can produce hydrogen by a reversible reduction of protons accumulated during fermentation to dihydrogen, a reaction which is catalysed by hydrogenases. Sequenced genomes provide completely new insights into the diversity of clostridial hydrogenases. Building on previous reports, we found that [FeFe] hydrogenases are...

  14. Flexibility in Anaerobic Metabolism as Revealed in a Mutant of Chlamydomonas reinhardtii Lacking Hydrogenase Activity

    Energy Technology Data Exchange (ETDEWEB)

    Dubini, A.; Mus, F.; Seibert, M.; Grossman, A. R.; Posewitz, M. C.

    2009-03-13

    The green alga Chlamydomonas reinhardtii has a network of fermentation pathways that become active when cells acclimate to anoxia. Hydrogenase activity is an important component of this metabolism, and we have compared metabolic and regulatory responses that accompany anaerobiosis in wild-type C. reinhardtii cells and a null mutant strain for the HYDEF gene (hydEF-1 mutant), which encodes an [FeFe] hydrogenase maturation protein. This mutant has no hydrogenase activity and exhibits elevated accumulation of succinate and diminished production of CO2 relative to the parental strain during dark, anaerobic metabolism. In the absence of hydrogenase activity, increased succinate accumulation suggests that the cells activate alternative pathways for pyruvate metabolism, which contribute to NAD(P)H reoxidation, and continued glycolysis and fermentation in the absence of O2. Fermentative succinate production potentially proceeds via the formation of malate, and increases in the abundance of mRNAs encoding two malateforming enzymes, pyruvate carboxylase and malic enzyme, are observed in the mutant relative to the parental strain following transfer of cells from oxic to anoxic conditions. Although C. reinhardtii has a single gene encoding pyruvate carboxylase, it has six genes encoding putative malic enzymes. Only one of the malic enzyme genes, MME4, shows a dramatic increase in expression (mRNA abundance) in the hydEF-1 mutant during anaerobiosis. Furthermore, there are marked increases in transcripts encoding fumarase and fumarate reductase, enzymes putatively required to convert malate to succinate. These results illustrate the marked metabolic flexibility of C. reinhardtii and contribute to the development of an informed model of anaerobic metabolism in this and potentially other algae.

  15. Electrocatalytic proton reduction by a model for [NiFeSe] hydrogenases.

    Science.gov (United States)

    Gezer, Gamze; Durán Jiménez, Dinesh; Siegler, Maxime A; Bouwman, Elisabeth

    2017-06-13

    Two new heterodinuclear nickel-iron complexes [Ni(pbSmSe)FeCpCO]PF6 and [Ni(xbSmSe)FeCpCO]PF6 were synthesized as mimics of the [NiFeSe] hydrogenase active site (HCp = cyclopentadiene; H2pbSmSe = 1,9-diselenol-3,7-dithia-2,2,8,8-tetramethylnonane; H2xbSmSe = 1,2,-bis(2-thiabutyl-3,3-dimethyl-4-selenol)benzene). The compounds were characterized by single crystal X-ray diffraction and cyclic voltammetry. X-ray structure determinations showed that in both NiFe complexes the nickel(ii) center is in a square-planar S2Se2 environment; the two selenolate donors are bridging to the iron(ii) center that is further coordinated to an η(5)-cyclopentadienyl group and a carbon monoxide ligand. Electrochemical studies showed that the complex [Ni(pbSmSe)FeCpCO]PF6 is an electrocatalyst for the production of H2 in DMF in the presence of acetic acid at -2.1 V vs. Fc(+)/Fc; a foot-of-the-wave (FOWA) analysis of the catalytic currents yielded an estimation of kobs of 24 s(-1).

  16. Genetic analysis of nifF and nifA and site-directed mutagenesis of nifE in Azotobacter vinelandii

    OpenAIRE

    Bennett, Lisa Tracy

    1989-01-01

    Nitrogenase-catalyzed nitrogen fixation is a biochemically and genetically complex process requiring the participation of a number of different nif (nitrogen fixation) gene products. The nifF (electron transport), nifA (nif gene regulation) and nifE (FeMo-cofactor biosynthesis) genes from Azotobacter vinelandii were genetically analyzed. The nucleotide sequence of the nifF gene, which encodes a flavodoxin, was determined. Specific mutation strains indicated that in A vineland...

  17. Replacing Electron Transport Cofactors with Hydrogenases

    KAUST Repository

    Laamarti, Rkia

    2016-12-01

    Enzymes have found applications in a broad range of industrial production processes. While high catalytic activity, selectivity and mild reaction conditions are attractive advantages of the biocatalysts, particularly costs arising from required cofactors pose a sever limitation. While cofactor-recycling systems are available, their use implies constraints for process set-up and conditions, which are a particular problem e.g. for solid-gas-phase reactions. Several oxidoreductases are able to directly exchange electrons with electrodes. Hence, the co-immobilization of both, an electron-utilizing and an electron-generating oxidoreductase on conductive nanoparticles should facilitate the direct electron flow from an enzymatic oxidation to a reduction reaction circumventing redox-cofactors requirements. In such a set-up, hydrogenases could generate and provide electrons directly form gaseous hydrogen. This thesis describes the co-immobilization of the oxygen tolerant hydrogenases from C. eutropha or C. metallidurans and cytochrome P450BM3 as test system. Conductive material in the form of carbon nanotubes (CNT) serves as a suitable support. A combination of the hydrogenase and the catalytic domain of P450BM3 immobilized on carbon nanotubes were tested for the oxidation of lauric acid in the presence of hydrogen instead of an electron-transport cofactor. The GC-MS analysis reveals the conversion of 4% of lauric acid (LA) into three products, which correspond to the hydroxylated lauric acid in three different positions with a total turnover (TON) of 34. The product distribution is similar to that obtained when using the wildtype P450BM3 with the nicotinamide adenine dinucleotide phosphate (NADPH) cofactor. Such electronic coupling couldn’t be achieved for the conversion of other substrates such as propane and cyclohexane, probably due to the high uncoupling rate within the heme-domain of cytochrome P450BM3 when unnatural substrates are introduced.

  18. [Fe]-hydrogenases in green algae: photo-fermentation and hydrogen evolution under sulfur deprivation

    Energy Technology Data Exchange (ETDEWEB)

    Winkler, M.; Hemschemeier, A.; Happe, T. [Botanisches Institut der Universitat Bonn (Germany); Gotor, C. [CSIC y Universidad de Sevilla (Spain). Instituto de Bioquimica Vegetal y Fotosintesis; Melis, A. [University of California, Berkeley, CA (United States). Department of Plant and Microbial Biology

    2002-12-01

    Recent studies indicate that [Fe]-hydrogenases and H{sub 2} metabolism are widely distributed among green algae. The enzymes are simple structured and catalyze H{sub 2} evolution with similar rates than the more complex [Fe]-hydrogenases from bacteria. Different green algal species developed diverse strategies to survive under sulfur deprivation. Chlamydomonas reinhardtii evolves large quantities of hydrogen gas in the absence of sulfur. In a sealed culture of C. reinhardtii, the photosynthetic O{sub 2} evolution rate drops below the rate of respiratory O{sub 2} consumption due to a reversible inhibition of photosystem II, thus leading to an intracellular anaerobiosis. The algal cells survive under these anaerobic conditions by switching their metabolism to a kind of photo-fermentation. Although possessing a functional [Fe]-hydrogenase gene, the cells of Scenedesmus obliquus produce no significant amounts of H{sub 2} under S-depleted conditions. Biochemical analyses indicate that S. obliquus decreases almost the complete metabolic activities while maintaining a low level of respiratory activity. (author)

  19. O2-stable membrane-bound [NiFe]hydrogenase from a newly isolated Citrobacter sp. S-77.

    Science.gov (United States)

    Eguchi, Shigenobu; Yoon, Ki-Seok; Ogo, Seiji

    2012-11-01

    Hydrogenases are of great interest due to their potential use in H(2)-based technology. However, most hydrogenases are highly sensitive to O(2), which have been the major bottleneck in hydrogenase studies. Here we report an O(2)-stable membrane-bound [NiFe]hydrogenase (MBH) purified from a newly isolated strain, S-77. According to the 16S rRNA gene sequence and phylogenetic analysis of the strain S-77, it belongs to the genus of Citrobacter. In vitro experiments using the cytoplasmic membrane of strain S-77 suggested that a cytochrome b acts as the physiological electron acceptor of the MBH. The purified MBH was composed of a dimer of heterodimers, consisting of two distinct subunits with the molecular weights of 58.5 and 38.5 kDa. The enzyme showed a specific activity for H(2)-oxidation of 661 U/mg, which is 35-fold greater than that for H(2)-production of 18.7 U/mg. Notably, the MBH showed a remarkable O(2)-stability, maintaining almost 95% of its original activity even after incubation for 30 h in air at 4°C. These results suggest that the O(2)-stable MBH may play an important role in the H(2)-metabolic pathway under the aerobic conditions of Citrobacter sp. S-77. This is the first report of the purification and biochemical characterization of an O(2)-stable MBH from the genus of Citrobacter.

  20. Hydrogen bioelectrooxidation on gold nanoparticle-based electrodes modified by Aquifex aeolicus hydrogenase: Application to hydrogen/oxygen enzymatic biofuel cells.

    Science.gov (United States)

    Monsalve, Karen; Roger, Magali; Gutierrez-Sanchez, Cristina; Ilbert, Marianne; Nitsche, Serge; Byrne-Kodjabachian, Deborah; Marchi, Valérie; Lojou, Elisabeth

    2015-12-01

    For the first time, gold nanoparticle-based electrodes have been used as platforms for efficient immobilization of the [NiFe] hydrogenase from the hyperthermophilic bacterium Aquifex aeolicus. AuNPs were characterized by electronic microscopy, dynamic light scattering and UV-Vis spectroscopy. Two sizes around 20.0±5.3 nm and 37.2±4.3 nm nm were synthesized. After thiol-based functionalization, the AuNPs were proved to allow direct H2 oxidation over a large range of temperatures. A high current density up to 1.85±0.15 mA·cm(-2) was reached at the smallest AuNPs, which is 170 times higher than the one recorded at the bare gold electrode. The catalytic current was especially studied as a function of the AuNP size and amount, and procedure for deposition. A synergetic effect between the AuNP porous deposit and the increase surface area was shown. Compared to previously used nanomaterials such as carbon nanofibers, the covalent grafting of the enzyme on the thiol-modified gold nanoparticles was shown to enhance the stability of the hydrogenase. This bioanode was finally coupled to a biocathode where BOD from Myrothecium verrucaria was immobilized on AuNP-based film. The performance of the so-mounted H2/O2 biofuel cell was evaluated, and a power density of 0.25 mW·cm(-2) was recorded.

  1. Transcription and Regulation of the Bidirectional Hydrogenase in the Cyanobacterium Nostoc sp. Strain PCC 7120▿

    Science.gov (United States)

    Sjöholm, Johannes; Oliveira, Paulo; Lindblad, Peter

    2007-01-01

    The filamentous, heterocystous cyanobacterium Nostoc sp. strain PCC 7120 (Anabaena sp. strain PCC 7120) possesses an uptake hydrogenase and a bidirectional enzyme, the latter being capable of catalyzing both H2 production and evolution. The completely sequenced genome of Nostoc sp. strain PCC 7120 reveals that the five structural genes encoding the bidirectional hydrogenase (hoxEFUYH) are separated in two clusters at a distance of approximately 8.8 kb. The transcription of the hox genes was examined under nitrogen-fixing conditions, and the results demonstrate that the cluster containing hoxE and hoxF can be transcribed as one polycistronic unit together with the open reading frame alr0750. The second cluster, containing hoxU, hoxY, and hoxH, is transcribed together with alr0763 and alr0765, located between the hox genes. Moreover, alr0760 and alr0761 form an additional larger operon. Nevertheless, Northern blot hybridizations revealed a rather complex transcription pattern in which the different hox genes are expressed differently. Transcriptional start points (TSPs) were identified 66 and 57 bp upstream from the start codon of alr0750 and hoxU, respectively. The transcriptions of the two clusters containing the hox genes are both induced under anaerobic conditions concomitantly with the induction of a higher level of hydrogenase activity. An additional TSP, within the annotated alr0760, 244 bp downstream from the suggested translation start codon, was identified. Electrophoretic mobility shift assays with purified LexA from Nostoc sp. strain PCC 7120 demonstrated specific interactions between the transcriptional regulator and both hox promoter regions. However, when LexA from Synechocystis sp. strain PCC 6803 was used, the purified protein interacted only with the promoter region of the alr0750-hoxE-hoxF operon. A search of the whole Nostoc sp. strain PCC 7120 genome demonstrated the presence of 216 putative LexA binding sites in total, including recA and rec

  2. Magneto-thermoelectric effects in NiFe thin films

    Energy Technology Data Exchange (ETDEWEB)

    Schmid, Maximilian

    2015-11-01

    In this thesis magneto-thermoelectric effects are investigated in a systematic way to separate the transverse spin Seebeck effect from other parasitic effects like the anomalous Nernst effect. In contrast to the first studies found in the literature, in NiFe thin films a contribution of the transverse spin Seebeck effect can be excluded. This surprising outcome was crosschecked in a variety of different sample layouts and collaborations with other universities to ensure the validity of these results. In general, this thesis solves a long time discussion about the existence of the transverse spin Seebeck effect in NiFe films and supports the importance of control measurements for the scientific community. Even if such ''negative'' results may not be the award winning ones, new discoveries should be treated with constructive criticism and be checked carefully by the scientific community.

  3. Magnetization dynamics in rare earth doped NiFe films

    Energy Technology Data Exchange (ETDEWEB)

    Kiessling, Matthias; Woltersdorf, Georg; Back, Christian [Institut fuer Experimentelle und Angewandte Physik, Universitaet Regensburg, D- 93040 Regensburg (Germany); Thiele, Jan-Ulrich; Schabes, Manfred [Hitachi Global Storage Technologies, 3403 Yerba Buena Road, San Jose, CA 95135 (United States)

    2007-07-01

    The influence of rare earth dopants on the damping parameter and the resulting possibility to control this parameter were investigated. In our experiments NiFe films were doped with Dysprosium, Holmium, Terbium, and Gadolinium. The magnetization dynamics of these rare earth doped films was mainly studied by means of ferromagnetic resonance (FMR) and network-analyzer ferromagnetic resonance. It is demonstrated that the doping of a NiFe film by a small amount of rare earth elements (Holmium, Terbium and Dysprosium) greatly effects its magnetic relaxation rate. This additional damping is proportional to the doping level. Compared to the pure NiFe film it is possible to increase the damping parameter of the magnetic film by two orders of magnitude. On the other hand Gadolinium as a dopant has no influence on the damping parameter. For small dopant concentrations the in and out-of-plane FMR measurements at various frequencies can be well described by the same damping parameter. This is expected for the Gilbert damping term in the equation of motion. Therefore the increased damping can be attributed to an increased rate of transfer of angular momentum from the spin system to the lattice.

  4. A threonine stabilizes the NiC and NiR catalytic intermediates of [NiFe]-hydrogenase.

    Science.gov (United States)

    Abou-Hamdan, Abbas; Ceccaldi, Pierre; Lebrette, Hugo; Gutiérrez-Sanz, Oscar; Richaud, Pierre; Cournac, Laurent; Guigliarelli, Bruno; De Lacey, Antonio L; Léger, Christophe; Volbeda, Anne; Burlat, Bénédicte; Dementin, Sébastien

    2015-03-27

    The heterodimeric [NiFe] hydrogenase from Desulfovibrio fructosovorans catalyzes the reversible oxidation of H2 into protons and electrons. The catalytic intermediates have been attributed to forms of the active site (NiSI, NiR, and NiC) detected using spectroscopic methods under potentiometric but non-catalytic conditions. Here, we produced variants by replacing the conserved Thr-18 residue in the small subunit with Ser, Val, Gln, Gly, or Asp, and we analyzed the effects of these mutations on the kinetic (H2 oxidation, H2 production, and H/D exchange), spectroscopic (IR, EPR), and structural properties of the enzyme. The mutations disrupt the H-bond network in the crystals and have a strong effect on H2 oxidation and H2 production turnover rates. However, the absence of correlation between activity and rate of H/D exchange in the series of variants suggests that the alcoholic group of Thr-18 is not necessarily a proton relay. Instead, the correlation between H2 oxidation and production activity and the detection of the NiC species in reduced samples confirms that NiC is a catalytic intermediate and suggests that Thr-18 is important to stabilize the local protein structure of the active site ensuring fast NiSI-NiC-NiR interconversions during H2 oxidation/production.

  5. Magnetic properties of electroplated nano/microgranular NiFe thin films for rf application

    NARCIS (Netherlands)

    Zhuang, Y.; Vroubel, M.; Rejaei, B.; Burghartz, J.N.; Attenborough, K.

    2005-01-01

    A granular NiFe thin film with large in-plane magnetic anisotropy and high ferromagnetic-resonance frequency developed for radio-frequency integrated circuit (IC) applications is presented. During the deposition, three-dimensional (3D) growth occurs, yielding NiFe grains (ϕ ∼ 1.0 μm). Nanonuclei (ϕ

  6. CO methanation over supported bimetallic Ni-Fe catalysts: From computational studies towards catalyst optimization

    DEFF Research Database (Denmark)

    Kustov, Arkadii; Frey, Anne Mette; Larsen, Kasper Emil

    2007-01-01

    DFT calculations combined with a computational screening method have previously shown that bimetallic Ni-Fe alloys should be more active than the traditional Ni-based catalyst for CO methanation. That was confirmed experimentally for a number of bimetallic Ni-Fe catalysts supported on MgAl2O4. He...

  7. Electrical, magnetic, and corrosion resistance properties of TiO2 nanotubes filled with NiFe2O4 quantum dots and Ni-Fe nanoalloy

    Science.gov (United States)

    Bahgat, Mohamed; Farghali, Ahmed A.; Moustafa, Ahmed F.; Khedr, Mohamed H.; Mohassab-Ahmed, Mohassab Y.

    2013-06-01

    This work was carried out as an integral part of a project aiming to improve the catalytic, electrical, magnetic, and mechanical properties of synthesized TiO2NTs filled with metal ferrites. TiO2 nanotubes in the anatase-phase (TiO2NTs) were prepared using a hydrothermal method followed by ion exchange and phase transformation. The obtained TiO2NTs were filled with NiFe2O4 quantum dots (QDs) and then reacted at 600 °C in a reducing atmosphere to produce TiO2NTs filled with Ni-Fe nanoalloy. The effect of the TiO2NTs' coating on the dissolution rate of Ni-Fe nanoalloy in 0.5 M HCl solution was monitored chemically using a weight-loss technique that was performed at different temperatures. The TiO2NTs' coating exhibited high protective performance and amazing corrosion resistance. The magnetic properties of the TiO2NTs filled with NiFe2O4 QDs and Ni-Fe nanoalloy compacts were analyzed by a vibrating sample magnetometer. The electrical conductivity-temperature dependence of anatase TiO2NTs, anatase TiO2NTs filled with NiFe2O4 quantum dots, anatase TiO2NTs filled with Ni-Fe nanoalloy, and NiFe2O4 was measured in the temperature range of 25-850 °C. The conductivity increased with temperature, indicating the semiconductor-like nature of the sample. During cooling, the conductivity retains values higher than that obtained during heating.

  8. Oxygen-tolerant hydrogenases in hydrogen-based technologies.

    Science.gov (United States)

    Friedrich, Bärbel; Fritsch, Johannes; Lenz, Oliver

    2011-06-01

    To develop a viable H2 technology, production of H2 has to be significantly enlarged by using renewable resources. One option of generating H2 is the photosynthetic conversion of sunlight and water directly to H2 and O2. Photosystems and hydrogenases are currently being exploited for the design of efficient H2-producing systems that require highly active and O2-tolerant biocatalysts. This communication focuses on two challenging features: hydrogenases that produce H2 in the presence of O2, and direct electron transfer between photosystem I (PS I) and hydrogenase. The latter is accomplished by connecting both modules through a protein fusion or a synthetic molecular wire. These are first steps toward a photosynthetic microbial cell or a semi-synthetic system that may be employed in future H2-based technologies.

  9. Maturation of [FeFe]-hydrogenases: Structures and mechanisms

    Energy Technology Data Exchange (ETDEWEB)

    Nicolet, Yvain; Fontecilla-Camps, Juan C. [Laboratoire de Cristallographie et Cristallogenese des Proteines, Institut de Biologie Structurale J.P. Ebel, Universite Joseph Fourier, Grenoble, CNRS, UMR 5075, CEA, DSV/IBS, 41 rue J. Horowitz F-38027 Grenoble cedex 1 (France); Fontecave, Marc [Laboratoire de Chimie et Biologie des Metaux; Universite Joseph Fourier, Grenoble, CNRS, UMR 5249, CEA, DSV/iRTSV, 17 rue des Martyrs F-38054 Grenoble cedex 9 (France); College de France, 11 place Marcelin-Berthelot 75231 Paris cedex 05 (France)

    2010-10-15

    Maturation of [FeFe]-hydrogenases, consisting in the synthesis and assembly of a di-iron center with a dithiolate bridging ligand as well as CO and CN ligands, depends on the concerted action of three metalloproteins, HydE, HydF and HydG. HydE and HydG are ''Radical-SAM'' enzymes involved in the synthesis of the ligands. HydF is proposed to function as a scaffold protein in which the di-iron center is assembled before being transferred to the hydrogenase. Here we review the current knowledge regarding the structure of the three maturases and the mechanisms of synthesis and assembly of the di-iron center of [FeFe]-hydrogenases. (author)

  10. Recombinant antibodies for specific detection of clostridial [Fe-Fe] hydrogenases

    Science.gov (United States)

    Mangayil, Rahul; Karp, Matti; Lamminmäki, Urpo; Santala, Ville

    2016-01-01

    Biological hydrogen production is based on activity of specific enzymes called hydrogenases. Hydrogenases are oxygen sensitive metalloenzymes containing Ni and/or Fe atoms at the active site, catalyzing reversible reduction of protons. Generally, [Fe-Fe] hydrogenases prefer proton reduction to molecular hydrogen, a potential energy carrier molecule that can be produced by bioprocesses in sustainable manner. Thus, monitoring tools have been developed to study the relationship between [Fe-Fe] hydrogenases and biohydrogen production in bioreactors at DNA and RNA levels. In the present study, novel molecular tools are introduced for quantitative monitoring of clostridial [Fe-Fe] hydrogenases at the protein level. Aerobic and anaerobic biopanning (for inactive and active [Fe-Fe] hydrogenase, respectively) of phage displayed single-chain variable fragment (scFv) antibody libraries aided in isolating nine potential scFvs. The enriched antibodies demonstrated high specificity towards Clostridium spp. [Fe-Fe] hydrogenases allowing detection from pure and mixed cultures. Additionally, the antibodies showed different binding characteristics towards hydrogenase catalytic states, providing a possible means for functional detection of clostridial [Fe-Fe] hydrogenases. From hydrogenase-antibody interaction studies we observed that though antibody binding reduced the enzyme catalytic activity, it facilitated to retain hydrogen evolution from oxygen exposed hydrogenases. PMID:27786270

  11. Recombinant antibodies for specific detection of clostridial [Fe-Fe] hydrogenases.

    Science.gov (United States)

    Mangayil, Rahul; Karp, Matti; Lamminmäki, Urpo; Santala, Ville

    2016-10-27

    Biological hydrogen production is based on activity of specific enzymes called hydrogenases. Hydrogenases are oxygen sensitive metalloenzymes containing Ni and/or Fe atoms at the active site, catalyzing reversible reduction of protons. Generally, [Fe-Fe] hydrogenases prefer proton reduction to molecular hydrogen, a potential energy carrier molecule that can be produced by bioprocesses in sustainable manner. Thus, monitoring tools have been developed to study the relationship between [Fe-Fe] hydrogenases and biohydrogen production in bioreactors at DNA and RNA levels. In the present study, novel molecular tools are introduced for quantitative monitoring of clostridial [Fe-Fe] hydrogenases at the protein level. Aerobic and anaerobic biopanning (for inactive and active [Fe-Fe] hydrogenase, respectively) of phage displayed single-chain variable fragment (scFv) antibody libraries aided in isolating nine potential scFvs. The enriched antibodies demonstrated high specificity towards Clostridium spp. [Fe-Fe] hydrogenases allowing detection from pure and mixed cultures. Additionally, the antibodies showed different binding characteristics towards hydrogenase catalytic states, providing a possible means for functional detection of clostridial [Fe-Fe] hydrogenases. From hydrogenase-antibody interaction studies we observed that though antibody binding reduced the enzyme catalytic activity, it facilitated to retain hydrogen evolution from oxygen exposed hydrogenases.

  12. Effect of sintering atmosphere on composition and properties of NiFe2O4 ceramic

    Institute of Scientific and Technical Information of China (English)

    田忠良; 张腾; 刘恺; 赖延清; 李劼

    2015-01-01

    NiFe2O4 ceramics were prepared in different sintering atmospheres. The phase compositions, microstructures and mechanical properties were studied. The results show that the stoichiometric compound NiFe2O4 cannot be obtained in vacuum or atmospheres with oxygen contents of 2×10−5, 2×10−4 and 2×10−3, respectively. All the samples sintered in above-mentioned atmospheres contain phases of NiFe2O4 and NiO. With increasing oxygen content, NiFe2O4 content in the ceramic increases, while NiO content appears a contrary trend. In vacuum, NiFe2O4 ceramic has average grain size of 3.94μm, and bending strength of 85.12 MPa. The changes of the phase composition and mechanical properties of NiFe2O4 based cermets are mainly caused by the alteration of their properties of NiFe2O4 ceramic.

  13. Effects of ultrasonic field in pulse electrodeposition of NiFe film on Cu substrate

    Energy Technology Data Exchange (ETDEWEB)

    Balachandran, R. [Faculty of Engineering, Multimedia University, Cyberjaya Campus, 63100, Cyberjaya (Malaysia); Yow, H.K. [Faculty of Engineering, Multimedia University, Cyberjaya Campus, 63100, Cyberjaya (Malaysia)], E-mail: hkyow@mmu.edu.my; Ong, B.H. [Faculty of Engineering, Multimedia University, Cyberjaya Campus, 63100, Cyberjaya (Malaysia); Manickam, R. [Electronics Faculty, Tyndale Education Group Pte Ltd., 188942 (Singapore); Saaminathan, V. [School of Material Science and Engineering, Nanyang Technological University, 639798 (Singapore); Tan, K.B. [Department of Chemistry, Universiti Putra Malaysia, Serdang, 43400 (Malaysia)

    2009-07-29

    NiFe film was pulse electrodeposited on conductive Cu substrate under galvanostatic mode in the presence of an ultrasonic field. The NiFe film electrodeposited was subjected to structural and surface analyses by X-ray diffraction, energy dispersive X-ray spectroscopy, surface profiling and scanning electron microscopy, respectively. The results show that the ultrasonic field has significantly improved the surface roughness, reduced the spherical grain size in the range from 490-575 nm to 90-150 nm, and increased the Ni content from 76.08% to 79.74% in the NiFe film electrodeposited.

  14. Interface-assisted magnetoresistance behavior for ultrathin NiFe films

    Energy Technology Data Exchange (ETDEWEB)

    Jiang, Shao-Long; Chen, Xi; Yang, Kang [Department of Materials Physics and Chemistry, University of Science and Technology Beijing, Beijing 100083 (China); Han, Gang [Institute for Advanced Materials and Technology, University of Science and Technology Beijing, Beijing 100083 (China); Teng, Jiao; Li, Xu-Jing; Yang, Guang; Liu, Qian-Qian; Liu, Yi-Wei [Department of Materials Physics and Chemistry, University of Science and Technology Beijing, Beijing 100083 (China); Ding, Lei [School of Materials and Chemical Engineering, Hainan University, Haikou 570228 (China); Yu, Guang-Hua, E-mail: ghyu@mater.ustb.edu.cn [Department of Materials Physics and Chemistry, University of Science and Technology Beijing, Beijing 100083 (China)

    2015-11-01

    Interface-assisted magnetoresistance (MR) behavior has been studied in Ta/MgO/NiFe/MgO/Ta multilayers by inserting a Mg metal layer between the NiFe layer and the top MgO layer. It is shown that MR ratio is about 31% larger than that in the films without Mg insertion. X-ray photoelectron spectroscopy and high resolution transmission electron microscope analyses show that the enhanced MR is primarily ascribed to effective control of chemical states at the NiFe/MgO interface and crystallization of the top MgO layer. - Highlights: • We studied magnetic and electric transport properties of ultrathin NiFe films. • Interface chemical states have strong influence on MR in NiFe films. • Crystallization of the top MgO layer has influence on MR in NiFe films.

  15. Formation of NiFe2O4/Expanded Graphite Nanocomposites with Superior Lithium Storage Properties

    Science.gov (United States)

    Xiao, Yinglin; Zai, Jiantao; Tian, Bingbing; Qian, Xuefeng

    2017-07-01

    A NiFe2O4/expanded graphite (NiFe2O4/EG) nanocomposite was prepared via a simple and inexpensive synthesis method. Its lithium storage properties were studied with the goal of applying it as an anode in a lithium-ion battery. The obtained nanocomposite exhibited a good cycle performance, with a capacity of 601 mAh g-1 at a current of 1 A g-1 after 800 cycles. This good performance may be attributed to the enhanced electrical conductivity and layered structure of the EG. Its high mechanical strength could postpone the disintegration of the nanocomposite structure, efficiently accommodate volume changes in the NiFe2O4-based anodes, and alleviate aggregation of NiFe2O4 nanoparticles.

  16. Evaluation of Young's Modulus and Residual Stress of NiFe Film by Microbridge Testing

    Institute of Scientific and Technical Information of China (English)

    Zhimin ZHOU; Yong ZHOU; Mingjun WANG; Chunsheng YANG; Ji'an CHEN; Wen DING; Xiaoyu GAO; Taihua ZHANG

    2006-01-01

    Microbridge testing was used to measure the Young's modulus and residual stress of metallic films. Samples of freestanding NiFe film microbridge were fabricated by microelectromechanical systems. Special ceramic shaft structure was designed to solve the problem of getting the load-deflection curve of NiFe film microbridge by the Nanoindenter XP system with normal Berkovich probe. Theoretical analysis of load-deflection curves of the microbridges was proposed to evaluate the Young's modulus and residual stress of the films simultaneously. The calculated results based on experimental measurements show that the average Young's modulus and residual stress for the electroplated NiFe films are 203.2 GPa and 333.0 MPa, respectively, while the Young's modulus measured by the Nano-hardness method is 209.6±11.8 GPa for the thick NiFe film with silicon substrate.

  17. Preparation of NiFe binary alloy nanocrystals for nonvolatile memory applications

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    In this work,an idea which applies binary alloy nanocrystal floating gate to nonvolatile memory application was introduced.The relationship between binary alloy’s work function and its composition was discussed theoretically.A nanocrystal floating gate structure with NiFe nanocrystals embedded in SiO2 dielectric layers was fabricated by magnetron sputtering.The micro-structure and composition deviation of the prepared NiFe nanocrystals were also investigated by TEM and EDS.

  18. Hydrogenase activity in aged, nonviable Desulfovibrio vulgaris cultures and its significance in anaerobic biocorrosion.

    Science.gov (United States)

    Chatelus, C; Carrier, P; Saignes, P; Libert, M F; Berlier, Y; Lespinat, P A; Fauque, G; Legall, J

    1987-07-01

    Batch cultures of Desulfovibrio vulgaris stored at 32 degrees C for 10 months have been found to retain 50% of the hydrogenase activity of a 1-day culture. The hydrogenase found in old cultures needs reducing conditions for its activation. Viable cell counts are negative after 6 months, showing that the hydrogenase activity does not depend on the presence of viable cells. These observations are of importance in the understanding of anaerobic biocorrosion of metals caused by depolarization phenomena.

  19. Oxygen-resistant hydrogenases and methods for designing and making same

    Science.gov (United States)

    King, Paul; Ghirardi, Maria Lucia; Seibert, Michael

    2014-03-04

    The invention provides oxygen-resistant iron-hydrogenases ([Fe]-hydrogenases) for use in the production of H.sub.2. Methods used in the design and engineering of the oxygen-resistant [Fe]-hydrogenases are disclosed, as are the methods of transforming and culturing appropriate host cells with the oxygen-resistant [Fe]-hydrogenases. Finally, the invention provides methods for utilizing the transformed, oxygen insensitive, host cells in the bulk production of H.sub.2 in a light catalyzed reaction having water as the reactant.

  20. Catalytic properties of the isolated diaphorase fragment of the NAD-reducing [NiFe]-hydrogenase from Ralstonia eutropha.

    Directory of Open Access Journals (Sweden)

    Lars Lauterbach

    Full Text Available The NAD+-reducing soluble hydrogenase (SH from Ralstonia eutropha H16 catalyzes the H₂-driven reduction of NAD+, as well as reverse electron transfer from NADH to H+, in the presence of O₂. It comprises six subunits, HoxHYFUI₂, and incorporates a [NiFe] H+/H₂ cycling catalytic centre, two non-covalently bound flavin mononucleotide (FMN groups and an iron-sulfur cluster relay for electron transfer. This study provides the first characterization of the diaphorase sub-complex made up of HoxF and HoxU. Sequence comparisons with the closely related peripheral subunits of Complex I in combination with UV/Vis spectroscopy and the quantification of the metal and FMN content revealed that HoxFU accommodates a [2Fe2S] cluster, FMN and a series of [4Fe4S] clusters. Protein film electrochemistry (PFE experiments show clear electrocatalytic activity for both NAD+ reduction and NADH oxidation with minimal overpotential relative to the potential of the NAD+/NADH couple. Michaelis-Menten constants of 56 µM and 197 µM were determined for NADH and NAD+, respectively. Catalysis in both directions is product inhibited with K(I values of around 0.2 mM. In PFE experiments, the electrocatalytic current was unaffected by O₂, however in aerobic solution assays, a moderate superoxide production rate of 54 nmol per mg of protein was observed, meaning that the formation of reactive oxygen species (ROS observed for the native SH can be attributed mainly to HoxFU. The results are discussed in terms of their implications for aerobic functioning of the SH and possible control mechanism for the direction of catalysis.

  1. Production and Application of a Soluble Hydrogenase from Pyrococcus furiosus

    Directory of Open Access Journals (Sweden)

    Chang-Hao Wu

    2015-01-01

    Full Text Available Hydrogen gas is a potential renewable alternative energy carrier that could be used in the future to help supplement humanity’s growing energy needs. Unfortunately, current industrial methods for hydrogen production are expensive or environmentally unfriendly. In recent years research has focused on biological mechanisms for hydrogen production and specifically on hydrogenases, the enzyme responsible for catalyzing the reduction of protons to generate hydrogen. In particular, a better understanding of this enzyme might allow us to generate hydrogen that does not use expensive metals, such as platinum, as catalysts. The soluble hydrogenase I (SHI from the hyperthermophile Pyrococcus furiosus, a member of the euryarchaeota, has been studied extensively and used in various biotechnological applications. This review summarizes the strategies used in engineering and characterizing three different forms of SHI and the properties of the recombinant enzymes. SHI has also been used in in vitro systems for hydrogen production and NADPH generation and these systems are also discussed.

  2. Overproduction of the membrane-bound [NiFe]-hydrogenase in Thermococcus kodakarensis and its effect on hydrogen production

    Directory of Open Access Journals (Sweden)

    Tamotsu eKanai

    2015-08-01

    Full Text Available The hyperthermophilic archaeon Thermococcus kodakarensis can utilize sugars or pyruvate for growth. In the absence of elemental sulfur, the electrons via oxidation of these substrates are accepted by protons, generating molecular hydrogen (H2. The hydrogenase responsible for this reaction is a membrane-bound [NiFe]-hydrogenase (Mbh. In this study, we have examined several possibilities to increase the protein levels of Mbh in T. kodakarensis by genetic engineering. Highest levels of intracellular Mbh levels were achieved when the promoter of the entire mbh operon (TK2080-TK2093 was exchanged to a strong constitutive promoter from the glutamate dehydrogenase gene (TK1431 (strain MHG1. When MHG1 was cultivated under continuous culture using pyruvate-based medium, a nearly 25 % higher specific hydrogen production rate (SHPR of 35.3 mmol H2 g-dcw-1 h-1 was observed at a dilution rate of 0.31 h-1. We also combined mbh overexpression using an even stronger constitutive promoter from the cell surface glycoprotein gene (TK0895 with disruption of the genes encoding the cytosolic hydrogenase (Hyh and an alanine aminotransferase (AlaAT, both of which are involved in hydrogen consumption (strain MAH1. At a dilution rate of 0.30 h-1, the SHPR was 36.2 mmol H2 g-dcw-1 h-1, corresponding to a 28 % increase compared to that of the host T. kodakarensis strain. Increasing the dilution rate to 0.83 h-1 resulted in a SHPR of 120 mmol H2 g-dcw-1 h-1, which is one of the highest production rates observed in microbial fermentation.

  3. Overproduction of the membrane-bound [NiFe]-hydrogenase in Thermococcus kodakarensis and its effect on hydrogen production.

    Science.gov (United States)

    Kanai, Tamotsu; Simons, Jan-Robert; Tsukamoto, Ryohei; Nakajima, Akihito; Omori, Yoshiyuki; Matsuoka, Ryoji; Beppu, Haruki; Imanaka, Tadayuki; Atomi, Haruyuki

    2015-01-01

    The hyperthermophilic archaeon Thermococcus kodakarensis can utilize sugars or pyruvate for growth. In the absence of elemental sulfur, the electrons via oxidation of these substrates are accepted by protons, generating molecular hydrogen (H2). The hydrogenase responsible for this reaction is a membrane-bound [NiFe]-hydrogenase (Mbh). In this study, we have examined several possibilities to increase the protein levels of Mbh in T. kodakarensis by genetic engineering. Highest levels of intracellular Mbh levels were achieved when the promoter of the entire mbh operon (TK2080-TK2093) was exchanged to a strong constitutive promoter from the glutamate dehydrogenase gene (TK1431) (strain MHG1). When MHG1 was cultivated under continuous culture conditions using pyruvate-based medium, a nearly 25% higher specific hydrogen production rate (SHPR) of 35.3 mmol H2 g-dcw(-1) h(-1) was observed at a dilution rate of 0.31 h(-1). We also combined mbh overexpression using an even stronger constitutive promoter from the cell surface glycoprotein gene (TK0895) with disruption of the genes encoding the cytosolic hydrogenase (Hyh) and an alanine aminotransferase (AlaAT), both of which are involved in hydrogen consumption (strain MAH1). At a dilution rate of 0.30 h(-1), the SHPR was 36.2 mmol H2 g-dcw(-1) h(-1), corresponding to a 28% increase compared to that of the host T. kodakarensis strain. Increasing the dilution rate to 0.83 h(-1) or 1.07 h(-1) resulted in a SHPR of 120 mmol H2 g-dcw(-1) h(-1), which is one of the highest production rates observed in microbial fermentation.

  4. [FeFe]-hydrogenase models assembled into vesicular structures.

    Science.gov (United States)

    Menzel, Kristin; Apfel, Ulf-Peter; Wolter, Nonio; Rüger, Ronny; Alpermann, Theodor; Steiniger, Frank; Gabel, Detlef; Förster, Stephan; Weigand, Wolfgang; Fahr, Alfred

    2014-03-01

    Compartmentalization is a major prerequisite for the origin of life on earth according to Wächtershäuser "Iron-Sulfur-World". The hypothesis is mainly based on an autocatalytic inorganic energy reproducing redox system consisting of iron and sulfur as requirement for the subsequent synthesis of complex organic structures. Here, we modified [FeFe]-hydrogenase models by means of covalent coupling to either oleic acid or the amphiphilic block copolymer polybutadiene-polyethyleneoxide (PB-PEO) and incorporated those into the membranes of vesicles composed of phospholipids (liposomes) or the unmodified amphiphilic polymer (polymersomes). We employed a [2Fe-2S] cluster as a hydrogenase model, since these structures are known to be suitable catalysts for the generation of H2 in the presence of weak acids. Successful incorporation was confirmed by spectrophotometric iron quantification and the vesicles formed were characterized by size determination (photon correlation spectroscopy (PCS)), and zeta potential as well as by cryo-transmission electron microscopy (Cryo-TEM). The modified models could be incorporated into liposomes or polymersomes up to molar proportions of 3.15% and 28%, respectively. Due to the immobilization in vesicular bilayers the [FeFe]-hydrogenase models can even exhibit catalytic action under the particular conditions of the intravesicular microenvironment. Our results suggest that the vesicular systems described may be applied as a nanoreactor for the reduction of encapsulated substances by generating hydrogen and thus as a minimal cell model.

  5. Wiring of Photosystem II to Hydrogenase for Photoelectrochemical Water Splitting.

    Science.gov (United States)

    Mersch, Dirk; Lee, Chong-Yong; Zhang, Jenny Zhenqi; Brinkert, Katharina; Fontecilla-Camps, Juan C; Rutherford, A William; Reisner, Erwin

    2015-07-08

    In natural photosynthesis, light is used for the production of chemical energy carriers to fuel biological activity. The re-engineering of natural photosynthetic pathways can provide inspiration for sustainable fuel production and insights for understanding the process itself. Here, we employ a semiartificial approach to study photobiological water splitting via a pathway unavailable to nature: the direct coupling of the water oxidation enzyme, photosystem II, to the H2 evolving enzyme, hydrogenase. Essential to this approach is the integration of the isolated enzymes into the artificial circuit of a photoelectrochemical cell. We therefore developed a tailor-made hierarchically structured indium-tin oxide electrode that gives rise to the excellent integration of both photosystem II and hydrogenase for performing the anodic and cathodic half-reactions, respectively. When connected together with the aid of an applied bias, the semiartificial cell demonstrated quantitative electron flow from photosystem II to the hydrogenase with the production of H2 and O2 being in the expected two-to-one ratio and a light-to-hydrogen conversion efficiency of 5.4% under low-intensity red-light irradiation. We thereby demonstrate efficient light-driven water splitting using a pathway inaccessible to biology and report on a widely applicable in vitro platform for the controlled coupling of enzymatic redox processes to meaningfully study photocatalytic reactions.

  6. Structural Characterization of the Novel and Thermal Stable Hydrogenases from the Purple Sulfur Bacteria Thiocapsa Roseopersicina and Lamprobacter Modestohalophilus

    Science.gov (United States)

    2011-08-01

    a molecules of hydrogenase from T. roseopersicina form a ring-shaped hexameric complex with D3 symmetry. It is assumed that the formation of such...Optimization of purification of the hydrogenase from L. modestohalophilus 8 3.4. Optimization of the purification of hydrogenase complex ...activity were pooled and concentrated by ultrafiltration . Methods of preparation of crystal and study of 3D structure of hydrogenase from T

  7. Enhanced Hydrogen Production by Co-cultures of Hydrogenase and Nitrogenase in Escherichia coli.

    Science.gov (United States)

    Lee, Hyun Jeong; Sekhon, Simranjeet Singh; Kim, Young Su; Park, Ju-Yong; Kim, Yang-Hoon; Min, Jiho

    2016-03-01

    Rhodobacter sphaeroides is a bacterium that can produce hydrogen by interaction with hydrogenase and nitrogenase. We report a hydrogen production system using co-cultivation of hydrogenase in liquid medium and immobilized nitrogenase in Escherichia coli. The recombinant plasmid has been constructed to analyze the effect of hydrogen production on the expression of hupSL hydrogenase and nifHDK nitrogenase isolated from R. sphaeroides. All recombinant E. coli strains were cultured anaerobically, and cells for nitrogenase were immobilized in agar gel, whereas cells for hydrogenase were supplemented on the nitrogenase agar gel. The hupSL hydrogenase has been observed to enhance hydrogen production and hydrogenase activity under co-culture with nifHDK nitrogenase. The maximum hydrogen production has been obtained at an agar gel concentration and a cell concentration for co-culture of 2 % and 6.4 × 10(8) CFU. Thus, co-culture of hupSL hydrogenase and nifHDK nitrogenase provides a promising route for enhancing the hydrogen production and hydrogenase activity.

  8. The [FeFe] hydrogenase of Nyctotherus ovalis has a chimeric origin

    NARCIS (Netherlands)

    Boxma, B.; Ricard, G.; Hoek, van A.H.A.M.; Severing, E.; Moon-van der Staay, S.Y.; Staay, van der G.W.M.; Alen, T.A.; Graaf, de R.M.; Cremers, G.; Kwantes, M.; McEwan, N.R.; Newbold, C.J.; Jouany, J.P.; Michalowski, T.; Pristas, P.; Huynen, M.A.; Hackstein, J.H.P.

    2007-01-01

    Background The hydrogenosomes of the anaerobic ciliate Nyctotherus ovalis show how mitochondria can evolve into hydrogenosomes because they possess a mitochondrial genome and parts of an electron-transport chain on the one hand, and a hydrogenase on the other hand. The hydrogenase permits direct

  9. The [FeFe] hydrogenase of Nyctotherus ovalis has a chimeric origin

    NARCIS (Netherlands)

    Boxma, B.; Ricard, G.; Hoek, A.H.A.M. van; Severing, E.; Moon-van der Staay, S.Y.; Staay, G.W.M. van der; Alen, T.A. van; Graaf, R.M. de; Cremers, G.; Kwantes, M.; McEwan, N.R.; Newbold, C.J.; Jouany, J.P.; Michalowski, T.; Pristas, P.; Huynen, M.A.; Hackstein, J.H.P.

    2007-01-01

    BACKGROUND: The hydrogenosomes of the anaerobic ciliate Nyctotherus ovalis show how mitochondria can evolve into hydrogenosomes because they possess a mitochondrial genome and parts of an electron-transport chain on the one hand, and a hydrogenase on the other hand. The hydrogenase permits direct

  10. Practical applications of hydrogenase I from Pyrococcus furiosus for NADPH generation and regeneration

    NARCIS (Netherlands)

    Mertens, R.; Greiner, L.; Ban, van den E.C.D.; Haaker, H.B.C.M.; Liese, A.

    2003-01-01

    The soluble hydrogenase I (H-2:NADP(+) oxidoreductase, EC 1.18.99.1) from the marine hyperthermophilic strain of the archaeon Pyrococcus furiosus was partially purified by anion-exchange chromatography. This P furiosus hydrogenase I preparation (PF H(2)ase I) has been used as biocatalyst in the enzy

  11. A hydrogenosomal [Fe]-hydrogenase from the anaerobic chytrid Neocallimastix sp. L2

    NARCIS (Netherlands)

    Voncken, Frank G.J.; Boxma, Brigitte; Hoek, Angela H.A.M. van; Akhmanova, Anna S.; Vogels, Godfried D.; Huynen, Martijn; Veenhuis, Marten; Hackstein, Johannes H.P.

    2002-01-01

    The presence of a [Fe]-hydrogenase in the hydrogenosomes of the anaerobic chytridiomycete fungus Neocallimastix sp. L2 has been demonstrated by immunocytochemistry, subcellular fractionation, Western-blotting and measurements of hydrogenase activity in the presence of various concentrations of carbo

  12. A hydrogenosomal [Fe]-hydrogenase from the anaerobic chytrid Neocallimastix sp L2

    NARCIS (Netherlands)

    Voncken, Frank G.J.; Boxma, Brigitte; Hoek, Angela H.A.M. van; Akhmanova, Anna S.; Vogels, Godfried D.; Huynen, Martijn; Veenhuis, Marten; Hackstein, Johannes H.P.

    2002-01-01

    The presence of a [Fe]-hydrogenase in the hydrogenosomes of the anaerobic chytridiomycete fungus Neocallimastix sp. L2 has been demonstrated by immunocytochemistry, subcellular fractionation, Western-blotting, and measurements of hydrogenase activity in the presence of various concentrations of carb

  13. New hypotheses for hydrogenase implication in the corrosion of mild steel

    Energy Technology Data Exchange (ETDEWEB)

    Mehanna, Maha; Basseguy, Regine; Delia, Marie-Line [Laboratoire de Genie Chimique (LGC) CNRS-INPT, 5 rue Paulin Talabot BP 1301, 31106 Toulouse (France); Girbal, Laurence; Demuez, Marie [Laboratoire d' Ingenierie des Systemes Biologiques et des Procedes (LISBP) CNRS-INSA, 135 Avenue de Rangueil, 31077 Toulouse (France); Bergel, Alain [Laboratoire de Genie Chimique (LGC) CNRS-INPT, 5 rue Paulin Talabot BP 1301, 31106 Toulouse (France)], E-mail: Alain.Bergel@ensiacet.fr

    2008-12-01

    The influence of [Fe]-hydrogenase from Clostridium acetobutylicum was studied on the anaerobic corrosion of mild steel. Two short-circuited mild steel electrodes were exposed to the same solution and hydrogenase was retained on the surface of only one electrode thanks to a dialysis membrane. The galvanic current and the electrode potential were measured as a function of time in order to monitor the difference in electrochemical behaviour induced by the presence of hydrogenase. A sharp potential decrease of around 500 mV was controlled by the deoxygenating phase. When hydrogenase was introduced after complete deoxygenation, significant heterogeneous corrosion was observed under the vivianite deposit on the electrode in contact with hydrogenase, while the other electrode only showed the vivianite deposit, which was analysed by MEB and EDX. The effect of hydrogenase was then confirmed by monitoring the free potential of single coupons exposed or not to the enzyme in a classical cell after complete deoxygenating. In both phosphate and Tris-HCl buffers, the presence of hydrogenase increased the free potential around 60 mV and induced marked general corrosion. It was concluded that [Fe]-hydrogenase acts in the absence of any final electron acceptor by catalysing direct proton reduction on the mild steel surface.

  14. Catalytic hydrodechlorination of monochloroacetic acid in wastewater using Ni-Fe bimetal prepared by ball milling.

    Science.gov (United States)

    Zhu, Hong; Xu, Fuyuan; Zhao, Jianzhuang; Jia, Linfang; Wu, Kunming

    2015-09-01

    Monochloroacetic acid (MCA) is a chemically stable and biologically toxic pollutant. It is often generated during the production of the pesticide dimethoate. Conventional wastewater treatment processes have difficulty degrading it. In this work, the dechlorination effects of Ni-Fe bimetal prepared using ball milling (BM) technology for the high concentrations of MCA in wastewater were examined. The MCA in aqueous solution was found to be degraded efficiently by the Ni-Fe bimetal. However, S-(methoxycarbonyl) methyl O, O-dimethyl phosphorodithioate (SMOPD) in wastewater, a by-product of the dimethoate production process, significantly inhibited the reductive dechlorination activity of Ni-Fe bimetal. Increasing the reaction temperature in the MCA wastewater enhanced the reduction activity of the Ni-Fe bimetal effectively. Oxygen was found to be unfavorable to dechlorination. Sealing the reaction to prevent oxidation was found to render the degradation process more efficient. The process retained over 88% efficiency after 10 treatment cycles with 50 g/L of Ni-Fe bimetal under field conditions.

  15. [FeFe]-Hydrogenase-Catalyzed H2 Production in a Photoelectrochemical Biofuel Cell

    Energy Technology Data Exchange (ETDEWEB)

    Hambourger, M.; Gervaldo, M.; Svedruzic, D.; King, P. W.; Gust, D.; Ghirardi, M.; Moore, A. L.; Moore, T. A.

    2008-01-01

    The Clostridium acetobutylicum [FeFe]-hydrogenase HydA has been investigated as a hydrogen production catalyst in a photoelectrochemical biofuel cell. Hydrogenase was adsorbed to pyrolytic graphite edge and carbon felt electrodes. Cyclic voltammograms of the immobilized hydrogenase films reveal cathodic proton reduction and anodic hydrogen oxidation, with a catalytic bias toward hydrogen evolution. When corrected for the electrochemically active surface area, the cathodic current densities are similar for both carbon electrodes, and 40% of those obtained with a platinum electrode. The high surface area carbon felt/hydrogenase electrode was subsequently used as the cathode in a photoelectrochemical biofuel cell. Under illumination, this device is able to oxidize a biofuel substrate and reduce protons to hydrogen. Similar photocurrents and hydrogen production rates were observed in the photoelectrochemical biofuel cell using either hydrogenase or platinum cathodes.

  16. Preparation, characterization, and antibacterial activity of NiFe2O4/PAMA/Ag-TiO2 nanocomposite

    Science.gov (United States)

    Allafchian, Alireza; Jalali, Seyed Amir Hossein; Bahramian, Hamid; Ahmadvand, Hossein

    2016-04-01

    We have described a facile fabrication of silver deposited on the TiO2, Poly Acrylonitrile Co Maleic Anhydride (PAMA) polymer and nickel ferrite composite (NiFe2O4/PAMA/Ag-TiO2) through a three-step procedure. A pre-synthesized NiFe2O4 was first coated with PAMA polymer and then Ag-TiO2 was deposited on the surface of PAMA polymer shell. After the characterization of this three-component composite by various techniques, such as FTIR, XRD, FESEM, BET, TEM and VSM, it was impregnated in standard antibiotic discs. The antibacterial activity of NiFe2O4/PAMA/Ag-TiO2 nanocomposite was investigated against some gram positive and gram negative bacteria by employing disc diffusion assay and then compared with that of naked NiFe2O4, NiFe2O4/Ag, AgNPs and NiFe2O4/PAMA. The results demonstrated that the AgNPs, when embedded in TiO2 and combined with NiFe2O4/PAMA, became an excellent antibacterial agent. The NiFe2O4/PAMA/Ag-TiO2 nanocomposite could be readily separated from water solution after the disinfection process by applying an external magnetic field.

  17. A study on electrodeposited NiFe1− alloy films

    Indian Academy of Sciences (India)

    M Bedir; Ö F Bakkaloğlu; İ H Karahan; M Öztaş

    2006-06-01

    NiFe1− (0.22 ≤ ≤ 0.62) alloy films were grown by electrodeposition technique. A shift in diffraction peaks of NiFe and Ni3Fe was detected with increasing Ni content. The highest positive magnetoresistance ratio was detected as 5% in Ni0.51Fe0.49. Positive and negative anisotropic magnetoresistance were observed in longitudinal and transverse geometries respectively. The highest anisotropic magnetoresistance ratio of 9.8% was also detected in Ni0.51Fe0.49. The angular variation of magnetoresistance was studied. Magnetisation loop curves show that NiFe alloy films have a linear decreasing anisotropy constant with increasing Ni deposit content and show a decreasing behavior of coercivity which indicates soft magnetic property with increasing Ni deposit content.

  18. Occurrence of H2-Uptake Hydrogenases in Bradyrhizobium sp. (Lupinus) and Their Expression in Nodules of Lupinus spp. and Ornithopus compressus1

    Science.gov (United States)

    Murillo, Jesús; Villa, Ana; Chamber, Manuel; Ruiz-Argüeso, Tomás

    1989-01-01

    Fifty-four strains of Bradyrhizobium sp. (Lupinus) from worldwide collections were screened by a colony hybridization method for the presence of DNA sequences homologous to the structural genes of the Bradyrhizobium japonicum hydrogenase. Twelve strains exhibited strong colony hybridization signals, and subsequent Southern blot hybridization experiments showed that they fell into two different groups on the basis of the pattern of EcoRI fragments containing the homology to the hup probe. All strains in the first group (UPM860, UPM861, and 750) expressed uptake hydrogenase activity in symbiosis with Lupinus albus, Lupinus angustifolius, Lupinus luteus, and Ornithopus compressus, but both the rate of H2 uptake by bacteroids and the relative efficiency of N2 fixation (RE = 1 - [H2 evolved in air/acetylene reduced]) by nodules were markedly affected by the legume host. L. angustifolius was the less permissive host for hydrogenase expression in symbiosis with the three strains (average RE = 0.76), and O. compressus was the more permissive (average RE = 1.0). None of the strains in the second group expressed hydrogenase activity in lupine nodules, and only one exhibited low H2-uptake activity in symbiosis with O. compressus. The inability of these putative Hup+ strains to induce hydrogenase activity in lupine nodules is discussed on the basis of the legume host effect. Among the 42 strains showing no homology to the B. japonicum hup-specific probe in the colony hybridization assay, 10 were examined in symbiosis with L. angustifolius. The average RE for these strains was 0.51. However, one strain, IM43B, exhibited high RE values (higher than 0.80) and high levels of hydrogenase activity in symbiosis with L. angustifolius, L. albus, and L. luteus. In Southern blot hybridization experiments, no homology was detected between the B. japonicum hup-specific DNA probe and total DNA from vegetative cells or bacteroids from strain IM43B even under low stringency hybridization

  19. Mutagenesis of nifE and nifN from Azotobacter vinelandii

    OpenAIRE

    Wilson, Mark Steven Michael

    1988-01-01

    The products of nifE and nifN from Azotobacter vinelandii, which are involved in the biosynthesis of the iron-molybdenum cofactor (FeMo-co) co) from nitrogenase, have been analyzed using a variety of mutagenic techniques. NifE was the object of several site-specific, amino acid substitutions that were designed to elicit information regarding metal cluster ligands, subunit-subunit interactions, and the proposed transfer of FeMo-co.from a nifEN-products complex to the apo-MoFe pr...

  20. In vitro hydrogen production by glucose dehydrogenase and hydrogenase

    Energy Technology Data Exchange (ETDEWEB)

    Woodward, J.; Mattingly, S.M. [Oak Ridge National Lab., TN (United States); Danson, M. [Univ. of Bath (United Kingdom)] [and others

    1996-07-01

    A new in vitro enzymatic pathway for the generation of molecular hydrogen from glucose has been demonstrated. The reaction is based on the oxidation of glucose by Thermoplasma acidophilum glucose dehydrogenase with the concomitant oxidation of NADPH by Pyrococcus furiosus hydrogenase. Stoichiometric yields of hydrogen were produced from glucose with the continuous recycling of cofactor. This simple system may provide a method for the biological production of hydrogen from renewable sources. In addition, the other product of this reaction, gluconic acid, is a high-value chemical commodity. 23 refs., 5 figs.

  1. In vitro hydrogen production by glucose dehydrogenase and hydrogenase

    Energy Technology Data Exchange (ETDEWEB)

    Woodward, J. [Oak Ridge National Lab., TN (United States)

    1996-10-01

    A new in vitro enzymatic pathway for the generation of molecular hydrogen from glucose has been demonstrated. The reaction is based upon the oxidation of glucose by Thermoplasma acidophilum glucose dehydrogenase with the concomitant oxidation of NADPH by Pyrococcus furiosus hydrogenase. Stoichiometric yields of hydrogen were produced from glucose with continuous cofactor recycle. This simple system may provide a method for the biological production of hydrogen from renewable sources. In addition, the other product of this reaction, gluconic acid, is a high-value commodity chemical.

  2. A synthetic system links FeFe-hydrogenases to essential E. coli sulfur metabolism

    Directory of Open Access Journals (Sweden)

    Grandl Gerald

    2011-05-01

    Full Text Available Abstract Background FeFe-hydrogenases are the most active class of H2-producing enzymes known in nature and may have important applications in clean H2 energy production. Many potential uses are currently complicated by a crucial weakness: the active sites of all known FeFe-hydrogenases are irreversibly inactivated by O2. Results We have developed a synthetic metabolic pathway in E. coli that links FeFe-hydrogenase activity to the production of the essential amino acid cysteine. Our design includes a complementary host strain whose endogenous redox pool is insulated from the synthetic metabolic pathway. Host viability on a selective medium requires hydrogenase expression, and moderate O2 levels eliminate growth. This pathway forms the basis for a genetic selection for O2 tolerance. Genetically selected hydrogenases did not show improved stability in O2 and in many cases had lost H2 production activity. The isolated mutations cluster significantly on charged surface residues, suggesting the evolution of binding surfaces that may accelerate hydrogenase electron transfer. Conclusions Rational design can optimize a fully heterologous three-component pathway to provide an essential metabolic flux while remaining insulated from the endogenous redox pool. We have developed a number of convenient in vivo assays to aid in the engineering of synthetic H2 metabolism. Our results also indicate a H2-independent redox activity in three different FeFe-hydrogenases, with implications for the future directed evolution of H2-activating catalysts.

  3. Influence of static and dynamic dipolar fields in bulk YIG/thin film NiFe systems probed via spin rectification effect

    Science.gov (United States)

    Soh, Wee Tee; Tay, Z. J.; Yakovlev, N. L.; Peng, Bin; Ong, C. K.

    2017-03-01

    The characteristics of the static and dynamic components of the dipolar fields originating from a bulk polycrystalline yttrium iron garnet (YIG) substrate are probed by depositing a NiFe (Permalloy) layer on it, which acts as a detector. By measuring dc voltages generated via spin rectification effect (SRE) within the NiFe layer under microwave excitation, we characterize the influence of dipolar fields from bulk YIG on the NiFe layer. It is found that the dynamic YIG dipolar fields modify the self-SRE of NiFe, driving its own rectification voltages within the NiFe layer, an effect we term as non-local SRE. This non-local SRE only occurs near the simultaneous resonance of both YIG and NiFe. On the other hand, the static dipolar field from YIG manifests itself as a negative anisotropy in the NiFe layer which shifts the latter's ferromagnetic resonance frequency.

  4. Inactivation of uptake hydrogenase leads to enhanced and sustained hydrogen production with high nitrogenase activity under high light exposure in the cyanobacterium Anabaena siamensis TISTR 8012

    Directory of Open Access Journals (Sweden)

    Khetkorn Wanthanee

    2012-10-01

    Full Text Available Abstract Background Biohydrogen from cyanobacteria has attracted public interest due to its potential as a renewable energy carrier produced from solar energy and water. Anabaena siamensis TISTR 8012, a novel strain isolated from rice paddy field in Thailand, has been identified as a promising cyanobacterial strain for use as a high-yield hydrogen producer attributed to the activities of two enzymes, nitrogenase and bidirectional hydrogenase. One main obstacle for high hydrogen production by A. siamensis is a light-driven hydrogen consumption catalyzed by the uptake hydrogenase. To overcome this and in order to enhance the potential for nitrogenase based hydrogen production, we engineered a hydrogen uptake deficient strain by interrupting hupS encoding the small subunit of the uptake hydrogenase. Results An engineered strain lacking a functional uptake hydrogenase (∆hupS produced about 4-folds more hydrogen than the wild type strain. Moreover, the ∆hupS strain showed long term, sustained hydrogen production under light exposure with 2–3 folds higher nitrogenase activity compared to the wild type. In addition, HupS inactivation had no major effects on cell growth and heterocyst differentiation. Gene expression analysis using RT-PCR indicates that electrons and ATP molecules required for hydrogen production in the ∆hupS strain may be obtained from the electron transport chain associated with the photosynthetic oxidation of water in the vegetative cells. The ∆hupS strain was found to compete well with the wild type up to 50 h in a mixed culture, thereafter the wild type started to grow on the relative expense of the ∆hupS strain. Conclusions Inactivation of hupS is an effective strategy for improving biohydrogen production, in rates and specifically in total yield, in nitrogen-fixing cultures of the cyanobacterium Anabaena siamensis TISTR 8012.

  5. Turning cellulose waste into electricity: hydrogen conversion by a hydrogenase electrode.

    Directory of Open Access Journals (Sweden)

    Sergey M Abramov

    Full Text Available Hydrogen-producing thermophilic cellulolytic microorganisms were isolated from cow faeces. Rates of cellulose hydrolysis and hydrogen formation were 0.2 mM L(-1 h(-1 and 1 mM L(-1 h(-1, respectively. An enzymatic fuel cell (EFC with a hydrogenase anode was used to oxidise hydrogen produced in a microbial bioreactor. The hydrogenase electrode was exposed for 38 days (912 h to a thermophilic fermentation medium. The hydrogenase activity remaining after continuous operation under load was 73% of the initial value.

  6. Improved hydrogen production by coupled systems of hydrogenase negative photosynthetic bacteria and fermentative bacteria in reverse micelles

    Energy Technology Data Exchange (ETDEWEB)

    Singh, Anita [Centre for Biotechnology, University of Allahabad, Allahabad 211002 (India); Misra, Krishna [Indo-Russian Center for Bioinformatics, Indian Institute of Information Technology, Allahabad 211011 (India)

    2008-11-15

    Significant improvement in biological hydrogen production is achieved by the use of coupled bacterial cells in reverse micellar systems. Two coupled systems (a) Rhodopseudomonas palustris CGA009/Citrobacter Y19, and (b) Rhodobacter sphaeroides 2.4.1/Citrobacter Y19 bacteria have been immobilized separately in aqueous pool of the reverse micelles fabricated by various surfactants (AOT, CBAC and SDS) and apolar organic solvents (benzene and isooctane). The gene for uptake hydrogenase enzyme has been manipulated further for hydrogen generation. Mutants deficient in uptake hydrogenase (Hup{sup -}) were obtained from R. palustris CGA009 and R. sphaeroides 2.4.1, and entrapped with Citrobacter Y19 in the reverse micellar systems. More than two fold increase in hydrogen production was obtained by the use of Hup{sup -} mutants instead of wild-type photosynthetic bacteria together with Citrobacter Y19. Addition of sodium dithionite, a reducing agent to AOT/H{sub 2}O/isooctane reverse micellar system with the coupled systems of wild-type photosynthetic bacteria and fermentative bacterium Y19 effected similar increase in hydrogen production rate as it is obtained by the use of mutants. CBAC/H{sub 2}O/isooctane reverse micellar system is used for the first time for hydrogen production and is as promising as AOT/H{sub 2}O/isooctane reverse micellar system. All reverse micellar systems of coupled bacterial cultures gave encouraging hydrogen production (rate as well as yield) compared to uncoupled bacterial culture. (author)

  7. Disruption of the Operon Encoding Ehb Hydrogenase Limits AnabolicCO2 Assimilation in the Archaeon Methanococcus maripaludis

    Energy Technology Data Exchange (ETDEWEB)

    Porat, Iris; Kim, Wonduck; Hendrickson, Erik L.; Xia, Qiangwei; Zhang, Yi; Wang, Tiansong; Taub, Fred; Moore, Brian C.; Anderson, IainJ.; Hackett, Murray; Leigh, John A.; Whitman, William B.

    2006-02-01

    Methanococcus maripaludis is a mesophilic archaeon thatreduces CO2 to methane with H2 or formate as an energy source. Itcontains two membrane-bound energy-conserving hydrogenases, Eha and Ehb.To determine therole of Ehb, a deletion in the ehb operon wasconstructed to yield the mutant, strain S40. Growth of S40 was severelyimpaired in minimal medium. Both acetate and yeast extract were necessaryto restore growth to nearly wild-type levels, suggesting that Ehb wasinvolved in multiple steps in carbon assimilation. However, nodifferences in the total hydrogenase specific activities were foundbetween the wild type and mutant in either cell extracts ormembrane-purified fractions. Methanogenesis by resting cells withpyruvate as the electron donor was also reduced by 30 percent in S40,suggesting a defect in pyruvate oxidation. CO dehydrogenase/acetylcoenzyme A (CoA) synthase and pyruvate oxidoreductase had higher specificactivities in the mutant, and genes encoding these enzymes, as well asAMP-forming acetyl-CoA synthetase, were expressed at increased levels.These observations support a role for Ehb in anabolic CO2 assimilation inmethanococci.

  8. Construction and use of a Cupriavidus necator H16 soluble hydrogenase promoter (PSH fusion to gfp (green fluorescent protein

    Directory of Open Access Journals (Sweden)

    Bat-Erdene Jugder

    2016-07-01

    Full Text Available Hydrogenases are metalloenzymes that reversibly catalyse the oxidation or production of molecular hydrogen (H2. Amongst a number of promising candidates for application in the oxidation of H2 is a soluble [Ni–Fe] uptake hydrogenase (SH produced by Cupriavidus necator H16. In the present study, molecular characterisation of the SH operon, responsible for functional SH synthesis, was investigated by developing a green fluorescent protein (GFP reporter system to characterise PSH promoter activity using several gene cloning approaches. A PSH promoter-gfp fusion was successfully constructed and inducible GFP expression driven by the PSH promoter under de-repressing conditions in heterotrophic growth media was demonstrated in the recombinant C. necator H16 cells. Here we report the first successful fluorescent reporter system to study PSH promoter activity in C. necator H16. The fusion construct allowed for the design of a simple screening assay to evaluate PSH activity. Furthermore, the constructed reporter system can serve as a model to develop a rapid fluorescent based reporter for subsequent small-scale process optimisation experiments for SH expression.

  9. Facile synthesis of inverse spinel NiFe2O4 nanocrystals and their superparamagnetic properties

    Directory of Open Access Journals (Sweden)

    Jie Tan

    2013-02-01

    Full Text Available Spinel NiFe2O4 nanocrystals have been obtained by means of a novel composite-hydroxide-salt-mediated approach, which is based on a reaction between metallic salt and metallic oxide in the solution of composite-hydroxide-salt eutectic at ~225 ºC and normal atmosphere without any organic dispersant or capping agent. The obtained products are characterized by an X-ray diffraction (XRD, a transmission electron microscopy (TEM and an alternating gradient magnetometer (AGM. The formation process of NiFe2O4 nanosheet is proposed to begin with a ‘‘dissolution-recrystallization’’ which is followed by an ‘‘Ostwald ripening’’ mechanism. The NiFe2O4 nano-octahedrons can be obtained through adjusting the reaction water content in the hydroxide melts at constant temperature. At 300 K, magnetic hysteresis loops at an applied field of 15 kOe show zero coercivity, indicating the superparamagnetic behavior of the as-prepared NiFe2O4 nanocrystals.

  10. Alloyed Ni-Fe nanoparticles as catalysts for NH3 decomposition

    DEFF Research Database (Denmark)

    Simonsen, Søren Bredmose; Chakraborty, Debasish; Chorkendorff, Ib

    2012-01-01

    temperatures and a low NH3-to-H2 ratio, the catalytic activity of the best Ni-Fe/Al2O3 catalyst was found to be comparable or even better to that of a more expensive Ru-based catalyst. Small Ni-Fe nanoparticle sizes are crucial for an optimal overall NH3 conversion because of a structural effect favoring...

  11. Facile synthesis of inverse spinel NiFe2O4 nanocrystals and their superparamagnetic properties

    Directory of Open Access Journals (Sweden)

    Jie Tan

    2012-01-01

    Full Text Available Spinel NiFe2O4 nanocrystals have been obtained by means of a novel composite-hydroxide-salt-mediated approach, which is based on a reaction between metallic salt and metallic oxide in the solution of composite-hydroxide-salt eutectic at ~225 ºC and normal atmosphere without any organic dispersant or capping agent. The obtained products are characterized by an X-ray diffraction (XRD, a transmission electron microscopy (TEM and an alternating gradient magnetometer (AGM. The formation process of NiFe2O4 nanosheet is proposed to begin with a ‘‘dissolution-recrystallization’’ which is followed by an ‘‘Ostwald ripening’’ mechanism. The NiFe2O4 nano-octahedrons can be obtained through adjusting the reaction water content in the hydroxide melts at constant temperature. At 300 K, magnetic hysteresis loops at an applied field of 15 kOe show zero coercivity, indicating the superparamagnetic behavior of the as-prepared NiFe2O4 nanocrystals.

  12. Physical and electrical characteristics of NiFe thin films using ultrasonic assisted pulse electrodeposition

    Science.gov (United States)

    Asa Deepthi, K.; Balachandran, R.; Ong, B. H.; Tan, K. B.; Wong, H. Y.; Yow, H. K.; Srimala, S.

    2016-01-01

    Nickel iron (NiFe) thin films were prepared on the copper substrate by ultrasonic assisted pulse electrodeposition under galvanostatic mode. Careful control of the thin films deposition is essential as the electrical properties of the films could be greatly affected, particularly if low quality films are produced. The preparation of NiFe/Cu thin films was aimed to reduce the grain size of NiFe particles, surface roughness and electrical resistivity of the copper substrates. Various parameters were systematically studied including current magnitude, deposition time and ultrasonic bath temperature. The optimized conditions to obtain NiFe permalloy, which subsequently applied to all investigated samples, were found at a current magnitude of 70 mA deposited for a duration of 2 min under ultrasonic bath temperature of 27 °C. The composition of NiFe permalloy was as close as Ni 80.71% and Fe 19.29% and the surface roughness was reduced from 12.76 nm to 2.25 nm. The films electrical resistivity was decreased nearly sevenfold from an initial value of 67.32 μΩ cm to 9.46 μΩ cm.

  13. Alloyed Ni-Fe nanoparticles as catalysts for NH3 decomposition

    DEFF Research Database (Denmark)

    Simonsen, Søren Bredmose; Chakraborty, Debasish; Chorkendorff, Ib;

    2012-01-01

    A rational design approach was used to develop an alloyed Ni-Fe/Al2O3 catalyst for decomposition of ammonia. The dependence of the catalytic activity is tested as a function of the Ni-to-Fe ratio, the type of Ni-Fe alloy phase, the metal loading and the type of oxide support. In the tests with high...

  14. Grain size effect on the Neel temperature and magnetic properties of nanocrystalline $NiFe{_2}O{_4}$ spinel

    OpenAIRE

    Chinnasamy, CN; Narayanasamy, A.; Ponpandian, N; Joseyphus, RJ; Jeyadevan, B.; Tohji, K.; Chattopadhyay, K.

    2002-01-01

    Nanocrystalline NiFe2O4 spinel ferrites with various grain sizes have been synthesized by ball milling the bulk NiFe2O4. The average grain sizes were estimated from the X-ray line broadening of the (3 1 1) reflection. The Neel temperatures of NiFe2O4 for various grain sizes were determined by magneto thermogravimetric method. The magnetic behaviour has been explained by combining the effects of changes in cation distribution on milling and finite size scaling. The shift in B-H loops has been ...

  15. Electrocatalytic mechanism of reversible hydrogen cycling by enzymes and distinctions between the major classes of hydrogenases

    OpenAIRE

    Hexter, Suzannah V.; Grey, Felix; Happe, Thomas; Climent, Victor; Armstrong, Fraser A.

    2012-01-01

    The extraordinary ability of Fe- and Ni-containing enzymes to catalyze rapid and efficient H+/H2 interconversion—a property otherwise exclusive to platinum metals—has been investigated in a series of experiments combining variable-temperature protein film voltammetry with mathematical modeling. The results highlight important differences between the catalytic performance of [FeFe]-hydrogenases and [NiFe]-hydrogenases and justify a simple model for reversible catalytic electron flow in enzymes...

  16. Aquifex aeolicus membrane hydrogenase for hydrogen biooxidation: Role of lipids and physiological partners in enzyme stability and activity

    Energy Technology Data Exchange (ETDEWEB)

    Infossi, Pascale; Lojou, Elisabeth; Giudici-Orticoni, Marie-Therese [Unite de Bioenergetique et Ingenierie des Proteines, UPR 9036, Institut de Microbiologie de la Mediterranee - CNRS, 31 Chemin Joseph Aiguier, 13402 Marseille Cedex 20 (France); Chauvin, Jean-Paul [Institut de Biologie du developpement de Marseille Luminy, UMR 6216, Parc Scientifique de Luminy, 163 Avenue de Luminy, BP 907, 13009 Marseille (France); Herbette, Gaetan [Spectropole FI 1739, Aix-Marseille Universite case 511, Faculte de St Jerome Avenue Escadrille Normandie Niemen, 13397 Marseille Cedex 20 (France); Brugna, Myriam [Unite de Bioenergetique et Ingenierie des Proteines, UPR 9036, Institut de Microbiologie de la Mediterranee - CNRS, 31 Chemin Joseph Aiguier, 13402 Marseille Cedex 20 (France); Universite de Provence, 3 Place Victor Hugo, 13331 Marseille Cedex 03 (France)

    2010-10-15

    Hydrogenase I from the hyperthermophilic bacterium Aquifex aeolicus is a good candidate for biotechnological devices thanks to its ability to oxidize hydrogen at high temperature, even in the presence of oxygen and CO. In order to enhance the enzyme stability and the catalytic efficiency, we investigated the hydrogen oxidation process with hydrogenase I embedded in a physiological-like environment. Hydrogenase I partners in the metabolic chain, namely membrane quinone and cytochrome b, were purified and fully characterized. The complex hydrogenase I-cytochrome b was inserted into liposomes. Surface Plasmon Resonance revealed that quinone took part in the stabilization of the complex. By use of molecular modelization and electrochemistry analysis, enzyme stability has been demonstrated to be stronger and enzymatic efficiency to be five times higher when hydrogenase is embedded into the liposomes. This result raises the possibility of using hydrogenases as biocatalysts in fuel cells. (author)

  17. Tyrosine, cysteine, and S-adenosyl methionine stimulate in vitro [FeFe] hydrogenase activation.

    Directory of Open Access Journals (Sweden)

    Jon M Kuchenreuther

    Full Text Available BACKGROUND: [FeFe] hydrogenases are metalloenzymes involved in the anaerobic metabolism of H(2. These proteins are distinguished by an active site cofactor known as the H-cluster. This unique [6Fe-6S] complex contains multiple non-protein moieties and requires several maturation enzymes for its assembly. The pathways and biochemical precursors for H-cluster biosynthesis have yet to be elucidated. PRINCIPAL FINDINGS: We report an in vitro maturation system in which, for the first time, chemical additives enhance [FeFe] hydrogenase activation, thus signifying in situ H-cluster biosynthesis. The maturation system is comprised of purified hydrogenase apoprotein; a dialyzed Escherichia coli cell lysate containing heterologous HydE, HydF, and HydG maturases; and exogenous small molecules. Following anaerobic incubation of the Chlamydomonas reinhardtii HydA1 apohydrogenase with S-adenosyl methionine (SAM, cysteine, tyrosine, iron, sulfide, and the non-purified maturases, hydrogenase activity increased 5-fold relative to incubations without the exogenous substrates. No conditions were identified in which addition of guanosine triphosphate (GTP improved hydrogenase maturation. SIGNIFICANCE: The in vitro system allows for direct investigation of [FeFe] hydrogenase activation. This work also provides a foundation for studying the biosynthetic mechanisms of H-cluster biosynthesis using solely purified enzymes and chemical additives.

  18. Studies of Hybrid Nano-Bio-System: Single-Walled Carbon Nanotubes and Hydrogenase

    Energy Technology Data Exchange (ETDEWEB)

    Svedruzic-Chang, D.; Blackburn, J. L.; McDonald, T. J.; Heben, M. J.; King, P. W.

    2008-01-01

    We have examined changes in single-walled carbon nanotubes (SWNT) optical signals upon addition of recombinant [FeFe] hydrogenases from Clostridium acetobutylicum or Chlamydomonas reinhardtii. We found evidence that novel and stable charge-transfer complexes are formed only under conditions of hydrogenase catalytic turnover. Formation of the complex sensitizes the nanotubes to the proton-to-hydrogen redox half-reaction. Thus, the experimental potential can be altered by changing the pH or molecular hydrogen concentration. In the presence of molecular hydrogen, hydrogenase mediates electron injection into the conduction band of semiconducting SWNT, which was observed as a quenching of the photoluminescence signals. Here, we will present recent Raman studies, which revealed that SWNTs in a complex with hydrogenase may undergo either oxidation or reduction, depending on the electronic structure of the SWNT and the oxidation state of the enzyme. In addition, we will describe our efforts to prepare stable, solubilized SWNT/hydrogenase complexes in the absence of detergent. This work shows that SWNT/hydrogenase complexes have potential applications as a component of an energy conversion device.

  19. Integration of an [FeFe]-hydrogenase into the anaerobic metabolism of Escherichia coli

    Science.gov (United States)

    Kelly, Ciarán L.; Pinske, Constanze; Murphy, Bonnie J.; Parkin, Alison; Armstrong, Fraser; Palmer, Tracy; Sargent, Frank

    2015-01-01

    Biohydrogen is a potentially useful product of microbial energy metabolism. One approach to engineering biohydrogen production in bacteria is the production of non-native hydrogenase activity in a host cell, for example Escherichia coli. In some microbes, hydrogenase enzymes are linked directly to central metabolism via diaphorase enzymes that utilise NAD+/NADH cofactors. In this work, it was hypothesised that heterologous production of an NAD+/NADH-linked hydrogenase could connect hydrogen production in an E. coli host directly to its central metabolism. To test this, a synthetic operon was designed and characterised encoding an apparently NADH-dependent, hydrogen-evolving [FeFe]-hydrogenase from Caldanaerobacter subterranus. The synthetic operon was stably integrated into the E. coli chromosome and shown to produce an active hydrogenase, however no H2 production was observed. Subsequently, it was found that heterologous co-production of a pyruvate::ferredoxin oxidoreductase and ferredoxin from Thermotoga maritima was found to be essential to drive H2 production by this system. This work provides genetic evidence that the Ca.subterranus [FeFe]-hydrogenase could be operating in vivo as an electron-confurcating enzyme. PMID:26839796

  20. Integration of an [FeFe]-hydrogenase into the anaerobic metabolism of Escherichia coli

    Directory of Open Access Journals (Sweden)

    Ciarán L. Kelly

    2015-12-01

    Full Text Available Biohydrogen is a potentially useful product of microbial energy metabolism. One approach to engineering biohydrogen production in bacteria is the production of non-native hydrogenase activity in a host cell, for example Escherichia coli. In some microbes, hydrogenase enzymes are linked directly to central metabolism via diaphorase enzymes that utilise NAD+/NADH cofactors. In this work, it was hypothesised that heterologous production of an NAD+/NADH-linked hydrogenase could connect hydrogen production in an E. coli host directly to its central metabolism. To test this, a synthetic operon was designed and characterised encoding an apparently NADH-dependent, hydrogen-evolving [FeFe]-hydrogenase from Caldanaerobacter subterranus. The synthetic operon was stably integrated into the E. coli chromosome and shown to produce an active hydrogenase, however no H2 production was observed. Subsequently, it was found that heterologous co-production of a pyruvate::ferredoxin oxidoreductase and ferredoxin from Thermotoga maritima was found to be essential to drive H2 production by this system. This work provides genetic evidence that the Ca.subterranus [FeFe]-hydrogenase could be operating in vivo as an electron-confurcating enzyme.

  1. Exchange bias and spin glass behavior in biphasic NiFe2O4/NiO thin films

    Science.gov (United States)

    Pebley, Andrew C.; Fuks, Preston E.; Pollock, Tresa M.; Gordon, Michael J.

    2016-12-01

    Magnetic exchange bias and coercivity of nanogranular NiFe2O4/NiO thin films, prepared using flow-stabilized microplasmas and post-deposition annealing, have been investigated as a function of ferrimagnet/antiferromagnet phase fraction, grain size, and temperature. Exchange bias (EB) and vertical shifts in hysteresis loops observed in the as-deposited and low-T annealed (≤600 °C) films were attributed to exchange coupling between nanocrystalline NiFe2O4 (~8-10 nm) and a structurally-disordered spin glass (SG)-like phase. At higher annealing temperature (850 °C), the observed EB was found to arise from exchange coupling between NiFe2O4 and NiO, rather than a SG phase, most likely due to reduction of structurally-disordered interfaces and a substantial increase in NiFe2O4 grain size (~26 nm).

  2. Variations in the saturation magnetization of nanosized NiFe2O4 particles on adsorption of carboxylic acids

    Directory of Open Access Journals (Sweden)

    Ryo Kurosawa

    2014-03-01

    Full Text Available This work investigated magnetization changes in NiFe2O4 nanoparticles induced by the adsorption of a series of carboxylic acids. The application of formic acid resulted in a significant 8.6% decrease in the magnetization of NiFe2O4 nanoparticles at 18,000 Oe. With increasing carbon bond number in the saturated carboxylic acids, reductions in the magnetization of NiFe2O4 nanoparticles became around 4%. All unsaturated carboxylic acids produced approximately equivalent reductions in the magnetization, regardless of their double bond content. Based on these results, the observed NiFe2O4 magnetization changes appear to depend on either the polarity or the molecular size of the carboxylic acids and are believed to be caused by canting or pinning of spins in the vicinity of particle surfaces following adsorption of the acids.

  3. Nickel recovery from electronic waste II electrodeposition of Ni and Ni-Fe alloys from diluted sulfate solutions.

    Science.gov (United States)

    Robotin, B; Ispas, A; Coman, V; Bund, A; Ilea, P

    2013-11-01

    This study focuses on the electrodeposition of Ni and Ni-Fe alloys from synthetic solutions similar to those obtained by the dissolution of electron gun (an electrical component of cathode ray tubes) waste. The influence of various parameters (pH, electrolyte composition, Ni(2+)/Fe(2+) ratio, current density) on the electrodeposition process was investigated. Scanning electron microscopy (SEM) and X-ray fluorescence analysis (XRFA) were used to provide information about the obtained deposits' thickness, morphology, and elemental composition. By controlling the experimental parameters, the composition of the Ni-Fe alloys can be tailored towards specific applications. Complementarily, the differences in the nucleation mechanisms for Ni, Fe and Ni-Fe deposition from sulfate solutions have been evaluated and discussed using cyclic voltammetry and potential step chronoamperometry. The obtained results suggest a progressive nucleation mechanism for Ni, while for Fe and Ni-Fe, the obtained data points are best fitted to an instantaneous nucleation model.

  4. Bamboo-like Te Nanotubes with Tailored Dimensions Synthesized from Segmental NiFe Nanowires as Sacrificial Templates

    Energy Technology Data Exchange (ETDEWEB)

    Suh, Ho Young; Jung, Hyun Sung; Myung, No Sang [University of California-Riverside, Riverside (United States); Hong, Kim In [KICET, Seoul (Korea, Republic of)

    2014-09-15

    Bamboo-like Te nanotubes were synthesized via the galvanic displacement reaction of NiFe nanowires with Ni-rich and Fe-rich segments. The thick and thin components of the synthesized Te nanotubes were converted from the Ni-rich and Fe-rich segments in the NiFe nanowires respectively. The dimensions of the Te nanotubes were controlled by employing sacrificial NiFe nanowires with tailored dimensions as the template for the galvanic displacement reaction. The segment lengths of the Te nanotubes were found to be dependent on those of the sacrificial NiFe nanowires. The galvanic displacement reaction was characterized by analyzing the open circuit potential and the corrosion resistance.

  5. Preparation of NiFe2O4 nanopowder via EDTA precursor and study of its properties

    Institute of Scientific and Technical Information of China (English)

    Amit B.Rajput; Subhenjit Hazra; Niharika B.Krishna; Pavani Chavali; Sneha Datla; Narendra Nath Ghosh

    2012-01-01

    An ethylene diamine tetra acetic acid (EDTA) precursor-based chemical method is reported for preparing single-phase nickel ferrite (NiFe2O4) nanopowder.The synthesized powder was characterized by thermogravimetric analysis,differential scanning calorimetric analysis,X-ray diffraction,particle size analysis and scanning electron microscopy.DC electrical resistivity and magnetic property of the synthesized NiFe2O4 were measured by using a two-probe method and a vibrating sample magnetometer respectively.The DC resistivity behaviour of the NiFe2O4 nanopowder was correlated with the change of microstructure,during sintering.This EDTA precursor-based method is capable of producing nanostructured single-phase NiFe2O4 powder at a comparatively low calcination temperature and offers the potential of a simple and cost-effective route,including the preparation of other ferrite nanoparticles.

  6. Decolorization and degradation of textile dyes with biosulfidogenic hydrogenases.

    Science.gov (United States)

    Mutambanengwe, C C Z; Togo, C A; Whiteley, C G

    2007-01-01

    Successful decolorization of azo dyes (Orange II, Amido Black 10, Reactive Black 5, and Reactive Red 120) and industrial textile dye influents and effluents with sulfate-reducing bacteria from within a biosulfidogenic reactor was achieved with decolorizations ranging from 96% to 49% over 144 h. Concomitant with the decrease in absorbance of the dye in the visible region (480-620 nm) was an increase in the absorbance at 280 nm, over 48 h, suggesting an increase in concentration of single aromatic amines. With an extended period of time there was a subsequent decrease in the absorbance at 280 nm indicating that the aromatic amines had been degraded. The anthraquinone dye, Reactive Blue 2, remained unchanged after 144 h of incubation in the biosulfidogenic reactor and was only rapidly decolored at 192 h, implying that certain factors are induced in the reactor to break down this non-azo dye. The fastest decolorization/degradation rates and highest hydrogenase enzyme production were observed with Orange II, while the slowest decolorization/degradation rate and least enzyme production were with Reactive Blue 2, suggesting that these processes are controlled, to a certain degree, by an enzymatic mechanism. With sulfate-reducing bacteria that had been cultured on a lactate medium, there was complete decolorization of both authentic dyes and industrial influents and effluents as monitored by the decrease of absorbance in the visible region (480-620 nm). There was, however, very little breakdown of the single aromatic compounds as the absorbance at 280 nm remained fairly significant. This supports the suggestion that, within the biosulfidogenic reactor, there are factors other than the identified hydrogenases that are responsible for degradation of the aromatic compounds.

  7. Thermal annealing and magnetic anisotropy of NiFe thin films on n{sup +}-Si for spintronic device applications

    Energy Technology Data Exchange (ETDEWEB)

    Lu, Q.H. [School of Physical Science and Technology, Lanzhou University, Lanzhou 730000 (China); Department of Physics, Semiconductor Photonics Research Center, Xiamen University, Xiamen 361005, People’s Republic of China (China); Gansu Key Laboratory of Sensor and Sensor Technology, Institute of Sensor Technology, Gansu Academy of Science, Lanzhou 730000 (China); Huang, R. [Department of Physics, Semiconductor Photonics Research Center, Xiamen University, Xiamen 361005, People’s Republic of China (China); Wang, L.S. [Department of Materials Science and Engineering, College of Materials, Xiamen University, Xiamen 361005 (China); Wu, Z.G., E-mail: zgwu@lzu.edu.cn [School of Physical Science and Technology, Lanzhou University, Lanzhou 730000 (China); Li, C., E-mail: lich@xmu.edu.cn [Department of Physics, Semiconductor Photonics Research Center, Xiamen University, Xiamen 361005, People’s Republic of China (China); Luo, Q. [Department of Materials Science and Engineering, College of Materials, Xiamen University, Xiamen 361005 (China); Zuo, S.Y. [School of Physical Science and Technology, Lanzhou University, Lanzhou 730000 (China); Li, J. [Department of Physics, Semiconductor Photonics Research Center, Xiamen University, Xiamen 361005, People’s Republic of China (China); Peng, D.L. [Department of Materials Science and Engineering, College of Materials, Xiamen University, Xiamen 361005 (China); Han, G.L. [Gansu Key Laboratory of Sensor and Sensor Technology, Institute of Sensor Technology, Gansu Academy of Science, Lanzhou 730000 (China); Yan, P.X. [School of Physical Science and Technology, Lanzhou University, Lanzhou 730000 (China)

    2015-11-15

    To ensure that the magnetic metal electrodes can meet the requirements of the spin injection, NiFe films prepared both on HfO{sub 2} dielectric layer and n{sup +}-Si directly by sputtering deposition, and treated by conventional furnace annealing and/or high vacuum magnetic field annealing were investigated. It was found that thermal annealing at 250 °C improved the crystalline quality and reduced surface roughness of the NiFe films, thus enhancing its saturation magnetization intensity. The 100 nm thick NiFe films had too large coercive force and saturation magnetization intensity in vertical direction to meet the requirements of Hanle curve detection. While, 30 nm thick NiFe films showed paramagnetic hysteresis loops in vertical direction, and the magnetization intensity of the sample after annealing at 250 °C for 30 min was less than 2% to the parallel when the external magnetic field was given between ±10 Oe. This was preferred to Hanle curve detection. The thin HfO{sub 2} dielectric layer between metal and Si partially suppressed the diffusion of Ni in NiFe into Si substrate and formation of NiSi, greatly enhancing the saturation magnetization intensity of the Al/NiFe/HfO{sub 2}/Si sample by thermal annealing. Those results suggest that Al/NiFe/HfO{sub 2}/Si structure, from the point view of magnetic electrodes, would be suitable for spin injection and detection applications. - Highlights: • The saturation magnetization intensity of NiFe thin-film was enhanced by thermal annealing. • A paramagnetic hysteresis loop of NiFe thin-film was observed in vertical direction. • The thin HfO{sub 2} dielectric layer between NiFe and Si partially suppressed the diffusion of Ni into Si.

  8. Dechlorination of chlorinated hydrocarbons by bimetallic Ni/Fe immobilized on polyethylene glycol-grafted microfiltration membranes under anoxic conditions.

    Science.gov (United States)

    Parshetti, Ganesh K; Doong, Ruey-an

    2012-01-01

    In this study, the dechlorination of chlorinated hydrocarbons including trichloroethylene (TCE), tetrachloroethylene (PCE) and carbon tetrachloride (CT) by bimetallic Ni/Fe nanoparticles immobilized on four different membranes was investigated under anoxic conditions. Effects of several parameters including the nature of membrane, initial concentration, pH value, and reaction temperature on the dechlorination efficiency were examined. The scanning electron microscopic images showed that the Ni/Fe nanoparticles were successfully immobilized inside the four membranes using polyethylene glycol as the cross-linker. The agglomeration of Ni/Fe were observed in poly(vinylidene fluoride), Millex GS and mixed cellulose ester membranes, while a relatively uniform distribution of Ni/Fe was found in nylon-66 membrane because of its hydrophilic nature. The immobilized Ni/Fe nanoparticles exhibited good reactivity towards the dechlorination of chlorinated hydrocarbons, and the pseudo-first-order rate constant for TCE dechlorination by Ni/Fe in nylon-66 were 3.7-11.7 times higher than those in other membranes. In addition, the dechlorination efficiency of chlorinated hydrocarbons followed the order TCE>PCE>CT. Ethane was the only end product for TCE and PCE dechlorination, while dichloromethane and methane were found to be the major products for CT dechlorination, clearly indicating the involvement of reactive hydrogen species in dechlorination. In addition, the initial rate constant for TCE dechlorination increased upon increasing initial TCE concentrations and the activation energy for TCE dechlorination by immobilized Ni/Fe was 34.9 kJ mol(-1), showing that the dechlorination of TCE by membrane-supported Ni/Fe nanoparticles is a surface-mediated reaction.

  9. [FeFe]-Hydrogenase Abundance and Diversity along a Vertical Redox Gradient in Great Salt Lake, USA

    Directory of Open Access Journals (Sweden)

    Eric S. Boyd

    2014-11-01

    Full Text Available The use of [FeFe]-hydrogenase enzymes for the biotechnological production of H2 or other reduced products has been limited by their sensitivity to oxygen (O2. Here, we apply a PCR-directed approach to determine the distribution, abundance, and diversity of hydA gene fragments along co-varying salinity and O2 gradients in a vertical water column of Great Salt Lake (GSL, UT. The distribution of hydA was constrained to water column transects that had high salt and relatively low O2 concentrations. Recovered HydA deduced amino acid sequences were enriched in hydrophilic amino acids relative to HydA from less saline environments. In addition, they harbored interesting variations in the amino acid environment of the complex H-cluster metalloenzyme active site and putative gas transfer channels that may be important for both H2 transfer and O2 susceptibility. A phylogenetic framework was created to infer the accessory cluster composition and quaternary structure of recovered HydA protein sequences based on phylogenetic relationships and the gene contexts of known complete HydA sequences. Numerous recovered HydA are predicted to harbor multiple N- and C-terminal accessory iron-sulfur cluster binding domains and are likely to exist as multisubunit complexes. This study indicates an important role for [FeFe]-hydrogenases in the functioning of the GSL ecosystem and provides new target genes and variants for use in identifying O2 tolerant enzymes for biotechnological applications.

  10. Nano-sized Si{sub 3}N{sub 4} reinforced NiFe nanocomposites by electroplating

    Energy Technology Data Exchange (ETDEWEB)

    Li Xiaochun; Li Zhiwei

    2003-10-15

    Permalloy NiFe/Si{sub 3}N{sub 4} nanocomposite layers were electroplated on a copper substrate. The volume fraction of nano-sized Si{sub 3}N{sub 4} particles in NiFe matrix was controlled by the addition of various percentages of Si{sub 3}N{sub 4} particles in the NiFe electrolyte. The nanocomposite layers of up to 400 {mu}m were analyzed by a scanning electron microscopy (SEM). Microhardness test was performed. With nano-sized Si{sub 3}N{sub 4} particles in the NiFe matrix, the microhardness of NiFe was improved. The samples were then annealed at 800 deg. C for about 20 h. The microhardness declined more with more Si{sub 3}N{sub 4} particles in the NiFe matrix. The analysis result from energy dispersive spectrometer (EDS) in the SEM showed that the hardness declination could be caused by the segregation of Si{sub 3}N{sub 4} in the NiFe matrix. Finally, this paper presents nanocomposite micromolds fabricated by electroplating onto polymer molds that were fabricated by micro-stereolithgraphy.

  11. Lightweight NiFe2O4 with controllable 3D network structure and enhanced microwave absorbing properties

    Science.gov (United States)

    Wang, Fen; Wang, Xing; Zhu, Jianfeng; Yang, Haibo; Kong, Xingang; Liu, Xiao

    2016-11-01

    3D network structure NiFe2O4 was successfully synthesized by a templated salt precipitation method using PMMA colloid crystal as templates. The morphology, phase composition and microwave absorbing properties of as-prepared samples were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), vector network analyzer (VNA), and so on. The results revealed that the 3D network structure was configurated with smooth spherical walls composed of NiFe2O4 nanocrystals and their pore diameters being in the range of 80–250 nm. The microwave absorption properties of the 3D network structure NiFe2O4 were crucially determined by the special structure. The synergy of intrinsic magnetic loss of magnetic NiFe2O4 and the interfacial polarization enhanced by 3D network structure and the interaction of multiple mechanisms endowed the sample with the feature of strong absorption, broad bandwidth and lightweight. There is more than one valley in the reflection loss curves and the maximum reflection loss is 27.5 dB with a bandwidth of 4 GHz. Moreover, the 3D network structure NiFe2O4 show a greater reflection loss with the same thickness comparing to the ordinary NiFe2O4 nanoparticles, which could achieve the feature of lightweight of the microwave absorbing materials.

  12. Synthesis and Characterization of NiFe2O4 Magnetic Nanoparticles by Combustion Method

    Institute of Scientific and Technical Information of China (English)

    M. Kooti; A. Naghdi Sedeh

    2013-01-01

    Magnetic nanoparticles of nickel ferrite (NiFe2O4) have been successfully synthesized by microwave-assisted combustion method using stable ferric and nickel salts as precursors and glycine as fuel.The as-synthesized samples were characterized by Fourier transform infrared spectroscopy (FT-IR),X-ray diffraction (XRD),transmission electron microscopy (TEM),vibrating sample magnetometer (VSM) and field emission scanning electron microscopy (FESEM).The effect of different dose of glycine on the structural parameters and magnetic properties of the prepared NiFe2O4 nanoparticles was also investigated.This study revealed that it was possible to produce larger size of nanoparticles with lower saturation magnetization by using higher dose of fuel.

  13. High-temperature Mossbauer spectroscopy of mechanically milled NiFe2O4

    DEFF Research Database (Denmark)

    Helgason, O.; Jiang, Jianzhong

    2002-01-01

    Oxide spinels, in particular those containing iron, often exhibit technically important electrical- and magnetic-properties. We report here on X-ray powder diffraction and Mossbauer studies of nanostructured NiFe2O4 particles prepared by high-energy ball milling from bulk NiFe2O4, which...... is an inverse spinel. The Mossbauer spectra were recorded in situ at different temperatures in the range of 300-850 K. The Mossbauer spectra of the milled samples show a broad distribution of magnetic hyperfine fields together with a paramagnetic state at room temperature. Initially, at 700 K the spectrum...... is mainly paramagnetic, but during the process of annealing, magnetic sextets emerge. The treatment results in a significant change in the B/A area ratio of the ferrite. The Neel temperature of the samples is estimated from the B(T) relation to be in the range of 800-850 K....

  14. Effects of Sodium Citrate Concentration on Electroless Ni-Fe Bath Stability and Deposition

    Science.gov (United States)

    Jung, Myung-Won; Kang, Sung K.; Lee, Jae-Ho

    2014-01-01

    In this research, electroless Ni-Fe bath stability and deposition characteristics were investigated for various sodium citrate concentrations. Complexing agents such as sodium citrate are one of the main components of such electroless plating baths. Since they could play various roles such as maintaining pH stability, preventing precipitation of metal salts, and reducing the concentrations of free metal ions, the concentration of complexing agents in the plating bath is an important parameter for electroless deposition processes. In this research, unstable baths were obtained for insufficient sodium citrate concentrations, and these phenomena were analyzed with ChemEQL. Moreover, the deposition characteristics of electroless Ni-Fe for under bump metallurgy diffusion barriers were also investigated using energy-dispersive spectroscopy and field-emission scanning electron microscopy.

  15. Breathing air to save energy--new insights into the ecophysiological role of high-affinity [NiFe]-hydrogenase in Streptomyces avermitilis.

    Science.gov (United States)

    Liot, Quentin; Constant, Philippe

    2016-02-01

    The Streptomyces avermitilis genome encodes a putative high-affinity [NiFe]-hydrogenase conferring the ability to oxidize tropospheric H2 in mature spores. Here, we used a combination of transcriptomic and mutagenesis approaches to shed light on the potential ecophysiological role of the enzyme. First, S. avermitilis was either exposed to low or hydrogenase-saturating levels of H2 to investigate the impact of H2 on spore transcriptome. In total, 1293 genes were differentially expressed, with 1127 and 166 showing lower and higher expression under elevated H2 concentration, respectively. High H2 exposure lowered the expression of the Sec protein secretion pathway and ATP-binding cassette-transporters, with increased expression of genes encoding proteins directing carbon metabolism toward sugar anabolism and lower expression of NADH dehydrogenase in the respiratory chain. Overall, the expression of relA responsible for the synthesis of the pleiotropic alarmone ppGpp decreased upon elevated H2 exposure, which likely explained the reduced expression of antibiotic synthesis and stress response genes. Finally, deletion of hhySL genes resulted in a loss of H2 uptake activity and a dramatic loss of viability in spores. We propose that H2 is restricted to support the seed bank of Streptomyces under a unique survival-mixotrophic energy mode and discuss important ecological implications of this finding.

  16. Hydrogenase Activity of Mineral-Associated and Suspended Populations of Desulfovibrio desulfuricans Essex 6

    Energy Technology Data Exchange (ETDEWEB)

    C.L. Reardon; T.S. Magnuson; E.S. Boyd; W.D. Leavitt; D.W. Reed; G.G. Geesey

    2014-02-01

    The interactions between sulfate-reducing microorganisms and iron oxides influence a number of important redox-sensitive biogeochemical processes including the formation of iron sulfides. Enzymes, such as hydrogenase which catalyze the reversible oxidation of molecular hydrogen, are known to mediate electron transfer to metals and may contribute to the formation and speciation of ferrous sulfides formed at the cell–mineral interface. In the present study, we compared the whole cell hydrogenase activity of Desulfovibrio desulfuricans strain Essex 6 growing as biofilms on hematite (hematite-associated) or as suspended populations using different metabolic pathways. Hematite-associated cells exhibited significantly greater hydrogenase activity than suspended populations during sulfate respiration but not during pyruvate fermentation. The enhanced activity of the hematite-associated, sulfate-grown cells appears to be dependent on iron availability rather than a general response to surface attachment since the activity of glass-associated cells did not differ from that of suspended populations. Hydrogenase activity of pyruvate-fermenting cells was stimulated by addition of iron as soluble Fe(II)Cl2 and, in the absence of added iron, both sulfate-reducing and pyruvate-fermenting cells displayed similar rates of hydrogenase activity. These data suggest that iron exerts a stronger influence on whole cell hydrogenase activity than either metabolic pathway or mode of growth. The location of hydrogenase to the cell envelope and the enhanced activity at the hematite surface in sulfate-reducing cells may influence the redox conditions that control the species of iron sulfides on the mineral surface.

  17. Characterization of NiFe oxyhydroxide electrocatalysts by integrated electronic structure calculations and spectroelectrochemistry

    Energy Technology Data Exchange (ETDEWEB)

    Goldsmith, Zachary K.; Harshan, Aparna K.; Gerken, James B.; Vörös, Márton; Galli, Giulia; Stahl, Shannon S.; Hammes-Schiffer, Sharon

    2017-03-06

    NiFe oxyhydroxide materials are highly active electrocatalysts for the oxygen evolution reaction (OER), an important process for carbon-neutral energy storage. Recent spectroscopic and computational studies increasingly support iron as the site of catalytic activity but differ with respect to the relevant iron redox state. A combination of hybrid periodic density functional theory calculations and spectroelectrochemical experiments elucidate the electronic structure and redox thermodynamics of Ni-only and mixed NiFe oxyhydroxide thin-film electrocatalysts. The UV/visible light absorbance of the Ni-only catalyst depends on the applied potential as metal ions in the film are oxidized before the onset of OER activity. In contrast, absorbance changes are negligible in a 25% Fe-doped catalyst up to the onset of OER activity. First-principles calculations of proton-coupled redox potentials and magnetizations reveal that the Ni-only system features oxidation of Ni2+ to Ni3+, followed by oxidation to a mixed Ni3+/4+ state at a potential coincident with the onset of OER activity. Calculations on the 25% Fedoped system show the catalyst is redox inert before the onset of catalysis, which coincides with the formation of Fe4+ and mixed Ni oxidation states. The calculations indicate that introduction of Fe dopants changes the character of the conduction band minimum from Ni-oxide in the Ni-only to predominantly Fe-oxide in the NiFe electrocatalyst. These findings provide a unified experimental and theoretical description of the electrochemical and optical properties of Ni and NiFe oxyhydroxide electrocatalysts and serve as an important benchmark for computational characterization of mixedmetal oxidation states in heterogeneous catalysts.

  18. Electrocatalytic oxygen evolution over supported small amorphous Ni-Fe nanoparticles in alkaline electrolyte.

    Science.gov (United States)

    Qiu, Yang; Xin, Le; Li, Wenzhen

    2014-07-08

    The electrocatalytic oxygen evolution reaction (OER) is a critical anode reaction often coupled with electron or photoelectron CO2 reduction and H2 evolution reactions at the cathode for renewable energy conversion and storage. However, the sluggish OER kinetics and the utilization of precious metal catalysts are key obstacles in the broad deployment of these energy technologies. Herein, inexpensive supported 4 nm Ni-Fe nanoparticles (NiyFe1-yOx/C) featuring amorphous structures have been prepared via a solution-phase nanocapsule method for active and durable OER electrocatalysts in alkaline electrolyte. The Ni-Fe nanoparticle catalyst containing 31% Fe (Ni0.69Fe0.31Ox/C) shows the highest activity, exhibiting a 280 mV overpotential at 10 mA cm(-2) (equivalent to 10% efficiency of solar-to-fuel conversion) and a Tafel slope of 30 mV dec(-1) in 1.0 M KOH solution. The achieved OER activity outperforms NiOx/C and commercial Ir/C catalysts and is close to the highest performance of crystalline Ni-Fe thin films reported in the literature. In addition, a Faradaic efficiency of 97% measured on Ni0.69Fe0.31Ox/C suggests that carbon support corrosion and further oxidation of nanoparticle catalysts are negligible during the electrocatalytic OER tests. Ni0.69Fe0.31Ox/C further demonstrates high stability as there is no apparent OER activity loss (based on a chronoamperometry test) or particle aggregation (based on TEM image observation) after a 6 h anodization test. The high efficiency and durability make these supported amorphous Ni-Fe nanoparticles potentially applicable in the (photo)electrochemical cells for water splitting to make H2 fuel or CO2 reduction to produce usable fuels and chemicals.

  19. Surface Structure and Catalytic Performance of Ni-Fe Catalyst for Low-Temperature CO Hydrogenation

    Directory of Open Access Journals (Sweden)

    Fanhui Meng

    2014-01-01

    Full Text Available Catalysts 16NixFe/Al2O3 (x is 0, 1, 2, 4, 6, 8 were prepared by incipient wetness impregnation method and the catalytic performance for the production of synthetic natural gas (SNG from CO hydrogenation in slurry-bed reactor were studied. The catalysts were characterized by BET, XRD, UV-Vis DRS, H2-TPR, CO-TPD, and XPS, and the results showed that the introduction of iron improved the dispersion of Ni species, weakened the interaction between Ni species and support and decreased the reduction temperature and that catalyst formed Ni-Fe alloy when the content of iron exceeded 2%. Experimental results revealed that the addition of iron to the catalyst can effectively improve the catalytic performance of low-temperature CO methanation. Catalyst 16Ni4Fe/Al2O3 with the iron content of 4% exhibited the best catalytic performance, the conversion of CO and the yield of CH4 reached 97.2% and 84.9%, respectively, and the high catalytic performance of Ni-Fe catalyst was related to the property of formed Ni-Fe alloy. Further increase of iron content led to enhancing the water gas shift reaction.

  20. Polymer stabilized Ni-Ag and Ni-Fe alloy nanoclusters: Structural and magnetic properties

    Energy Technology Data Exchange (ETDEWEB)

    Kabir, L.; Mandal, A.R. [Department of Physics, Visva-Bharati, Santiniketan-731 235 (India); Mandal, S.K., E-mail: sk_mandal@hotmail.co [Department of Physics, Visva-Bharati, Santiniketan-731 235 (India)

    2010-04-15

    We report here the structural and magnetic behaviors of nickel-silver (Ni-Ag) and nickel-iron (Ni-Fe) nanoclusters stabilized with polymer (polypyrrole). High resolution transmission electron microscopy (HRTEM) indicates Ni-Ag nanoclusters to stabilize in core-shell configuration while that of Ni-Fe nanoclusters in a mixed type of geometry. Structural characterizations by X-ray diffraction (XRD) reveal the possibility of alloying in such bimetallic nanoclusters to some extent even at temperatures much lower than that of bulk alloying. Electron paramagnetic resonance (EPR) spectra clearly reveal two different absorption behaviors: one is ascribed to non-isolated Ni{sup 2+} clusters surrounded by either silver or iron giving rise to a broad signal, other (very narrow signal) being due to the isolated superparamagnetic Ni{sup 2+} clusters or bimetallic alloy nanoclusters. Results obtained for Ni-Ag and Ni-Fe nanoclusters have been further compared with the behavior exhibited by pure Ni nanoclusters in polypyrrole host. Temperature dependent studies (at 300 and 77 K) of EPR parameters, e.g. linewidth, g-value, line shape and signal intensity indicating the significant influence of surrounding paramagnetic silver or ferromagnetic iron within polymer host on the EPR spectra have been presented.

  1. NiFe2O4/activated carbon nanocomposite as magnetic material from petcoke

    Science.gov (United States)

    Briceño, Sarah; Brämer-Escamilla, W.; Silva, P.; García, J.; Del Castillo, H.; Villarroel, M.; Rodriguez, J. P.; Ramos, M. A.; Morales, R.; Diaz, Y.

    2014-06-01

    Nickel ferrite (NiFe2O4) was supported on activated carbon (AC) from petroleum coke (petcoke). Potassium hydroxide (KOH) was employed with petcoke to produce activated carbon. NiFe2O4 were synthesized using PEG-Oleic acid assisted hydrothermal method. The structural and magnetic properties were determined using thermogravimetric and differential thermal analysis (TGA-DTA), X-ray diffraction (XRD), Fourier Transform Infrared (IR-FT), surface area (BET), scanning electron microscopy (SEM), transmission electron microscopy (TEM) and vibrating sample magnetometry (VSM). XRD analysis revealed the cubic spinel structure and ferrite phase with high crystallinity. IR-FT studies showed that chemical modification promoted the formation of surface oxygen functionalities. Morphological investigation by SEM showed conglomerates of spherical nanoparticles with an average particle size of 72 nm and TEM showed the formation of NiFe2O4/carbon nanofibers. Chemical modification and activation temperature of 800 °C prior to activation dramatically increased the BET surface area of the resulting activated carbon to 842.4 m2/g while the sulfur content was reduced from 6 to 1%. Magnetic properties of nanoparticles show strong dependence on the particle size.

  2. Magnetic properties of Ni-Fe nanowire arrays: effect of template material and deposition conditions

    Energy Technology Data Exchange (ETDEWEB)

    Singleton, John [Los Alamos National Laboratory; Aravamudhan, Shyan [U OF SOUTH FL; Goddard, Paul A [U OF OXFORD; Bhansali, Shekhar [U OF SOUTH FL

    2008-01-01

    The objective of this work is to study the magnetic properties of arrays of Ni-Fe nanowires electrodeposited in different template materials such as porous silicon, polycarbonate and alumina. Magnetic properties were studied as a function of template material, applied magnetic field (parallel and perpendicular) during deposition, wire length, as well as magnetic field orientation during measurement. The results show that application of magnetic field during deposition strongly influences the c-axis preferred orientation growth of Ni-Fe nanowires. The samples with magnetic field perpendicular to template plane during deposition exhibits strong perpendicular anisotropy with greatly enhanced coercivity and squareness ratio, particularly in Ni-Fe nanowires deposited in polycarbonate templates. In case of polycarbonate template, as magnetic field during deposition increases, both coercivity and squareness ratio also increase. The wire length dependence was also measured for polycarbonate templates. As wire length increases, coercivity and squarness ratio decrease, but saturation field increases. Such magnetic behavior (dependence on template material, magnetic field, wire length) can be qualitatively explained by preferential growth phenomena, dipolar interactions among nanowires, and perpendicular shape anisotropy in individual nanowires.

  3. Multilayer Ni/Fe thin films as oxygen evolution catalysts for solar fuel production

    Science.gov (United States)

    Biset-Peiró, M.; Murcia-López, S.; Fàbrega, C.; Morante, J. R.; Andreu, T.

    2017-03-01

    The slow kinetics and high overpotential of the oxygen evolution reaction is one of the main limiting factors to achieve the minimum required performances of the so-called photoelectrochemical water splitting systems. An oxygen evolution catalyst (OEC) becomes essential in order to perform this process with higher efficiency. Herein, we report the physical, optical and electrochemical characterization of multilayer Ni/Fe thin films as earth-abundant OEC, to avoid the use of platinum group metals (PGM). Uniform films of thicknesses ranging from 1 to 10 nm were fabricated by sequential and alternate thermal evaporation of Ni and Fe. It was found that the successive deposition allows the fabrication of a Ni terminated surface that does not need activation due to the Fe underlayer. The lowest overpotential achieved for NiFe was 370 mV at 10 mA cm‑2 and a Tafel slope of 37 mV dec‑1 with 1 nm thickness and 95% transmittance. Finally, NiFe OEC was implemented on top of Mo:BiVO4 photoanodes which resulted in a reduction of the open circuit potential of 0.2 V and up to five fold increase of the oxidation efficiency at 0.7 VRHE. The results presented facilitate the practical implementation of BiVO4 photoanodes in tandem configuration for bias free photoassisted water splitting.

  4. Elemental redistribution in a nanocrystalline Ni-Fe alloy induced by high-pressure torsion

    Energy Technology Data Exchange (ETDEWEB)

    Ni, S. [School of Aerospace, Mechanical and Mechatronic Engineering, University of Sydney, Sydney, NSW 2006 (Australia); Sha, G., E-mail: gang.sha@sydney.edu.au [Australian Centre for Microscopy and Microanalysis, University of Sydney, Sydney, NSW 2006 (Australia); Wang, Y.B. [School of Aerospace, Mechanical and Mechatronic Engineering, The University of Sydney, Sydney, NSW 2006 (Australia); Liao, X.Z., E-mail: xiaozhou.liao@sydney.edu.au [School of Aerospace, Mechanical and Mechatronic Engineering, University of Sydney, Sydney, NSW 2006 (Australia); Alhajeri, S.N. [Department of Manufacturing Engineering, College of Technological Studies, PAAET, Shuwaikh 70654 (Kuwait); Li, H.Q. [Los Alamos National Laboratory, Los Alamos, NM 87545 (United States); Zhu, Y.T. [Department of Materials Science and Engineering, North Carolina State University, Raleigh, NC 27659-7919 (United States); Langdon, T.G. [Materials Research Group, School of Engineering Sciences, University of Southampton, Southampton SO17 1BJ (United Kingdom); Departments of Aerospace and Mechanical Engineering and Materials Science, University of Southern California, Los Angeles, CA 90089-1453 (United States); Ringer, S.P. [Australian Centre for Microscopy and Microanalysis, University of Sydney, Sydney, NSW 2006 (Australia)

    2011-09-25

    Highlights: {center_dot} Elemental distribution of a nc Ni-Fe alloy before and after high-pressure torsion. {center_dot} The supersaturated Ni-Fe solid solution was stable under HPT. {center_dot} C and S atoms further segregated to the remaining GBs during grain growth. {center_dot} GB diffusion and the motion of defects facilitate the elemental redistribution. - Abstract: An electrochemically deposited nanocrystalline supersaturated face-centred-cubic Ni-21 at.% Fe alloy with an initial average grain size of {approx}21 nm was processed using high-pressure torsion (HPT) that resulted in grain growth via grain rotation and coalescence to an average grain size of {approx}53 nm. Atom probe tomography investigations revealed that the supersaturated Ni-Fe solid solution was stable under HPT and that C and S atoms, which are the major impurities in the material and segregated to the grain boundaries (GBs) of the as-deposited material, migrated from disappearing GBs to the remaining GBs during HPT. We propose that the elemental redistribution was facilitated by GB diffusion and the motion of a large volume of HPT-induced defects at the GB regions during the grain growth process. This elemental redistribution process is different from other HPT-induced elemental redistribution processes reported in the literature.

  5. New insights into [FeFe] hydrogenase activation and maturase function.

    Directory of Open Access Journals (Sweden)

    Jon M Kuchenreuther

    Full Text Available [FeFe] hydrogenases catalyze H(2 production using the H-cluster, an iron-sulfur cofactor that contains carbon monoxide (CO, cyanide (CN(-, and a dithiolate bridging ligand. The HydE, HydF, and HydG maturases assist in assembling the H-cluster and maturing hydrogenases into their catalytically active form. Characterization of these maturases and in vitro hydrogenase activation methods have helped elucidate steps in the H-cluster biosynthetic pathway such as the HydG-catalyzed generation of the CO and CN(- ligands from free tyrosine. We have refined our cell-free approach for H-cluster synthesis and hydrogenase maturation by using separately expressed and purified HydE, HydF, and HydG. In this report, we illustrate how substrates and protein constituents influence hydrogenase activation, and for the first time, we show that each maturase can function catalytically during the maturation process. With precise control over the biomolecular components, we also provide evidence for H-cluster synthesis in the absence of either HydE or HydF, and we further show that hydrogenase activation can occur without exogenous tyrosine. Given these findings, we suggest a new reaction sequence for the [FeFe] hydrogenase maturation pathway. In our model, HydG independently synthesizes an iron-based compound with CO and CN(- ligands that is a precursor to the H-cluster [2Fe](H subunit, and which we have termed HydG-co. We further propose that HydF is a transferase that stabilizes HydG-co and also shuttles the complete [2Fe](H subcluster to the hydrogenase, a translocation process that may be catalyzed by HydE. In summary, this report describes the first example of reconstructing the [FeFe] hydrogenase maturation pathway using purified maturases and subsequently utilizing this in vitro system to better understand the roles of HydE, HydF, and HydG.

  6. Optimization of NiFe2O4/rGO composite electrode for lithium-ion batteries

    Science.gov (United States)

    Li, Chen; Wang, Xia; Li, Shandong; Li, Qiang; Xu, Jie; Liu, Xiaomin; Liu, Changkun; Xu, Yuanhong; Liu, Jingquan; Li, Hongliang; Guo, Peizhi; Zhao, Xiu Song

    2017-09-01

    The combination of carbon compositing and the proper choice of binders in one system offer an effective strategy for improving electrode performance for lithium ion batteries (LIBs). Here, we focus on the optimization of reduced graphene oxide content in NiFe2O4/reduced graphene oxide (abbreviated to NiFe2O4/rGO) composites and the proper choice of binders to enhance the cycling stability of the NiFe2O4 electrode. The NiFe2O4/rGO composites were fabricated by a hydrothermal-annealing method, in which the mean size of spinel NiFe2O4 nanoparticles was approximately 20 nm. When tested as anode materials for LIBs, the NiFe2O4/rGO electrodes with carboxymethylcellulose (CMC) binder exhibited excellent lithium-storage performance including high reversible capacity, good cycling durability and high-rate capability. The capacity could be retained as high as 1105 mAh g-1 at a current density of 100 mA g-1 for over 50 cycles, even cycled at higher current density of 1000 mA g-1, a capacity of 800 mAh g-1can be obtained, whereas the electrode with the polyvinylidene fluoride (PVDF) binder suffered from rapid capacity decay under the same test conditions. As a result, the NiFe2O4/rGO composites with CMC binder electrode in this work are promising as anodes for high-performance LIBs, resulting from the synergistic effect of optimal graphene content and proper choice of binder.

  7. Cooperativity and Dynamics Increase the Performance of NiFe Dry Reforming Catalysts.

    Science.gov (United States)

    Kim, Sung Min; Abdala, Paula Macarena; Margossian, Tigran; Hosseini, Davood; Foppa, Lucas; Armutlulu, Andac; van Beek, Wouter; Comas-Vives, Aleix; Copéret, Christophe; Müller, Christoph

    2017-02-08

    The dry reforming of methane (DRM), i.e., the reaction of methane and CO2 to form a synthesis gas, converts two major greenhouse gases into a useful chemical feedstock. In this work, we probe the effect and role of Fe in bimetallic NiFe dry reforming catalysts. To this end, monometallic Ni, Fe, and bimetallic Ni-Fe catalysts supported on a MgxAlyOz matrix derived via a hydrotalcite-like precursor were synthesized. Importantly, the textural features of the catalysts, i.e., the specific surface area (172-178 m(2)/gcat), pore volume (0.51-0.66 cm(3)/gcat), and particle size (5.4-5.8 nm) were kept constant. Bimetallic, Ni4Fe1 with Ni/(Ni + Fe) = 0.8, showed the highest activity and stability, whereas rapid deactivation and a low catalytic activity were observed for monometallic Ni and Fe catalysts, respectively. XRD, Raman, TPO, and TEM analysis confirmed that the deactivation of monometallic Ni catalysts was in large due to the formation of graphitic carbon. The promoting effect of Fe in bimetallic Ni-Fe was elucidated by combining operando XRD and XAS analyses and energy-dispersive X-ray spectroscopy complemented with density functional theory calculations. Under dry reforming conditions, Fe is oxidized partially to FeO leading to a partial dealloying and formation of a Ni-richer NiFe alloy. Fe migrates leading to the formation of FeO preferentially at the surface. Experiments in an inert helium atmosphere confirm that FeO reacts via a redox mechanism with carbon deposits forming CO, whereby the reduced Fe restores the original Ni-Fe alloy. Owing to the high activity of the material and the absence of any XRD signature of FeO, it is very likely that FeO is formed as small domains of a few atom layer thickness covering a fraction of the surface of the Ni-rich particles, ensuring a close proximity of the carbon removal (FeO) and methane activation (Ni) sites.

  8. Amperometric hydrogen peroxide and glucose biosensor based on NiFe2/ordered mesoporous carbon nanocomposites.

    Science.gov (United States)

    Xiang, Dong; Yin, Longwei; Ma, Jingyun; Guo, Enyan; Li, Qun; Li, Zhaoqiang; Liu, Kegao

    2015-01-21

    Nanocomposites of NiFex embedded in ordered mesoporous carbon (OMC) (x = 0, 1, 2) were prepared by a wet impregnation and hydrogen reduction process and were used to construct electrochemical biosensors for the amperometric detection of hydrogen peroxide (H2O2) or glucose. The NiFe2/OMC nanocomposites were demonstrated to have a large surface area, suitable mesoporous channels, many edge-plane-like defective sites, and a good distribution of alloyed nanoparticles. The NiFe2/OMC and Nafion modified glass carbon electrode (GCE) exhibited excellent electrocatalytic activities toward the reduction of H2O2 as well. By utilizing it as a bioplatform, GOx (glucose oxidase) cross-linked with Nafion was immobilized on the surface of the electrode for the construction of an amperometric glucose biosensor. Our results indicated that the amperometric hydrogen peroxide biosensor (NiFe2/OMC + Nafion + GCE) showed good analytical performances in term of a high sensitivity of 4.29 μA mM(-1) cm(-2), wide linearity from 6.2 to 42,710 μM and a low detection limit of 0.24 μM at a signal-to-noise ratio of 3 (S/N = 3). This biosensor exhibited excellent selectivity, high stability and negligible interference for the detection of H2O2. In addition, the immobilized enzyme on NiFe2/OMC + Nafion + GCE, retaining its bioactivity, exhibited a reversible two-proton and two-electron transfer reaction, a fast heterogeneous electron transfer rate and an effective Michaelis-Menten constant (K) (3.18 mM). The GOx + NiFe2/OMC + Nafion + GCE could be used to detect glucose based on the oxidation of glucose catalyzed by GOx and exhibited a wide detection range of 48.6-12,500 μM with a high sensitivity of 6.9 μA mM(-1) cm(-2) and a low detection limit of 2.7 μM (S/N = 3). The enzymic biosensor maintained a high selectivity and stability features, and shows great promise for application in the detection of glucose.

  9. Characterization of the oxygen tolerance of a hydrogenase linked to a carbon monoxide oxidation pathway in Rubrivivax gelatinosus.

    Science.gov (United States)

    Maness, Pin-Ching; Smolinski, Sharon; Dillon, Anne C; Heben, Michael J; Weaver, Paul F

    2002-06-01

    A hydrogenase linked to the carbon monoxide oxidation pathway in Rubrivivax gelatinosus displays tolerance to O2. When either whole-cell or membrane-free partially purified hydrogenase was stirred in full air (21% O2, 79% N2), its H2 evolution activity exhibited a half-life of 20 or 6 h, respectively, as determined by an anaerobic assay using reduced methyl viologen. When the partially purified hydrogenase was stirred in an atmosphere containing either 3.3 or 13% O2 for 15 min and evaluated by a hydrogen-deuterium (H-D) exchange assay, nearly 80 or 60% of its isotopic exchange rate was retained, respectively. When this enzyme suspension was subsequently returned to an anaerobic atmosphere, more than 90% of the H-D exchange activity was recovered, reflecting the reversibility of this hydrogenase toward O2 inactivation. Like most hydrogenases, the CO-linked hydrogenase was extremely sensitive to CO, with 50% inhibition occurring at 3.9 microM dissolved CO. Hydrogen production from the CO-linked hydrogenase was detected when ferredoxins of a prokaryotic source were the immediate electron mediator, provided they were photoreduced by spinach thylakoid membranes containing active water-splitting activity. Based on its appreciable tolerance to O2, potential applications of this hydrogenase are discussed.

  10. 大豆和花生根瘤菌氢酶的研究%Studies on Hydrogenase from Rhizobium japonicum and Rhizobium arachis

    Institute of Scientific and Technical Information of China (English)

    许良树; 张凤章; 龙敏南; 曾定; 黄河清; 刘月英; 刘广发

    2001-01-01

    be detected in the fractions with the highest specific activity of hydrogenase from R. japonicum. R. arachis L8-3 strains(hup+) expressed high hydrogenase activity when grow autotrophicully in mineral salt-vitamins medium and gas mixture containing H2 and CO2. It is evident that H2 uptake by hydrogenase significantly increased the dinitrogen fixation either in free-living cultures or in symbiotic nodules. A genomic library of R. arachis L8-3(hup+) has been constructed. Four clones(pZ-27, pZ-55, pZ-60, pZ-61) containing hup gene were screened by PCR and hydrogenase probe. The fragment of insert DNA of recombinant cosmid pZ-55 was about 19. 6 kb. When the cosmid pZ-55 was transferred into the hup- strain of R. arachis and hup- strain of R. japonicum by triparental matting, the transconjugants showed high levels of H2 uptake activities in free-living state. Inoculating test was carried out in the field by using the transconjugant containing pZ-55. The result indicated that H2-uptake activity of nodules inoculated with hup+ transconjugant was 4 fold high than those of nodule inoculated with recipient (hup-). The total nitrogen content of peanut leaves and seed increased by 17%0 and8.9 % respectively. The peanut yield inoculated with transconjugant increased by 9.6 %0 in comparison with that inoculated with recipient.

  11. Non-local detection of spin dynamics via spin rectification effect in yttrium iron garnet/SiO2/NiFe trilayers near simultaneous ferromagnetic resonance

    Science.gov (United States)

    Soh, Wee Tee; Peng, Bin; Ong, C. K.

    2015-08-01

    The spin rectification effect (SRE), a phenomenon that generates dc voltages from ac microwave fields incident onto a conducting ferromagnet, has attracted widespread attention due to its high sensitivity to ferromagnetic resonance (FMR) as well as its relevance to spintronics. Here, we report the non-local detection of yttrium iron garnet (YIG) spin dynamics by measuring SRE voltages from an adjacent conducting NiFe layer up to 200 nm thick. In particular, we detect, within the NiFe layer, SRE voltages stemming from magnetostatic surface spin waves (MSSWs) of the adjacent bulk YIG which are excited by a shorted coaxial probe. These non-local SRE voltages within the NiFe layer that originates from YIG MSSWs are present even in 200 nm-thick NiFe films with a 50 nm thick SiO2 spacer between NiFe and YIG, thus strongly ruling out the mechanism of spin-pumping induced inverse spin Hall effect in NiFe as the source of these voltages. This long-range influence of YIG dynamics is suggested to be mediated by dynamic fields generated from YIG spin precession near YIG/NiFe interface, which interacts with NiFe spins near the simultaneous resonance of both spins, to generate a non-local SRE voltage within the NiFe layer.

  12. Non-local detection of spin dynamics via spin rectification effect in yttrium iron garnet/SiO2/NiFe trilayers near simultaneous ferromagnetic resonance

    Directory of Open Access Journals (Sweden)

    Wee Tee Soh

    2015-08-01

    Full Text Available The spin rectification effect (SRE, a phenomenon that generates dc voltages from ac microwave fields incident onto a conducting ferromagnet, has attracted widespread attention due to its high sensitivity to ferromagnetic resonance (FMR as well as its relevance to spintronics. Here, we report the non-local detection of yttrium iron garnet (YIG spin dynamics by measuring SRE voltages from an adjacent conducting NiFe layer up to 200 nm thick. In particular, we detect, within the NiFe layer, SRE voltages stemming from magnetostatic surface spin waves (MSSWs of the adjacent bulk YIG which are excited by a shorted coaxial probe. These non-local SRE voltages within the NiFe layer that originates from YIG MSSWs are present even in 200 nm-thick NiFe films with a 50 nm thick SiO2 spacer between NiFe and YIG, thus strongly ruling out the mechanism of spin-pumping induced inverse spin Hall effect in NiFe as the source of these voltages. This long-range influence of YIG dynamics is suggested to be mediated by dynamic fields generated from YIG spin precession near YIG/NiFe interface, which interacts with NiFe spins near the simultaneous resonance of both spins, to generate a non-local SRE voltage within the NiFe layer.

  13. Non-local detection of spin dynamics via spin rectification effect in yttrium iron garnet/SiO{sub 2}/NiFe trilayers near simultaneous ferromagnetic resonance

    Energy Technology Data Exchange (ETDEWEB)

    Soh, Wee Tee, E-mail: a0046479@u.nus.edu; Ong, C. K. [Center for Superconducting and Magnetic Materials, Department of Physics, National University of Singapore, 2 Science Drive 3, Singapore 117551 (Singapore); Peng, Bin [State Key Laboratory of Electronic Thin Films and Integrated Devices, University of Electronic Science and Technology of China, Chengdu 610054 (China)

    2015-08-15

    The spin rectification effect (SRE), a phenomenon that generates dc voltages from ac microwave fields incident onto a conducting ferromagnet, has attracted widespread attention due to its high sensitivity to ferromagnetic resonance (FMR) as well as its relevance to spintronics. Here, we report the non-local detection of yttrium iron garnet (YIG) spin dynamics by measuring SRE voltages from an adjacent conducting NiFe layer up to 200 nm thick. In particular, we detect, within the NiFe layer, SRE voltages stemming from magnetostatic surface spin waves (MSSWs) of the adjacent bulk YIG which are excited by a shorted coaxial probe. These non-local SRE voltages within the NiFe layer that originates from YIG MSSWs are present even in 200 nm-thick NiFe films with a 50 nm thick SiO{sub 2} spacer between NiFe and YIG, thus strongly ruling out the mechanism of spin-pumping induced inverse spin Hall effect in NiFe as the source of these voltages. This long-range influence of YIG dynamics is suggested to be mediated by dynamic fields generated from YIG spin precession near YIG/NiFe interface, which interacts with NiFe spins near the simultaneous resonance of both spins, to generate a non-local SRE voltage within the NiFe layer.

  14. Hydrogen production by a hyperthermophilic membrane-bound hydrogenase in water-soluble nanolipoprotein particles.

    Science.gov (United States)

    Baker, Sarah E; Hopkins, Robert C; Blanchette, Craig D; Walsworth, Vicki L; Sumbad, Rhoda; Fischer, Nicholas O; Kuhn, Edward A; Coleman, Matt; Chromy, Brett A; Létant, Sonia E; Hoeprich, Paul D; Adams, Michael W W; Henderson, Paul T

    2009-06-10

    Hydrogenases constitute a promising class of enzymes for ex vivo hydrogen production. Implementation of such applications is currently hindered by oxygen sensitivity and, in the case of membrane-bound hydrogenases (MBHs), poor water solubility. Nanolipoprotein particles (NLPs) formed from apolipoproteins and phospholipids offer a novel means of incorporating MBHs into a well-defined water-soluble matrix that maintains the enzymatic activity and is amenable to incorporation into more complex architectures. We report the synthesis, hydrogen-evolving activity, and physical characterization of the first MBH-NLP assembly. This may ultimately lead to the development of biomimetic hydrogen-production devices.

  15. Hydrogen Production by a Hyperthermophilic Membrane-Bound Hydrogenase in Soluble Nanolipoprotein Particles

    Energy Technology Data Exchange (ETDEWEB)

    Baker, S E; Hopkins, R C; Blanchette, C; Walsworth, V; Sumbad, R; Fischer, N; Kuhn, E; Coleman, M; Chromy, B; Letant, S; Hoeprich, P; Adams, M W; Henderson, P T

    2008-10-22

    Hydrogenases constitute a promising class of enzymes for ex vivo hydrogen production. Implementation of such applications is currently hindered by oxygen sensitivity and, in the case of membrane-bound hydrogenases (MBH), poor water solubility. Nanolipoprotein particles (NLPs), formed from apolipoproteins and phospholipids, offer a novel means to incorporate MBH into in a well-defined water-soluble matrix that maintains the enzymatic activity and is amenable to incorporation into more complex architectures. We report the synthesis, hydrogen-evolving activity and physical characterization of the first MBH-NLP assembly. This may ultimately lead to the development of biomimetic hydrogen production devices.

  16. High-Quality Draft Genome Sequence of Thermocrinis jamiesonii GBS1T Isolated from Great Boiling Spring, Nevada.

    Science.gov (United States)

    Ganji, Rakesh; Murugapiran, Senthil K; Ong, John C; Manoharan, Namritha; Huntemann, Marcel; Clum, Alicia; Pillay, Manoj; Palaniappan, Krishnaveni; Varghese, Neha; Mikhailova, Natalia; Stamatis, Dimitrios; Reddy, T B K; Ngan, Chew Yee; Daum, Chris; Duffy, Kecia; Shapiro, Nicole; Markowitz, Victor; Ivanova, Natalia; Kyrpides, Nikos; Woyke, Tanja; Dodsworth, Jeremy A; Hedlund, Brian P

    2016-10-20

    The draft genome of Thermocrinis jamiesonii GBS1(T) is 1,315,625 bp in 10 contigs and encodes 1,463 predicted genes. The presence of sox genes and various glycoside hydrolases and the absence of uptake NiFe hydrogenases (hyaB) are consistent with a requirement for thiosulfate and suggest the ability to use carbohydrate polymers.

  17. Formation of magnetic microstructure of the nanosized NiFe2O4 synthesized via solid-state reaction

    Directory of Open Access Journals (Sweden)

    Žák T.

    2012-01-01

    Full Text Available Magnetic NiFe2O4 structure formation was studied through structural, compositional and magnetic characterization of obtained reaction products of a simple, high yielding and low-cost solid-state reaction. Initial annealing of the starting oxides mixture at 700ºC did not allow us to observe formation of the desired magnetic phase. In contrast, subsequent thermomagnetic measurements up to 800ºC indicated the considerable increase of the magnetic moment, which can be reasonably assigned to the changes in phase composition and formation of magnetic NiFe2O4 structure during the heating cycle of measurements. Nanosized NiFe2O4 phase formation has been confirmed by the following XRD and MS phase analyses and its nanocrystalline structure by XRD and SEM/TEM techniques. The obtained hysteresis loop taken after TM measurements suggest the increased volume of magnetically active material and thus additionally support the previous findings.

  18. Novel synthesis of Ni-ferrite (NiFe2O4) electrode material for supercapacitor applications

    Science.gov (United States)

    Venkatachalam, V.; Jayavel, R.

    2015-06-01

    Novel nanocrystalline NiFe2O4 has been synthesized through combustion route using citric acid as a fuel. Phase of the synthesized material was analyzed using powder X-ray diffraction. The XRD study revealed the formation of spinel phase cubic NiFe2O4 with high crystallinity. The average crystallite size of NiFe2O4 nanomaterial was calculated from scherrer equation. The electrochemical properties were realized by cyclic voltammetry, chronopotentiometry and electrochemical impedance spectroscopy. The electrode material shows a maximum specific capacitance of 454 F/g with pseudocapacitive behavior. High capacitance retention of electrode material over 1000 continuous charging-discharging cycles suggests its excellent electrochemical stability. The results revealed that the nickel ferrite electrode is a potential candidate for energy storage applications in supercapacitor.

  19. Remediation of polybrominated diphenyl ethers in soil using Ni/Fe bimetallic nanoparticles: Influencing factors, kinetics and mechanism

    Energy Technology Data Exchange (ETDEWEB)

    Xie, Yingying [School of Chemistry and Environment, South China Normal University, Guangzhou 51006 (China); Guangdong Technology Research Center for Ecological Management and Remediation of Urban Water System, Guangzhou 51006 (China); Fang, Zhanqiang, E-mail: zhqfang@scnu.edu.cn [School of Chemistry and Environment, South China Normal University, Guangzhou 51006 (China); Guangdong Technology Research Center for Ecological Management and Remediation of Urban Water System, Guangzhou 51006 (China); Cheng, Wen [School of Chemistry and Environment, South China Normal University, Guangzhou 51006 (China); Guangdong Technology Research Center for Ecological Management and Remediation of Urban Water System, Guangzhou 51006 (China); Tsang, Pokeung Eric [Department of Science and Environmental Studies, The Hong Kong Institute of Education, Hong Kong 00852 (China); Zhao, Dongye [Environmental Engineering Program, Department of Civil Engineering, Auburn University, Auburn, AL 36849 (United States)

    2014-07-01

    Polybrominated diphenyl ethers (PBDEs) are commonly used as additive flame retardants in all kinds of electronic products. PBDEs are now ubiquitous in the environment, with soil as a major sink, especially in e-waste recycling sites. This study investigated the degradation of decabromodiphenyl ether (BDE209) in a spiked soil using Ni/Fe bimetallic nanoparticles. The results indicated that Ni/Fe bimetallic nanoparticles are able to degrade BDE209 in soil at ambient temperature and the removal efficiency can reach 72% when an initial pH of 5.6 and at a Ni/Fe dosage of 0.03 g/g. A declining trend in degradation was noticed with decreasing Ni loading and increasing of initial BDE209 concentration. The degradation products of BDE209 were analyzed by GC-MS, which showed that the degradation of BDE209 was a process of stepwise debromination from nBr to (n − 1)Br. And a possible debromination pathway was proposed. At last, the degradation process was analyzed as two-step mechanism, mass transfer and reaction. This current study shows the potential ability of Ni/Fe nanoparticles to be used for removal of PBDEs in contaminated soil. - Highlights: • Ni/Fe bimetallic nanoparticles could effectively degradate BDE209 in soil. • The effects of various factors on remediation of BDE209 in soil using Ni/Fe were considered. • The degradation of BDE209 was a process of stepwise debromination from nBr to (n − 1)Br. • A possible debromination pathway and mechanism about removal of BDE209 in soil were proposed.

  20. Large enhancement of magnetoresistance in NiFe film with MgO layers sandwiched after annealing

    Energy Technology Data Exchange (ETDEWEB)

    Zhao, Zhi-Duo; Li, Ming-Hua [Department of Materials Physics and Chemistry, University of Science and Technology Beijing, Beijing 100083 (China); Zhao, Chong-Jun, E-mail: zhaocj@bj-nmc.cn [Beijing Nmc Co., Ltd., Beijing 100084 (China); Department of Mechanical Engineering, Tsinghua University, Beijing 100084 (China); Yang, Guang; Zhang, Jing-Yan; Jiang, Shao-Long; Zhao, Yun-Chi; Cui, Xiao-Peng [Department of Materials Physics and Chemistry, University of Science and Technology Beijing, Beijing 100083 (China); Yu, Guang-Hua, E-mail: ghyu@mater.ustb.edu.cn [Department of Materials Physics and Chemistry, University of Science and Technology Beijing, Beijing 100083 (China)

    2014-12-01

    Highlights: • Ta/MgO/Ni{sub 81}Fe{sub 19}/MgO/Ta and Ta/Ni{sub 81}Fe{sub 19}/Ta were prepared by magnetron sputtering. • The electronic transport properties were studied. • The microstructure of NiFe film (100 nm) with MgO sandwiched was measured by HRTEM. • The average grain size and the crystallinity were determined from X-ray diffraction studies. • Δρ and ρ for samples as function of the NiFe thickness were analyzed. - Abstract: A large enhancement of magnetoresistance (MR) up to 6.0% has been observed in NiFe sandwiched by MgO layers, which is 50% larger than the highest MR (4%) in bulk materials. The great improvement of MR derives from the slight increase in corresponding resistivity change Δρ and the great decrease in resistivity ρ. The enhancement of Δρ is attributed to the strengthened spin-dependent scattering of the interfacial conductive electrons but the contribution will be slight with the increase in NiFe thickness. The main contribution is from the significant decrease in ρ, originating from confinement effect on electrons from well-formed oxide/metal interfaces after annealing. Surprisingly, this effect still exists when the NiFe thickness reaches 100 nm. Meanwhile, the oxide (MgO) layers inserted prevents the atomic interdiffusion of Ta and NiFe at the interfaces, which decreases the thickness of magnetic dead layer.

  1. Structural and magnetic properties of NiFe 2O 4-SnO 2 nanocomposite

    Science.gov (United States)

    Albuquerque, A. S.; Ardisson, J. D.; Macedo, W. A. A.; Plivelic, T. S.; Torriani, I. L.; Larrea J., J.; Saitovitch, E. B.

    2004-05-01

    The structural and magnetic properties of the NiFe 2O 4-SnO 2 composite, obtained by ball-milling during different times, were investigated by X-ray diffraction, small-angle X-ray scattering, Mössbauer spectroscopy and vibrating sample magnetometry. The results showed the reduction of the crystalline particle size and modification in the nature of the system interfaces as a consequence of the mechanical treatment. Specimens with smaller particles displayed strong superparamagnetism. Large variation of the hysteresis loops for the different milling times was observed.

  2. Direct observation of domain walls in NiFe films using high-resolution Lorentz microscopy

    Science.gov (United States)

    Wong, Bunsen Y.; Laughlin, David E.

    1996-04-01

    A novel approach to observe the interaction between magnetic domain wall and nanoscale microstructural features is demonstrated. The method is based on Focault mode Lorentz microscopy and utilizes a Gatan energy image filter to provide additional magnification. A postexperimental image processing technique was applied to separate lattice diffraction from that induced by magnetic domains. The effect of NiFe thickness on the width of a 180° Néel wall has been studied. It was found that the thickness dependence has a similar profile to the theoretically predicted trend but the actual wall thickness is smaller than the calculated values.

  3. Effect of La on Partial Oxidation of Ethanol to Hydrogen over Ni/Fe Catalysts

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    The partial oxidation of ethanol to hydrogen was investigated over Ni/Fe/La catalysts prepared by the co-precipitation method. The effects of introduction of La promoter and the reaction temperature on the catalytic performance were studied. It was found that the introduction of La into Ni/Fe catalysts is helpful to increase the selectivity to hydrogen and the stability of the catalysts. The results of XRD and XPS characterization show that the structure of the catalyst was changed during the reaction. The existence of LaFeO3 species is possibly the main reason of the increase of the catalyst stability.

  4. KARAKTERISTIK MAGNETIK LAPISAN TIPIS NI-FE SEBAGAI FLAT CORE FLUX GATE SENSOR

    Directory of Open Access Journals (Sweden)

    B. Purnama

    2014-08-01

    Full Text Available ABSTRAKKarakteristik magnetik lapisan tipis Ni-Fe hasil elektrodeposisi sebagai flat core flux gate sensor didiskusikan pada makalah ini.  Lapisan tipis Ni-Fe dideposisi di atas substrat printed circuit board (PCB dengan variasi komposisi Ni dan rapat arus J.  Untuk memperoleh kualitas lapisan tipis yang baik, zat aditif vanily digunakan untuk mengurangi laju deposisi untuk keseluruhan ekperimen.  Hasil menunjukkan modifikasi karakteristik magnetik hasil vibrating sample magnetometer karena perubahan parameter penumbuhannya. Koersif field (Hc meningkat dengan kenaikan prosentase komposisi Ni pada lapisan tipis yang terbentuk.  Sedangkan Hc tidak membentuk pola tertentu dengan modifikasi apat arus J. Akhirnya magnetisasi jenuh maksimum mS = 0.72 emu/gr diperoleh untuk prosentase Ni = 51%, dan mS = 0.43 emu/gr untuk J = 5 mA/cm2.  Klarifikasi dengan STM, hal ini akibat perubahan struktur mikroskopik lapisan tipis Ni-Fe.ABSTRACTThis paper discusses magnetic characteristic of Ni-Fe permalloy thin film produced by electro deposited as flat core flux gate sensor.  Ni-Fe thin films were deposited on printed circuit board (PCB substrates with variation of composition in Ni percentage and current density J.  In order to obtain high quality thin films, an additive ingredient of vanily was used to reduce rate deposition for whole experimental procedure. Experiment results showed that modified of magnetic characteristic evaluated by vibrating sample magnetometer was caused by the modified of deposition parameters.  Coersive field (Hc increased with the increase of percentage of Ni composition, while the Hc did not form any particular pattern by modifying current density J. Finally, maximum saturated magnetization mS = 0.72 emu/gr was attained for percentage Ni of 51% and mS = 0.43 emu/gr for J = 5 mA/cm2. Clarification by using STM showed that this was caused by the change of microscopic structure in Ni-Fe thin films.

  5. KARAKTERISTIK MAGNETIK LAPISAN TIPIS NI-FE SEBAGAI FLAT CORE FLUX GATE SENSOR

    Directory of Open Access Journals (Sweden)

    B. Purnama

    2013-07-01

    Full Text Available ABSTRAKKarakteristik magnetik lapisan tipis Ni-Fe hasil elektrodeposisi sebagai flat core flux gate sensor didiskusikan pada makalah ini.  Lapisan tipis Ni-Fe dideposisi di atas substrat printed circuit board (PCB dengan variasi komposisi Ni dan rapat arus J.  Untuk memperoleh kualitas lapisan tipis yang baik, zat aditif vanily digunakan untuk mengurangi laju deposisi untuk keseluruhan ekperimen.  Hasil menunjukkan modifikasi karakteristik magnetik hasil vibrating sample magnetometer karena perubahan parameter penumbuhannya. Koersif field (Hc meningkat dengan kenaikan prosentase komposisi Ni pada lapisan tipis yang terbentuk.  Sedangkan Hc tidak membentuk pola tertentu dengan modifikasi apat arus J. Akhirnya magnetisasi jenuh maksimum mS = 0.72 emu/gr diperoleh untuk prosentase Ni = 51%, dan mS = 0.43 emu/gr untuk J = 5 mA/cm2.  Klarifikasi dengan STM, hal ini akibat perubahan struktur mikroskopik lapisan tipis Ni-Fe.ABSTRACTThis paper discusses magnetic characteristic of Ni-Fe permalloy thin film produced by electro deposited as flat core flux gate sensor.  Ni-Fe thin films were deposited on printed circuit board (PCB substrates with variation of composition in Ni percentage and current density J.  In order to obtain high quality thin films, an additive ingredient of vanily was used to reduce rate deposition for whole experimental procedure. Experiment results showed that modified of magnetic characteristic evaluated by vibrating sample magnetometer was caused by the modified of deposition parameters.  Coersive field (Hc increased with the increase of percentage of Ni composition, while the Hc did not form any particular pattern by modifying current density J. Finally, maximum saturated magnetization mS = 0.72 emu/gr was attained for percentage Ni of 51% and mS = 0.43 emu/gr for J = 5 mA/cm2. Clarification by using STM showed that this was caused by the change of microscopic structure in Ni-Fe thin films.

  6. The energetics and electronic origins for atomic long- and short-range order in Ni-Fe invar alloys

    Energy Technology Data Exchange (ETDEWEB)

    Johnson, D.D. [Sandia National Labs., Albuquerque, NM (United States); Shelton, W.A. [Oak Ridge National Lab., TN (United States)

    1996-12-31

    States of magnetic and compositional order are strongly coupled in many magnetic alloys, with Ni-Fe Invar being the most celebrated example. Results of an electronic-based method that addresses compositional and magnetic disorder, as well as atomic short-range order and energetics, are discussed. This allows a system-dependent microscopic understanding of the interplay of chemical, magnetic, and displacive effects, and a direct comparison to diffuse scattering experiments. Discussion is in context of total-energy calculations for various magnetic states in chemically disordered and ordered Ni- Fe alloys, emphasizing the importance of exchange-splitting and the implication for phase stability in Ni-Fe system.

  7. Microstructure, AC impedance and DC electrical conductivity characteristics of NiFe2-xGdxO4 (x = 0, 0.05 and 0.075

    Directory of Open Access Journals (Sweden)

    K. Kamala Bharathi

    2012-03-01

    Full Text Available The structure and electrical characteristics of Gd doped Ni ferrite materials, namely NiFe1.95Gd0.05O4 and NiFe1.925Gd0.075O4, are reported to demonstrate their improved electrical properties compared to that of pure NiFe2O4. NiFe1.95Gd0.05O4 and NiFe1.925Gd0.075O4 compounds crystallize in the cubic inverse spinel phase with a very small amount of GdFeO3 additional phase while pure NiFe2O4 crystallize in inverse spinel phase without any impurity phase. The back scattered electron imaging analysis indicate the primary and secondary formation in NiFe1.95Gd0.05O4 and NiFe1.925Gd0.075O4 compounds. Atomic force microscopy measurements indicate that the bulk grains are ∼2-5 micron size while the grain boundaries are thin compared to bulk grains. Impedance spectroscopic analysis at different temperature indicates the different relaxation mechanisms and their variation with temperature, bulk grain and grain-boundary contributions to the electrical conductivity (Rg and capacitance (Cg of these materials. The conductivity in pure NiFeO4 is found to be predominantly due to intrinsic bulk contribution (Rg=213 kΩ and Cg=4.5 x 10-8 F. In the case of NiFe1.95Gd0.05O4 and NiFe1.925Gd0.075O4 compounds, grain and grain-boundary contributions to the conductivity are clearly observed. The DC conductivity values (at 300 K of NiFe2O4, NiFe1.95Gd0.05O4 and NiFe1.925Gd0.075O4 compounds are found to be 1.06 x 10-7 Ω-1 cm-1, 5.73 x 10-8 Ω-1 cm-1 and 1.28 x 10-8 Ω-1 cm-1 respectively.

  8. A cell-free microtiter plate screen for improved [FeFe] hydrogenases.

    Directory of Open Access Journals (Sweden)

    James A Stapleton

    Full Text Available BACKGROUND: [FeFe] hydrogenase enzymes catalyze the production and dissociation of H(2, a potential renewable fuel. Attempts to exploit these catalysts in engineered systems have been hindered by the biotechnologically inconvenient properties of the natural enzymes, including their extreme oxygen sensitivity. Directed evolution has been used to improve the characteristics of a range of natural catalysts, but has been largely unsuccessful for [FeFe] hydrogenases because of a lack of convenient screening platforms. METHODOLOGY/PRINCIPAL FINDINGS: Here we describe an in vitro screening technology for oxygen-tolerant and highly active [FeFe] hydrogenases. Despite the complexity of the protocol, we demonstrate a level of reproducibility that allows moderately improved mutants to be isolated. We have used the platform to identify a mutant of the Chlamydomonas reinhardtii [FeFe] hydrogenase HydA1 with a specific activity approximately 4 times that of the wild-type enzyme. CONCLUSIONS/SIGNIFICANCE: Our results demonstrate the feasibility of using the screen presented here for large-scale efforts to identify improved biocatalysts for energy applications. The system is based on our ability to activate these complex enzymes in E. coli cell extracts, which allows unhindered access to the protein maturation and assay environment.

  9. Photocatalytic hydrogen production from a simple water-soluble [FeFe]-hydrogenase model system.

    Science.gov (United States)

    Cao, Wei-Ning; Wang, Feng; Wang, Hong-Yan; Chen, Bin; Feng, Ke; Tung, Chen-Ho; Wu, Li-Zhu

    2012-08-21

    Combined with a simple water soluble [FeFe]-hydrogenase mimic 1, Ru(bpy)(3)(2+) and ascorbic acid enable hydrogen production photocatalytically. More than 88 equivalents of H(2) were achieved in water, which is much better than that obtained in an organic solvent or a mixture of organic solvent and water.

  10. Enzymatic recovery of platinum (IV) from industrial wastewater using a biosulphidogenic hydrogenase

    CSIR Research Space (South Africa)

    Rashamuse, KJ

    2008-04-01

    Full Text Available this treatment process suggesting that platinum sulphide was not formed and supporting the argument that the increased amount (78%) of platinum removal from the industrial wastewater by the growing SRB cells was due to more hydrogenase/cytochrome c3 enzyme...

  11. Supersolar Ni/Fe production in the Type IIP SN 2012ec

    CERN Document Server

    Jerkstrand, Anders; Sollerman, Jesper; Inserra, Cosimo; Fraser, Morgan; Spyromilio, Jason; Fransson, Claes; Chen, Ting-Wan; Barbarino, Cristina; Dall'Ora, Massimo; Botticella, Maria Teresa; Della Valle, Massimo; Gal-Yam, Avishay; Valenti, Stefano; Maguire, Kate; Mazzali, Paolo; Tomasella, Lina

    2014-01-01

    SN 2012ec is a Type IIP supernova (SN) with a progenitor detection and comprehensive photospheric-phase observational coverage. Here, we present Very Large Telescope and PESSTO observations of this SN in the nebular phase. We model the nebular [O I] 6300, 6364 lines and find their strength to suggest a progenitor main-sequence mass of 13-15 Msun. SN 2012ec is unique among hydrogen-rich SNe in showing a distinct and unblended line of stable nickel [Ni II] 7378. This line is produced by 58Ni, a nuclear burning ash whose abundance is a sensitive tracer of explosive burning conditions. Using spectral synthesis modelling, we use the relative strengths of [Ni II] 7378 and [Fe II] 7155 (the progenitor of which is 56Ni) to derive a Ni/Fe production ratio of 0.19pm0.07, which is a factor 3.5pm1.2 times the solar value. High production of stable nickel is confirmed by a strong [Ni II] 1.939 micron line. This is the third reported case of a core-collapse supernova producing a Ni/Fe ratio far above the solar value, which...

  12. Anionic Regulated NiFe (Oxy)Sulfide Electrocatalysts for Water Oxidation.

    Science.gov (United States)

    Li, Bo-Quan; Zhang, Shu-Yuan; Tang, Cheng; Cui, Xiaoyang; Zhang, Qiang

    2017-07-01

    The construction of active sites with intrinsic oxygen evolution reaction (OER) is of great significance to overcome the limited efficiency of abundant sustainable energy devices such as fuel cells, rechargeable metal-air batteries, and in water splitting. Anionic regulation of electrocatalysts by modulating the electronic structure of active sites significantly promotes OER performance. To prove the concept, NiFeS electrocatalysts are fabricated with gradual variation of atomic ratio of S:O. With the rise of S content, the overpotential for water oxidation exhibits a volcano plot under anionic regulation. The optimized NiFeS-2 electrocatalyst under anionic regulation possesses the lowest OER overpotential of 286 mV at 10 mA cm(-2) and the fastest kinetics being 56.3 mV dec(-1) to date. The anionic regulation methodology not only serves as an effective strategy to construct superb OER electrocatalysts, but also enlightens a new point of view for the in-depth understanding of electrocatalysis at the electronic and atomic level. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Constraints on explosive silicon burning in core-collapse supernovae from measured Ni/Fe ratios

    CERN Document Server

    Jerkstrand, A; Magkotsios, G; Sim, S A; Fransson, C; Spyromilio, J; Heger, A; Müller, B; Sollerman, J; Smartt, S J

    2015-01-01

    Measurements of explosive nucleosynthesis yields in core-collapse supernovae provide tests for explosion models. We investigate constraints on explosive conditions derivable from measured amounts of nickel and iron after radioactive decays using nucleosynthesis networks with parameterized thermodynamic trajectories. The Ni/Fe ratio is for most regimes dominated by the production ratio of 58Ni/(54Fe + 56Ni), which tends to grow with higher neutron excess and with higher entropy. For SN 2012ec, a supernova that produced a Ni/Fe ratio of $3.4\\pm1.2$ times solar, we find that burning of a fuel with neutron excess $\\eta \\approx 6\\times 10^{-3}$ is required. Unless the progenitor metallicity is over 5 times solar, the only layer in the progenitor with such a neutron excess is the silicon shell. Supernovae producing large amounts of stable nickel thus suggest that this deep-lying layer can be, at least partially, ejected in the explosion. We find that common spherically symmetric models of $M_{\\rm ZAMS} \\lesssim 13$...

  14. Remediation of polybrominated diphenyl ethers in soil using Ni/Fe bimetallic nanoparticles: influencing factors, kinetics and mechanism.

    Science.gov (United States)

    Xie, Yingying; Fang, Zhanqiang; Cheng, Wen; Tsang, Pokeung Eric; Zhao, Dongye

    2014-07-01

    Polybrominated diphenyl ethers (PBDEs) are commonly used as additive flame retardants in all kinds of electronic products. PBDEs are now ubiquitous in the environment, with soil as a major sink, especially in e-waste recycling sites. This study investigated the degradation of decabromodiphenyl ether (BDE209) in a spiked soil using Ni/Fe bimetallic nanoparticles. The results indicated that Ni/Fe bimetallic nanoparticles are able to degrade BDE209 in soil at ambient temperature and the removal efficiency can reach 72% when an initial pH of 5.6 and at a Ni/Fe dosage of 0.03 g/g. A declining trend in degradation was noticed with decreasing Ni loading and increasing of initial BDE209 concentration. The degradation products of BDE209 were analyzed by GC-MS, which showed that the degradation of BDE209 was a process of stepwise debromination from nBr to (n-1)Br. And a possible debromination pathway was proposed. At last, the degradation process was analyzed as two-step mechanism, mass transfer and reaction. This current study shows the potential ability of Ni/Fe nanoparticles to be used for removal of PBDEs in contaminated soil. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. Photosynthetic electron partitioning between [FeFe]-hydrogenase and ferredoxin:NADP⁺-oxidoreductase (FNR) enzymes in vitro

    National Research Council Canada - National Science Library

    Iftach Yacoby; Sergii Pochekailov; Hila Toporik; Maria L. Ghirardi; Paul W. King; Shuguang Zhang

    2011-01-01

    .... To elucidate the basis for competition, we bioengineered a ferredoxin-hydrogenase fusion and characterized hydrogen production kinetics in the presence of Fd, ferredoxin:NADP⁺-oxidoreductase (FNR), and NADP...

  16. A redox hydrogel protects the O2 -sensitive [FeFe]-hydrogenase from Chlamydomonas reinhardtii from oxidative damage.

    Science.gov (United States)

    Oughli, Alaa Alsheikh; Conzuelo, Felipe; Winkler, Martin; Happe, Thomas; Lubitz, Wolfgang; Schuhmann, Wolfgang; Rüdiger, Olaf; Plumeré, Nicolas

    2015-10-12

    The integration of sensitive catalysts in redox matrices opens up the possibility for their protection from deactivating molecules such as O2 . [FeFe]-hydrogenases are enzymes catalyzing H2 oxidation/production which are irreversibly deactivated by O2 . Therefore, their use under aerobic conditions has never been achieved. Integration of such hydrogenases in viologen-modified hydrogel films allows the enzyme to maintain catalytic current for H2 oxidation in the presence of O2 , demonstrating a protection mechanism independent of reactivation processes. Within the hydrogel, electrons from the hydrogenase-catalyzed H2 oxidation are shuttled to the hydrogel-solution interface for O2 reduction. Hence, the harmful O2 molecules do not reach the hydrogenase. We illustrate the potential applications of this protection concept with a biofuel cell under H2 /O2 mixed feed.

  17. Research data supporting "Photoelectrochemical H2 Evolution with a Hydrogenase Immobilized on a TiO2-protected Silicon Electrode"

    OpenAIRE

    Lee, Chong-Yong; Park, Hyun S.; Fontecilla-Camps, Juan C.; Reisner, Erwin

    2016-01-01

    Experimental Raw data supporting publication: Photoelectrochemical H2 Evolution with a Hydrogenase Immobilized on a TiO2-protected Silicon Electrode This research data supports “Photoelectrochemical H2 Evolution with a Hydrogenase Immobilized on a TiO2-protected Silicon Electrode" which has been published in “Angewandte Chemie international edition (English)”. This work was supported by the BBSRC [grant number BB/K010220/1].

  18. Effect of MnO2 on properties of NiFe2O4 spinel based inert anode

    Institute of Scientific and Technical Information of China (English)

    XI; Jin-hui; YAO; Guang-chun; LIU; Yi-han; ZHANG; Xiao-ming

    2005-01-01

    In order to improve the properties of NiFe2O4 spinel based inert anode, some additive MnO2 were added to raw materials. NiFe2O4 spinel with MnO2 was made by solid-phase reaction at 1200℃for 6 h. XRD were carried out and the effects of MnO2 on density, conductivity and corrosion resistance were measured. XRD shows when MnO2 was added no new phases exist and MnO2 and NiFe2O4 formed solid solution; Mn4+ replaced parts of Fe3+ and the sample still had the structure of NiFe2O4 spinel. The crystal lattice of NiFe2 O4 spinel became aberrated when MnO2 was added, which can promote sintering, and improve density. Because Mn4+ replaces parts of Fe3+ and produces conduction electron, which can improve conductivity.The corrosion resistance of the samples was enhanced. When MnO2 is 1.0%, the sample's corrosion rate is 1/5 of that of the sample without MnO2. The reason is that Al2 O3 in the melt reacts with Mn4+ in the sample to produce MnAl2O4. MnAl2 O4 forms a dense protecting coat, which can prevent melt from eroding further.Because the key problem with inert anodes is anode corrosion, so we consider the optimal amount of MnO2 is 1.0%.

  19. Purification and characterization of the [NiFe]-hydrogenase of Shewanella oneidensis MR-1.

    Science.gov (United States)

    Shi, Liang; Belchik, Sara M; Plymale, Andrew E; Heald, Steve; Dohnalkova, Alice C; Sybirna, Kateryna; Bottin, Hervé; Squier, Thomas C; Zachara, John M; Fredrickson, James K

    2011-08-15

    Shewanella oneidensis MR-1 possesses a periplasmic [NiFe]-hydrogenase (MR-1 [NiFe]-H(2)ase) that has been implicated in H(2) production and oxidation as well as technetium [Tc(VII)] reduction. To characterize the roles of MR-1 [NiFe]-H(2)ase in these proposed reactions, the genes encoding both subunits of MR-1 [NiFe]-H(2)ase were cloned and then expressed in an MR-1 mutant without hyaB and hydA genes. Expression of recombinant MR-1 [NiFe]-H(2)ase in trans restored the mutant's ability to produce H(2) at 37% of that for the wild type. Following purification, MR-1 [NiFe]-H(2)ase coupled H(2) oxidation to reduction of Tc(VII)O(4)(-) and methyl viologen. Change of the buffers used affected MR-1 [NiFe]-H(2)ase-mediated reduction of Tc(VII)O(4)(-) but not methyl viologen. Under the conditions tested, all Tc(VII)O(4)(-) used was reduced in Tris buffer, while in HEPES buffer, only 20% of Tc(VII)O(4)(-) was reduced. The reduced products were soluble in Tris buffer but insoluble in HEPES buffer. Transmission electron microscopy analysis revealed that Tc precipitates reduced in HEPES buffer were aggregates of crystallites with diameters of ∼5 nm. Measurements with X-ray absorption near-edge spectroscopy revealed that the reduction products were a mixture of Tc(IV) and Tc(V) in Tris buffer but only Tc(IV) in HEPES buffer. Measurements with extended X-ray adsorption fine structure showed that while the Tc bonding environment in Tris buffer could not be determined, the Tc(IV) product in HEPES buffer was very similar to Tc(IV)O(2)·nH(2)O, which was also the product of Tc(VII)O(4)(-) reduction by MR-1 cells. These results shows for the first time that MR-1 [NiFe]-H(2)ase catalyzes Tc(VII)O(4)(-) reduction directly by coupling to H(2) oxidation.

  20. Impact of the chemicals, essential for the purification process of strict Fe-hydrogenase, on the corrosion of mild steel.

    Science.gov (United States)

    Rouvre, Ingrid; Gauquelin, Charles; Meynial-Salles, Isabelle; Basseguy, Régine

    2016-06-01

    The influence of additional chemical molecules, necessary for the purification process of [Fe]-hydrogenase from Clostridium acetobutylicum, was studied on the anaerobic corrosion of mild steel. At the end of the purification process, the pure [Fe-Fe]-hydrogenase was recovered in a Tris-HCl medium containing three other chemicals at low concentration: DTT, dithionite and desthiobiotin. Firstly, mild steel coupons were exposed in parallel to a 0.1 M pH7 Tris-HCl medium with or without pure hydrogenase. The results showed that hydrogenase and the additional molecules were in competition, and the electrochemical response could not be attributed solely to hydrogenase. Then, solutions with additional chemicals of different compositions were studied electrochemically. DTT polluted the electrochemical signal by increasing the Eoc by 35 mV 24 h after the injection of 300 μL of control solutions with DTT, whereas it drastically decreased the corrosion rate by increasing the charge transfer resistance (Rct 10 times the initial value). Thus, DTT was shown to have a strong antagonistic effect on corrosion and was removed from the purification process. An optimal composition of the medium was selected (0.5 mM dithionite, 7.5 mM desthiobiotin) that simultaneously allowed a high activity of hydrogenase and a lower impact on the electrochemical response for corrosion tests.

  1. Study on sintering technique of NiFe2O4/SiCp used as matrix of inert anodes in aluminium electrolysis

    Institute of Scientific and Technical Information of China (English)

    ZHANG; Shu-ting; YAO; Guang-chun; LIU; Yi-han

    2005-01-01

    In order to improve deficiencies of NiFe2O4 spinel used as matrix of inert anode in aluminium electrolysis, NiFe2O4/SiCp were prepared by the solid state reaction for the first time. Microstructural changes were observed by scanning electronic microscope and phase was determined with X-ray detector. Effect of sintering temperature and times on density, porosity and microstructure were researched, and the reasons that caused the difference were discussed deeply. At the same time the thermodynamical compatibility of NiFe2O4 and SiC was proved under 1200℃ by DTA.The results showed that the microstructure was more homogeneous when the sintering temperature reached 1 180℃and the density attained their maximum about 6 h sintering. The appropriate sintering technique of NiFe2 O4/SiCw composite materials was 1180℃× 6 h.

  2. Mesoscale Compositionally Modulated Nanocrystalline Ni-Fe Electrodeposits for Nanopatterning Applications

    Directory of Open Access Journals (Sweden)

    P. Egberts

    2008-01-01

    Full Text Available A considerable range of surface nanostructures can be fabricated by the selective dissolution of elements or phases from metallic alloys. Selectively etched electrodeposited multilayers may find useful application in optoelectronic and MEMS devices. One issue with electrodeposited multilayers is that the fine-scale multilayer structure can often exhibit significant waviness if the band layer spacing is on the same order of magnitude as the grain size. In the present study, the mean grain size was reduced to below 10 nm in a compositionally modulated Ni-Fe alloy. Preferential etching on the electroform cross-section resulted in highly uniform and directional surface channels. The evolution of this nanopatterned surface morphology was characterized by atomic force microscopy and directional roughness parameters were obtained.

  3. Electrical manipulation of ferromagnetic NiFe by antiferromagnetic IrMn

    Science.gov (United States)

    Tshitoyan, V.; Ciccarelli, C.; Mihai, A. P.; Ali, M.; Irvine, A. C.; Moore, T. A.; Jungwirth, T.; Ferguson, A. J.

    2015-12-01

    We demonstrate that an antiferromagnet can be employed for a highly efficient electrical manipulation of a ferromagnet. In our study, we use an electrical detection technique of the ferromagnetic resonance driven by an in-plane ac current in a NiFe/IrMn bilayer. At room temperature, we observe antidampinglike spin torque acting on the NiFe ferromagnet, generated by an in-plane current driven through the IrMn antiferromagnet. A large enhancement of the torque, characterized by an effective spin-Hall angle exceeding most heavy transition metals, correlates with the presence of the exchange-bias field at the NiFe/IrMn interface. It highlights that, in addition to the strong spin-orbit coupling, the antiferromagnetic order in IrMn governs the observed phenomenon.

  4. Superparamagnetic behavior and AC-losses in NiFe2O4 nanoparticles

    Science.gov (United States)

    Yelenich, O. V.; Solopan, S. O.; Kolodiazhnyi, T. V.; Dzyublyuk, V. V.; Tovstolytkin, A. I.; Belous, A. G.

    2013-06-01

    Crystallographic, microstructural and magnetic properties of NiFe2O4 nanoparticles synthesized by precipitation from nonaqueous solutions have been studied in the work. The transmission electron microscopy studies reveal particle sizes ˜5 nm for the as-prepared particles which increase up to ˜20 nm upon annealing at 500 °C. Quasistatic magnetic measurements show superparamagnetic behavior with blocking temperature below room temperature for both the as-prepared and annealed particles. Characteristic magnetic parameters of the particles including average magnetic moment of an individual nanoparticle and effective anisotropy constant have been determined. The specific loss power which is released on the exposure of an ensemble of synthesized particles to an electromagnetic field is calculated and measured experimentally.

  5. A Ni-Fe Layered Double Hydroxide-Carbon Nanotube Complex for Water Oxidation

    CERN Document Server

    Gong, Ming; Wang, Hailiang; Liang, Yongye; Wu, Justin Zachary; Zhou, Jigang; Wang, Jian; Regier, Tom; Wei, Fei; Dai, Hongjie

    2013-01-01

    Highly active, durable and cost-effective electrocatalysts for water oxidation to evolve oxygen gas hold a key to a range of renewable energy solutions including water splitting and rechargeable metal-air batteries. Here, we report the synthesis of ultrathin nickel iron layered double hydroxide nanoplates on mildly oxidized multi-walled carbon nanotubes. Incorporation of Fe into the nickel hydroxide induced the formation of NiFe-layered double hydroxide. The nanoplates were covalently attached to a network of nanotubes, affording excellent electrical wiring to the nanoplates. The ultra-thin Ni-Fe layered double hydroxide nanoplates/carbon nanotube complex was found to exhibit unusually high electro-catalytic activity and stability for oxygen evolution and outperformed commercial precious metal Ir catalysts.

  6. [NiFe] dithiolene diphosphine complex for hydrogen gas activation: a Theoretic Insight

    CERN Document Server

    GuYan, Jing

    2015-01-01

    A diphosphino-nickel-iron dithiolene complex, [Ni(bdt)(dppf)] (bdt = 1,2-benzenedithiolate, dppf = 1,1-bis(diphenylphosphino)ferrocene), has been recently found to be reasonably active on proton reduction to dihydrogen (J. Am. Chem. Soc. 2015, 137, 1109). Interestingly, this exceptional complex was found to be also reactive towards dihydrogen activation as indicated by the electrochemical investigation. However, a pure nickel dithiolene diphosphine theoretical mode, excluding the contributions from iron moiety, was applied to attribute the experimental catalytic observation. We have re-visited the theoretical approach in details for this [NiFe] catalyst and compared it with the non-active nickel dithiolene diphosphine complexes. We found that both nickel and iron moieties in this newly developed complex were imperative for the observed catalytic per-formance, particularly towards the activation of dihydrogen.

  7. Characterization of nanostructured spinel NiFe2O4 obtained by soft mechanochemical synthesis

    Directory of Open Access Journals (Sweden)

    Lazarević Z.Ž.

    2012-01-01

    Full Text Available Powdery nickel ferrite, NiFe2O4 has been obtained by soft mechanochemical synthesis in a planetary ball mill. Ni(OH2 and Fe(OH3 are used as initial compounds. This mixture was mechanically activated for 25h, uniaxial pressed and sintered at 1100°C for 2h. The phase composition of the sintered sample was analyzed by X-ray diffraction (XRD, energy dispersive spectrometer (EDS and Raman spectroscopy. Morphologies were examined by scanning electron microscopy (SEM. The electrical DC/resistivity/conductivity at different temperatures was measured using a Source Meter Keithley 2410. An Impedance/Gain-Phase Analyzer (HP-4194 was used to measure the impedance spectra (100Hz - 10MHz at different temperatures. [Projekat Ministarstva nauke Republike Srbije, br. III 45003 i br. III 45015

  8. Comparison of carbon materials as electrodes for enzyme electrocatalysis: hydrogenase as a case study.

    Science.gov (United States)

    Quinson, Jonathan; Hidalgo, Ricardo; Ash, Philip A; Dillon, Frank; Grobert, Nicole; Vincent, Kylie A

    2014-01-01

    We present a study of electrocatalysis by an enzyme adsorbed on a range of carbon materials, with different size, surface area, morphology and graphitic structure, which are either commercially available or prepared via simple, established protocols. We choose as our model enzyme the hydrogenase I from E. coli (Hyd-1), which is an active catalyst for H2 oxidation, is relatively robust and has been demonstrated in H2 fuel cells and H2-driven chemical synthesis. The carbon materials were characterised according to their surface area, surface morphology and graphitic character, and we use the electrocatalytic H2 oxidation current for Hyd-1 adsorbed on these materials to evaluate their effectiveness as enzyme electrodes. Here, we show that a variety of carbon materials are suitable for adsorbing hydrogenases in an electroactive configuration. This unified study provides insight into selection and design of carbon materials for study of redox enzymes and different applications of enzyme electrocatalysis.

  9. Isolation, purification and characterization of the hydrogen evolution promoting factor of hydrogenase of Spirulina platensis

    Science.gov (United States)

    Gu, Tian-Qing; Zhang, Hui-Miao; Sun, Shi-Hua

    1996-03-01

    A component (s-factor) with obvious promoting effect on hydrogen evolution of hydrogenase has been isolated and extracted from a cell-free preparation of Spirulina platensis. The effect of the s-factor in the reaction system is similar to that of Na2S2O4, but is coupled with light. The s-factor has the maximum absorption peak at 620 nm in the oxidized state, at 590 nm in the reduced state. The partially purified s-factor showed two bands by SDS-PAGE and is distinctly different from phycocyanin, which has no change of oxidized state and reduced state absorption spectra, and also has no promoting effect on hydrogenase of Spirulina platensis under the light.

  10. Mechanism of O2 diffusion and reduction in FeFe hydrogenases

    Science.gov (United States)

    Kubas, Adam; Orain, Christophe; de Sancho, David; Saujet, Laure; Sensi, Matteo; Gauquelin, Charles; Meynial-Salles, Isabelle; Soucaille, Philippe; Bottin, Hervé; Baffert, Carole; Fourmond, Vincent; Best, Robert B.; Blumberger, Jochen; Léger, Christophe

    2017-01-01

    FeFe hydrogenases are the most efficient H2-producing enzymes. However, inactivation by O2 remains an obstacle that prevents them being used in many biotechnological devices. Here, we combine electrochemistry, site-directed mutagenesis, molecular dynamics and quantum chemical calculations to uncover the molecular mechanism of O2 diffusion within the enzyme and its reactions at the active site. We propose that the partial reversibility of the reaction with O2 results from the four-electron reduction of O2 to water. The third electron/proton transfer step is the bottleneck for water production, competing with formation of a highly reactive OH radical and hydroxylated cysteine. The rapid delivery of electrons and protons to the active site is therefore crucial to prevent the accumulation of these aggressive species during prolonged O2 exposure. These findings should provide important clues for the design of hydrogenase mutants with increased resistance to oxidative damage.

  11. Purification and Characterization of [NiFe]-Hydrogenase of Shewanella oneidensis MR-1

    Energy Technology Data Exchange (ETDEWEB)

    Shi, Liang; Belchik, Sara M.; Plymale, Andrew E.; Heald, Steve M.; Dohnalkova, Alice; Sybirna, Kateryna; Bottin, Herve; Squier, Thomas C.; Zachara, John M.; Fredrickson, Jim K.

    2011-08-02

    The γ-proteobacterium Shewanella oneidensis MR-1 possesses a periplasmic [NiFe]-hydrogenase (MR-1 [NiFe]-H2ase) that was implicated in both H2 production and oxidation as well as technetium [Tc(VII)] reduction. To characterize the roles of MR-1 [NiFe]-H2ase in these proposed reactions, the genes encoding both subunits of MR-1 [NiFe]-H2ase were cloned into a protein expression vector. The resulting plasmid was transformed into a MR-1 mutant deficient in H2 formation. Expression of MR-1 [NiFe]-H2ase in trans restored the mutant’s ability to produce H2 at 37% of that for wild type. Following expression, MR-1 [NiFe]-H2ase was purified to near homogeneity. The purified MR-1 [NiFe]-H2ase could couple H2 oxidation to reduction of Tc(VII) and methyl viologen directly. Change of the buffers used affected MR-1 [NiFe]-H2ase-mediated Tc(VII) but not methyl viologen reductions. Under the conditions tested, Tc(VII) reduction was complete in Tris buffer but not in HEPES buffer. The reduced Tc(IV) was soluble in Tris buffer but insoluble in HEPES buffer. Transmission electron microscopy analysis revealed that Tc(IV) precipitates formed in HEPES buffer were packed with crystallites. Although X-ray absorption near-edge spectroscopy measurements confirmed that the reduction products found in both buffers were Tc(IV), extended X-ray adsorption fine-structure measurements revealed that these products were very different. While the product in Tris buffer could not be determined, the Tc(IV) product in HEPES buffer was very similar to Tc(IV)O2•nH2O. These results shows for the first time that MR-1 [NiFe]-H2ase is a bidirectional enzyme that catalyzes both H2 formation and oxidation as well as Tc(VII) reduction directly by coupling H2 oxidation.

  12. Lyophilization protects [FeFe]-hydrogenases against O2-induced H-cluster degradation

    OpenAIRE

    Jens Noth; Ramona Kositzki; Kathrin Klein; Martin Winkler; Michael Haumann; Thomas Happe

    2015-01-01

    Nature has developed an impressive repertoire of metal-based enzymes that perform complex chemical reactions under moderate conditions. Catalysts that produce molecular hydrogen (H2) are particularly promising for renewable energy applications. Unfortunately, natural and chemical H2-catalysts are often irreversibly degraded by molecular oxygen (O2). Here we present a straightforward procedure based on freeze-drying (lyophilization), that turns [FeFe]-hydrogenases, which are excellent H2-produ...

  13. Optimized Expression and Purification for High-Activity Preparations of Algal [FeFe]-Hydrogenase

    Energy Technology Data Exchange (ETDEWEB)

    Yacoby, I.; Tegler, L. T.; Pochekailov, S.; Zhang, S.; King, P. W.

    2012-04-01

    Recombinant expression and purification of metallo-enzymes, including hydrogenases, at high-yields is challenging due to complex, and enzyme specific, post-translational maturation processes. Low fidelities of maturation result in preparations containing a significant fraction of inactive, apo-protein that are not suitable for biophysical or crystallographic studies. We describe the construction, overexpression and high-yield purification of a fusion protein consisting of the algal [2Fe2S]-ferredoxin PetF (Fd) and [FeFe]-hydrogenase HydA1. The maturation of Fd-HydA1 was optimized through improvements in culture conditions and media components used for expression. We also demonstrated that fusion of Fd to the N-terminus of HydA1, in comparison to the C-terminus, led to increased expression levels that were 4-fold higher. Together, these improvements led to enhanced HydA1 activity and improved yield after purification. The strong binding-affinity of Fd for DEAE allowed for two-step purification by ion exchange and StrepTactin affinity chromatography. In addition, the incorporation of a TEV protease site in the Fd-HydA1 linker allowed for the proteolytic removal of Fd after DEAE step, and purification of HydA1 alone by StrepTactin. In combination, this process resulted in HydA1 purification yields of 5 mg L{sup -1} of culture from E. coli with specific activities of 1000 U (U = 1 {micro}mol hydrogen evolved mg{sup -1} min{sup -1}). The [FeFe]-hydrogenases are highly efficient enzymes and their catalytic sites provide model structures for synthetic efforts to develop robust hydrogen activation catalysts. In order to characterize their structure-function properties in greater detail, and to use hydrogenases for biotechnological applications, reliable methods for rapid, high-yield expression and purification are required.

  14. The key role of biochar in the rapid removal of decabromodiphenyl ether from aqueous solution by biochar-supported Ni/Fe bimetallic nanoparticles

    Science.gov (United States)

    Yi, Yunqiang; Wu, Juan; Wei, Yufen; Fang, Zhanqiang; Tsang, Eric Pokeung

    2017-07-01

    Some problems exist in the current remediation of polybrominated diphenyl ethers (PBDEs) from aqueous solution by using iron-based nanoparticles. Our efforts have contributed to the synthesis of biochar-supported Ni/Fe bimetallic nanoparticle composites (BC@Ni/Fe). Under the optimum operating parameters of BC@Ni/Fe, the morphologic analysis revealed that biochar effectively solved the agglomeration of Ni/Fe nanoparticles and the removal efficiency of BDE209 obtained by BC@Ni/Fe (91.29%) was seven times higher than the sum of biochar (2.55%) and Ni/Fe (11.22%) in 10 min. The degradation products of BDE209 in the solution and absorbed on the BC@Ni/Fe were analyzed with gas chromatography-mass spectroscopy, which indicated that the degradation of BDE209 was mainly a process of stepwise debromination. Meanwhile, compared with Ni/Fe nanoparticles, the adsorption ability of the by-products of BDE209 by BC@Ni/Fe was greater, to a certain extent, which reduced the additional environmental burden. In addition, the concentration of nickle ion leaching from the Ni/Fe nanoparticles was 3.09 mg/L; conversely, the concentration of nickle leaching from BC@Ni/Fe was not detected. This excellent performance in our study indicates a possible means to enhance the reactivity and reduce the secondary risks of Ni/Fe nanoparticles.

  15. Synthesis, structure and reactivity of Ni site models of [NiFeSe] hydrogenases.

    Science.gov (United States)

    Wombwell, Claire; Reisner, Erwin

    2014-03-21

    A series of structural models of the Ni centre in [NiFeSe] hydrogenases has been developed which exhibits key structural features of the Ni site in the H2 cycling enzyme. Specifically, two complexes with a hydrogenase-analogous four-coordinate 'NiS3Se' primary coordination sphere and complexes with a 'NiS2Se2' and a 'NiS4' core are reported. The reactivity of the complexes towards oxygen and protons shows some relevance to the chemistry of [NiFeSe] hydrogenases. Exposure of a 'NiS3Se' complex to atmospheric oxygen results in the oxidation of the selenolate group in the complex to a diselenide, which is released from the nickel site. Oxidation of the selenolate ligand on Ni occurs approximately four times faster than oxidation with the analogous sulfur complex. Reaction of the complexes with one equivalent of HBF4 results in protonation of the monodentate chalcogenolate and the release of this ligand from the metal centre as a thiol or selenol. Unrelated to their biomimetic nature, the complexes serve also as molecular precursors to modify electrodes with Ni-S-Se containing particles by electrochemical deposition. The activated electrodes evolve H2 in pH neutral water with an electrocatalytic onset potential of -0.6 V and a current density of 15 μA cm(-2) at -0.75 V vs. NHE.

  16. Proton Reduction Using a Hydrogenase-Modified Nanoporous Black Silicon Photoelectrode

    Energy Technology Data Exchange (ETDEWEB)

    Zhao, Yixin; Anderson, Nicholas C.; Ratzloff, Michael W.; Mulder, David W.; Zhu, Kai; Turner, John A.; Neale, Nathan R.; King, Paul W.; Branz, Howard M.

    2016-06-15

    Metalloenzymes featuring earth-abundant metal-based cores exhibit rates for catalytic processes such as hydrogen evolution comparable to those of noble metals. Realizing these superb catalytic properties in artificial systems is challenging owing to the difficulty of effectively interfacing metalloenzymes with an electrode surface in a manner that supports efficient charge-transfer. Here, we demonstrate that a nanoporous 'black' silicon (b-Si) photocathode provides a unique interface for binding an adsorbed [FeFe]-hydrogenase enzyme ([FeFe]-H2ase). The resulting [FeFe]-H2ase/b-Si photoelectrode displays a 280 mV more positive onset potential for hydrogen generation than bare b-Si without hydrogenase, similar to that observed for a b-Si/Pt photoelectrode at the same light intensity. Additionally, we show that this H2ase/b-Si electrode exhibits a turnover frequency of >/=1300 s-1 and a turnover number above 107 and sustains current densities of at least 1 mA/cm2 based on the actual surface area of the electrode (not the smaller projected geometric area), orders of magnitude greater than that observed for previous enzyme-catalyzed electrodes. While the long-term stability of hydrogenase on the b-Si surface remains too low for practical applications, this work extends the proof-of-concept that biologically derived metalloenzymes can be interfaced with inorganic substrates to support technologically relevant current densities.

  17. A study on the electrodeposition of NiFe alloy thin films using chronocoulometry and electrochemical quartz crystal microgravimetry

    CERN Document Server

    Myung, N S

    2001-01-01

    Ni, Fe and NiFe alloy thin films were electrodeposited at a polycrystalline Au surface using a range of electrolytes and potentials. Coulometry and EQCM were used for real-time monitoring of electroplating efficiency of the Ni and Fe. The plating efficiency of NiFe alloy thin films was computed with the aid of ICP spectrometry. In general, plating efficiency increased to a steady value with deposition time. Plating efficiency of Fe was lower than that of Ni at -0.85 and -1.0 V but the efficiency approached to the similar plateau value to that of Ni at more negative potentials. The films with higher content of Fe showed different stripping behavior from the ones with higher content of Ni. Finally, compositional data and real-time plating efficiency are presented for films electrodeposited using a range of electrolytes and potentials.

  18. Synthesis and characterization of multilayered BaTiO3/NiFe2O4 thin films

    Directory of Open Access Journals (Sweden)

    Branimir Bajac

    2013-03-01

    Full Text Available Presented research was focused on the fabrication of multiferroic thin film structures, composed of ferrielectric barium titanate perovskite phase and magnetostrictive nickel ferrite spinel phase. The applicability of different, solution based, deposition techniques (film growth from solution, dip coating and spin coating for thefabrication of multilayered BaTiO3 /NiFe2O4 thin films was investigated. It was shown that only spin coating produces films of desired nanostructure, thickness and smooth and crackfree surfaces.

  19. Effects of Ni/Fe bimetallic nanoparticles on phytotoxicity and translocation of polybrominated diphenyl ethers in contaminated soil.

    Science.gov (United States)

    Wu, Juan; Xie, Yingying; Fang, Zhanqiang; Cheng, Wen; Tsang, Pokeung Eric

    2016-11-01

    In vivo studies of the interactions of polybrominated diphenyl ethers (PBDEs) in plants have generally focused on uptake, translocation, metabolism and accumulation, but there were limited reports about the phytotoxicity and translocation of PBDEs in contaminated soil with the effects of nanoparticles. In this study, the effects of Ni/Fe bimetallic nanoparticles on translocation of polybrominated diphenyl ethers (PBDEs) in contaminated soil and its phytotoxicity to Chinese cabbage were investigated by soil culture experiments. The results showed that the plant biomass, germination rate, and shoot and root lengths of treated soil (S-5) increased by 0.0044 g, 15%, and 5 and 6 mm, respectively, compared with untreated soil (S-2B). The average Ni and Fe contents of the edible parts(stem and leaf) of the S-5 sample, which contained 0.03 g/g Ni/Fe and 10 mg/kg BDE209, were measured at 1.71 and 184 mg/kg, respectively. The superoxide dismutase, peroxidase and catalase activities in the S-5 sample decreased by 12%, 6.1% and 5.9%, respectively, while compared with the S-2B sample. In all treatments, the contents of BDE209 and the total PBDEs in sample S-5 were lowest, suggesting that the fresh Ni/Fe nanoparticles had higher toxicity than that of the aged nanoparticles. And the lower brominated PBDEs (tri-to nona-) were detected in samples, indicating uptake, debromination and/or metabolism of PBDEs existed in plants. The phytotoxicity and translocation of BDE209 in the contaminated soil decreased as a result of the effects of the Ni/Fe bimetallic nanoparticles. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Identification, cloning and heterologous expression of active [NiFe]-hydrogenase 2 from Citrobacter sp. SG in Escherichia coli.

    Science.gov (United States)

    Maier, Johannes A H; Ragozin, Sergey; Jeltsch, Albert

    2015-04-10

    Hydrogen (H2) is a potential alternative energy carrier which only produces water and heat upon combustion. Today, industrial hydrogen production mainly uses thermochemical processes based on fossil fuels or electrolysis of water. Therefore, biotechnological approaches to produce H2 from biomass are an interesting alternative. We introduce here a novel direct hydrogen measurement system using a semiconducting device specific for hydrogen detection. Using this device, a bacterium producing considerable amounts of hydrogen under aerobic cultivation was isolated and identified by 16S ribosomal DNA sequencing as Citrobacter sp. The enzyme responsible for the observed hydrogenase activity was partially purified by 3 chromatographic purification steps and could be identified by peptide mass fingerprinting to be a type 2 [NiFe]-hydrogenase. Expression of the [NiFe]-hydrogenase 2 containing operon from Citrobacter sp. SG in Escherichia coli allowed recombinant hydrogen production. The [NiFe]-hydrogenase 2 identified here may be useful for biotechnological hydrogen production. We speculate that the expression of the hydrogenase in Citrobacter may be an adaptation to growth in acidic conditions.

  1. Magnetostriction of fcc(110 single-crystal films of Ni-Fe, Ni, and Co under rotating magnetic fields

    Directory of Open Access Journals (Sweden)

    Ohtani Taiki

    2014-07-01

    Full Text Available Ni-Fe, Ni, and Co(110 single-crystal films with uniaxial magnetic anisotropies are prepared on MgO(110 substrates by radio-frequency magnetron sputtering. The magnetostriction behavior under rotating magnetic fields is investigated. The Ni-Fe film shows waveforms consisting of a mixture of sinusoidal and triangular shapes under fields lower than 200 Oe. The peak of sinusoidal shape is observed when the field is applied along the easy magnetization axis, whereas that of triangular shape appears when the field is applied along the hard axis. With increasing the field from 200 to 300 Oe, the waveform changes to a usual sinusoidal shape. The waveform variation is related to the difference between the directions of uniaxial magnetic anisotropy and magnetization of magnetically unsaturated film. Waveforms consisting of sinusoidal and triangular shapes are also observed for the Ni and the Co films under low rotating fields. The threshold magnetic field where the shape changes to sinusoidal increases in the order of Ni-Fe < Ni < Co. The waveform is influenced by the symmetry and the strength of magnetic anisotropy.

  2. An innovative cloning platform enables large-scale production and maturation of an oxygen-tolerant [NiFe]-hydrogenase from Cupriavidus necator in Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Johannes Schiffels

    Full Text Available Expression of multiple heterologous genes in a dedicated host is a prerequisite for approaches in synthetic biology, spanning from the production of recombinant multiprotein complexes to the transfer of tailor-made metabolic pathways. Such attempts are often exacerbated, due in most cases to a lack of proper directional, robust and readily accessible genetic tools. Here, we introduce an innovative system for cloning and expression of multiple genes in Escherichia coli BL21 (DE3. Using the novel methodology, genes are equipped with individual promoters and terminators and subsequently assembled. The resulting multiple gene cassettes may either be placed in one vector or alternatively distributed among a set of compatible plasmids. We demonstrate the effectiveness of the developed tool by production and maturation of the NAD(+reducing soluble [NiFe]-hydrogenase (SH from Cupriavidus necator H16 (formerly Ralstonia eutropha H16 in E. coli BL21Star™ (DE3. The SH (encoded in hoxFUYHI was successfully matured by co-expression of a dedicated set of auxiliary genes, comprising seven hyp genes (hypC1D1E1A2B2F2X along with hoxW, which encodes a specific endopeptidase. Deletion of genes involved in SH maturation reduced maturation efficiency substantially. Further addition of hoxN1, encoding a high-affinity nickel permease from C. necator, considerably increased maturation efficiency in E. coli. Carefully balanced growth conditions enabled hydrogenase production at high cell-densities, scoring mg·(Liter culture(-1 yields of purified functional SH. Specific activities of up to 7.2±1.15 U·mg(-1 were obtained in cell-free extracts, which is in the range of the highest activities ever determined in C. necator extracts. The recombinant enzyme was isolated in equal purity and stability as previously achieved with the native form, yielding ultrapure preparations with anaerobic specific activities of up to 230 U·mg(-1. Owing to the combinatorial power

  3. An innovative cloning platform enables large-scale production and maturation of an oxygen-tolerant [NiFe]-hydrogenase from Cupriavidus necator in Escherichia coli.

    Science.gov (United States)

    Schiffels, Johannes; Pinkenburg, Olaf; Schelden, Maximilian; Aboulnaga, El-Hussiny A A; Baumann, Marcus E M; Selmer, Thorsten

    2013-01-01

    Expression of multiple heterologous genes in a dedicated host is a prerequisite for approaches in synthetic biology, spanning from the production of recombinant multiprotein complexes to the transfer of tailor-made metabolic pathways. Such attempts are often exacerbated, due in most cases to a lack of proper directional, robust and readily accessible genetic tools. Here, we introduce an innovative system for cloning and expression of multiple genes in Escherichia coli BL21 (DE3). Using the novel methodology, genes are equipped with individual promoters and terminators and subsequently assembled. The resulting multiple gene cassettes may either be placed in one vector or alternatively distributed among a set of compatible plasmids. We demonstrate the effectiveness of the developed tool by production and maturation of the NAD(+)reducing soluble [NiFe]-hydrogenase (SH) from Cupriavidus necator H16 (formerly Ralstonia eutropha H16) in E. coli BL21Star™ (DE3). The SH (encoded in hoxFUYHI) was successfully matured by co-expression of a dedicated set of auxiliary genes, comprising seven hyp genes (hypC1D1E1A2B2F2X) along with hoxW, which encodes a specific endopeptidase. Deletion of genes involved in SH maturation reduced maturation efficiency substantially. Further addition of hoxN1, encoding a high-affinity nickel permease from C. necator, considerably increased maturation efficiency in E. coli. Carefully balanced growth conditions enabled hydrogenase production at high cell-densities, scoring mg·(Liter culture)(-1) yields of purified functional SH. Specific activities of up to 7.2±1.15 U·mg(-1) were obtained in cell-free extracts, which is in the range of the highest activities ever determined in C. necator extracts. The recombinant enzyme was isolated in equal purity and stability as previously achieved with the native form, yielding ultrapure preparations with anaerobic specific activities of up to 230 U·mg(-1). Owing to the combinatorial power exhibited by the

  4. Preliminary testing of NiFe2O4-NiO as ceramic matrix of cermet inert anode in aluminum electrolysis

    Institute of Scientific and Technical Information of China (English)

    秦庆伟; 赖延清; 肖劲; 李劼; 刘业翔

    2003-01-01

    Sintered samples of nickel ferrite-nickel oxide ceramic, usually used as the ceramic phase of cermet inert anode in aluminum electrolysis, were prepared and characterized. The solubilities of NiFe2O4-NiO ceramics were measured using an equilibration technique in the Na3AlF6-10%AlF3-5%CaF2-5 %Al2O3 melts at 960 ℃. Electrical resistivity was also measured for NiFe2O4-NiO ceramic samples prepared using the usual ceramic technique as function of temperature and content of NiO with an improved pyroconductivity test device, consisting of a specially constructed closed furnace and a Potentiostat/Galvanostat, based on the conventional direct current four-probe technique. Results show that, under the experimental conditions, the solubility of Fe from NiFe2O4 is 0.06% and Ni from NiFe2O4 is 0.008%. The solubility of Fe and Ni from NiFe2O4-NiO ceramic is inversely related to each other. The solubility of Ni increases but overall solubility of NiFe2O4-NiO ceramics decreases with increasing NiO content. The studied ceramic samples have a semiconductor behavior where electrical resistivity ρ decreases with increasing temperature and the resistivity ρ increases with increasing porosity. The resistivity ρ of NiFe2O4-NiO ceramic shows a minimum with increasing the content of NiO at various temperatures.

  5. One-pot fabrication of NiFe2O4 nanoparticles on α-Ni(OH)2 nanosheet for enhanced water oxidation

    Science.gov (United States)

    Chen, Hong; Yan, Junqing; Wu, Huan; Zhang, Yunxia; Liu, Shengzhong (Frank)

    2016-08-01

    Water splitting has been intensively investigated as a promising solution to resolve the future environmental and energy crises. The oxygen evolution reaction (OER) of the photo- and electric field-induced water splitting limits the development of other reactions, including hydrogen evolution reaction (HER). Fe, Ni and NiFe (hydro) oxide-based catalysts are generally acknowledged among the best candidates of OER catalysts for water splitting. Herein, we developed a one-pot simple hydrothermal process to assemble NiFe2O4 nanoparticles onto the α-Ni(OH)2 nanosheets. The first formed NiFe2O4 under high temperature and pressure environment induces and assists the α-Ni(OH)2 formation without any further additives, because the distance between the neighboring Ni atoms in the cubic NiFe2O4 is similar to that in the α-Ni(OH)2 {003} facets. We have synthesized a series of NiFe2O4/α-Ni(OH)2 compounds and find that the overpotential decreases with the increase of Ni(OH)2 content while the OER kinetics stays unchanged, suggesting that Ni(OH)2 plays a major role in overpotential while NiFe2O4 mainly affects the OER kinetics. The obtained NiFe2O4/α-Ni(OH)2 compounds is also found to be a promising co-catalyst for the photocatalytic water oxidation. In fact, it is even more active than the noble PtOx with acceptable stability for the oxygen generation.

  6. Controllable synthesis and enhanced microwave absorbing properties of Fe3O4/NiFe2O4/Ni heterostructure porous rods

    Science.gov (United States)

    Li, Yana; Wu, Tong; Jin, Keying; Qian, Yao; Qian, Naxin; Jiang, Kedan; Wu, Wenhua; Tong, Guoxiu

    2016-11-01

    We developed a coordinated self-assembly/precipitate transfer/sintering method that allows the controllable synthesis of Fe3O4/NiFe2O4/Ni heterostructure porous rods (HPRs). A series of characterizations confirms that changing [Ni2+] can effectively control the crystal size, internal strain, composition, textural characteristics, and properties of HPRs. Molar percentages of Ni and NiFe2O4 in HPRs increase with [Ni2+] in various Boltzmann function modes. Saturation magnetization Ms and coercivity Hc show U-shaped change trends because of crystal size, composition, and interface magnetic coupling. High magnetic loss is maintained after decorating NiFe2O4 and Ni on the surface of Fe3O4 PRs. Controlling the NiFe2O4 interface layers and Ni content can improve impedance matching and dielectric losses, thereby leading to lighter weight, stronger absorption, and broader absorption band of Fe3O4/NiFe2O4/Ni HPRs than Fe3O4 PRs. An optimum EM wave absorbing property was exhibited by Fe3O4/NiFe2O4/Ni HPRs formed at [Ni2+] = 0.05 M. The maximum reflection loss (RL) reaches -58.4 dB at 13.68 GHz, which corresponds to a 2.1 mm matching thickness. The absorbing bandwidth (RL ≤ -20 dB) reaches 14.4 GHz with the sample thickness at 1.6-2.4 and 2.8-10.0 mm. These excellent properties verify that Fe3O4/NiFe2O4/Ni HPRs are promising candidates for new and effective absorptive materials.

  7. Removal of Methyl Orange Dye from Aqueous Solutions using NiFe2O4 Nanoparticles: Equilibrium and Kinetic Studies

    Directory of Open Access Journals (Sweden)

    S Sobhanardakani

    2016-09-01

    Full Text Available Background and Objective: Dyes are important pollutants that lead to producing serious hazards to human, other animals and organisms. Dyes are not biodegradable by aerobic treatment processes. Therefore, their removal from industrial effluents before discharging into the environment requires extreme and great attention. The aim of this research was to evaluate removal efficacy of methyl orange dye from aqueous solutions using NiFe2O4 nanoparticles. Materials and Methods: This study was an empirical investigation in which NiFe2O4 nanoparticles were synthesized by co-precipitation method and were used as an adsorbent for the removal of methyl orange from aqueous solution. NiFe2O4 nanoparticles were characterized using X-Ray Diffraction (XRD, Transmission Electronic Microscopy (TEM, pHpzc and SEM-EDX elemental analysis methods. Experiments were conducted discontinuously using 20 mL methyl orange solution of 40 mg/L. The effect of variables such as pH (2-8, amount of adsorbent (0.009-0.07 g and contact time (2-70 min on the efficacy of dye removal was studied. Finally, experimental data were compared by Langmuir, Freundlich, and Temkin isotherms and pseudo-first-order and pseudo-second-order kinetic models. Results: TEM images showed that the NiFe2O4 nanoparticles had spherical shapes with the size of 12 nm. The results indicated that removal efficiency increased up to 0.04 g adsorbent and 20 min contact time. The optimum pH for methyl range removal was 2. Moreover, under these conditions, the adsorption process followed the Langmuir adsorption isotherm with a correlation coefficient of 0.995 and pseudo-second-order kinetic model with a correlation coefficient of 0.999. Also, the maximum adsorption capacity of the prepared adsorbent was 135 (mg/g for Langmuir isotherm. Conclusion: The NiFe2O4 nanoparticles are effective and available adsorbents for the removal of methyl orange from industrial wastewater.

  8. High-performance hydrogen production and oxidation electrodes with hydrogenase supported on metallic single-wall carbon nanotube networks.

    Science.gov (United States)

    Svedružić, Draženka; Blackburn, Jeffrey L; Tenent, Robert C; Rocha, John-David R; Vinzant, Todd B; Heben, Michael J; King, Paul W

    2011-03-30

    We studied the electrocatalytic activity of an [FeFe]-hydrogenase from Clostridium acetobutylicum (CaH2ase) immobilized on single-wall carbon nanotube (SWNT) networks. SWNT networks were prepared on carbon cloth by ultrasonic spraying of suspensions with predetermined ratios of metallic and semiconducting nanotubes. Current densities for both proton reduction and hydrogen oxidation electrocatalytic activities were at least 1 order of magnitude higher when hydrogenase was immobilized onto SWNT networks with high metallic tube (m-SWNT) content in comparison to hydrogenase supported on networks with low metallic tube content or when SWNTs were absent. We conclude that the increase in electrocatalytic activities in the presence of SWNTs was mainly due to the m-SWNT fraction and can be attributed to (i) substantial increases in the active electrode surface area, and (ii) improved electronic coupling between CaH2ase redox-active sites and the electrode surface.

  9. Guiding Principles of Hydrogenase Catalysis Instigated and Clarified by Protein Film Electrochemistry.

    Science.gov (United States)

    Armstrong, Fraser A; Evans, Rhiannon M; Hexter, Suzannah V; Murphy, Bonnie J; Roessler, Maxie M; Wulff, Philip

    2016-05-17

    Protein film electrochemistry (PFE) is providing cutting-edge insight into the chemical principles underpinning biological hydrogen. Attached to an electrode, many enzymes exhibit "reversible" electrocatalytic behavior, meaning that a catalyzed redox reaction appears reversible or quasi-reversible when viewed by cyclic voltammetry. This efficiency is most relevant for enzymes that are inspiring advances in renewable energy, such as hydrogen-activating and CO2-reducing enzymes. Exploiting the rich repertoire of available instrumental methods, PFE experiments yield both a general snapshot and fine detail, all from tiny samples of enzyme. The dynamic electrochemical investigations blaze new trails and add exquisite detail to the information gained from structural and spectroscopic studies. This Account describes recent investigations of hydrogenases carried out in Oxford, including ideas initiated with PFE and followed through with complementary techniques, all contributing to an eventual complete picture of fast and efficient H2 activation without Pt. By immobilization of an enzyme on an electrode, catalytic electron flow and the chemistry controlling it can be addressed at the touch of a button. The buried nature of the active site means that structures that have been determined by crystallography or spectroscopy are likely to be protected, retained, and fully relevant in a PFE experiment. An electrocatalysis model formulated for the PFE of immobilized enzymes predicts interesting behavior and gives insight into why some hydrogenases are H2 producers and others are H2 oxidizers. Immobilization also allows for easy addition and removal of inhibitors along with precise potential control, one interesting outcome being that formaldehyde forms a reversible complex with reduced [FeFe]-hydrogenases, thereby providing insight into the order of electron and proton transfers. Experiments on O2-tolerant [NiFe]-hydrogenases show that O2 behaves like a reversible inhibitor: it

  10. Degradation of aniline by heterogeneous Fenton's reaction using a Ni-Fe oxalate complex catalyst.

    Science.gov (United States)

    Liu, Yucan; Zhang, Guangming; Fang, Shunyan; Chong, Shan; Zhu, Jia

    2016-11-01

    A Ni-Fe oxalate complex catalyst was synthesized and characterized by means of Brunauer-Emmet-Teller (BET) method, scanning electron microscope (SEM) and X-ray photo-electron spectroscopy (XPS). The catalyst showed good catalytic activity for aniline degradation by heterogeneous Fenton's reaction, in which the synergetic index was 9.3. The effects of reaction temperature, catalyst dosage, hydrogen peroxide concentration and initial pH were investigated. Under the optimum conditions (T = 293 K, catalyst dosage = 0.2 g/L, H2O2 concentration = 4 mmol/L and initial pH = 5.4), 100% aniline could be removed within 35 min, and approximately 88% deamination efficiency was achieved in 60 min. The aniline degradation process followed the pseudo-first-order kinetic (k = 0.177 min(-1)) with activation energy (Ea) of 49.4 kJ mol(-1). Aniline could be removed in a broad initial pH (3-8) due to the excellent pH-tolerance property of the catalyst. The detected ammonium ion indicated that deamination occurred during aniline degradation. It was proposed that deamination synchronized with aniline removal, and aniline was attacked by free radicals to generate benzoquinonimine and phenol. This system is promising for the removal of aniline from water.

  11. Anodic behaviour of oxidised Ni-Fe alloys in cryolite-alumina melts

    Energy Technology Data Exchange (ETDEWEB)

    Chapman, Vivien, E-mail: v.singleton@student.unsw.edu.a [Centre for Electrochemical and Mineral Processing, School of Chemical Engineering, University of New South Wales, Sydney 2052 (Australia); Welch, Barry J. [Welbank Consulting Ltd., PO Box 207, Whitianga 3542 (New Zealand); Skyllas-Kazacos, Maria [Centre for Electrochemical and Mineral Processing, School of Chemical Engineering, University of New South Wales, Sydney 2052 (Australia)

    2011-01-01

    Nickel-iron alloys have been identified as promising inert anode candidates for the Hall-Heroult process. In this study, binary Ni-Fe alloys of various compositions were subjected to short-term galvanostatic electrolysis in a cryolite-alumina bath at 960 {sup o}C. Prior to electrolysis, the anodes were oxidised at 800 {sup o}C for 48 h, forming a protective scale. Fe{sub 2}O{sub 3}, Ni{sub x}Fe{sub 3-x}O{sub 4} and Ni{sub x}Fe{sub 1-x}O were identified as the major scale components using a combination of X-ray diffraction (XRD) analysis and energy dispersive X-ray spectroscopy (EDX). Anodes having Ni content of 50-65 wt% performed adequately during short-term electrolysis, operating at a steady potential of 3-3.5 V vs. AlF{sub 3}/Al. Overall, it was found that the pre-formed oxide scale was effective in reducing anode wear and fluoridation. In the absence of a pre-formed scale, anodes were shown to undergo appreciable internal corrosion and/or passivation due to metal fluoride formation. Analysis of the anodes following electrolysis was performed using XRD and electron microprobe analysis (EPMA).

  12. Hydrogenase activity in Azospirillum brasilense is inhibited by nitrite, nitric oxide, carbon monoxide, and acetylene

    Energy Technology Data Exchange (ETDEWEB)

    Tibelius, K.H.; Knowles, R.

    1984-10-01

    Nitrite, NO, CO, and C/sub 2/H/sub 2/ inhibited O/sub 2/-dependent H/sub 2/ uptake (H/sup 3/H oxidation) in denitrifying Azospirillum brasilense Sp7 grown anaerobically on N/sub 2/O or NO/sub 3//sup -/. The apparent K/sub i/ values for inhibition of O/sub 2/-dependent H/sub 2/ uptake were 20 ..mu..M for NO/sub 2//sup -/, 0.4 ..mu..M for NO, 28 ..mu..M for CO, and 88 ..mu..M for C/sub 2/H/sub 2/. These inhibitors also affected methylene blue-dependent H/sub 2/ uptake, presumably by acting directly on the hydrogenase. Nitrite and NO inhibited H/sub 2/ uptake irreversibly, whereas inhibition due to CO was easily reversed by repeatedly evacuating and backfilling with N/sub 2/. The C/sub 2/H/sub 2/ inhibition was not readily reversed, partly due to difficulty in removing the last traces of this gas from solution. The NO/sub 2//sup -/ inhibition of malate-dependent respiration was readily reversed by repeatedly washing the cells, in contrast to the effect of NO/sub 2//sup -/ on H/sub 2/-dependent respiration. These results suggest that the low hydrogenase activities observed in NO/sub 3//sup -/-grown cultures of A. brasilense may be due to the irreversible inhibition of hydrogenase by NO/sub 2//sup -/ and NO produced by NO/sub 3//sup -/ reduction.

  13. Analysis of hydrogenase 1 levels reveals an intimate link between carbon and hydrogen metabolism in Escherichia coli K-12.

    Science.gov (United States)

    Pinske, Constanze; McDowall, Jennifer S; Sargent, Frank; Sawers, R Gary

    2012-03-01

    Two of the three [NiFe]-hydrogenases (Hyd) of Escherichia coli have a hydrogen-uptake function in anaerobic metabolism. While Hyd-2 is maximally synthesized when the bacterium grows by fumarate respiration, Hyd-1 synthesis shows a correlation with fermentation of sugar substrates. In an attempt to advance our knowledge on the physiological function of Hyd-1 during fermentative growth, we examined Hyd-1 activity and levels in various derivatives of E. coli K-12 MC4100 with specific defects in sugar utilization. MC4100 lacks a functional fructose phosphotransferase system (PTS) and therefore grows more slowly under anaerobic conditions in rich medium in the presence of d-fructose compared with d-glucose. Growth in the presence of fructose resulted in an approximately 10-fold increase in Hyd-1 levels in comparison with growth under the same conditions with glucose. This increase in the amount of Hyd-1 was not due to regulation at the transcriptional level. Reintroduction of a functional fruBKA-encoded fructose PTS into MC4100 restored growth on d-fructose and reduced Hyd-1 levels to those observed after growth on d-glucose. Reducing the rate of glucose uptake by introducing a mutation in the gene encoding the cAMP receptor protein, or consumption through glycolysis, by introducing a mutation in phosphoglucose isomerase, increased Hyd-1 levels during growth on glucose. These results suggest that the ability to oxidize hydrogen by Hyd-1 shows a strong correlation with the rate of carbon flow through glycolysis and provides a direct link between hydrogen, carbon and energy metabolism.

  14. Microstructural evolution of NiFe2O4-10NiO powder prepared by high temperature solid state reaction

    Institute of Scientific and Technical Information of China (English)

    ZHANG Lei; ZHOU Ke-chao; LI Zhi-you; YANG Wen-jie

    2006-01-01

    The NiFe2O4-10NiO powder for inert anode of aluminium electrolysis was prepared by high temperature solid state reaction. The microstructural evolution from the raw materials NiO and Fe2O3 to the NiFe2O4-10NiO powder was studied by SEM. The results show that the domain structure making up of the agglomerate particles of Fe2O3 remains after high temperature solid state reaction, and the diffusion of Ni2+ into Fe2O3 structure is the control step of the reaction process. A microstructure with compact structure and fine grain inside the particle results from the sintering of NiFe2O4-10NiO powder.

  15. 浑球红细菌(Rhodobacter sphaeroides)中氢化酶正调节基因hupR的克隆及功能分析%Isolation and Analysis of hupR Gene Required for the Expression of Hydrogenase in Rhodobacter sphaeroides

    Institute of Scientific and Technical Information of China (English)

    徐冬青; 吴永强

    2001-01-01

    Cosmid 1 containing the hup genes isolated from the photosynthetic b acterium Rhodobacter sphaeroides was studied. The hupR gene from cosmid 1 was cl oned and sequenced (EMBL accession number AJ243734). It encoded a 54.031 kD prot ein homologous to transcriptional regulators belonging to the superfamily of two -component regulatory systems. The HupR protein was overexpressed in Escheric hia coli in the form of His6-tagged HupR. The cloned hupR gene could res tore hydrog enase activity in R. Sphaeroides hupR mutants and activate hupSL gene transcription.%从光合细菌Rhodobacter sphaeroides基因文库中分离出含有氢化酶基因簇(hup)的粘粒cosmid 1后, 亚克隆了R. sphaeroides的氢化酶调节基因hupR, 测定了hupR的核苷酸序列, 并完成了氢化酶基因簇的部分物理图谱. 实验结果表明, hupR基因全长1 476 bp, 编码的HupR蛋白分子量约为54.031 kD (EMBL接受号: AJ243734). 与R. capsulatus中HupR相比, 同源性高达73%. 同源性比较结果表明, 它属于双组分调节系统中受体蛋白. hupR基因在E. coli中进行了体外表达, 纯化后测定得到的HupR蛋白的分子量大小与hupR基因推测的分子量大小一致. 通过双交换, 将卡那霉素抗性基因插入hupR基因, 获得丧失氢化酶活性的hupR-的突变株, KR5和KR7. hupS∷lacZ融合基因在野生型中的转录表达量是在该突变株中的7~9倍. 将hupR基因置于弱启动子pfru下游, 构建了质粒pNRC3, 并将其导入hupR-的突变株, 可使突变株重新获得氢化酶活性. 以上结果说明, HupR蛋白对氢化酶的转录表达起着正调节作用. 在HupR蛋白的磷酸化区域进行定点和缺失突变, 不影响HupR激活氢化酶基因的表达, 推测HupR蛋白是在非磷酸化的状态下起调节作用的.

  16. Distal [FeS]-Cluster Coordination in [NiFe]-Hydrogenase Facilitates Intermolecular Electron Transfer.

    Science.gov (United States)

    Petrenko, Alexander; Stein, Matthias

    2017-01-05

    Biohydrogen is a versatile energy carrier for the generation of electric energy from renewable sources. Hydrogenases can be used in enzymatic fuel cells to oxidize dihydrogen. The rate of electron transfer (ET) at the anodic side between the [NiFe]-hydrogenase enzyme distal iron-sulfur cluster and the electrode surface can be described by the Marcus equation. All parameters for the Marcus equation are accessible from Density Functional Theory (DFT) calculations. The distal cubane FeS-cluster has a three-cysteine and one-histidine coordination [Fe₄S₄](His)(Cys)₃ first ligation sphere. The reorganization energy (inner- and outer-sphere) is almost unchanged upon a histidine-to-cysteine substitution. Differences in rates of electron transfer between the wild-type enzyme and an all-cysteine mutant can be rationalized by a diminished electronic coupling between the donor and acceptor molecules in the [Fe₄S₄](Cys)₄ case. The fast and efficient electron transfer from the distal iron-sulfur cluster is realized by a fine-tuned protein environment, which facilitates the flow of electrons. This study enables the design and control of electron transfer rates and pathways by protein engineering.

  17. Electrochemistry of Simple Organometallic Models of Iron-Iron Hydrogenases in Organic Solvent and Water.

    Science.gov (United States)

    Gloaguen, Frederic

    2016-01-19

    Synthetic models of the active site of iron-iron hydrogenases are currently the subjects of numerous studies aimed at developing H2-production catalysts based on cheap and abundant materials. In this context, the present report offers an electrochemist's view of the catalysis of proton reduction by simple binuclear iron(I) thiolate complexes. Although these complexes probably do not follow a biocatalytic pathway, we analyze and discuss the interplay between the reduction potential and basicity and how these antagonist properties impact the mechanisms of proton-coupled electron transfer to the metal centers. This question is central to any consideration of the activity at the molecular level of hydrogenases and related enzymes. In a second part, special attention is paid to iron thiolate complexes holding rigid and unsaturated bridging ligands. The complexes that enjoy mild reduction potentials and stabilized reduced forms are promising iron-based catalysts for the photodriven evolution of H2 in organic solvents and, more importantly, in water.

  18. Hydride bridge in [NiFe]-hydrogenase observed by nuclear resonance vibrational spectroscopy

    Science.gov (United States)

    Ogata, Hideaki; Krämer, Tobias; Wang, Hongxin; Schilter, David; Pelmenschikov, Vladimir; van Gastel, Maurice; Neese, Frank; Rauchfuss, Thomas B.; Gee, Leland B.; Scott, Aubrey D.; Yoda, Yoshitaka; Tanaka, Yoshihito; Lubitz, Wolfgang; Cramer, Stephen P.

    2015-08-01

    The metabolism of many anaerobes relies on [NiFe]-hydrogenases, whose characterization when bound to substrates has proven non-trivial. Presented here is direct evidence for a hydride bridge in the active site of the 57Fe-labelled fully reduced Ni-R form of Desulfovibrio vulgaris Miyazaki F [NiFe]-hydrogenase. A unique `wagging' mode involving H- motion perpendicular to the Ni(μ-H)57Fe plane was studied using 57Fe-specific nuclear resonance vibrational spectroscopy and density functional theory (DFT) calculations. On Ni(μ-D)57Fe deuteride substitution, this wagging causes a characteristic perturbation of Fe-CO/CN bands. Spectra have been interpreted by comparison with Ni(μ-H/D)57Fe enzyme mimics [(dppe)Ni(μ-pdt)(μ-H/D)57Fe(CO)3]+ and DFT calculations, which collectively indicate a low-spin Ni(II)(μ-H)Fe(II) core for Ni-R, with H- binding Ni more tightly than Fe. The present methodology is also relevant to characterizing Fe-H moieties in other important natural and synthetic catalysts.

  19. Optimization of detecting hydrogenase activity for acidogenic fermentation of activated sludge

    Institute of Scientific and Technical Information of China (English)

    ZHENG Guo-chen; HE Jun-guo; LI Jian-zheng; AJAY Kumar Jha; ZHANG Li-guo

    2010-01-01

    In order to evaluate the hydrogen-producing efficiency of anaerobic activated sludge in Anaerobic Baffled Reactor(ABR)fermentation processes,the optimal conditions for hydrogen producing hydrogenase method on methyl viologen(MV)assay was used to detect the hydrogen production activity of the activated sludge.The most favorable parameters such as 0.6 mL sodium acetate buffer(pH 5.0),100 μL lysozyme,0.2 mL sodium di bromoethane(9.0 mmol/L)and 0.7 mmol/L iron added into 1 mL activated sludge(2.66~26.64 gMLVSS/L)were found.Furthermore,reaction temperature and culture time were detected as 40 ℃ and 30 min respectively.Sodium thiosulfate and sodium sulfides were taken as the reducing agent while trichloroacetic acid as terminator.Under the MV optimal conditions,micro-texic Dimethyl sulfoxide(DMSO)get higher security and better accuracy.The sensitivity of the detection methods(DMSO as electron carrier)was increased by more than30%.The results show that the optimal conditions can be applied to measure hydrogenase activity correlating with its specific hydrogen production rate in a hydrogen-producing anaerobic activated sludge system.

  20. Using in vitro maturation and cell-free expression to explore [FeFe] hydrogenase activation and protein scaffolding requirements

    Energy Technology Data Exchange (ETDEWEB)

    Swartz, James [Stanford Univ., CA (United States)

    2017-01-25

    Final Project Report describing work to elucidate mechanisms for the activation of [FeFe]-hydrogenases and to explore the impact of the polypeptide scaffolding on the function of the Fe-S redox and catalytic centers with emphasis on improving oxygen tolerance.

  1. Patterns of [FeFe] hydrogenase diversity in the gut microbial communities of lignocellulose-feeding higher termites.

    Science.gov (United States)

    Ballor, Nicholas R; Leadbetter, Jared R

    2012-08-01

    Hydrogen is the central free intermediate in the degradation of wood by termite gut microbes and can reach concentrations exceeding those measured for any other biological system. Degenerate primers targeting the largest family of [FeFe] hydrogenases observed in a termite gut metagenome have been used to explore the evolution and representation of these enzymes in termites. Sequences were cloned from the guts of the higher termites Amitermes sp. strain Cost010, Amitermes sp. strain JT2, Gnathamitermes sp. strain JT5, Microcerotermes sp. strain Cost008, Nasutitermes sp. strain Cost003, and Rhyncotermes sp. strain Cost004. Each gut sample harbored a more rich and evenly distributed population of hydrogenase sequences than observed previously in the guts of lower termites and Cryptocercus punctulatus. This accentuates the physiological importance of hydrogen for higher termite gut ecosystems and may reflect an increased metabolic burden, or metabolic opportunity, created by a lack of gut protozoa. The sequences were phylogenetically distinct from previously sequenced [FeFe] hydrogenases. Phylogenetic and UniFrac comparisons revealed congruence between host phylogeny and hydrogenase sequence library clustering patterns. This may reflect the combined influences of the stable intimate relationship of gut microbes with their host and environmental alterations in the gut that have occurred over the course of termite evolution. These results accentuate the physiological importance of hydrogen to termite gut ecosystems.

  2. First-principles study of ferroelectricity induced by p-d hybridization in ferrimagnetic NiFe2O4

    Science.gov (United States)

    Jong, Un-Gi; Yu, Chol-Jun; Park, Yong-Su; Ri, Chong-Suk

    2016-09-01

    We investigate the ferrimagnetism and ferroelectricity of bulk NiFe2O4 with tetragonal P41 22 symmetry by means of density functional theory calculations using generalized gradient approximation + Hubbard U approach. Special attention is paid to finding the most energetically favorable configuration on magnetic ordering and further calculating the reliable spontaneous electric polarization. With the fully optimized crystalline structure of the most stable configuration, the spontaneous polarization is obtained to be 23 μC/cm2 along the z direction, which originates from the hybridization between the 3d states of the Fe3+ cation and the 2p states of oxygen induced by Jahn-Teller effect.

  3. Thermo-therapeutic applications of chitosan- and PEG-coated NiFe2O4 nanoparticles

    Science.gov (United States)

    Manjura Hoque, S.; Tariq, Mehrin; Liba, S. I.; Salehin, F.; Mahmood, Z. H.; Khan, M. N. I.; Chattopadhayay, K.; Islam, Rafiqul; Akhter, S.

    2016-07-01

    The paper reports the thermo-therapeutic applications of chitosan- and PEG-coated nickel ferrite (NiFe2O4) nanoparticles. In this study NiFe2O4 nanoparticles were synthesized by the co-precipitation method, tuning the particle size through heat treatment in the temperature range from 200-800 °C for 3 h. XRD and TEM analysis revealed that the the ultrafine nanoparticles were of size 2-58 nm. Crystallinity of the NiFe2O4 nanoparticles in the as-dried condition with the particle size ˜2-3 nm was confirmed from the presence of a lattice fringe in the HRTEM image. VSM measurements showed that a superparamagnetic/ferromagnetic transition occurs with increasing particle size, which was further confirmed by Mössbauer spectroscopy. The nickel ferrite nanoparticles with optimum particle size of 10 nm were then coated with materials commonly used for biomedical applications, i.e. chitosan and PEG, to form homogeneous suspensions. The hydrodynamic diameter and the polydispersity index (PDI) were analyzed by dynamic light scattering at the physiological temperature of 37 °C and found to be 187 nm and 0.21 for chitosan-coated nanoparticles and 285 nm and 0.32 for PEG-coated ones. The specific loss power of rf induction heating by the set-up for hyperthermia and r 2 relaxivity by the nuclear magnetic resonance were determined. The results of induction heating measurements showed that the temperature attained by the nanoparticles of size 10 nm and concentration of about 20 mg ml-1 was >70 °C (for chitosan) and >64 °C (for PEG). It has been demonstrated that the required temperature for hyperthermia heating could be tuned by tuning the particle size, shape and magnetization and the concentration of solution. For other potential biomedical applications of the NiFe2O4 nanoparticle solution, e.g. magnetic resonance imaging, the NMR studies yielded the T 1 and T 2 relaxivities as 0.348 and 89 mM-1 s-1 respectively. The fact that the T 2 relaxivity is orders of magnitude higher

  4. Metagenomic and PCR-Based Diversity Surveys of [FeFe]-Hydrogenases Combined with Isolation of Alkaliphilic Hydrogen-Producing Bacteria from the Serpentinite-Hosted Prony Hydrothermal Field, New Caledonia

    Science.gov (United States)

    Mei, Nan; Postec, Anne; Monnin, Christophe; Pelletier, Bernard; Payri, Claude E.; Ménez, Bénédicte; Frouin, Eléonore; Ollivier, Bernard; Erauso, Gaël; Quéméneur, Marianne

    2016-01-01

    High amounts of hydrogen are emitted in the serpentinite-hosted hydrothermal field of the Prony Bay (PHF, New Caledonia), where high-pH (~11), low-temperature (< 40°C), and low-salinity fluids are discharged in both intertidal and shallow submarine environments. In this study, we investigated the diversity and distribution of potentially hydrogen-producing bacteria in Prony hyperalkaline springs by using metagenomic analyses and different PCR-amplified DNA sequencing methods. The retrieved sequences of hydA genes, encoding the catalytic subunit of [FeFe]-hydrogenases and, used as a molecular marker of hydrogen-producing bacteria, were mainly related to those of Firmicutes and clustered into two distinct groups depending on sampling locations. Intertidal samples were dominated by new hydA sequences related to uncultured Firmicutes retrieved from paddy soils, while submarine samples were dominated by diverse hydA sequences affiliated with anaerobic and/or thermophilic submarine Firmicutes pertaining to the orders Thermoanaerobacterales or Clostridiales. The novelty and diversity of these [FeFe]-hydrogenases may reflect the unique environmental conditions prevailing in the PHF (i.e., high-pH, low-salt, mesothermic fluids). In addition, novel alkaliphilic hydrogen-producing Firmicutes (Clostridiales and Bacillales) were successfully isolated from both intertidal and submarine PHF chimney samples. Both molecular and cultivation-based data demonstrated the ability of Firmicutes originating from serpentinite-hosted environments to produce hydrogen by fermentation, potentially contributing to the molecular hydrogen balance in situ. PMID:27625634

  5. Spectroelectrochemical characterization of the active site of the [FeFe] hydrogenase HydA1 from Chlamydomonas reinhardtii.

    Science.gov (United States)

    Silakov, Alexey; Kamp, Christina; Reijerse, Eduard; Happe, Thomas; Lubitz, Wolfgang

    2009-08-25

    Hydrogenases catalyze the reversible oxidation of molecular hydrogen. The active site of the [FeFe] hydrogenases (H-cluster) contains a catalytically active binuclear subcluster ([2Fe](H)) connected to a "cubane" [4Fe4S](H) subcluster. Here we present an IR spectroelectrochemical study of the [FeFe] hydrogenase HydA1 isolated from the green alga Chlamydomonas reinhardtii. The enzyme shows IR bands similar to those observed for bacterial [FeFe] hydrogenases. They are assigned to the stretching vibrations of the CN(-) and CO ligands on both irons of the [2Fe](H) subcluster. By following changes in frequencies of the IR bands during electrochemical titrations, two one-electron redox processes of the active enzyme could be distinguished. The reduction of the oxidized state (H(ox)) occurred at a midpoint potential of -400 mV vs NHE (H(ox)/H(red) transition) and relates to a change of the formal oxidation state of the binuclear subcluster. A subsequent reduction (H(red)/H(sred) transition) was determined to have a midpoint potential of -460 mV vs NHE. On the basis of the IR spectra, it is suggested that the oxidation state of the binuclear subcluster does not change in this transition. Tentatively, a reduction of the [4Fe4S](H) cluster has been proposed. In contrast to the bacterial [FeFe] hydrogenases, where the bridging CO ligand becomes terminal when going from H(ox) to H(red), in HydA1 the bridging CO is present in both the H(ox) and H(red) state. The removal of the bridging CO moiety has been observed in the H(red) to H(sred) transition. The significance of this result for the hydrogen conversion mechanism of this class of enzymes is discussed.

  6. 海马齿甜菜碱醛脱氢酶基因克隆、高效表达及酶学特性分析%Cloning, Expression, and Enzymatic Characteristics of Betaine Aldehyde De-hydrogenase Gene inSesuvium portulacastrum L.

    Institute of Scientific and Technical Information of China (English)

    喻时周; 杨成龙; 郭建春; 段瑞军

    2016-01-01

    在许多渗透调节剂中,甜菜碱是最理想的有机小分子渗透调节物质。甜菜碱在植物体内大量积累不会带来危害,同时能提高植物对环境胁迫的抗性。将海马齿中克隆到的甜菜碱醛脱氢酶基因构建到表达载体pET-28a(+)上,获得重组载体pET-SpBADH并将其成功地转化到BL21(DE3)中得到重组工程菌,经IPTG诱导能高效表达55 kD目的蛋白,表达量可以达到301µg mL–1。酶学特征分析表明,该蛋白最适pH值为7.2,在偏碱条件下能维持较高的催化活性; SpBADH蛋白对高温敏感,且温度对催化活性影响较大,超过55℃时酶活性只有20%,最适酶催化活性温度为37℃;而有机小分子醇类对酶的催化活性有保护作用,可以通过自身特征维持酶催化活性的微环境。%Among many osmotic materials, glycine betaine is a best organic micro-molecular, and functionally works for osmotic regulation in plants, which is non-toxic to plant growth. A lot of glycine betaine accumulatedin plant can enhance the resistance of plants to environmental stresses. In the study, a full-length sequence of betaine aldehyde dehydrogenase gene fromSesuvium portulacastrum was ligated with the vector pET-[28a](+), named pET-SpBADH, and successfully transformed into BL21(DE3) to obtain the corresponding recombinant engineering bacteria, which could highly express 55 kD protein induced by IPTG, with the expression level to 301 µg mL–1. The purified protein was obtained, showing the optimum pH value of 7.2, and maintain high catalytic activity the enzyme under slightly alkaline conditions. SpBADH protein very sensitive to high temperature effected the enzyme activity, with the optimum temperature to 37℃. The enzyme activity was only 20% when temperature was over 55℃. The small organic molecules of the reveral compounds of alcohol had a protective effect on the catalytic activity of the enzyme. The microenvironment of catalytic activity could be

  7. Amorphous-crystalline Ni-Fe powder mixture: Hydrogenation and annealing effects on microstructure and electrical and magnetic properties

    Directory of Open Access Journals (Sweden)

    Milinčić Radovan

    2016-01-01

    Full Text Available The hydrogenation of a crystalline Ni-Fe (80 wt.% Ni, 20 wt.% Fe powder mixture leads to the formation of a mixture of Face Centered Cubic (FCC-Ni and FCC-Fe phase nanocrystals embedded in an amorphous matrix. The magnetic susceptibility of the nanostructured powder is 2.1 times higher than that of the as-produced crystalline mixture. Heating in the temperature range 420-590 K causes structural relaxation in the hydrogenated powder, resulting in an increase of the magnetic susceptibility and a decrease of the electrical resistivity. During the heating procedure, the reorientation of magnetic domains in nickel and iron takes place in the temperature range 580-650 K and 790-850 K, respectively. In the pressed sample from the powder mixture, the crystallization of the amorphous phase of nickel and its FCC lattice crystalline grain growth occurs in the temperature range 620-873 K causing a decrease in the magnetic susceptibility of the nickel FCC phase and a sudden drop in the electrical resistivity. Prolonged heating of the mixed powders at 873K results in the formation of a Ni-Fe solid solution with higher magnetic susceptibility than the starting mixture. [Projekat Ministarstva nauke Republike Srbije, br. 172057

  8. A superlattice of alternately stacked Ni-Fe hydroxide nanosheets and graphene for efficient splitting of water.

    Science.gov (United States)

    Ma, Wei; Ma, Renzhi; Wang, Chengxiang; Liang, Jianbo; Liu, Xiaohe; Zhou, Kechao; Sasaki, Takayoshi

    2015-02-24

    Cost-effective electrocatalysts based on nonprecious metals for efficient water splitting are crucial for various technological applications represented by fuel cell. Here, 3d transition metal layered double hydroxides (LDHs) with varied contents of Ni and Fe were successfully synthesized through a homogeneous precipitation. The exfoliated Ni-Fe LDH nanosheets were heteroassembled with graphene oxide (GO) as well as reduced graphene oxide (rGO) into superlattice-like hybrids, in which two kinds of oppositely charged nanosheets are stacked face-to-face in alternating sequence. Heterostructured composites of Ni2/3Fe1/3 LDH nanosheets and GO (Ni2/3Fe1/3-GO) exhibited an excellent oxygen evolution reaction (OER) efficiency with a small overpotential of about 0.23 V and Tafel slope of 42 mV/decade. The activity was further improved via the combination of Ni2/3Fe1/3 LDH nanosheets with more conductive rGO (Ni2/3Fe1/3-rGO) to achieve an overpotential as low as 0.21 V and Tafel plot of 40 mV/decade. The catalytic activity was enhanced with an increased Fe content in the bimetallic Ni-Fe system. Moreover, the composite catalysts were found to be effective for hydrogen evolution reaction. An electrolyzer cell powered by a single AA battery of 1.5 V was demonstrated by using the bifunctional catalysts.

  9. EFFECT OF MnO2 ADDITIVE ON PERFORMANCES OF NiFe2O4 SPINEL BASED INERT ANODE

    Institute of Scientific and Technical Information of China (English)

    W.L. Jiao; L. Zhang; G. C. Yao

    2005-01-01

    The NiFe2O4 inert anode is synthesized by high-temperature solid-state reaction method using NiO and Fe2O3 as main raw materials and adding MnO2 powder as additive. Archimedes method using water immersion technique is used to measure the sintering performances of samples. The static thermal corrosion rates of samples are measured by weight loss. SEM is employed for the observation of material microstructure, and phase structure of the sample surface after corrosion is determined by XRD. The experimental results indicate that a suitable MnO2additive content is 2%, while the sintering performance is the best, and the static thermal corrosion rate is the lowest. Because of MnO2 dopant enriching at crystal boundary, the corrosion reaction of molten salt to crystal grain creates Mn2AlO4 phase, which is denser than NiFe2O4phase, and prevents the cryolite molten salt to penetrate into the inert anode, thus reducing the corrosion.

  10. Biochar supported Ni/Fe bimetallic nanoparticles to remove 1,1,1-trichloroethane under various reaction conditions.

    Science.gov (United States)

    Li, Hui; Qiu, Yue-Feng; Wang, Xiao-Li; Yang, Jie; Yu, Yun-Jiang; Chen, Ya-Qin; Liu, Yong-di

    2017-02-01

    In this study, Ni/Fe nanoparticles supported by biochar to stimulate the reduction of 1,1,1-trichloroethane (1,1,1-TCA) in groundwater remediation was investigated. In order to enhance the reactivity of ZVI (zero valent iron) nanoparticles, surface modification of ZVI was performed using nickel and biochar. The removal efficiency of 1,1,1-TCA increased from 42.3% to 99.3% as the biochar-to-Ni/Fe mass ratio increased from 0 to 1.0. However a higher biochar-to-Ni/Fe ratio showed little difference in the 1,1,1-TCA degradation efficiency. In the presence of Ni, atomic hydrogen generated by ZVI corrosion could be absorbed in the metal additive's lattice and then produce a hydride-like species (H) that represented the primary redox-active entity. The effects of various factors were evaluated, including pH, humic acid (HA) and inorganic matters (Cl(-), CO3(2-), HCO3(-), NO3(-) and SO4(2-)). The degradation of 1,1,1-TCA was greatly affected by pH. The presence of Cl(-), CO3(2-), HCO3(-) and SO4(2-) had negligible effects, but NO3(-) and HA showed a significant inhibitory effects on 1,1,1-TCA degradation. In conclusion, biochar supported Ni/Fe nanoparticles could be highly effective for 1,1,1-TCA degradation. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. High performance NiFe layered double hydroxide for methyl orange dye and Cr(VI) adsorption.

    Science.gov (United States)

    Lu, Yi; Jiang, Bin; Fang, Liang; Ling, Faling; Gao, Jiemei; Wu, Fang; Zhang, Xihua

    2016-06-01

    The NiFe layered double hydroxides (LDHs) with different mole ratio of Ni/Fe (4:1, 3:1, 7:3 and 1:1) were prepared by a simple coprecipitation method. The adsorption performance were evaluated by the removal of methyl orange (MO) dye and hexavalent chromium(VI) heavy metal ion. It is found that Ni4Fe1-LDH can remove more than 92% of MO in 10 min at the 10 mg/L MO initial concentration, and 97% of Cr(VI) in 1 h at 4 mg/L Cr2O7(2-) initial concentration. The saturated adsorption capacity of Ni4Fe1-LDH is found to be as large as 205.76 mg/g for MO and 26.78 mg/g for Cr(VI). The adsorption behavior of this new adsorbent is fitted well with Langmuir isotherm and the pseudo-second-order kinetic model, indicative of a monolayer and chemical adsorption that synergistically originates from exchangeable anions mechanism and layer charge density. Due to the excellent removal capacity of MO and Cr(VI), the NiFe-LDHs could be a promising adsorbent for wastewater treatment.

  12. Fabrication and evaluation of the thin NiFe supported solid oxide fuel cell by co-firing method

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Kyeong Hyun; Kim, Haekyoung [School of Materials Science and Engineering, Yeungnam University, Gyeongsan 712-749 (Korea, Republic of); Park, Young Min [Fuel Cell Project, Research Institute of Industrial Science and Technology, Pohang 790-330 (Korea, Republic of)

    2010-12-15

    Metal supported solid oxide fuel cells (SOFCs) are one of the most promising candidates for power plants as well as portable applications due to their good mechanical and thermal properties. In this study, the thin NiFe supported SOFC, which consisted of thin metal support (NiFe, {proportional_to}70 {mu}m), anode functional layer (Ni-YSZ, {proportional_to}30 {mu}m), electrolyte (YSZ, {proportional_to}15 {mu}m), and cathode (LSCF, {proportional_to}30 {mu}m), was fabricated through tape casting and co-firing method. The cell showed 1.05-1.1 V of open circuit voltage and 1.4 W cm{sup -2} of maximum power density at 800 C with the ohmic resistance (R{sub ohm}) of 0.12 {omega} cm{sup 2} and the polarization resistance (R{sub p}) of 0.38 {omega} cm{sup 2}. The high performance and the successful measurement of thin metal supported cell showed the possibility for mobile applications through the large area cell fabrications. (author)

  13. Comparison of N(2) Fixation and Yields in Cajanus cajan between Hydrogenase-Positive and Hydrogenase-Negative Rhizobia by In Situ Acetylene Reduction Assays and Direct N Partitioning.

    Science.gov (United States)

    La Favre, J S; Focht, D D

    1983-08-01

    Pigeon peas [Cajanus cajan (L.) Millsp.] were grown in soil columns containing (15)N-enriched organic matter. Seasonal N(2) fixation activity was determined by periodically assaying plants for reduction of C(2)H(2). N(2) fixation rose sharply from the first assay period at 51 days after planting to a peak of activity between floral initiation and fruit set. N(2) fixation (acetylene reduction) activity dropped concomitantly with pod maturation but recovered after pod harvests. Analysis of (15)N content of plant shoots revealed that approximately 91 to 94% of plant N was derived from N(2) fixation. The effect of inoculation with hydrogenase-positive and hydrogenase-negative rhizobia was examined. Pigeon peas inoculated with strain P132 (hydrogenase-positive) yielded significantly more total shoot N than other inoculated or uninoculated treatments. However, two other hydrogenase-positive strains did not yield significantly more total shoot N than a hydrogenase-negative strain. The extent of nodulation by inoculum strains compared to indigenous rhizobia was determined by typing nodules according to intrinsic antibiotic resistance of the inoculum strains. The inoculum strains were detected in almost all typed nodules of inoculated plants.Gas samples were taken from soil columns several times during the growth cycle of the plants. H(2) was never detected, even in columns containing pigeon peas inoculated with hydrogenase-negative rhizobia. This was attributed to H(2) consumption by soil bacteria. Estimation of N(2) fixation by acetylene reduction activity was closest to the direct (15)N method when ethylene concentrations in the gas headspace (between the column lid and soil surface) were extrapolated to include the soil pore space as opposed solely to measurement in the headspace. There was an 8-fold difference between the two acetylene reduction assay methods of estimation. Based on a planting density of 15,000 plants per hectare, the direct (15)N fixation rates ranged

  14. Hydrothermal synthesis and properties of NiFe2O4@BaTiO3 composites with well-matched interface

    Directory of Open Access Journals (Sweden)

    Jian-Ping Zhou, Li Lv, Qian Liu, Yu-Xiang Zhang and Peng Liu

    2012-01-01

    Full Text Available NiFe2O4@BaTiO3 multiferroic composite particles were produced by a simple hydrothermal method in two steps: preparing NiFe2O4 nanoparticles and then synthesizing core-shell nanocomposites. Multiferroic composite ceramics were sintered from these powders. X-ray diffraction, Raman scattering and energy dispersive x-ray analyses indicated that the core-shell composites with a NiFe2O4 core and BaTiO3 shell were formed in the hydrothermal environment. Different types of sharp interfaces were self-assembled owing to the minimization of direct elastic energy. The saturation magnetization of the composites linearly increased with the NiFe2O4 content while the dielectric constant decreased. A dielectric peak appeared at around 460 °C because of the oxygen vacancies in the BaTiO3 ceramics. It resulted in an enhancement of magnetic permeability in the composites, indicating magnetoelectric coupling that was also observed by direct magnetoelectric measurements.

  15. Science Letters:Dechlorination mechanism of 2,4-dichlorophenol by Ni/Fe nanoparticles in the presence of humic acid

    Institute of Scientific and Technical Information of China (English)

    Jing-jing WO; Zhen ZHANG; Xin-hua XU

    2009-01-01

    To understand the feasibility of its application to the in situ remediation of contaminated groundwater, the dechlori-nation of 2,4-dichlorophenol (2,4-DCP) by Ni/Fe nanoparticles in the presence of humic acid (HA) was investigated. We found that, as high performance liquid chromatography (HPLC) was used, the 2,4-DCP was first quickly reduced to o-chlorophenol (o-CP) andp-chlorophenol (p-CP), and then reduced to phenol as the final product. Our experimental results indicated that HA had an adverse effect on the dechlorination of 2,4-DCP by Ni/Fe nanoparticles, as the HA concentration increased, the removal rate decreased evidently. It also demonstrated that 2,4-DCP was reduced more easily to o-CP than to p-CP, and that the sequence of the tendency in dechlorination of intermediates was p-CP>o-CP. Transmission electron microscope (TEM) showed that HA could act as an adsorbate to compete reactive sites on the surface of Ni/Fe nanoparticles to decrease the dechlorination rate. Also we con-cluded that the dechlorination reaction of 2,4-DCP over Ni/Fe nanoparticles progressed through catalytic reductive dechlorination.

  16. Preparation of Magnetic Photocatalyst TiO2 Supported on NiFe2O4 and Effect of Magnetic Carrier on Photocatalytic Activity

    Institute of Scientific and Technical Information of China (English)

    XU Shi-Hong; FENG Dao-Lun; LI Deng-Xin; SHANGGUAN Wen-Feng

    2008-01-01

    A magnetic photocatalyst TiO2/NiFe2O4(TN)with typical ferromagnetic hysteresis was prepared by a sol-gel method,which is easy to be separated from a slurry-type photoreactor under the application of an external magnetic field,being one of promising photocatalysts for wastewater treatment.The analysis of XRD indicated that the highly dispersed NiFe2O4 nanoparticles prevented the formation of futile phase to some extent.A transmission elec.tron microscope(TEM)was used to characterize the structure of the photocatalyst,indicating that the NiFe2O4 nanoparticles highly dispersed among TiO2 nanoparticles.The prepared photocatalyst showed high photocatalytic activity for the degradation of methyl orange in water.The degradation results revealed that the NiFe2O4 nanoparticles played the role of recombination centre of photogenerated electrons and holes for the TN photocatalyst.which gave rise to the decrease in photocatalytic activity.Moreover,the experiment on recycled use of TN demonstrated a good repeatability of the photocatalytic activity.

  17. Facile hybridization of Ni@Fe2O3 superparticles with functionalized reduced graphene oxide and its application as anode material in lithium-ion batteries.

    Science.gov (United States)

    Backert, Gregor; Oschmann, Bernd; Tahir, Muhammad Nawaz; Mueller, Franziska; Lieberwirth, Ingo; Balke, Benjamin; Tremel, Wolfgang; Passerini, Stefano; Zentel, Rudolf

    2016-09-15

    In our present work we developed a novel graphene wrapping approach of Ni@Fe2O3 superparticles, which can be extended as a concept approach for other nanomaterials as well. It uses sulfonated reduced graphene oxide, but avoids thermal treatments and use of toxic agents like hydrazine for its reduction. The modification of graphene oxide is achieved by the introduction of sulfate groups accompanied with reduction and elimination reactions, due to the treatment with oleum. The successful wrapping of nanoparticles is proven by energy dispersive X-ray spectroscopy, high-resolution transmission electron microscopy and Raman spectroscopy. The developed composite material shows strongly improved performance as anode material in lithium-ion batteries (compared to unwrapped Ni@Fe2O3) as it offers a reversible capacity of 1051mAhg(-1) after 40 cycles at C/20, compared with 460mAhg(-1) for unwrapped Ni@Fe2O3. The C rate capability is also improved by the wrapping approach, as specific capacities for wrapped particles are about twice of those offered by unwrapped particles. Additionally, the benefit for the use of the advanced superparticle morphology is demonstrated by comparing wrapped Ni@Fe2O3 particles with wrapped Fe2O3 nanorice.

  18. Chapter Eight - Structural Characterization of Poised States in the Oxygen Sensitive Hydrogenases and Nitrogenases

    Energy Technology Data Exchange (ETDEWEB)

    King, Paul W [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Mulder, David W [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Artz, Jacob H. [Washington State University; Zadvornyy, Oleg A. [Washington State University; Peters, John W. [Washington State University

    2017-08-21

    The crystallization of FeS cluster-containing proteins has been challenging due to their oxygen sensitivity, and yet these enzymes are involved in many critical catalytic reactions. The last few years have seen a wealth of innovative experiments designed to elucidate not just structural but mechanistic insights into FeS cluster enzymes. Here, we focus on the crystallization of hydrogenases, which catalyze the reversible reduction of protons to hydrogen, and nitrogenases, which reduce dinitrogen to ammonia. A specific focus is given to the different experimental parameters and strategies that are used to trap distinct enzyme states, specifically, oxidants, reductants, and gas-treatments. Other themes presented here include the recent use of Cryo-EM, and how coupling various spectroscopies to crystallization is opening up new approaches for structural and mechanistic analysis.

  19. Following [FeFe] Hydrogenase Active Site Intermediates by Time-Resolved Mid-IR Spectroscopy.

    Science.gov (United States)

    Mirmohades, Mohammad; Adamska-Venkatesh, Agnieszka; Sommer, Constanze; Reijerse, Edward; Lomoth, Reiner; Lubitz, Wolfgang; Hammarström, Leif

    2016-08-18

    Time-resolved nanosecond mid-infrared spectroscopy is for the first time employed to study the [FeFe] hydrogenase from Chlamydomonas reinhardtii and to investigate relevant intermediates of the enzyme active site. An actinic 355 nm, 10 ns laser flash triggered photodissociation of a carbonyl group from the CO-inhibited state Hox-CO to form the state Hox, which is an intermediate of the catalytic proton reduction cycle. Time-resolved infrared spectroscopy allowed us to directly follow the subsequent rebinding of the carbonyl, re-forming Hox-CO, and determine the reaction half-life to be t1/2 ≈ 13 ± 5 ms at room temperature. This gives direct information on the dynamics of CO inhibition of the enzyme.

  20. Improved production of biohydrogen in light-powered Escherichia coli by co-expression of proteorhodopsin and heterologous hydrogenase.

    Science.gov (United States)

    Kim, Jaoon Y H; Jo, Byung Hoon; Jo, Younghwa; Cha, Hyung Joon

    2012-01-04

    Solar energy is the ultimate energy source on the Earth. The conversion of solar energy into fuels and energy sources can be an ideal solution to address energy problems. The recent discovery of proteorhodopsin in uncultured marine γ-proteobacteria has made it possible to construct recombinant Escherichia coli with the function of light-driven proton pumps. Protons that translocate across membranes by proteorhodopsin generate a proton motive force for ATP synthesis by ATPase. Excess protons can also be substrates for hydrogen (H(2)) production by hydrogenase in the periplasmic space. In the present work, we investigated the effect of the co-expression of proteorhodopsin and hydrogenase on H(2) production yield under light conditions. Recombinant E. coli BL21(DE3) co-expressing proteorhodopsin and [NiFe]-hydrogenase from Hydrogenovibrio marinus produced ~1.3-fold more H(2) in the presence of exogenous retinal than in the absence of retinal under light conditions (70 μmole photon/(m2·s)). We also observed the synergistic effect of proteorhodopsin with endogenous retinal on H(2) production (~1.3-fold more) with a dual plasmid system compared to the strain with a single plasmid for the sole expression of hydrogenase. The increase of light intensity from 70 to 130 μmole photon/(m(2)·s) led to an increase (~1.8-fold) in H(2) production from 287.3 to 525.7 mL H(2)/L-culture in the culture of recombinant E. coli co-expressing hydrogenase and proteorhodopsin in conjunction with endogenous retinal. The conversion efficiency of light energy to H(2) achieved in this study was ~3.4%. Here, we report for the first time the potential application of proteorhodopsin for the production of biohydrogen, a promising alternative fuel. We showed that H(2) production was enhanced by the co-expression of proteorhodopsin and [NiFe]-hydrogenase in recombinant E. coli BL21(DE3) in a light intensity-dependent manner. These results demonstrate that E. coli can be applied as light

  1. Improved production of biohydrogen in light-powered Escherichia coli by co-expression of proteorhodopsin and heterologous hydrogenase

    Directory of Open Access Journals (Sweden)

    Kim Jaoon YH

    2012-01-01

    Full Text Available Abstract Background Solar energy is the ultimate energy source on the Earth. The conversion of solar energy into fuels and energy sources can be an ideal solution to address energy problems. The recent discovery of proteorhodopsin in uncultured marine γ-proteobacteria has made it possible to construct recombinant Escherichia coli with the function of light-driven proton pumps. Protons that translocate across membranes by proteorhodopsin generate a proton motive force for ATP synthesis by ATPase. Excess protons can also be substrates for hydrogen (H2 production by hydrogenase in the periplasmic space. In the present work, we investigated the effect of the co-expression of proteorhodopsin and hydrogenase on H2 production yield under light conditions. Results Recombinant E. coli BL21(DE3 co-expressing proteorhodopsin and [NiFe]-hydrogenase from Hydrogenovibrio marinus produced ~1.3-fold more H2 in the presence of exogenous retinal than in the absence of retinal under light conditions (70 μmole photon/(m2·s. We also observed the synergistic effect of proteorhodopsin with endogenous retinal on H2 production (~1.3-fold more with a dual plasmid system compared to the strain with a single plasmid for the sole expression of hydrogenase. The increase of light intensity from 70 to 130 μmole photon/(m2·s led to an increase (~1.8-fold in H2 production from 287.3 to 525.7 mL H2/L-culture in the culture of recombinant E. coli co-expressing hydrogenase and proteorhodopsin in conjunction with endogenous retinal. The conversion efficiency of light energy to H2 achieved in this study was ~3.4%. Conclusion Here, we report for the first time the potential application of proteorhodopsin for the production of biohydrogen, a promising alternative fuel. We showed that H2 production was enhanced by the co-expression of proteorhodopsin and [NiFe]-hydrogenase in recombinant E. coli BL21(DE3 in a light intensity-dependent manner. These results demonstrate that E. coli

  2. Isolation and characterization of the small subunit of the uptake hydrogenase from the cyanobacterium Nostoc punctiforme.

    Science.gov (United States)

    Raleiras, Patrícia; Kellers, Petra; Lindblad, Peter; Styring, Stenbjörn; Magnuson, Ann

    2013-06-21

    In nitrogen-fixing cyanobacteria, hydrogen evolution is associated with hydrogenases and nitrogenase, making these enzymes interesting targets for genetic engineering aimed at increased hydrogen production. Nostoc punctiforme ATCC 29133 is a filamentous cyanobacterium that expresses the uptake hydrogenase HupSL in heterocysts under nitrogen-fixing conditions. Little is known about the structural and biophysical properties of HupSL. The small subunit, HupS, has been postulated to contain three iron-sulfur clusters, but the details regarding their nature have been unclear due to unusual cluster binding motifs in the amino acid sequence. We now report the cloning and heterologous expression of Nostoc punctiforme HupS as a fusion protein, f-HupS. We have characterized the anaerobically purified protein by UV-visible and EPR spectroscopies. Our results show that f-HupS contains three iron-sulfur clusters. UV-visible absorption of f-HupS has bands ∼340 and 420 nm, typical for iron-sulfur clusters. The EPR spectrum of the oxidized f-HupS shows a narrow g = 2.023 resonance, characteristic of a low-spin (S = ½) [3Fe-4S] cluster. The reduced f-HupS presents complex EPR spectra with overlapping resonances centered on g = 1.94, g = 1.91, and g = 1.88, typical of low-spin (S = ½) [4Fe-4S] clusters. Analysis of the spectroscopic data allowed us to distinguish between two species attributable to two distinct [4Fe-4S] clusters, in addition to the [3Fe-4S] cluster. This indicates that f-HupS binds [4Fe-4S] clusters despite the presence of unusual coordinating amino acids. Furthermore, our expression and purification of what seems to be an intact HupS protein allows future studies on the significance of ligand nature on redox properties of the iron-sulfur clusters of HupS.

  3. Interplay between out-of-plane anisotropic L1{sub 1}-type CoPt and in-plane anisotropic NiFe layers in CoPt/NiFe exchange springs

    Energy Technology Data Exchange (ETDEWEB)

    Saravanan, P. [Department of Physics, National Taiwan University, Taipei 106, Taiwan (China); Defence Metallurgical Research Laboratory, Hyderabad 500058 (India); Hsu, Jen-Hwa, E-mail: jhhsu@phys.ntu.edu.tw; Tsai, C. L. [Department of Physics, National Taiwan University, Taipei 106, Taiwan (China); Tsai, C. Y.; Lin, Y. H. [Graduate Institute of Materials Science and Engineering, National Taiwan University, Taipei 106, Taiwan (China); Kuo, C. Y.; Wu, J.-C. [Department of Physics, National Chang Hua University of Education, Chang Hua 50000, Taiwan (China); Lee, C.-M. [Graduate School of Materials Science, National Yunlin University of Science and Technology, Douliou 64002, Taiwan (China)

    2014-06-28

    Films of L1{sub 1}-type CoPt/NiFe exchange springs were grown with different NiFe (Permalloy) layer thickness (t{sub NiFe} = 0–10 nm). X-ray diffraction analysis reveals that the characteristic peak position of NiFe(111) is not affected by the CoPt-layer—confirming the absence of any inter-diffusion between the CoPt and NiFe layers. Magnetic studies indicate that the magnetization orientation of NiFe layer can be tuned through varying t{sub NiFe} and the perpendicular magnetic anisotropy of L1{sub 1}-type CoPt/NiFe films cannot sustain for t{sub NiFe} larger than 3.0 nm due to the existence of exchange interaction at the interface of L1{sub 1}-CoPt and NiFe layers. Magnetic force microscopy analysis on the as-grown samples shows the changes in morphology from maze-like domains with good contrast to hazy domains when t{sub NiFe} ≥ 3.0 nm. The three-dimensional micro-magnetic simulation results demonstrate that the magnetization orientation in NiFe layer is not uniform, which continuously increases from the interface to the top of NiFe layer. Furthermore, the tilt angle of the topmost NiFe layers can be changed over a very wide range from a small number to about 75° by varying t{sub NiFe} from 1 to 10 nm. It is worth noting that there is an abrupt change in the magnetization direction at the interface, for all the t{sub NiFe} investigated. The results of present study demonstrate that the tunable tilted exchange springs can be realized with L1{sub 1}-type CoPt/NiFe bilayers for future applications in three-axis magnetic sensors or advanced spintronic devices demanding inclined magnetic anisotropy.

  4. Structural, magnetic, dielectric, and electrical properties of NiFe2O4 spinel ferrite nanoparticles prepared by honey-mediated sol-gel combustion

    Science.gov (United States)

    Yadav, Raghvendra Singh; Kuřitka, Ivo; Vilcakova, Jarmila; Havlica, Jaromir; Masilko, Jiri; Kalina, Lukas; Tkacz, Jakub; Enev, Vojtěch; Hajdúchová, Miroslava

    2017-08-01

    In this study, NiFe2O4 nanoparticles were synthesized using a honey-mediated sol-gel combustion method. The synthesized nanoparticles and samples annealed at 800 °C and 1100 °C were characterized by X-ray diffraction (XRD), field emission-scanning electron microscopy (FE-SEM), Raman spectroscopy, X-ray photoelectron spectroscopy (XPS), and vibrating sample magnetometry (VSM). XRD and Raman spectroscopy confirmed the formation of a cubic spinel ferrite structure. FE-SEM demonstrated the octahedral morphology of the NiFe2O4 spinel ferrite nanoparticles with sizes ranging from 10 to 70 nm. Quantitative analysis based on XPS suggested a mixed spinel structure comprising NiFe2O4 nanoparticles. XPS analysis determined occupation formulae of (Ni0.212+ Fe0.443+)[Ni0.792+ Fe1.563+]O4 and (Ni0.232+ Fe0.503+)[Ni0.772+ Fe1.503+]O4, for the as-prepared NiFe2O4 nanoparticles and those annealed at 1100 °C, respectively. Magnetic measurements showed that the saturation magnetization increased with the crystallite size from 32.3 emu/g (20 nm) to 49.9 emu/g (163 nm), whereas the coercivity decreased with the crystallite size from 162 Oe (20 nm) to 47 Oe (163 nm). Furthermore, the dielectric constant, dielectric loss tangent, and AC conductivity of the NiFe2O4 nanoparticles were dependent on the frequency (1-107 Hz) and grain size. The influence of the grain size was also observed by modulus spectroscopy based on the Cole-Cole plot.

  5. Formate dehydrogenases and hydrogenases in syntrophic propionate-oxidizing communities : gene analysis and transcritional profiling

    NARCIS (Netherlands)

    Worm, P.

    2010-01-01

    Many places on earth are without oxygen (anaerobic) such as rice paddy fields, swamps and sediments of freshwater lakes and oceans. When oxygen, nitrate or other electron acceptors are not present, organic material is degraded to carbon dioxide and methane by mixed microbial species that each hav

  6. Symbiotic Legume Nodules Employ Both Rhizobial Exo- and Endo-Hydrogenases to Recycle Hydrogen Produced by Nitrogen Fixation

    OpenAIRE

    Ciccolella, Christopher O.; Raynard, Nathan A.; John H-M Mei; Derek C Church; Ludwig, Robert A.

    2010-01-01

    BACKGROUND: In symbiotic legume nodules, endosymbiotic rhizobia (bacteroids) fix atmospheric N(2), an ATP-dependent catalytic process yielding stoichiometric ammonium and hydrogen gas (H(2)). While in most legume nodules this H(2) is quantitatively evolved, which loss drains metabolic energy, certain bacteroid strains employ uptake hydrogenase activity and thus evolve little or no H(2). Rather, endogenous H(2) is efficiently respired at the expense of O(2), driving oxidative phosphorylation, ...

  7. International Hydrogenase Conference (7th) Held at the University of Reading on August 24th to 29th 2004.

    Science.gov (United States)

    2004-08-19

    Homologous/heterologous over-expression in Clostridium p68 acetobutylicum and characterization of purified clostridial and algal Fe-only hydrogenases with...yield from fermentative anaerobes can vary from 1.61-2.36 mol H2/mol glucose in Clostridium species to 3.3 mol H,/mol glucose from the extreme...and purification system functional in Clostridium acetobutylicum . C. acetobutylicum ATCC 824 was selected for the homologous over-expression of its Fe

  8. Identification of a Catalytic Iron-Hydride at the H-Cluster of [FeFe]-Hydrogenase

    Energy Technology Data Exchange (ETDEWEB)

    Mulder, David W.; Guo, Yisong; Ratzloff, Michael W.; King, Paul W.

    2017-01-11

    Hydrogenases couple electrochemical potential to the reversible chemical transformation of H2 and protons, yet the reaction mechanism and composition of intermediates are not fully understood. In this Communication we describe the biophysical properties of a hydride-bound state (Hhyd) of the [FeFe]-hydrogenase from Chlamydomonas reinhardtii. The catalytic H-cluster of [FeFe]-hydrogenase consists of a [4Fe-4S] subcluster ([4Fe-4S]H) linked by a cysteine thiol to an azadithiolate-bridged 2Fe subcluster ([2Fe]H) with CO and CN- ligands. Mossbauer analysis and density functional theory (DFT) calculations show that Hhyd consists of a reduced [4Fe-4S]H+ coupled to a diferrous [2Fe]H with a terminally bound Fe-hydride. The existence of the Fe-hydride in Hhyd was demonstrated by an unusually low Mossbauer isomer shift of the distal Fe of the [2Fe]H subcluster. A DFT model of Hhyd shows that the Fe-hydride is part of a H-bonding network with the nearby bridging azadithiolate to facilitate fast proton exchange and catalytic turnover.

  9. Using Gas Chromatography/Isotope Ratio Mass Spectrometry to Determine the Fractionation Factor for H2 Production by Hydrogenases

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Hui; Ghandi, H.; Shi, Liang; Kreuzer, Helen W.; Ostrom, Nathaniel; Hegg, Eric L.

    2012-01-15

    Hydrogenases catalyze the reversible formation of H2, and they are key enzymes in the biological cycling of H2. H isotopes should be a very useful tool in quantifying proton trafficking in biological H2 production processes, but there are several obstacles that have thus far limited the use of this tool. In this manuscript, we describe a new method that overcomes some of these barriers and is specifically designed to measure isotopic fractionation during enzyme-catalyzed H2 evolution. A key feature of this technique is that purified hydrogenases are employed, allowing precise control over the reaction conditions and therefore a high level of precision. A custom-designed high-throughput gas chromatography-isotope ratio mass spectrometer is employed to measure the isotope ratio of the H2. Using this method, we determined that the fractionation factor of H2 production by the [NiFe]-hydrogenase from Desulfivibrio fructosovran is 0.27. This result indicates that, as expected, protons are highly favored over deuterons during H2 evolution. Potential applications of this new method are discussed.

  10. Electrokinetic properties of PMAA functionalized NiFe2O4 nanoparticles synthesized by thermal plasma route

    Science.gov (United States)

    Bhosale, Shivaji V.; Mhaske, Pravin; Kanhe, N.; Navale, A. B.; Bhoraskar, S. V.; Mathe, V. L.; Bhatt, S. K.

    2014-04-01

    The magnetic nickel ferrite (NiFe2O4) nanoparticles with an average size of 30nm were synthesised by Transferred arc DC Thermal Plasma route. The synthesized nickel ferrite nanoparticles were characterized by TEM and FTIR techniques. The synthesized nickel ferrite nanoparticles were further functionalized with PMAA (polymethacrylic acid) by self emulsion polymerization method and subsequently were characterized by FTIR and Zeta Analyzer. The variation of zeta potential with pH was systematically studied for both PMAA functionalized (PNFO) and uncoated nickel ferrite nanoparticles (NFO). The IEP (isoelectric points) for PNFO and NFO was determined from the graph of zeta potential vs pH. It was observed that the IEP for NFO was at 7.20 and for PNFO it was 2.52. The decrease in IEP of PNFO was attributed to the COOH functional group of PMAA.

  11. Excitation and tailoring of diffractive spin-wave beams in NiFe using nonuniform microwave antennas

    Science.gov (United States)

    Körner, H. S.; Stigloher, J.; Back, C. H.

    2017-09-01

    We experimentally demonstrate by time-resolved scanning magneto-optical Kerr microscopy the possibility to locally excite multiple spin-wave beams in the dipolar-dominated regime in metallic NiFe films. For this purpose we employ differently shaped nonuniform microwave antennas consisting of several coplanar waveguide sections different in size, thereby adapting an approach for the generation of spin-wave beams in the exchange-dominated regime suggested by Gruszecki et al. [Sci. Rep. 6, 22367 (2016), 10.1038/srep22367]. The occurring spin-wave beams are diffractive and we show that the width of the beam and its widening as it propagates can be tailored by the shape and the length of the nonuniformity. Moreover, the propagation direction of the diffractive beams can be manipulated by changing the bias field direction.

  12. The characteristic of deformability and quantitative description of the microstructure of hot-deformed Ni-Fe superalloy

    Directory of Open Access Journals (Sweden)

    Ducki K. J.

    2017-03-01

    Full Text Available The paper presents the results of research concerning the influence of hot plastic working parameters on the deformability and microstructure of a Ni-Fe superalloy. The research was performed on a torsion plastometer in the range of temperatures of 900-1150°C, at a strain rates 0.1 and 1.0 s-1. Plastic properties of the alloy were characterized by the worked out flow curves and the temperature relationships of flow stress and strain limit. The structural inspections were performed on microsections taken from plastometric samples after so-called “freezing”. The stereological parameters as the recrystallized grain size, inhomogenity and grain shape have been determined. Functional relations between the Zener-Hollomon parameter and the maximum yield stress and the average grain area have been developed and the activation energy for hot working has been estimated.

  13. Effect of cobalt substitution on structural, electrical and magnetic properties of NiFe2O4

    Directory of Open Access Journals (Sweden)

    Kannipamula Vijaya Babu

    2017-03-01

    Full Text Available Nickel ferrite and cobalt substituted nickel ferrite were synthesized by sol-gel method using citric acid as chelating reagent. X-ray diffraction revealed the formation of nanocrystalline particles having spinel structure with space group Fd3m. FTIR spectra showed fundamental absorption bands in the range 400–1300 cm-1, related to iron ions on both octahedral and tetrahedral sites which are typical for ferrite structure. SEM analyses confirmed that the particles are agglomerated with an average size of about 1.5 µm. The ferrite powders were pressed at 5 MPa and sintered at 1200 °C for 5 h. Conductivities, impedance analyses and dielectric properties of NiFe2O4 and Ni0.7Co0.3Fe2O4 ceramics were investigated over the frequency range of 20 Hz to 1 MHz at room temperature. The obtained experimental results are in good agreement with the reported values.

  14. Plasticity of nanocrystalline alloys with chemical order: on the strength and ductility of nanocrystalline Ni-Fe.

    Science.gov (United States)

    Schäfer, Jonathan; Albe, Karsten

    2013-01-01

    Plastic deformation and alloying of nanocrystalline Ni-Fe is studied by means of atomic scale computer simulations. By using a combination of Monte-Carlo and molecular dynamics methods we find that solutes have an ordering tendency even if grain sizes are in the nanometer regime, where the phase field of the ordered state is widened as compared to larger grain sizes. Tensile testing of disordered structures with various elemental distributions and the simultaneous analysis of intragranular defects reveal that solid solution strengthening is absent for the studied grain sizes. The composition and relaxation state of the grain boundary control the strength of the material, which is also found for ordered structures (L12), where dislocation activity is suppressed.

  15. Hydrothermal synthesis of NiFe$_2$O$_4$ nano-particles: structural, morphological, optical, electrical and magnetic properties

    Indian Academy of Sciences (India)

    K CHANDRA BABU NAIDU; W MADHURI

    2017-04-01

    NiFe$_2$O$_4$ nano-crystallites with an average diameter of 8.9 nm are synthesized via hydrothermal method.The single-phase spinel structure is confirmed from X-ray diffractograms. Morphology is analysed by transmissionand field emission scanning electron microscopes. High specific surface area of 55.7 m$^2$ g$^{−1}$ is obtained for nano-particles. The M–H loop and M–T curve behaviours are investigated by vibrating sample magnetometry.The optical band gap energy is estimated from the UV–visible spectrum. In addition, the frequency dependence of dielectric properties is investigated. Cole–Cole plots are drawn to study electrical conduction mechanism and thekind of relaxation—Debye or non-Debye type. Low a.c. conductivity and low magnetic losses are noticed at 5 MHz frequency, which are suitable for microwave device applications.

  16. Shear-Coupled Grain Growth and Texture Development in a Nanocrystalline Ni-Fe Alloy during Cold Rolling

    Science.gov (United States)

    Li, Li; Ungár, Tamás; Toth, Laszlo S.; Skrotzki, Werner; Wang, Yan Dong; Ren, Yang; Choo, Hahn; Fogarassy, Zsolt; Zhou, X. T.; Liaw, Peter K.

    2016-12-01

    The evolution of texture, grain size, grain shape, dislocation, and twin density has been determined by synchrotron X-ray diffraction and line profile analysis in a nanocrystalline Ni-Fe alloy after cold rolling along different directions related to the initial fiber and the long axis of grains. The texture evolution has been simulated by the Taylor-type relaxed-constraints viscoplastic polycrystal model. The simulations were based on the activity of partial dislocations in correlation with the experimental results of dislocation density determination. The concept of stress-induced shear coupling is supported and strengthened by both the texture simulations and the experimentally determined evolution of the microstructure parameters. Grain growth and texture evolution are shown to proceed by the shear coupling mechanism supported by dislocation activity as long as the grain size is not smaller than about 20 nm.

  17. Microfabrication of magnetostrictive beams based on NiFe film doped with B and Mo for integrated sensor systems

    KAUST Repository

    Alfadhel, Ahmed

    2012-03-09

    This paper reports the development of integrated micro-sensors consisting of 1 -µm-thick magnetostrictive cantilevers or bridges with 500 µm in length and conducting interrogation elements. The thin films are fabricated by sputter deposition of NiFe doped with B and Mo, and the magnetic properties are enhanced by field annealing, resulting in a coercivity of 2.4 Oe. In operation, an alternating current applied to the interrogation elements magnetizes the magnetostrictive structures. The longitudinal resonant frequency is detected as an impedance change of the interrogation elements. The magnetostrictive micro-beams provide high resonant frequencies—2.95 MHz for the cantilever and 5.46 MHz for the bridge—which can be exploited to develop sensors of high sensitivity.

  18. Hydrogen-producing microflora and Fe-Fe hydrogenase diversities in seaweed bed associated with marine hot springs of Kalianda, Indonesia.

    Science.gov (United States)

    Xu, Shou-Ying; He, Pei-Qing; Dewi, Seswita-Zilda; Zhang, Xue-Lei; Ekowati, Chasanah; Liu, Tong-Jun; Huang, Xiao-Hang

    2013-05-01

    Microbial fermentation is a promising technology for hydrogen (H(2)) production. H(2) producers in marine geothermal environments are thermophilic and halotolerant. However, no one has surveyed an environment specifically for thermophilic bacteria that produce H(2) through Fe-Fe hydrogenases (H(2)ase). Using heterotrophic medium, several microflora from a seaweed bed associated with marine hot springs were enriched and analyzed for H(2) production. A H(2)-producing microflora was obtained from Sargassum sp., 16S rRNA genes and Fe-Fe H(2)ase diversities of this enrichment were also analyzed. Based on 16S rRNA genes analysis, 10 phylotypes were found in the H(2)-producing microflora showing 90.0-99.5 % identities to known species, and belonged to Clostridia, Gammaproteobacteria, and Bacillales. Clostridia were the most abundant group, and three Clostridia phylotypes were most related to known H(2) producers such as Anaerovorax odorimutans (94.0 % identity), Clostridium papyrosolvens (98.4 % identity), and Clostridium tepidiprofundi (93.1 % identity). For Fe-Fe H(2)ases, seven phylotypes were obtained, showing 63-97 % identities to known Fe-Fe H(2)ases, and fell into four distinct clusters. Phylotypes HW55-3 and HM55-1 belonged to thermophilic and salt-tolerant H(2)-producing Clostridia, Halothermothrix orenii-like Fe-Fe H(2)ases (80 % identity), and cellulolytic H(2)-producing Clostridia, C. papyrosolvens-like Fe-Fe H(2)ases (97 % identity), respectively. The results of both 16S rRNA genes and Fe-Fe H(2)ases surveys suggested that the thermophilic and halotolerant H(2)-producing microflora in seaweed bed of hot spring area represented previously unknown H(2) producers, and have potential application for H(2) production.

  19. Molecular Dynamics Study of the Proposed Proton Transport Pathways in [FeFe]-Hydrogenase

    Energy Technology Data Exchange (ETDEWEB)

    Ginovska-Pangovska, Bojana; Ho, Ming-Hsun; Linehan, John C.; Cheng, Yuhui; Dupuis, Michel; Raugei, Simone; Shaw, Wendy J.

    2014-01-15

    Possible proton channels in Clostridium pasteurianum [FeFe]-hydrogenase were investigated with molecular dynamics simulations. This study was undertaken to discern proposed channels, compare their properties, evaluate the functional channel, and to provide insight into the features of an active proton channel. Our simulations suggest that protons are not transported through water wires. Instead, a five-residue motif (E282, S319, E279, HOH, C299) was found to be the likely channel, consistent with experimental observations. This channel connects the surface of the enzyme and the di-thiomethylamine bridge of the catalytic H-cluster, permitting the transport of protons. The channel was found to have a persistent hydrogen bonded core (residues E279 to S319), with less persistent hydrogen bonds at the ends of the channel. The hydrogen bond occupancy in this network was found to be sensitive to the protonation state of the residues in the channel, with different protonation states enhancing or stabilizing hydrogen bonding in different regions of the network. Single site mutations to non-hydrogen bonding residues break the hydrogen bonding network at that residue, consistent with experimental observations showing catalyst inactivation. In many cases, these mutations alter the hydrogen bonding in other regions of the channel which may be equally important in catalytic failure. A correlation between the protein dynamics near the proton channel and the redox partner binding regions was also found as a function of protonation state. The likely mechanism of proton movement in [FeFe]-hydrogenases is discussed based on the structural analysis presented here. This work was funded by the DOE Office of Science Early Career Research Program through the Office of Basic Energy Sciences. Computational resources were provided at W. R. Wiley Environmental Molecular Science Laboratory (EMSL), a national scientific user facility sponsored by the Department of Energy’s Office of

  20. Microoxic Niches within the Thylakoid Stroma of Air-Grown Chlamydomonas reinhardtii Protect [FeFe]-Hydrogenase and Support Hydrogen Production under Fully Aerobic Environment1[OPEN

    Science.gov (United States)

    Liran, Oded; Milrad, Yuval; Eilenberg, Haviva; Weiner, Iddo

    2016-01-01

    Photosynthetic hydrogen production in the microalga Chlamydomonas reinhardtii is catalyzed by two [FeFe]-hydrogenase isoforms, HydA1 and HydA2, both irreversibly inactivated upon a few seconds exposure to atmospheric oxygen. Until recently, it was thought that hydrogenase is not active in air-grown microalgal cells. In contrast, we show that the entire pool of cellular [FeFe]-hydrogenase remains active in air-grown cells due to efficient scavenging of oxygen. Using membrane inlet mass spectrometry, 18O2 isotope, and various inhibitors, we were able to dissect the various oxygen uptake mechanisms. We found that both chlororespiration, catalyzed by plastid terminal oxidase, and Mehler reactions, catalyzed by photosystem I and Flavodiiron proteins, significantly contribute to oxygen uptake rate. This rate is considerably enhanced with increasing light, thus forming local anaerobic niches at the proximity of the stromal face of the thylakoid membrane. Furthermore, we found that in transition to high light, the hydrogen production rate is significantly enhanced for a short duration (100 s), thus indicating that [FeFe]-hydrogenase functions as an immediate sink for surplus electrons in aerobic as well as in anaerobic environments. In summary, we show that an anaerobic locality in the chloroplast preserves [FeFe]-hydrogenase activity and supports continuous hydrogen production in air-grown microalgal cells. PMID:27443604

  1. GenBank blastx search result: AK060481 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK060481 001-012-G11 AF319635.1 Thermococcus litoralis putative dihidrodipicolinate synthase (dapA), Ni-Fe... hydrogenase maturation protein HypC (hypC), Ni-Fe hydrogenase maturation protein Hyp

  2. GenBank blastx search result: AK060557 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK060557 001-022-D11 AF319635.1 Thermococcus litoralis putative dihidrodipicolinate synthase (dapA), Ni-Fe... hydrogenase maturation protein HypC (hypC), Ni-Fe hydrogenase maturation protein Hyp

  3. GenBank blastx search result: AK119212 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK119212 001-041-E06 AF319635.1 Thermococcus litoralis putative dihidrodipicolinate synthase (dapA), Ni-Fe... hydrogenase maturation protein HypC (hypC), Ni-Fe hydrogenase maturation protein Hyp

  4. GenBank blastx search result: AK059062 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK059062 001-021-G03 AF319635.1 Thermococcus litoralis putative dihidrodipicolinate synthase (dapA), Ni-Fe... hydrogenase maturation protein HypC (hypC), Ni-Fe hydrogenase maturation protein Hyp

  5. GenBank blastx search result: AK060835 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK060835 001-034-C09 AF319635.1 Thermococcus litoralis putative dihidrodipicolinate synthase (dapA), Ni-Fe... hydrogenase maturation protein HypC (hypC), Ni-Fe hydrogenase maturation protein Hyp

  6. GenBank blastx search result: AK060537 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK060537 001-021-D11 AF319635.1 Thermococcus litoralis putative dihidrodipicolinate synthase (dapA), Ni-Fe... hydrogenase maturation protein HypC (hypC), Ni-Fe hydrogenase maturation protein Hyp

  7. Effect of adding methods of metallic phase on microstructure and thermal shock resistance of Ni/(90NiFe2O4-10NiO) cermets

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Ball mixing and electroless plating were respectively used as the adding methods of metallic phase to prepare Ni/(90NiFe2O4-10NiO) cermets for the inert anode in aluminum electrolysis. The microstructure and thermal shock resistance of cermet samples were studied. The results show that, for the samples prepared by ball mixing method, aggregation of metallic phase is found in either the green blocks or sintered samples and the extent of aggregation increases with the increase of metal content. For 6.5Ni/(90NiFe2O4-10NiO) cermets prepared with electroless plating method, the homogeneous and fine metallic particles are found in either the green compacts or sintered samples, but the relative density and thermal shock residual strength decrease by 3% and 28%-58% respectively, compared with samples prepared with ball mixing method.

  8. Structural, dielectric and magnetic properties of NiFe2O4 prepared via sol-gel auto-combustion method

    Science.gov (United States)

    Sun, Li; Zhang, Ru; Wang, Zhenduo; Ju, Lin; Cao, Ensi; Zhang, Yongjia

    2017-01-01

    Nickelferrite (NiFe2O4)powders were synthesized via sol-gel auto-combustion method and the corresponding temperature dependence of microstructure, dielectric and magnetic properties have been investigated. Results of XRD and SEM indicate that the NiFe2O4 samples exhibit a typical single phase spinel structure and a uniform particle distribution. The dielectric constant and dielectric loss measurements show strong frequency dependence of all the samples. The peak observed in frequency dependence of dielectric loss measurements shifts to higher frequency with the increasing sintering temperature, indicating a Debye-like dielectric relaxation. The remanent magnetization increases with the increasing grain size while the coercivity is just the opposite. The saturation magnetization can achieve 50 emu/g when the sintering temperature is more than 1000 °C, and the lowest coercivity (159.49 Oe) was observed in the NFO sample sintered at 1300 °C for 2 h.

  9. Raman Spectroscopy of Charge Transfer Interactions Between Single Wall Carbon Nanotubes and [FeFe] Hydrogenase

    Energy Technology Data Exchange (ETDEWEB)

    Blackburn, J. L. Svedruzic, D.; McDonald, T. J.; Kim, Y. H.; King, P. W.; Heben, M. J.

    2008-01-01

    We report a Raman spectroscopy study of charge transfer interactions in complexes formed by single-walled carbon nanotubes (SWNTs) and [FeFe] hydrogenase I (CaHydI) from Clostridium acetobutylicum. The choice of Raman excitation wavelength and sample preparation conditions allows differences to be observed for complexes involving metallic (m) and semiconducting (s) species. Adsorbed CaHydI can reversibly inject electronic charge into the LUMOs of s-SWNTs, while charge can be injected and removed from m-SWNTs at lower potentials just above the Fermi energy. Time-dependent enzymatic assays demonstrated that the reduced and oxidized forms of CaHydI are deactivated by oxygen, but at rates that varied by an order of magnitude. The time evolution of the oxidative decay of the CaHydI activity reveals different time constants when complexed with m-SWNTs and s-SWNTs. The correlation of enzymatic assays with time-dependent Raman spectroscopy provides a novel method by which the charge transfer interactions may be investigated in the various SWNT-CaHydI complexes. Surprisingly, an oxidized form of CaHydI is apparently more resistant to oxygen deactivation when complexed to m-SWNTs rather than s-SWNTs.

  10. Biomimetic peptide-based models of [FeFe]-hydrogenases: utilization of phosphine-containing peptides.

    Science.gov (United States)

    Roy, Souvik; Nguyen, Thuy-Ai D; Gan, Lu; Jones, Anne K

    2015-09-07

    Two synthetic strategies for incorporating diiron analogues of [FeFe]-hydrogenases into short peptides via phosphine functional groups are described. First, utilizing the amine side chain of lysine as an anchor, phosphine carboxylic acids can be coupled via amide formation to resin-bound peptides. Second, artificial, phosphine-containing amino acids can be directly incorporated into peptides via solution phase peptide synthesis. The second approach is demonstrated using three amino acids each with a different phosphine substituent (diphenyl, diisopropyl, and diethyl phosphine). In total, five distinct monophosphine-substituted, diiron model complexes were prepared by reaction of the phosphine-peptides with diiron hexacarbonyl precursors, either (μ-pdt)Fe2(CO)6 or (μ-bdt)Fe2(CO)6 (pdt = propane-1,3-dithiolate, bdt = benzene-1,2-dithiolate). Formation of the complexes was confirmed by UV/Vis, FTIR and (31)P NMR spectroscopy. Electrocatalysis by these complexes is reported in the presence of acetic acid in mixed aqueous-organic solutions. Addition of water results in enhancement of the catalytic rates.

  11. Hydrogen production by the naked active site of the di-iron hydrogenases in water.

    Science.gov (United States)

    Zipoli, Federico; Car, Roberto; Cohen, Morrel H; Selloni, Annabella

    2009-10-01

    We explored the reactivity of the active center of the [FeFe]-hydrogenases detached from the enzyme and immersed in acidified water by first-principles Car-Parrinello molecular-dynamics simulations. We focused on the identification of the structures that are stable and metastable in acidified water and on their activity for hydrogen production. Our calculations revealed that the naked active center could be an efficient catalyst provided that electrons are transferred to the cluster. We found that both bridging and terminal isomers are present at equilibrium and that the bridging configuration is essential for efficient hydrogen production. The formation of the hydrogen molecule occurs via sequential protonations of the distal iron and of the N-atom of the S-CH(2)-NH-CH(2)-S chelating group. H(2) desorption does not involve a significant energy barrier, making the process very efficient at room temperature. We established that the bottleneck in the reaction is the direct proton transfer from water to the vacant site of the distal iron. Moreover, we found that even if the terminal isomer is present at the equilibrium, its strong local hydrophobicity prevents poisoning of the cluster.

  12. Hydrogen-Activation Mechanism of [Fe] Hydrogenase Revealed by Multi-Scale Modeling

    CERN Document Server

    Finkelmann, Arndt Robert; Reiher, Markus

    2014-01-01

    When investigating the mode of hydrogen activation by [Fe] hydrogenases, not only the chemical reactivity at the active site is of importance but also the large-scale conformational change between the so-called open and closed conformations, which leads to a special spatial arrangement of substrate and iron cofactor. To study H2 activation, a complete model of the solvated and cofactor-bound enzyme in complex with the substrate methenyl-H4MPT+ was constructed. Both the closed and open conformations were simulated with classical molecular dynamics on the 100 ns time scale. Quantum-mechanics/molecular-mechanics calculations on snapshots then revealed the features of the active site that enable the facile H2 cleavage. The hydroxyl group of the pyridinol ligand can easily be deprotonated. With the deprotonated hydroxyl group and the structural arrangement in the closed conformation, H2 coordinated to the Fe center is subject to an ionic and orbital push-pull effect and can be rapidly cleaved with a concerted hydr...

  13. Application of mixed colloidal magnetic fluid of single domain Fe3O4 and NiFe2O4 ferrite nanoparticles in audio speaker

    Directory of Open Access Journals (Sweden)

    S. D. Kemkar

    2017-01-01

    Full Text Available Ferrofluids are stable suspensions of colloidal ferrimagnetic particles in suitable non – magnetic carrier liquids. They have attracted a lot of attention from scientists and engineers due to their many interesting properties and applications in various branches of engineering. The present work reports the performance of colloidal fluid of single domain nanoparticles of NiFe2O4 and Fe3O4. The thermal properties and its dynamics on magnetization as well as its effect on thermal conductivity on the colloidal fluid are studied here. Advantages of the increased thermal conductivity and optimization of magnetization of mixed colloidal fluid is used to extract the heat from voice coil. Nanoparticles of 21 nm of Fe3O4 and 12 nm of NiFe2O4 are used for mixed colloidal fluid. The suspension of particles is achieved by coating the nanoparticles with mono-corboxylic group on both the types of particles. The higher size (21 nm of Fe3O4 and 12 nm of NiFe2O4 particles are taken for synthesizing colloidal fluid, to have magnetic property of mixed colloidal liquid at elevated temperature of voice coil of speaker (Higher sized particles gives better magnetization. Oil is used as a carrier. Mixed magnetic colloidal fluid is used as a medium for damping so that noise is reduced at higher temperature of voice coil.

  14. Investigation of electronic and local structural changes during lithium uptake and release of nano-crystalline NiFe2O4 by X-ray absorption spectroscopy

    Science.gov (United States)

    Zhou, Dong; Permien, Stefan; Rana, Jatinkumar; Krengel, Markus; Sun, Fu; Schumacher, Gerhard; Bensch, Wolfgang; Banhart, John

    2017-02-01

    Nano-crystalline NiFe2O4 particles were synthesized and used as active electrode material for a lithium ion battery that showed a high discharge capacity of 1534 mAh g-1 and charge capacity of 1170 mAh g-1 during the 1st cycle. X-ray absorption spectroscopy including XANES and EXAFS were used to investigate electronic and local structural changes of NiFe2O4 during the 1st lithiation and de-lithiation process. As lithium is inserted into the structure, tetrahedral site Fe3+ ions are reduced to Fe2+ and moved from tetrahedral sites to empty octahedral sites, while Ni2+ ions are unaffected. As a consequence, the matrix spinel structure collapses and transforms to an intermediate rock-salt monoxide phase. Meanwhile, the inserted Li is partially consumed by the formation of SEI and other side reactions during the conversion reaction. With further lithiation, the monoxide phase is reduced to highly disordered metallic Fe/Ni nanoparticles with a number of nearest neighbors of 6.0(8) and 8.1(4) for Fe and Ni, respectively. During subsequent de-lithiation, the metal particles are individually re-oxidized to Fe2O3 and NiO phases instead to the original NiFe2O4 spinel phase.

  15. Irreversibility in room temperature current-voltage characteristics of NiFe2O4 nanoparticles: A signature of electrical memory effect

    Science.gov (United States)

    Dey, P.; Debnath, Rajesh; Singh, Swati; Mandal, S. K.; Roy, J. N.

    2017-01-01

    Room temperature I-V characteristics study, both in presence and absence of magnetic field (1800 Oe), has been performed on NiFe2O4 nanoparticles, having different particle size (V~14, 21 and 31 nm). Our experiments on these nanoparticles provide evidences for: (1) electrical irreversibility or hysteretic behaviour; (2) positive magnetoresistance and (3) magnetic field dependent electrical irreversibility or hysteresis in the sample. "Hysteretic" nature of I-V curve reveals the existence of electrical memory effect in the sample. Significantly, such hysteresis has been found to be tuned by magnetic field. In order to explain the observed electrical irreversibility, we have proposed a phenomenological model on the light of induced polarization in the sample. Both the positive magnetoresistance and the observed magnetic field dependence of electrical irreversibility have been explained through magnetostriction phenomenon. Interestingly, such effects are found to get reduced with increasing particle size. For NiFe2O4 nanoparticles having V=31 nm, we did not observe any irreversibility effect. This feature has been attributed to the enhanced grain surface effect that in turn gives rise to the residual polarization and hence electrical memory effect in NiFe2O4 nanoparticles, having small nanoscopic particle size.

  16. Effect of cathode vibration and heat treatment on electromagnetic properties of flake-shaped diatomite coated with Ni-Fe alloy by electroplating

    Science.gov (United States)

    Lan, Mingming; Li, Huiqin; Huang, Weihua; Xu, Guangyin; Li, Yan

    2015-03-01

    In this paper, flake-shaped diatomite particles were used as forming templates for the fabrication of the ferromagnetic functional fillers by way of electroplating Ni-Fe alloy method. The effects of cathode vibration frequency on the content of Ni-Fe alloy in the coating and the surface morphologies of the coatings were evaluated. The electromagnetic properties of the coated diatomite particles before and after heat treatment were also investigated in detail. The results show that the core-shell flake-shaped diatomite particles with high content of Ni-Fe alloy and good surface qualities of the coatings can be obtained by adjusting cathode vibration frequency. The coated diatomite particles with heat treatment filled paraffin wax composites exhibit a superior microwave absorbing and electromagnetic properties compared to the non-heat treated samples. Additionally, the peaks of reflection loss are found to be able to shift to lower frequency by the heat treatment process, which indicates the heat treatment can adjust microwave absorbing frequency band.

  17. Formate hydrogenlyase in the hyperthermophilic archaeon, Thermococcus litoralis

    Directory of Open Access Journals (Sweden)

    Rákhely Gábor

    2008-06-01

    Full Text Available Abstract Background Thermococcus litoralis is a heterotrophic facultative sulfur dependent hyperthermophilic Archaeon, which was isolated from a shallow submarine thermal spring. It has been successfully used in a two-stage fermentation system, where various keratinaceous wastes of animal origin were converted to biohydrogen. In this system T. litoralis performed better than its close relative, P. furiosus. Therefore, new alternative enzymes involved in peptide and hydrogen metabolism were assumed in T. litoralis. Results An about 10.5 kb long genomic region was isolated and sequenced from Thermococcus litoralis. In silico analysis revealed that the region contained a putative operon consisting of eight genes: the fdhAB genes coding for a formate dehydrogenase and the mhyCDEFGH genes encoding a [NiFe] hydrogenase belonging to the group of the H2-evolving, energy-conserving, membrane-bound hydrogenases. Reverse transcription linked quantitative Real-Time PCR and Western blotting experiments showed that the expression of the fdh-mhy operon was up-regulated during fermentative growth on peptides and down-regulated in cells cultivated in the presence of sulfur. Immunoblotting and protein separation experiments performed on cell fractions indicated that the formate dehydrogenase part of the complex is associated to the membrane-bound [NiFe] hydrogenase. Conclusion The formate dehydrogenase together with the membrane-bound [NiFe] hydrogenase formed a formate hydrogenlyase (formate dehydrogenase coupled hydrogenase, FDH-MHY complex. The expression data suggested that its physiological role is linked to the removal of formate likely generated during anaerobic peptide fermentation.

  18. Multiphase and Double-Layer NiFe2O4@NiO-Hollow-Nanosphere-Decorated Reduced Graphene Oxide Composite Powders Prepared by Spray Pyrolysis Applying Nanoscale Kirkendall Diffusion.

    Science.gov (United States)

    Park, Gi Dae; Cho, Jung Sang; Kang, Yun Chan

    2015-08-05

    Multicomponent metal oxide hollow-nanosphere decorated reduced graphene oxide (rGO) composite powders are prepared by spray pyrolysis with nanoscale Kirkendall diffusion. The double-layer NiFe2O4@NiO-hollow-nanosphere decorated rGO composite powders are prepared using the first target material. The NiFe-alloy-nanopowder decorated rGO powders are prepared as an intermediate product by post-treatment under the reducing atmosphere of the NiFe2O4/NiO-decorated rGO composite powders obtained by spray pyrolysis. The different diffusion rates of Ni (83 pm for Ni(2+)) and Fe (76 pm for Fe(2+), 65 pm for Fe(3+)) cations with different radii during nanoscale Kirkendall diffusion result in multiphase and double-layer NiFe2O4@NiO hollow nanospheres. The mean size of the hollow NiFe2O4@NiO nanospheres decorated uniformly within crumpled rGO is 14 nm. The first discharge capacities of the nanosphere-decorated rGO composite powders with filled NiFe2O4/NiO and hollow NiFe2O4@NiO at a current density of 1 A g(-1) are 1168 and 1319 mA h g(-1), respectively. Their discharge capacities for the 100th cycle are 597 and 951 mA h g(-1), respectively. The discharge capacity of the NiFe2O4@NiO-hollow-nanosphere-decorated rGO composite powders at the high current density of 4 A g(-1) for the 400th cycle is 789 mA h g(-1).

  19. Gas exchange in the filamentous cyanobacterium Nostoc punctiforme strain ATCC 29133 and Its hydrogenase-deficient mutant strain NHM5.

    Science.gov (United States)

    Lindberg, Pia; Lindblad, Peter; Cournac, Laurent

    2004-04-01

    Nostoc punctiforme ATCC 29133 is a nitrogen-fixing, heterocystous cyanobacterium of symbiotic origin. During nitrogen fixation, it produces molecular hydrogen (H(2)), which is recaptured by an uptake hydrogenase. Gas exchange in cultures of N. punctiforme ATCC 29133 and its hydrogenase-free mutant strain NHM5 was studied. Exchange of O(2), CO(2), N(2), and H(2) was followed simultaneously with a mass spectrometer in cultures grown under nitrogen-fixing conditions. Isotopic tracing was used to separate evolution and uptake of CO(2) and O(2). The amount of H(2) produced per molecule of N(2) fixed was found to vary with light conditions, high light giving a greater increase in H(2) production than N(2) fixation. The ratio under low light and high light was approximately 1.4 and 6.1 molecules of H(2) produced per molecule of N(2) fixed, respectively. Incubation under high light for a longer time, until the culture was depleted of CO(2), caused a decrease in the nitrogen fixation rate. At the same time, hydrogen production in the hydrogenase-deficient strain was increased from an initial rate of approximately 6 micro mol (mg of chlorophyll a)(-1) h(-1) to 9 micro mol (mg of chlorophyll a)(-1) h(-1) after about 50 min. A light-stimulated hydrogen-deuterium exchange activity stemming from the nitrogenase was observed in the two strains. The present findings are important for understanding this nitrogenase-based system, aiming at photobiological hydrogen production, as we have identified the conditions under which the energy flow through the nitrogenase can be directed towards hydrogen production rather than nitrogen fixation.

  20. Role of the Azadithiolate Cofactor in Models for the [FeFe]-Hydrogenase: Novel Structures and Catalytic Implications

    OpenAIRE

    Olsen, Matthew T.; Thomas B. Rauchfuss; Wilson, Scott R.

    2010-01-01

    The report summarizes studies on the redox behavior of synthetic models for the [FeFe]-hydrogenases, consisting of diiron dithiolato carbonyl complexes bearing the amine cofactor and its N-benzyl derivative. Of specific interest are the causes of the low reactivity of oxidized models toward H2, which contrasts with the high activity of these enzymes for H2 oxidation. The redox and acid-base properties of the model complexes [Fe2[(SCH2)2NR](CO)3(dppv)(PMe3)]+ ([2]+ for R = H and [2′]+ for R = ...

  1. [FeFe]-hydrogenase maturation: insights into the role HydE plays in dithiomethylamine biosynthesis.

    Science.gov (United States)

    Betz, Jeremiah N; Boswell, Nicholas W; Fugate, Corey J; Holliday, Gemma L; Akiva, Eyal; Scott, Anna G; Babbitt, Patricia C; Peters, John W; Shepard, Eric M; Broderick, Joan B

    2015-03-10

    HydE and HydG are radical S-adenosyl-l-methionine enzymes required for the maturation of [FeFe]-hydrogenase (HydA) and produce the nonprotein organic ligands characteristic of its unique catalytic cluster. The catalytic cluster of HydA (the H-cluster) is a typical [4Fe-4S] cubane bridged to a 2Fe-subcluster that contains two carbon monoxides, three cyanides, and a bridging dithiomethylamine as ligands. While recent studies have shed light on the nature of diatomic ligand biosynthesis by HydG, little information exists on the function of HydE. Herein, we present biochemical, spectroscopic, bioinformatic, and molecular modeling data that together map the active site and provide significant insight into the role of HydE in H-cluster biosynthesis. Electron paramagnetic resonance and UV-visible spectroscopic studies demonstrate that reconstituted HydE binds two [4Fe-4S] clusters and copurifies with S-adenosyl-l-methionine. Incorporation of deuterium from D2O into 5'-deoxyadenosine, the cleavage product of S-adenosyl-l-methionine, coupled with molecular docking experiments suggests that the HydE substrate contains a thiol functional group. This information, along with HydE sequence similarity and genome context networks, has allowed us to redefine the presumed mechanism for HydE away from BioB-like sulfur insertion chemistry; these data collectively suggest that the source of the sulfur atoms in the dithiomethylamine bridge of the H-cluster is likely derived from HydE's thiol containing substrate.

  2. Redox reactions of [FeFe]-hydrogenase models containing an internal amine and a pendant phosphine.

    Science.gov (United States)

    Zheng, Dehua; Wang, Mei; Chen, Lin; Wang, Ning; Sun, Licheng

    2014-02-03

    A diiron dithiolate complex with a pendant phosphine coordinated to one of the iron centers, [(μ-SCH2)2N(CH2C6H4-o-PPh2){Fe2(CO)5}] (1), was prepared and structurally characterized. The pendant phosphine is dissociated together with a CO ligand in the presence of excess PMe3, to afford [(μ-SCH2)2N(CH2C6H4-o-PPh2){Fe(CO)2(PMe3)}2] (2). Redox reactions of 2 and related complexes were studied in detail by in situ IR spectroscopy. A series of new Fe(II)Fe(I) ([3](+) and [6](+)), Fe(II)Fe(II) ([4](2+)), and Fe(I)Fe(I) (5) complexes relevant to Hox, Hox(CO), and Hred states of the [FeFe]-hydrogenase active site were detected. Among these complexes, the molecular structures of the diferrous complex [4](2+) with the internal amine and the pendant phosphine co-coordinated to the same iron center and the triphosphine diiron complex 5 were determined by X-ray crystallography. To make a comparison, the redox reactions of an analogous complex, [(μ-SCH2)2N(CH2C6H5){Fe(CO)2(PMe3)}2] (7), were also investigated by in situ IR spectroscopy in the absence or presence of extrinsic PPh3, which has no influence on the oxidation reaction of 7. The pendant phosphine in the second coordination sphere makes the redox reaction of 2 different from that of its analogue 7.

  3. Effects of particle composition and environmental parameters on catalytic hydrodechlorination of trichloroethylene by nanoscale bimetallic Ni-Fe.

    Science.gov (United States)

    Wei, Jianjun; Qian, Yajing; Liu, Wenjuan; Wang, Lutao; Ge, Yijie; Zhang, Jianghao; Yu, Jiang; Ma, Xingmao

    2014-05-01

    Catalytic nickel was successfully incorporated into nanoscale iron to enhance its dechlorination efficiency for trichloroethylene (TCE), one of the most commonly detected chlorinated organic compounds in groundwater. Ethane was the predominant product. The greatest dechlorination efficiency was achieved at 22 molar percent of nickel. This nanoscale Ni-Fe is poorly ordered and inhomogeneous; iron dissolution occurred whereas nickel was relatively stable during the 24-hr reaction. The morphological characterization provided significant new insights on the mechanism of catalytic hydrodechlorination by bimetallic nanoparticles. TCE degradation and ethane production rates were greatly affected by environmental parameters such as solution pH, temperature and common groundwater ions. Both rate constants decreased and then increased over the pH range of 6.5 to 8.0, with the minimum value occurring at pH 7.5. TCE degradation rate constant showed an increasing trend over the temperature range of 10 to 25°C. However, ethane production rate constant increased and then decreased over the range, with the maximum value occurring at 20°C. Most salts in the solution appeared to enhance the reaction in the first half hour but overall they displayed an inhibitory effect. Combined ions showed a similar effect as individual salts.

  4. Nitrogen–doped graphitized carbon shell encapsulated NiFe nanoparticles: A highly durable oxygen evolution catalyst

    Energy Technology Data Exchange (ETDEWEB)

    Du, Lei; Luo, Langli; Feng, Zhenxing; Engelhard, M. H.; Xie, Xiaohong; Han, Binghong; Sun, Junming; Zhang, Jianghao; Yin, Geping; Wang, Chongmin; Wang, Yong; Shao, Yuyan

    2017-09-01

    Oxygen evolution reaction (OER) plays a crucial role in various energy conversion devices such as water electrolyzers and metal–air batteries. Precious metal catalysts such as Ir, Ru and their oxides are usually used for enhanced reaction kinetics but are limited by their scarce resource. The challenges associated with alternative non–precious metal catalysts such as transition metal oxides and (oxy)hydroxides etc. are their low electronic conductivity and poor durability. Here, we report OER catalysts of NiFe nanoparticles encapsulated by nitrogen–doped graphitized carbon shells derived from bimetallic metal–organic frameworks (MOFs) precursors. The optimal OER catalyst shows excellent activity (360 mV overpotential at 10 mA cm–2GEO) and durability (no obvious degradation after 20 000 cycles). The electron-donation from Fe and tuned electronic structure of metal cores by Ni are revealed to be primary contributors to the enhanced OER activity. We further demonstrated that the structure and morphology of encapsulating carbon shells, which are the key factors influencing the durability, are facilely controlled by chemical state of precursors. Severe metal particle growth probably caused by oxidation of carbon shells and encapsulated nanoparticles is believed to the main mechanism for activity degradation in these catalysts.

  5. Nitrogen-doped graphitized carbon shell encapsulated NiFe nanoparticles: A highly durable oxygen evolution catalyst

    Energy Technology Data Exchange (ETDEWEB)

    Du, Lei; Luo, Langli; Feng, Zhenxing; Engelhard, Mark H.; Xie, Xiaohong; Han, Binghong; Sun, Junming; Zhang, Jianghao; Yin, Geping; Wang, Chongmin; Wang, Yong; Shao, Yuyan

    2017-09-01

    Oxygen evolution reaction (OER) plays a crucial role in various energy conversion devices such as water electrolyzers and metal–air batteries. Precious metal catalysts such as Ir, Ru and their oxides are usually used for enhancing reaction kinetics but are limited by their scarce resource. The challenges associated with alternative non–precious metal catalysts such as transition metal oxides and (oxy)hydroxides etc. are their low electronic conductivity and durability. Herein, we report a highly active (360 mV overpotential at 10 mA cm–2GEO) and durable (no degradation after 20000 cycles) OER catalyst derived from bimetallic metal–organic frameworks (MOFs) precursors. This catalyst consists of NiFe nanoparticles encapsulated by nitrogen–doped graphitized carbon shells. The electron−donation/deviation from Fe and tuned electronic structure of metal cores by Ni are revealed to be primary contributors to the enhanced OER activity, whereas N concentration contributes negligibly. We further demonstrated that the structure and morphology of encapsulating carbon shells, which are the key factors influencing the durability, are facilely controlled by the chemical state of precursors.

  6. The effects of fuel type in synthesis of NiFe2O4 nanoparticles by microwave assisted combustion method

    Science.gov (United States)

    Karcıoğlu Karakaş, Zeynep; Boncukçuoğlu, Recep; Karakaş, İbrahim H.

    2016-04-01

    In this study, it was investigated the effects of the used fuels on structural, morphological and magnetic properties of nanoparticles in nanoparticle synthesis with microwave assisted combustion method with an important method in quick, simple and low cost at synthesis of the nanoparticles. In this aim, glycine, urea and citric acid were used as fuel, respectively. The synthesised nanoparticles were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), Brunauer-Emmet-Teller surface area (BET), and vibrating sample magnetometry (VSM) techniques. We observed that fuel type is quite effective on magnetic properties and surface properties of the nanoparticles. X-ray difractograms of the obtained nanoparticles were compared with standard powder diffraction cards of NiFe2O4 (JCPDS Card Number 54-0964). The results demonstrated that difractograms are fully compatible with standard reflection peaks. According to the results of the XRD analysis, the highest crystallinity was observed at nanoparticles synthesized with glycine. The results demonstrated that the nanoparticles prepared with urea has the highest surface area. The micrographs of SEM showed that all of the nanoparticles have nano-crystalline behaviour and particles indication cubic shape. VSM analysis demonstrated that the type of fuel used for synthesis is highly effective a parameter on magnetic properties of nanoparticles.

  7. Effect of additive on corrosion resistance of NiFe2O4 ceramics as inert anodes

    Institute of Scientific and Technical Information of China (English)

    XI Jin-hui; XIE Ying-jie; YAO Guang-chun; LIU Yi-han

    2008-01-01

    In order to improve the corrosion resistance of NiFe2O4 ceramics as inert anode, additive V2O5 was added to raw materials NiO and Fe2O3. The inert anodes of nickel-ferrite ceramics were prepared by powder metallurgic method and the static corrosion rate in Na3AlF6-Al2O3 was determined by mass loss measurement. The effect of V2O5 on sintering property and corrosion resistance was studied. The results show that V2O5 can promote the grain to develop completely and improve sintering property. EDS results show the reaction product Ni2FeVO6 distributes along the grain boundary. The corrosion tests show that V2O5 is beneficial to improving corrosion resistance remarkably. The reasons that V2O5 can improve the corrosion resistance must be V2O5 promoting the gains to develop completely and Ni2FeVO6 distributes along the grain boundary. The stable structure can control the chemical dissolution of ceramics anode and the reinforced grain boundary can control the grain-boundary corrosion rate.

  8. Calculation of the free energy of NiFe2O4 nanopoarticles by Monte Carlo simulation

    Science.gov (United States)

    Zhou, Chenggang; Landau, D. P.

    2005-03-01

    Magnetic properties of nanoparticles are of great current interest in light of possible applications to high density magnetic storage media. Finite size and surface effect are important for magnetic nanoparticles and differentiate them from their bulk counterparts. We use Monte Carlo simulation to study a model of NiFe2O4 nanopoarticles proposed by Kodama and Berkowitz [1]. The Hamiltonian of the nanoparticle contains superexchanges between magnetic ions modeled by Heisenberg spins, and surface/bulk anisotropy terms. A continuous version of the Wang- Landau algorithm [2] is used to calculate the joint density of states ρ(M, E) efficiently. From ρ(M, E), we can directly evaluate the free energy of the particle, and many other physical quantities. A hysteresis loop for particles with surface disorder and surface anisotropy is observed, in agreement with previous studies [1]. We found that such a hysteresis loop is the result of interplay between surface disorder and surface anisotropy. Compared with micromagnetic modeling, our Monte Carlo simulation treats the thermodynamic effects properly and is capable of calculating physical quantities at all temperatures and magnetic fields with very limited CPU time. [1] R. H. Kodama, et. al. Phys. Rev. Lett. 77, 394 (1996); Phys. Rev. B 59, 6321 (1999). [2] C. Zhou, et al., in preparation.

  9. A flexible alkaline rechargeable Ni/Fe battery based on graphene foam/carbon nanotubes hybrid film.

    Science.gov (United States)

    Liu, Jilei; Chen, Minghua; Zhang, Lili; Jiang, Jian; Yan, Jiaxu; Huang, Yizhong; Lin, Jianyi; Fan, Hong Jin; Shen, Ze Xiang

    2014-12-10

    The development of portable and wearable electronics has promoted increasing demand for high-performance power sources with high energy/power density, low cost, lightweight, as well as ultrathin and flexible features. Here, a new type of flexible Ni/Fe cell is designed and fabricated by employing Ni(OH)2 nanosheets and porous Fe2O3 nanorods grown on lightweight graphene foam (GF)/carbon nanotubes (CNTs) hybrid films as electrodes. The assembled f-Ni/Fe cells are able to deliver high energy/power densities (100.7 Wh/kg at 287 W/kg and 70.9 Wh/kg at 1.4 kW/kg, based on the total mass of active materials) and outstanding cycling stabilities (retention 89.1% after 1000 charge/discharge cycles). Benefiting from the use of ultralight and thin GF/CNTs hybrid films as current collectors, our f-Ni/Fe cell can exhibit a volumetric energy density of 16.6 Wh/l (based on the total volume of full cell), which is comparable to that of thin film battery and better than that of typical commercial supercapacitors. Moreover, the f-Ni/Fe cells can retain the electrochemical performance with repeated bendings. These features endow our f-Ni/Fe cells a highly promising candidate for next generation flexible energy storage systems.

  10. Escherichia coli K-12 survives anaerobic exposure at pH 2 without RpoS, Gad, or hydrogenases, but shows sensitivity to autoclaved broth products.

    Directory of Open Access Journals (Sweden)

    Daniel P Riggins

    Full Text Available Escherichia coli and other enteric bacteria survive exposure to extreme acid (pH 2 or lower in gastric fluid. Aerated cultures survive via regulons expressing glutamate decarboxylase (Gad, activated by RpoS, cyclopropane fatty acid synthase (Cfa and others. But extreme-acid survival is rarely tested under low oxygen, a condition found in the stomach and the intestinal tract. We observed survival of E. coli K-12 W3110 at pH 1.2-pH 2.0, conducting all manipulations (overnight culture at pH 5.5, extreme-acid exposure, dilution and plating in a glove box excluding oxygen (10% H2, 5% CO2, balance N2. With dissolved O2 concentrations maintained below 6 µM, survival at pH 2 required Cfa but did not require GadC, RpoS, or hydrogenases. Extreme-acid survival in broth (containing tryptone and yeast extract was diminished in media that had been autoclaved compared to media that had been filtered. The effect of autoclaved media on extreme-acid survival was most pronounced when oxygen was excluded. Exposure to H2O2 during extreme-acid treatment increased the death rate slightly for W3110 and to a greater extent for the rpoS deletion strain. Survival at pH 2 was increased in strains lacking the anaerobic regulator fnr. During anaerobic growth at pH 5.5, strains deleted for fnr showed enhanced transcription of acid-survival genes gadB, cfa, and hdeA, as well as catalase (katE. We show that E. coli cultured under oxygen exclusion (<6 µM O2 requires mechanisms different from those of aerated cultures. Extreme acid survival is more sensitive to autoclave products under oxygen exclusion.

  11. Performance of NiFe2O4-SiO2-TiO2 Magnetic Photocatalyst for the Effective Photocatalytic Reduction of Cr(VI in Aqueous Solutions

    Directory of Open Access Journals (Sweden)

    Mike O. Ojemaye

    2017-01-01

    Full Text Available Investigation into the reduction of Cr(VI in aqueous solution was carried out through some batch photocatalytic studies. The photocatalysts used were silica coated nickel ferrite nanoparticles (NiFe2O4-SiO2, nickel ferrite titanium dioxide (NiFe2O4-TiO2, nickel ferrite silica titanium dioxide (NiFe2O4-SiO2-TiO2, and titanium dioxide (TiO2. The characterization of the materials prepared via stepwise synthesis using coprecipitation and sol-gel methods were carried out with the aid of X-ray diffraction (XRD, transmission electron microscopy (TEM, scanning electron microscopy (SEM, Fourier transform infrared (FTIR spectroscopy, thermal gravimetric analysis (TGA, and vibrating sample magnetometry (VSM. The reduction efficiency was studied as a function of pH, photocatalyst dose, and contact time. The effects of silica interlayer between the magnetic photocatalyst materials reveal that reduction efficiency of NiFe2O4-SiO2-TiO2 towards Cr(VI was higher than that of NiFe2O4-TiO2. However, TiO2 was observed to have the highest reduction efficiency at all batch photocatalytic experiments. Kinetics study shows that photocatalytic reduction of Cr(VI obeyed Langmuir-Hinshelwood model and first-order rate kinetics. Regenerability study also suggested that the photocatalyst materials can be reused.

  12. Improved purification, crystallization and crystallographic study of Hyd-2-type [NiFe]-hydrogenase from Citrobacter sp. S-77.

    Science.gov (United States)

    Muhd Noor, Noor Dina; Nishikawa, Koji; Nishihara, Hirofumi; Yoon, Ki Seok; Ogo, Seiji; Higuchi, Yoshiki

    2016-01-01

    The purification procedure of Hyd-2-type [NiFe]-hydrogenase from Citrobacter sp. S-77 was improved by applying treatment with trypsin before chromatography. Purified protein samples both with and without trypsin treatment were successfully crystallized using the sitting-drop vapour-diffusion method with polyethylene glycol as a precipitant. Both crystals belonged to space group P21, with unit-cell parameters a = 63.90, b = 118.89, c = 96.70 Å, β = 100.61° for the protein subjected to trypsin treatment and a = 65.38, b = 121.45, c = 98.63 Å, β = 102.29° for the sample not treated with trypsin. The crystal obtained from the trypsin-treated protein diffracted to 1.60 Å resolution, which is considerably better than the 2.00 Å resolution obtained without trypsin treatment. The [NiFe]-hydrogenase from Citrobacter sp. S-77 retained catalytic activity with some amount of O2, indicating that it has clear O2 tolerance.

  13. Hydrogen-induced activation of the [NiFe]-hydrogenase from Allochromatium vinosum as studied by stopped-flow infrared spectroscopy.

    NARCIS (Netherlands)

    Kurkin, S.; George, S.J.; Thorneley, R.N.; Albracht, S.P.J.

    2004-01-01

    The reaction between hydrogen and the [NiFe]-hydrogenase from Allochromatium Vinosum in its inactive form has been studied by stopped-flow infrared spectroscopy. The data, for the first time, clearly show that at room temperature enzyme in the unready state, either oxidized or reduced, does not

  14. Influence of pH and ionic strength on electrostatic properties of ferredoxin, FNR, and hydrogenase and the rate constants of their interaction

    Science.gov (United States)

    Diakonova, A. N.; Khrushchev, S. S.; Kovalenko, I. B.; Riznichenko, G. Yu; Rubin, A. B.

    2016-10-01

    Ferredoxin (Fd) protein transfers electrons from photosystem I (PSI) to ferredoxin:NADP+-reductase (FNR) in the photosynthetic electron transport chain, as well as other metabolic pathways. In some photosynthetic organisms including cyanobacteria and green unicellular algae under anaerobic conditions Fd transfers electrons not only to FNR but also to hydrogenase—an enzyme which catalyzes reduction of atomic hydrogen to H2. One of the questions posed by this competitive relationship between proteins is which characteristics of thylakoid stroma media allow switching of the electron flow between the linear path PSI-Fd-FNR-NADP+ and the path PSI-Fd-hydrogenase-H2. The study was conducted using direct multiparticle simulation approach. In this method protein molecules are considered as individual objects that experience Brownian motion and electrostatic interaction with the surrounding media and each other. Using the model we studied the effects of pH and ionic strength (I) upon complex formation between ferredoxin and FNR and ferredoxin and hydrogenase. We showed that the rate constant of Fd-FNR complex formation is constant in a wide range of physiologically significant pH values. Therefore it can be argued that regulation of FNR activity doesn’t involve pH changes in stroma. On the other hand, in the model rate constant of Fd-hydrogenase interaction dramatically depends upon pH: in the range 7-9 it increases threefold. It may seem that because hydrogenase reduces protons it should be more active when pH is acidic. Apparently, regulation of hydrogenase’s affinity to both her reaction partners (H+ and Fd) is carried out by changes in its electrostatic properties. In the dark, the protein is inactive and in the light it is activated and starts to interact with both Fd and H+. Therefore, we can conclude that in chloroplasts the rate of hydrogen production is regulated by pH through the changes in the affinity between hydrogenase and ferredoxin.

  15. Nickel binding and [NiFe]-hydrogenase maturation by the metallochaperone SlyD with a single metal-binding site in Escherichia coli.

    Science.gov (United States)

    Kaluarachchi, Harini; Altenstein, Matthias; Sugumar, Sonia R; Balbach, Jochen; Zamble, Deborah B; Haupt, Caroline

    2012-03-16

    SlyD (sensitive to lysis D) is a nickel metallochaperone involved in the maturation of [NiFe]-hydrogenases in Escherichia coli (E. coli) and specifically contributes to the nickel delivery step during enzyme biosynthesis. This protein contains a C-terminal metal-binding domain that is rich in potential metal-binding residues that enable SlyD to bind multiple nickel ions with high affinity. The SlyD homolog from Thermus thermophilus does not contain the extended cysteine- and histidine-rich C-terminal tail of the E. coli protein, yet it binds a single Ni(II) ion tightly. To investigate whether a single metal-binding motif can functionally replace the full-length domain, we generated a truncation of E. coli SlyD, SlyD155. Ni(II) binding to SlyD155 was investigated by using isothermal titration calorimetry, NMR and electrospray ionization mass spectrometry measurements. This in vitro characterization revealed that SlyD155 contains a single metal-binding motif with high affinity for nickel. Structural characterization by X-ray absorption spectroscopy and NMR indicated that nickel was coordinated in an octahedral geometry with at least two histidines as ligands. Heterodimerization between SlyD and another hydrogenase accessory protein, HypB, is essential for optimal hydrogenase maturation and was confirmed for SlyD155 via cross-linking experiments and NMR titrations, as were conserved chaperone and peptidyl-prolyl isomerase activities. Although these properties of SlyD are preserved in the truncated version, it does not modulate nickel binding to HypB in vitro or contribute to the maturation of [NiFe]-hydrogenases in vivo, unlike the full-length protein. This study highlights the importance of the unusual metal-binding domain of E. coli SlyD in hydrogenase biogenesis.

  16. Site saturation mutagenesis demonstrates a central role for cysteine 298 as proton donor to the catalytic site in CaHydA [FeFe]-hydrogenase.

    Directory of Open Access Journals (Sweden)

    Simone Morra

    Full Text Available [FeFe]-hydrogenases reversibly catalyse molecular hydrogen evolution by reduction of two protons. Proton supply to the catalytic site (H-cluster is essential for enzymatic activity. Cysteine 298 is a highly conserved residue in all [FeFe]-hydrogenases; moreover C298 is structurally very close to the H-cluster and it is important for hydrogenase activity. Here, the function of C298 in catalysis was investigated in detail by means of site saturation mutagenesis, simultaneously studying the effect of C298 replacement with all other 19 amino acids and selecting for mutants with high retained activity. We demonstrated that efficient enzymatic turnover was maintained only when C298 was replaced by aspartic acid, despite the structural diversity between the two residues. Purified CaHydA C298D does not show any significant structural difference in terms of secondary structure and iron incorporation, demonstrating that the mutation does not affect the overall protein fold. C298D retains the hydrogen evolution activity with a decrease of k(cat only by 2-fold at pH 8.0 and it caused a shift of the optimum pH from 8.0 to 7.0. Moreover, the oxygen inactivation rate was not affected demonstrating that the mutation does not influence O(2 diffusion to the active site or its reactivity with the H-cluster. Our results clearly demonstrate that, in order to maintain the catalytic efficiency and the high turnover number typical of [FeFe] hydrogenases, the highly conserved C298 can be replaced only by another ionisable residue with similar steric hindrance, giving evidence of its involvement in the catalytic function of [FeFe]-hydrogenases in agreement with an essential role in proton transfer to the active site.

  17. Diseño de producto de una red anti cínifes portable y auto plegable en material textil

    OpenAIRE

    2016-01-01

    Monet es una red anti cínifes que previene las enfermedades transmitidas por los mismos. El producto consta de una malla protectora, una carcasa, una estructura de montaje y un sistema de auto plegado, que es su mayor aportación y diferenciación del resto de mosquiteras. Está diseñado para ocupar el mínimo espacio y ser lo más ligera posible. Aportando unas grandes calidades y prestaciones mediante materiales reciclados. Monet es reparable, todas sus piezas son sustituibles ...

  18. Hydrogen from Water in a Novel Recombinant Cyanobacterial System

    Energy Technology Data Exchange (ETDEWEB)

    Weyman, Philip D [J. Craig Venter Institute; Smith, Hamillton O.

    2014-12-03

    Photobiological processes are attractive routes to renewable H2 production. With the input of solar energy, photosynthetic microbes such as cyanobacteria and green algae carry out oxygenic photosynthesis, using sunlight energy to extract protons and high energy electrons from water. These protons and high energy electrons can be fed to a hydrogenase system yielding H2. However, most hydrogen-evolving hydrogenases are inhibited by O2, which is an inherent byproduct of oxygenic photosynthesis. The rate of H2 production is thus limited. Certain photosynthetic bacteria are reported to have an O2-tolerant evolving hydrogenase, yet these microbes do not split water, and require other more expensive feedstocks. To overcome these difficulties, the goal of this work has been to construct novel microbial hybrids by genetically transferring O2-tolerant hydrogenases from other bacteria into a class of photosynthetic bacteria called cyanobacteria. These hybrid organisms will use the photosynthetic machinery of the cyanobacterial hosts to perform the water-oxidation reaction with the input of solar energy, and couple the resulting protons and high energy electrons to the O2-tolerant bacterial hydrogenase, all within the same microbe (Fig. 1). The ultimate goal of this work has been to overcome the sensitivity of the hydrogenase enzyme to O2 and address one of the key technological hurdles to cost-effective photobiological H2 production which currently limits the production of hydrogen in algal systems. In pursuit of this goal, work on this project has successfully completed many subtasks leading to a greatly increased understanding of the complicated [NiFe]-hydrogenase enzymes. At the beginning of this project, [NiFe] hydrogenases had never been successfully moved across wide species barriers and had never been heterologously expressed in cyanobacteria. Furthermore, the idea that whole, functional genes could be extracted from complicated, mixed-sequence meta-genomes was not

  19. The effects of the dimethylether bridging moiety in the H-cluster of the Clostridium pasteurianum hydrogenase on the mechanism of H{sub 2} production: A quantum mechanics/molecular mechanics study

    Energy Technology Data Exchange (ETDEWEB)

    Trohalaki, Steven [Air Force Research Laboratory, Materials and Manufacturing Directorate, Wright-Patterson Air Force Base, OH 45433 (United States); General Dynamics Information Technology, Inc. (United States); Pachter, Ruth [Air Force Research Laboratory, Materials and Manufacturing Directorate, Wright-Patterson Air Force Base, OH 45433 (United States)

    2010-12-15

    [Fe-Fe]-hydrogenases are naturally occurring metalloenzymes that catalyze the reversible production of H{sub 2} from two protons and two electrons. [Fe-Fe]-hydrogenases found in two species -Clostridium pasteurianum (CpI) and Desulfovibrio desulfuricans (DdH) - were shown with x-ray crystallography to have active sites that are very similar, although several atoms that bridge the dithiolate ligand were unresolved. In earlier work, we employed density functional theory (DFT) within a QM/MM method to investigate two previously proposed mechanisms of hydrogen production by DdH and CpI hydrogenases. In one mechanism (I), a CO ligand bridging two Fe atoms in the active site rotates to a terminal position while in the other (II) the CO bridge remains intact throughout the catalytic cycle. We previously assumed that the active sites for the two hydrogenases were identical; each had a dimethylamine bridging moiety, whose basicity is important for Mechanism II. Our overall conclusion, taking into consideration an energy comparison for the two mechanisms and activation energies for the CO-unbridging step in Mechanism I, was that Mechanism II was favored for both hydrogenases. In this paper, we extend our previous work to show that Mechanism II is favored over Mechanism I even if the bridging moiety in CpI hydrogenase is dimethylether, a significantly weaker base than dimethylamine, providing further support for Mechanism II even though experimental verification of the bridging moiety for the CpI H-cluster is lacking. (author)

  20. Multiscale modeling of the active site of [Fe] hydrogenase: the H₂ binding site in open and closed protein conformations.

    Science.gov (United States)

    Hedegård, Erik Donovan; Kongsted, Jacob; Ryde, Ulf

    2015-05-18

    A series of QM/MM optimizations of the full protein of [Fe] hydrogenase were performed. The FeGP cofactor has been optimized in the water-bound resting state (1), with a side-on bound dihydrogen (2), or as a hydride intermediate (3). For inclusion of H4MPT in the closed structure, advanced multiscale modeling appears to be necessary, especially to obtain reliable distances between CH-H4MPT(+) and the dihydrogen (H2) or hydride (H(-)) ligand in the FeGP cofactor. Inclusion of the full protein is further important for the relative energies of the two intermediates 2 and 3. We finally find that hydride transfer from 3 has a significantly higher barrier than found in previous studies neglecting the full protein environment.

  1. Influence of carboxylic acid type on microstructure and magnetic properties of polymeric complex sol-gel driven NiFe2O4

    Science.gov (United States)

    Hessien, M. M.; Mostafa, Nasser Y.; Abd-Elkader, Omar H.

    2016-01-01

    Citric, oxalic and tartaric acids were used for synthesis of NiFe2O4 using polymeric complex precursor route. The dry precursor gels were calcined at various temperatures (400-1100 °C) for 2 h. All carboxylic acids produce iron-deficient NiFe2O4 with considerable amount of α-Fe2O3 at 400 °C. Increase in the annealing temperature caused reaction of α-Fe2O3 with iron-deficient ferrite phase. The amount of initially formed α-Fe2O3 is directly correlated with stability constant and inversely correlated with the decomposition temperature of Fe(III) carboxylate precursors. In case of tartaric acid precursor, single phase of the ferrite was obtained at 450 °C. However, in case of oxalic acid and citric acid precursors, single phase ferrite was obtained at 550 °C and 700 °C, respectively. The lattice parameters were increased with increasing annealing temperature and with decreasing the amount of α-Fe2O3. Maximum saturation magnetization (55 emu/g) was achieved using tartaric acid precursor annealed at 1100 °C.

  2. Effect of Yb2O3 doping on the grain boundary of NiFe2O4-10NiO-based cermets after sintering

    Institute of Scientific and Technical Information of China (English)

    Han-bing He

    2015-01-01

    xYb2O3–15(20Ni–Cu)/(85−x)(NiFe2O4–10NiO) (x=0, 0.25, 0.5, 0.75, 1.0, 2.0, and 10.0) cermets for aluminum electrolysis were prepared to investigate the effect of Yb2O3 doping on the grain boundary of the cermets after sintering. The results showed that each interface was very clear and that with increasing Yb2O3 content, most of the Yb was evenly distributed at the grain boundary. Moreover, according to the phase composition and microstructural analysis by X-ray diffraction (XRD), scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM/EDX), and electron probe microanalysis (EPMA), YbFeO3 was produced along the grain boundary. The YbFeO3 was concluded to not only have formed from the interaction between the NiFe2O4 or Fe2O3 component and Yb2O3 at the grain boundary of the cermets, but also from the decomposition of NiFe2O4 into NiO and Fe2O3 and the subsequent reaction of Fe2O3 with Yb2O3. Thus, the pro-duction of YbFeO3 resulted in a cermet with high relative density, good electrical conductivity, and good corrosion resistance.

  3. First principles calculation of stable structure and adhesive strength of plated Ni/Fe(100) or Cu/Fe(100) interfaces

    Institute of Scientific and Technical Information of China (English)

    Ryota NAKANISHI; Koji SUEOKA; Seiji SHIBA; Makoto HINO; Koji MURAKAMI; Ken MURAOKA

    2009-01-01

    A study the with first principles calculation of the interfaces of the Ni layer or Cu layer on the Fe(100) surface formed with metal plating was performed. Ni or Cu atoms were shown to adopt the corresponding position to the bcc structure of the Fe(100) substrate. Other calculations showed that the interfaces of Ni (5 atomic layers)/Fe(100) (5 layers) or Cu (5 atomic layers)/Fe(100) (5 layers) had square lattices. The orientation relationship of Ni/Fe(100) interface corresponds to fcc-Ni(100)//bcc-Fe(100), Ni[011]//Fe[010], and Similar results were obtained for Cu/Fe(100) interfaces. This structure was supported by TEM analysis of plated Ni layer on Fe(100) surfaces. The adhesion strength of the Ni/Fe(100) interface evaluated by first principles calculation was higher than that of the Cu/Fe(100) interface. The experimental results of Hull cell iron plated with Ni or Cu supported the results of the calculation. These results indicate that the first principles calculation, which deals with the ideal interface at the atomic scale, has the potential to evaluate the adhesion strength of metallic material interfaces.

  4. Synthesis and characterization of the NiFe2O4@TEOS-TPS@Ag nanocomposite and investigation of its antibacterial activity

    Science.gov (United States)

    Allafchian, Ali R.; Jalali, S. A. H.; Amiri, R.; Shahabadi, Sh.

    2016-11-01

    In this study, the NiFe2O4 was embedded in (3-mercaptopropyl) trimethoxysilane (TPS) and tetraethyl orthosilicate (TEOS) using the sol-gel method. These compounds were used as the support of Ag nanoparticles (Ag NPs). The NiFe2O4@TEOS-TPS@Ag nanocomposites were obtained with the development of bonding between the silver atoms of Ag NPs and the sulfur atoms of TPS molecule. Field emission scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FT-IR) were used for the characterization of the Ag nanocomposites. Also, the magnetic properties of these nanocomposites were studied by using a vibrating sample magnetometer (VSM) technique. The disk diffusion, minimum inhibition concentration (MIC) and minimum bactericidal concentrations (MBC) tests were used for the investigation of the antibacterial effect of this nanocomposite against bacterial strains. The synthesized nanocomposite presented high reusability and good antibacterial activity against gram-positive and gram-negative bacteria. Remarkably, this nanocomposite could be easily removed from the disinfected media by magnetic decantation.

  5. Temperature dependence of magnetization and anisotropy in uniaxial NiFe2O4 nanomagnets: Deviation from the Callen-Callen power law

    Science.gov (United States)

    Chatterjee, Biplab K.; Ghosh, C. K.; Chattopadhyay, K. K.

    2014-10-01

    The thermal variation of magnetic anisotropy (K) and saturation magnetization (MS) for uniaxial nickel ferrite (NiFe2O4) nanomagnets are investigated. Major magnetic hysteresis loops are measured for the sample at temperatures over the range 5-280 K using a vibrating sample magnetometer. The high-field regimes of the hysteresis loops are modeled using the law of approach to saturation, based on the assumption that at sufficiently high field only direct rotation of spin-moment take place, with an additional forced magnetization term that is linear with applied field. The uniaxial anisotropy constant K is calculated from the fitting of the data to the theoretical equation. As temperature increases from 5 K to 280 K, a 49% reduction of K, accompanied by an 85% diminution of MS is observed. Remarkably, K is linearly proportional to MS2.6 in the whole temperature range violating the existing theoretical model by Callen and Callen. The unusual power-law behavior for the NiFe2O4 uniaxial nanomagnets is ascribed to the non-negligible contributions from inter-sublattice pair interactions, Neel surface anisotropy, and higher order anisotropies. A complete realization of the unusual anisotropy-magnetization scaling behavior for nanoscale two-sublattice magnetic materials require a major modification of the existing theory by considering the exact mechanism of each contributions to the effective anisotropy.

  6. Synthesis of Zno-Nife2o4 Magnetic Nanocomposites by Simple Solvothermal Method for Photocatalytic Dye Degradation Under Solar Light

    Directory of Open Access Journals (Sweden)

    Rahmayeni

    2016-06-01

    Full Text Available ZnO-NiFe2O4 magnetic nanocomposite photocatalysts were successfully synthesized by simple solvothermal method using nitric salts as starting materials and ethanol as solvent. The synthesized samples were characterized by XRD, FESEM-EDX, TEM, DRS-UV-vis, VSM, and FTIR instruments. Photocatalytic activity of the samples was evaluated by dye degradation under solar light irradiation. The results show that the diffraction peaks of nanocomposites correspond to the hexagonal wurzite of ZnO and spinal cubic structure of NiFe2O4. The microstructure studies revealed that nanocomposites were dominated by granular-like of ZnO nanoparticles were synthesized at 180°C and rod-like at 200°C. The estimated band gap value of ZnO-NiFe2O4 (1:0.01 nanocomposites is 2.78 eV which is lower than ZnO. The magnetic saturation (Ms result showed a super paramagnetic behavior for nanocomposites. It was found that the photocatalytic activity of synthesized nanocomposites were higher than ZnO and NiFe2O4.

  7. Powerful Fermentative Hydrogen Evolution of Photosynthate in the Cyanobacterium Lyngbya aestuarii BL J Mediated by a Bidirectional Hydrogenase

    Directory of Open Access Journals (Sweden)

    Ankita eKothari

    2014-12-01

    Full Text Available Cyanobacteria are considered good models for biohydrogen production because they are relatively simple organisms with a demonstrable ability to generate H2 under certain physiological conditions. However, most produce only little H2, revert readily to H2 consumption, and suffer from hydrogenase sensitivity to O2. Strains of the cyanobacteria Lyngbya aestuarii and Microcoleus chthonoplastes obtained from marine intertidal cyanobacterial mats were recently found to display much better H2 production potential. Because of their ecological origin in environments that become quickly anoxic in the dark, we hypothesized that this differential ability may have evolved to serve a role in the fermentation of the photosynthate. Here we show that, when forced to ferment internal substrate, these cyanobacteria display desirable characteristics of physiological H2 production. Among them, the strain L. aestuarii BL J had the fastest specific rates and attained the highest H2 concentrations during fermentation of photosynthate, which proceeded via a mixed-acid fermentation pathway to yield acetate, ethanol, lactate, H2, CO2 and pyruvate. Contrary to expectations, the H2 yield per mole of glucose was only average compared to that of other cyanobacteria. Thermodynamic analyses point to the use of electron donors more electronegative than NAD(PH in Lyngbya hydrogenases as the basis for its strong H2 production ability. In any event, the high specific rates and H2 concentrations coupled with the lack of reversibility of the enzyme, at the expense of internal, photosynthetically generated reductants, makes L. aestuarii BL J and/or its enzymes, a potentially feasible platform for large-scale H2 production.

  8. Size-controlled synthesis of NiFe2O4 nanospheres via a PEG assisted hydrothermal route and their catalytic properties in oxidation of alcohols by periodic acid

    Science.gov (United States)

    Paul, Bappi; Purkayastha, Debraj Dhar; Dhar, Siddhartha Sankar

    2016-05-01

    A novel and facile approach for synthesis of spinel nickel ferrites (NiFe2O4) nanoparticles (NPs) employing homogeneous chemical precipitation followed by hydrothermal heating is reported. The synthesis involves use of tributylamine (TBA) as a hydroxylating agent in synthesis of nickel ferrites. Polyethylene glycol (PEG) 4000 was used as surfactant. As-synthesized NiFe2O4 NPs were characterized by powder X-ray diffraction (XRD), transmission electron microscopy (TEM), N2 adsorption-desorption isotherm (BET) and vibrating sample magnetometry (VSM). The XRD pattern revealed formation of cubic face-centered NiFe2O4 and TEM image showed spherical particles of sizes 2-10 nm. These NiFe2O4 NPs were used as magnetically recoverable catalyst in oxidation of cyclic alcohols to their corresponding aldehydes by periodic acid. This eco-friendly procedure affords products in very high yield and selectivity. The reusability of the catalyst is proved to be noteworthy as the material exhibits no significant changes in its catalytic activity even after five cycles of reuse.

  9. The Catalytic Reductive Dechlorination of Ni/Fe Nano-alloy to Trichloro Ethylene(TCE)in Water%纳米Ni/Fe合金对水中三氯乙烯(TCE)的催化还原脱氯

    Institute of Scientific and Technical Information of China (English)

    张卫华; 张春华; 陈保国

    2016-01-01

    利用纳米Ni/Fe合金粉末对水中TCE进行催化还原脱氯研究,结果表明,纳米Ni/Fe合金对水中TCE具有较好的脱氯作用.在Ni含量5wt%、初始pH值为6.5、Ni/Fe粉末投加量为0.4g/L的条件下,TCE的脱氯率在60min内达72%;合金中较高Ni含量、较低初始pH、较大的粉末投加量均有利于脱氯反应;脱氯产物为丁烷、丁烯和乙烷等,可推测该反应是发生在双金属合金表面的加氢脱氯.%Dechlorination rate of TCE is up to 72%within 60 minutes under conditions of 5 wt%Ni,initial pH value 6.5 and 0.4g/L Ni/Fe powder. With the higher Ni content,the lower initial pH value and the more Ni/Fe powder,the Ni/Fe nano-alloy powder can better dechlorinate TCE in the water and produce butane,butene and ethane. This hy-drodechlorination occurrs on the surface of Ni/Fe alloy.

  10. Contributing factors in the improvement of cellulosic H2 production in Clostridium thermocellum/Thermoanaerobacterium co-cultures.

    Science.gov (United States)

    Wang, Mingyu; Zhao, Qi; Li, Ling; Niu, Kangle; Li, Yi; Wang, Fangzhong; Jiang, Baojie; Liu, Kuimei; Jiang, Yi; Fang, Xu

    2016-10-01

    Lignocellulosic biohydrogen is a promising renewable energy source that could be a potential alternative to the unsustainable fossil fuel-based energy. Biohydrogen production could be performed by Clostridium thermocellum that is the fastest known cellulose-degrading bacterium. Previous investigations have shown that the co-culture of C. thermocellum JN4 and a non-cellulolytic bacterium Thermoanaerobacterium thermosaccharolyticum GD17 produces more hydrogen than the C. thermocellum JN4 mono-culture, but the mechanism of this improvement is unknown. In this work, we carried out genomic and evolutionary analysis of hydrogenase-coding genes in C. thermocellum and T. thermosaccharolyticum, identifying one Ech-type [NiFe] hydrogenase complex in each species, and, respectively, five and four monomeric or multimeric [FeFe] hydrogenases in the two species. Further transcriptional analysis showed hydrogenase-coding genes in C. thermocellum are regulated by carbon sources, while hydrogenase-coding genes in T. thermosaccharolyticum are not. However, comparison between transcriptional abundance of hydrogenase-coding genes in mono- and co-cultures showed the co-culturing condition leads to transcriptional changes of hydrogenase-coding genes in T. thermosaccharolyticum but not C. thermocellum. Further metabolic analysis showed T. thermosaccharolyticum produces H2 at a rate 4-12-fold higher than C. thermocellum. These findings lead to the suggestion that the improvement of H2 production in the co-culture over mono-culture should be attributed to changes in T. thermosaccharolyticum but not C. thermocellum. Further suggestions can be made that C. thermocellum and T. thermosaccharolyticum perform highly specialized tasks in the co-culture, and optimization of the co-culture for more lignocellulosic biohydrogen production should be focused on the improvement of the non-cellulolytic bacterium.

  11. Expression of Uptake Hydrogenase and Molybdenum Nitrogenase in Rhodobacter capsulatus Is Coregulated by the RegB-RegA Two-Component Regulatory System

    OpenAIRE

    2000-01-01

    Purple photosynthetic bacteria are capable of generating cellular energy from several sources, including photosynthesis, respiration, and H2 oxidation. Under nutrient-limiting conditions, cellular energy can be used to assimilate carbon and nitrogen. This study provides the first evidence of a molecular link for the coregulation of nitrogenase and hydrogenase biosynthesis in an anoxygenic photosynthetic bacterium. We demonstrated that molybdenum nitrogenase biosynthesis is under the control o...

  12. Synthesis, characterization and DFT studies of 1, 1′-Bis(diphenylphosphino)ferrocene substituted diiron complexes: Bioinspired [FeFe] hydrogenase model complexes

    Indian Academy of Sciences (India)

    Sandeep Kaur-Ghumaan; A Sreenithya; Raghavan B Sunoj

    2015-03-01

    The reaction of [Fe2(CO)6(-toluene-3, 4-benzenedithiolate)] 1 and bidentate diphosphine, 1, 1′-bis(diphenylphosphino)ferrocene (dppf) has been studied. New complexes obtained have been characterized by various spectroscopic techniques as bioinspired models of the iron hydrogenase active site. The crystal structure of [Fe2(CO)5(1-dppfO)(-toluene-3, 4-benzenedithiolate)] 4 is reported.

  13. Bi2MoO6/Ni-Fe LDH复合材料的制备及可见光催化性能%Preparation and Visible Light Responsive Photocatalytic Activity of Bi2MoO6/Ni-Fe LDH Composites

    Institute of Scientific and Technical Information of China (English)

    曲婷; 黄强; 赵振波

    2015-01-01

    采用水热法和共沉淀法结合制备Bi2MoO6/Ni-Fe LDH复合材料,通过XRD、FT-IR、SEM、TEM、XPS和N2物理吸附等对样品的结构和形貌进行表征。以甲基橙、亚甲基蓝、罗丹明 B和苯酚为目标降解物,在可见光下进行复合材料的光催化性能测试,以降解甲基橙溶液为例研究复合材料的光催化反应机理。结果表明,复合材料的BET比表面积随着Ni-Fe LDH含量的增加而增大,光催化活性明显提高。Bi2MoO6/Ni-Fe LDH复合材料中Ni-Fe LDH的含量为4.5%时具有最好的光催化效果,可见光照射60 min,甲基橙的降解率达91%,较Bi2MoO6和Ni-Fe LDH分别提高52%和16%。Bi2MoO6/Ni-Fe LDH复合材料具有良好的稳定性,循环使用5次,甲基橙(MO)的降解率为88%。复合材料光催化降解甲基橙反应遵循一级反应动力学。%Bi2MoO6/Ni-Fe LDH composites were prepared by hydrothermal method and co-precipitation. The morphology and structure of the sample were characterized by XRD, FT-IR, SEM, TEM, XPS and N2-physisorption. Photocatalytic degradation activity and mechanism of the samples were investigated by the photocatalytic degrada-tion of methyl orange (MO), methylene blue, butyl rhodamine B and phenol under visible light irradiation. The re-sults showed that BET specific surface area of the composites increased with the LDH content increase. Photocata-lytic degradation activity of MO under visible irradiation exhibited significant enhancement. After visible light ir-radiation for 60 min, the Bi2MoO6/Ni-Fe LDH composites with LDH content of 4.5wt% showed the highest degra-dation rate of 91%, higher than that of Bi2MoO6and Ni-Fe LDH by 52% and 16%, respectively. And the composites photocatalytic degradation followed first-order reaction kinetics. The composites decolorizing rate still remained 88% after 5 times recycle, showing high catalytic stability.

  14. Cell-free H-cluster synthesis and [FeFe] hydrogenase activation: all five CO and CN⁻ ligands derive from tyrosine.

    Directory of Open Access Journals (Sweden)

    Jon M Kuchenreuther

    Full Text Available [FeFe] hydrogenases are promising catalysts for producing hydrogen as a sustainable fuel and chemical feedstock, and they also serve as paradigms for biomimetic hydrogen-evolving compounds. Hydrogen formation is catalyzed by the H-cluster, a unique iron-based cofactor requiring three carbon monoxide (CO and two cyanide (CN⁻ ligands as well as a dithiolate bridge. Three accessory proteins (HydE, HydF, and HydG are presumably responsible for assembling and installing the H-cluster, yet their precise roles and the biosynthetic pathway have yet to be fully defined. In this report, we describe effective cell-free methods for investigating H-cluster synthesis and [FeFe] hydrogenase activation. Combining isotopic labeling with FTIR spectroscopy, we conclusively show that each of the CO and CN⁻ ligands derive respectively from the carboxylate and amino substituents of tyrosine. Such in vitro systems with reconstituted pathways comprise a versatile approach for studying biosynthetic mechanisms, and this work marks a significant step towards an understanding of both the protein-protein interactions and complex reactions required for H-cluster assembly and hydrogenase maturation.

  15. Identification of an Isothiocyanate on the HypEF Complex Suggests a Route for Efficient Cyanyl-Group Channeling during [NiFe]-Hydrogenase Cofactor Generation.

    Directory of Open Access Journals (Sweden)

    Sven T Stripp

    Full Text Available [NiFe]-hydrogenases catalyze uptake and evolution of H2 in a wide range of microorganisms. The enzyme is characterized by an inorganic nickel/ iron cofactor, the latter of which carries carbon monoxide and cyanide ligands. In vivo generation of these ligands requires a number of auxiliary proteins, the so-called Hyp family. Initially, HypF binds and activates the precursor metabolite carbamoyl phosphate. HypF catalyzes removal of phosphate and transfers the carbamate group to HypE. In an ATP-dependent condensation reaction, the C-terminal cysteinyl residue of HypE is modified to what has been interpreted as thiocyanate. This group is the direct precursor of the cyanide ligands of the [NiFe]-hydrogenase active site cofactor. We present a FT-IR analysis of HypE and HypF as isolated from E. coli. We follow the HypF-catalyzed cyanation of HypE in vitro and screen for the influence of carbamoyl phosphate and ATP. To elucidate on the differences between HypE and the HypEF complex, spectro-electrochemistry was used to map the vibrational Stark effect of naturally cyanated HypE. The IR signature of HypE could ultimately be assigned to isothiocyanate (-N=C=S rather than thiocyanate (-S-C≡N. This has important implications for cyanyl-group channeling during [NiFe]-hydrogenase cofactor generation.

  16. Effect of the culture media optimization, pH and temperature on the biohydrogen production and the hydrogenase activities by Klebsiella pneumoniae ECU-15.

    Science.gov (United States)

    Xiao, Yan; Zhang, Xu; Zhu, Minglong; Tan, Wensong

    2013-06-01

    The low yield of the biohydrogen production is the main constraint for its industrialization process. In order to improve its production, medium compositions of the hydrogen fermentation by Klebsiella pneumoniae ECU-15 were optimized through the response surface methodology (RSM). Experimental results showed that the optimum hydrogen production of 5363.8 ml/L was obtained when the concentration of glucose, the ammonium sulfate and the trace elements were 35.62 g/L, 2.78 g/L and 23.15 ml/L at temperature 37.0°C, pH 6.0. H2 evolving hydrogenase was greatly enhanced by the optimization of the medium compositions. The activity of H2 evolving hydrogenase increased with the temperature, and decreased with the pH, while the activity of the uptake hydrogenase increased with the temperature and the pH. So the biohydrogen production process of the K. pneumoniae ECU-15 was the comprehensive results of the evolution hydrogen process and the uptake hydrogen process.

  17. Electroanalytical Assessment of the Effect of Ni:Fe Stoichiometry and Architectural Expression on the Bifunctional Activity of Nanoscale NiyFe1-yOx.

    Science.gov (United States)

    Ko, Jesse S; Chervin, Christopher N; Vila, Mallory N; DeSario, Paul A; Parker, Joseph F; Long, Jeffrey W; Rolison, Debra R

    2017-09-19

    Electrocatalysis of the oxygen evolution reaction (OER) and oxygen reduction reaction (ORR) was assessed for a series of Ni-substituted ferrites (NiyFe1-yOx, where y = 0.1 to 0.9) as expressed in porous, high-surface-area forms (ambigel and aerogel nanoarchitectures). We then correlate electrocatalytic activity with Ni:Fe stoichiometry as a function of surface area, crystallite size, and free volume. In order to ensure in-series comparisons, calcination at 350 °C/air was necessary to crystallize the respective NiyFe1-yOx nanoarchitectures, which index to the inverse spinel structure for Fe-rich materials (y ≤ 0.33), rock salt for the most Ni-rich material (y = 0.9), and biphasic for intermediate stoichiometry (0.5 ≤ y ≤ 0.67). In the intermediate Ni:Fe stoichiometric range (0.33 ≤ y ≤ 0.67), the OER current density at 390 mV increases monotonically with increasing Ni content and increasing surface area, but with different working curves for ambigels versus aerogels. At a common stoichiometry within this range, ambigels and aerogels yield comparable OER performance, but do so by expressing larger crystallite size (ambigel) versus higher surface area (aerogel). Effective OER activity can be achieved without requiring supercritical-fluid extraction as long as moderately high surface area, porous materials can be prepared. We find improved OER performance (η decreases from 390 to 373 mV) for Ni0.67Fe0.33Ox aerogel heat-treated at 300 °C/Ar, owing to an increase in crystallite size (2.7 to 4.1 nm). For the ORR, electrocatalytic activity favors Fe-rich NiyFe1-yOx materials; however, as the Ni-content increases beyond y = 0.5, a two-electron reduction pathway is still exhibited, demonstrating that bifunctional OER and ORR activity may be possible by choosing a nickel ferrite nanoarchitecture that provides high OER activity with sufficient ORR activity. Assessing the catalytic activity requires an appreciation of the multivariate interplay among Ni:Fe

  18. Characterization of the urease gene cluster from Rhizobium leguminosarum bv. viciae.

    Science.gov (United States)

    Toffanin, Annita; Cadahia, Esther; Imperial, Juan; Ruiz-Argüeso, Tomás; Palacios, Manuel

    2002-04-01

    Moderate levels of urease activity (ca. 300 mU mg(-1)) were detected in Rhizobium leguminosarum bv. viciae UPM791 vegetative cells. This activity did not require urea for induction and was partially repressed by the addition of ammonium into the medium. Lower levels of urease activity (ca. 100 mU mg(-1)) were detected also in pea bacteroids. A DNA region of ca. 9 kb containing the urease structural genes ( ureA, ureB and ureC), accessory genes ( ureD, ureE, ureF, and ureG), and five additional ORFs ( orf83, orf135, orf207, orf223, and orf287) encoding proteins of unknown function was sequenced. Three of these ORFs ( orf83, orf135 and orf207) have a homologous counterpart in a gene cluster from Sinorhizobium meliloti, reported to be involved in urease and hydrogenase activities. R. leguminosarum mutant strains carrying Tn 5 insertions within this region exhibited a urease-negative phenotype, but induced wild-type levels of hydrogenase and nitrogenase activities in bacteroids. orf287 encodes a potential transmembrane protein with a C-terminal GGDEF domain. A mutant affected in orf287 exhibited normal levels of urease activity in culture cells. Experiments aimed at cross-complementing Ni-binding proteins required for urease and hydrogenase synthesis (UreE and HypB, respectively) indicated that these two proteins are not functionally interchangeable in R. leguminosarum.

  19. Fractionation of Sulfur Isotopes by Desulfovibrio vulgaris Mutants Lacking Periplasmic Hydrogenases or the Type I Tetraheme Cytochrome c3

    Science.gov (United States)

    Sim, M.; Ono, S.; Bosak, T.

    2012-12-01

    A large fraction of anaerobic mineralization of organic compounds relies on microbial sulfate reduction. Sulfur isotope fractionation by these microbes has been widely used to trace the biogeochemical cycling of sulfur and carbon, but intracellular mechanisms behind the wide range of fractionations observed in nature and cultures are not fully understood. In this study, we investigated the influence of electron transport chain components on the fractionation of sulfur isotopes by culturing Desulfovibrio vulgaris Hildenborough mutants lacking hydrogenases or type I tetraheme cytochrome c3 (Tp1-c3). The mutants were grown both in batch and continuous cultures. All tested mutants grew on lactate or pyruvate as the sole carbon and energy sources, generating sulfide. Mutants lacking cytoplasmic and periplasmic hydrogenases exhibited similar growth physiologies and sulfur isotope fractionations to their parent strains. On the other hand, a mutant lacking Tp1-c3 (ΔcycA) fractionated the 34S/32S ratio more than the wild type, evolving H2 in the headspace and exhibiting a lower specific respiration rate. In the presence of high concentrations of pyruvate, the growth of ΔcycA relied largely on fermentation rather than sulfate reduction, even when sulfate was abundant, producing the largest sulfur isotope effect observed in this study. Differences between sulfur isotope fractionation by ΔcycA and the wild type highlight the effect of electron transfer chains on the magnitude of sulfur isotope fractionation. Because Tp1-c3 is known to exclusively shuttle electrons from periplasmic hydrogenases to transmembrane complexes, electron transfers in the absence of Tp1-c3 should bypass the periplasmic hydrogen cycling, and the loss of reducing equivalents in the form of H2 can impair the flow of electrons from organic acids to sulfur, increasing isotope fractionation. Larger fractionation by ΔcycA can inform interpretations of sulfur isotope data at an environmental scale as well

  20. Gene

    Data.gov (United States)

    U.S. Department of Health & Human Services — Gene integrates information from a wide range of species. A record may include nomenclature, Reference Sequences (RefSeqs), maps, pathways, variations, phenotypes,...

  1. Mesoporous Silica Matrix as a Tool for Minimizing Dipolar Interactions in NiFe2O4 and ZnFe2O4 Nanoparticles

    Directory of Open Access Journals (Sweden)

    Maider Virumbrales

    2017-06-01

    Full Text Available NiFe2O4 and ZnFe2O4 nanoparticles have been prepared encased in the MCM (Mobile Composition of Matter type matrix. Their magnetic behavior has been studied and compared with that corresponding to particles of the same composition and of a similar size (prepared and embedded in amorphous silica or as bare particles. This study has allowed elucidation of the role exerted by the matrix and interparticle interactions in the magnetic behavior of each ferrite system. Thus, very different superparamagnetic behavior has been found in ferrite particles of similar size depending on the surrounding media. Also, the obtained results clearly provide evidence of the vastly different magnetic behavior for each ferrite system.

  2. Magnetic field induced assembling of nanoparticles in ferrofluidic liquid thin films based on NiFe1-Fe2O4

    Indian Academy of Sciences (India)

    V S Abraham; S Swapna Nair; S Rajesh; U S Sajeev; M R Anantharaman

    2004-04-01

    Ultra fine precursors for ferrofluid synthesis, belonging to the series, NiFe1-Fe2O4 (where `’ varies from `0’ to `0.6’ in steps of 0.1), were synthesized. Ferrofluids based on these fine particles were prepared with oleic acid as surfactant and kerosene as carrier. Ferrofluidic thin films were made on glass substrates and magnetic field induced laser transmission was studied. The pattern exhibited by the films under the influence of a magnetic field was observed with the help of a CCD camera. The analysis of results confirms the chain formation of particles in the presence of an applied magnetic field and their saturation at higher applied fields.

  3. The NiFe2O4-MgFe2O4 series as electrode materials for electrochemical reduction of NOx

    DEFF Research Database (Denmark)

    Bræstrup, Frantz Radzik; Kammer Hansen, Kent

    2009-01-01

    -three-electrode setup in the temperature range of 400–600 °C. Cyclic voltammetry and electrochemical impedance spectroscopy were used to characterize the electrochemical behavior in 1% NO and 10% O2. Measurements show that NiFe2O4 has relatively high cathodic activity in both NO and O2, whereas MgFe2O4 shows much...... higher activity in NO compared to O2. MgFe2O4 was also measured with cyclic voltammetry in 1% NO2 and different gas mixtures of NO and O2 at 300 and 400 °C. Results show that the cathodic activities (−0.6 V) are relatively high with current ratios, , ranging from 10.1–167.7 and with a maximum at 400 °C...

  4. Results from 100 h electrolysis testing of NiFe2O4 based cermet as inert anode in aluminum reduction

    Institute of Scientific and Technical Information of China (English)

    LAI Yan-qing; TIAN Zhong-liang; LI Jie; YE Shao-long; LI Xin-zheng; LIU Ye-xiang

    2006-01-01

    A 100 mm diameter cup-shaped inert anode for aluminum electrolysis consisting of cermet 17Ni/83(10NiO-90NiFe2O4) was prepared and the operating performance was evaluated in a laboratory cell with the electrolyte CR2.3 and Al2O3 concentration 7.43% (mass fraction). The results indicate that no major operational difficulties are encountered during the testing which lasts for 101.5 h and the inert anode exhibits good general performances. The steady-state average concentration of impurity Ni in the bath is close to the solubility, however, the Fe concentration is lower than its solubility. The contents of the main contaminants for aluminum produced are Ni 0.128 8%, Fe 1.007 4%. The corrosion rate of inert anode under electrolysis conditions based on the content of impurity Ni in metal aluminum is approximately 8.51 mm/a.

  5. The NiFe2O4-MgFe2O4 series as electrode materials for electrochemical reduction of NOx

    DEFF Research Database (Denmark)

    Bræstrup, Frantz Radzik; Kammer Hansen, Kent

    2009-01-01

    -three-electrode setup in the temperature range of 400–600 °C. Cyclic voltammetry and electrochemical impedance spectroscopy were used to characterize the electrochemical behavior in 1% NO and 10% O2. Measurements show that NiFe2O4 has relatively high cathodic activity in both NO and O2, whereas MgFe2O4 shows much...... higher activity in NO compared to O2. MgFe2O4 was also measured with cyclic voltammetry in 1% NO2 and different gas mixtures of NO and O2 at 300 and 400 °C. Results show that the cathodic activities (−0.6 V) are relatively high with current ratios, , ranging from 10.1–167.7 and with a maximum at 400 °C...

  6. Hierachical Ni@Fe2O3 superparticles through epitaxial growth of γ-Fe2O3 nanorods on in situ formed Ni nanoplates

    Science.gov (United States)

    Tahir, Muhammad Nawaz; Herzberger, Jana; Natalio, Filipe; Köhler, Oskar; Branscheid, Robert; Mugnaioli, Enrico; Ksenofontov, Vadim; Panthöfer, Martin; Kolb, Ute; Frey, Holger; Tremel, Wolfgang

    2016-05-01

    One endeavour of nanochemistry is the bottom-up synthesis of functional mesoscale structures from basic building blocks. We report a one-pot wet chemical synthesis of Ni@γ-Fe2O3 superparticles containing Ni cores densely covered with highly oriented γ-Fe2O3 (maghemite) nanorods (NRs) by controlled reduction/decomposition of nickel acetate (Ni(ac)2) and Fe(CO)5. Automated diffraction tomography (ADT) of the Ni-Fe2O3 interface in combination with Mössbauer spectroscopy showed that selective and oriented growth of the γ-Fe2O3 nanorods on the Ni core is facilitated through the formation of a Fe0.05Ni0.95 alloy and the appearance of superstructure features that may reduce strain at the Ni-Fe2O3 interface. The common orientation of the maghemite nanorods on the Ni core of the superparticles leads to a greatly enhanced magnetization. After functionalization with a catechol-functional polyethylene glycol (C-PEG) ligand the Ni@γ-Fe2O3 superparticles were dispersible in water.One endeavour of nanochemistry is the bottom-up synthesis of functional mesoscale structures from basic building blocks. We report a one-pot wet chemical synthesis of Ni@γ-Fe2O3 superparticles containing Ni cores densely covered with highly oriented γ-Fe2O3 (maghemite) nanorods (NRs) by controlled reduction/decomposition of nickel acetate (Ni(ac)2) and Fe(CO)5. Automated diffraction tomography (ADT) of the Ni-Fe2O3 interface in combination with Mössbauer spectroscopy showed that selective and oriented growth of the γ-Fe2O3 nanorods on the Ni core is facilitated through the formation of a Fe0.05Ni0.95 alloy and the appearance of superstructure features that may reduce strain at the Ni-Fe2O3 interface. The common orientation of the maghemite nanorods on the Ni core of the superparticles leads to a greatly enhanced magnetization. After functionalization with a catechol-functional polyethylene glycol (C-PEG) ligand the Ni@γ-Fe2O3 superparticles were dispersible in water. Electronic supplementary

  7. Direct selenylation of mixed Ni/Fe metal-organic frameworks to NiFe-Se/C nanorods for overall water splitting

    Science.gov (United States)

    Xu, Bo; Yang, He; Yuan, Lincheng; Sun, Yiqiang; Chen, Zhiming; Li, Cuncheng

    2017-10-01

    Development of low-cost, highly active bifunctional catalyst for efficient overall water splitting based on earth-abundant metals is still a great challenging task. In this work, we report a NiFe-Se/C composite nanorod as efficient non-precious-metal electrochemical catalyst derived from direct selenylation of a mixed Ni/Fe metal-organic framework. The as-obtained catalyst requires low overpotential to drive 10 mA cm-2 for HER (160 mV) and OER (240 mV) in 1.0 M KOH, respectively, and its catalytic activity is maintained for at least 20 h. Moreover, water electrolysis using this catalyst achieves high water splitting current density of 10 mA cm-2 at cell voltage of 1.68 V.

  8. Magnetic and electrical characterization of nickel-rich NiFe thin films synthesized by atomic layer deposition and subsequent thermal reduction

    Science.gov (United States)

    Espejo, A. P.; Zierold, R.; Gooth, J.; Dendooven, J.; Detavernier, C.; Escrig, J.; Nielsch, K.

    2016-08-01

    Nickel-rich NiFe thin films (Ni92Fe8, Ni89Fe11 and Ni83Fe17) were prepared by combining atomic layer deposition (ALD) with a subsequent thermal reduction process. In order to obtain Ni x Fe1-x O y films, one ALD supercycle was performed according to the following sequence: m NiCp2/O3, with m = 1, 2 or 3, followed by one FeCp2/O3 cycle. The supercycle was repeated n times. The thermal reduction process in hydrogen atmosphere was investigated by in situ x-ray diffraction studies as a function of temperature. The metallic nickel iron alloy thin films were investigated and characterized with respect to crystallinity, morphology, resistivity, and magnetism. As proof-of-concept magnetic properties of an array of Ni83Fe17, close to the perfect Permalloy stoichiometry, nanotubes and an isolated tube were investigated.

  9. Models of the Ni-L and Ni-SIa States of the [NiFe]-Hydrogenase Active Site.

    Science.gov (United States)

    Chambers, Geoffrey M; Huynh, Mioy T; Li, Yulong; Hammes-Schiffer, Sharon; Rauchfuss, Thomas B; Reijerse, Edward; Lubitz, Wolfgang

    2016-01-19

    A new class of synthetic models for the active site of [NiFe]-hydrogenases are described. The Ni(I/II)(SCys)2 and Fe(II)(CN)2CO sites are represented with (RC5H4)Ni(I/II) and Fe(II)(diphos)(CO) modules, where diphos = 1,2-C2H4(PPh2)2(dppe) or cis-1,2-C2H2(PPh2)2(dppv). The two bridging thiolate ligands are represented by CH2(CH2S)2(2-) (pdt(2-)), Me2C(CH2S)2(2-) (Me2pdt(2-)), and (C6H5S)2(2-). The reaction of Fe(pdt)(CO)2(dppe) and [(C5H5)3Ni2]BF4 affords [(C5H5)Ni(pdt)Fe(dppe)(CO)]BF4 ([1a]BF4). Monocarbonyl [1a]BF4 features an S = 0 Ni(II)Fe(II) center with five-coordinated iron, as proposed for the Ni-SIa state of the enzyme. One-electron reduction of [1a](+) affords the S = 1/2 derivative [1a](0), which, according to density functional theory (DFT) calculations and electron paramagnetic resonance and Mössbauer spectroscopies, is best described as a Ni(I)Fe(II) compound. The Ni(I)Fe(II) assignment matches that for the Ni-L state in [NiFe]-hydrogenase, unlike recently reported Ni(II)Fe(I)-based models. Compound [1a](0) reacts with strong acids to liberate 0.5 equiv of H2 and regenerate [1a](+), indicating that H2 evolution is catalyzed by [1a](0). DFT calculations were used to investigate the pathway for H2 evolution and revealed that the mechanism can proceed through two isomers of [1a](0) that differ in the stereochemistry of the Fe(dppe)CO center. Calculations suggest that protonation of [1a](0) (both isomers) affords Ni(III)-H-Fe(II) intermediates, which represent mimics of the Ni-C state of the enzyme.

  10. Investigation of hydrogenase molecular marker to optimize hydrogen production from organic wastes and effluents of agro-food industries [abstract

    Directory of Open Access Journals (Sweden)

    Hamilton, C.

    2010-01-01

    Full Text Available In recent years policy makers have started looking for alternatives to fossil fuels, not only to counter the threat of global warming, but also to reduce the risk of overdependence on imported oil and gas supplies. By contrast with hydrocarbon fuels, hydrogen (H2, whether burned directly or used in fuel cells, is intrinsically a clean energy vector with near zero emission. However the main current method of producing hydrogen, steam reforming of methane, involves the release of large quantities of greenhouse gases. So although hydrogen already accounts for around 2% of world consumption of energy, its more widespread adoption is limited by several challenges. Therefore new processes are investigated, especially those using renewable raw material, e.g. woods and organic wastes, and/or involving microorganisms. Indeed, for some algae and bacteria, the generation of molecular hydrogen is an essential part of their energy metabolism. The approach with the greatest commercial potential is fermentative hydrogen generation (dark fermentation by bacteria from the Clostridium genus. This biological process, as a part of the methane-producing anaerobic digestion process, is very promising since it allows the production of hydrogen from a wide variety of renewable resources such as carbohydrate waste from the agricultural and agro-food industries or processed urban waste and sewage. To date most publications on hydrogen production by Clostridium strains have focused on the effects of operating parameters (such as temperature, pH, dilution rate, etc.. We now need to extend this knowledge by identifying and monitoring the various different metabolic agents involved in high H2 activity. Consequently the aim of this research at the CWBI in the University of Liege is to investigate the role of [Fe] hydrogenases, the key enzymes that remove excess electrons accumulating during fermentation. Clostridium butyricum CWBI1009, the strain used for these investigations

  11. Interfacial characteristics and multiferroic properties of ion-doped BiFeO3/NiFe2O4 thin films

    Science.gov (United States)

    Guo, Meiyou; Tan, Guoqiang; Zheng, Yujuan; Liu, Wenlong; Ren, Huijun; Xia, Ao

    2017-05-01

    Multi-ion doped BiFeO3/NiFe2O4 bilayered thin films were successfully prepared on fluorine-doped SnO2/glass (SnO2:F) substrates by sol-gel method. The crystalline structure, leakage current, interfacial characteristics, and multiferroic properties were investigated in detail. The results of Rietveld refinement showed that the structure of BSrSFMC layer is transformed from rhombohedral to tetragonal structure by the means of ion-doping. The difference of leakage current density of the BSrSFMC/NiFe2O4 (NFO) bilayered films of the -40 V to 40 V and 40 V to -40 V are 0.32 × 10-5 and 1.13 × 10-5 A/cm2, respectively. It was observed that there are obvious interface effects between BSrSFMC and NFO layers, which will cause the accumulation of space charges and the establishment of built-in internal electric field (EI) at the interface. Therefore, different EI directions will affect the dipoles reversal and migration of carriers in the BSrSFMC layer, which will result in different values of transient current with the same applied voltage in the opposite directions. The larger coercive field (Ec ˜ 750 kV/cm) of BSrSFMC/NFO film indicated that there is a tensile stress at the interface between BSrSFMC and NFO layers, making the polarization difficult. These results showed that the above interesting phenomena of the J-V are closely related to the interface effects between the layer of BiFeO3 and NiFe2O4.

  12. Experimental Investigations on Pulsed Nd:YAG Laser Welding of C17300 Copper-Beryllium and 49Ni-Fe Soft Magnetic Alloys

    Science.gov (United States)

    Mousavi, S. A. A. Akbari; Ebrahimzadeh, H.

    2011-01-01

    Copper-beryllium and soft magnetic alloys must be joined in electrical and electro-mechanical applications. There is a high difference in melting temperatures of these alloys which cause to make the joining process very difficult. In addition, copper-beryllium alloys are of age hardenable alloys and precipitations can brittle the weld. 49Ni-Fe alloy is very hot crack sensitive. Moreover, these alloys have different heat transfer coefficients and reflection of laser beam in laser welding process. Therefore, the control of welding parameters on the formation of adequate weld puddle composition is very difficult. Laser welding is an advanced technique for joining of dissimilar materials since it can precisely control and adjust the welding parameters. In this study, a 100W Nd:YAG pulsed laser machine was used for joining 49Ni-Fe soft magnetic to C17300 copper-beryllium alloys. Welding of samples was carried out autogenously by changing the pulse duration, diameter of beam, welding speed, voltage and frequency. The spacing between samples was set to almost zero. The ample were butt welded. It was required to apply high voltage in this study due to high reflection coefficient of copper alloys. Metallography, SEM analysis, XRD and microhardness measurement was used for survey of results. The results show that the weld strength depends upon the chemical composition of the joints. To change the wells composition and heat input of the welds, it was attempted to deviate the laser focus away from the weld centerline. The best strength was achieved by deviation of the laser beam away about 0.1mm from the weld centerline. The result shows no intermetallic compounds if the laser beam is deviated away from the joint.

  13. Exploring the microbially-mediated soil H2 sink: A lab-based study of the physiology and related H2 consumption of isolates from the Harvard Forest

    Science.gov (United States)

    Rao, D.; Meredith, L. K.; Bosak, T.; Hansel, C. M.; Ono, S.; Prinn, R. G.

    2012-12-01

    Atmospheric hydrogen (H2) is a secondary greenhouse gas because it attenuates the removal of methane (CH4) from the atmosphere. The largest and most uncertain term in the H2 biogeochemical cycle, microbe-mediated soil uptake, is responsible for about 80% of Earth's tropospheric H2 sink. Recently, the first H2-oxidizing soil microorganisms were discovered (genus Streptomyces) whose low-threshold, high-affinity NiFe-hydrogenase functions at ambient H2 levels (approx. 530 ppb). To better understand the ecological function of this hydrogenase, we conducted a controlled laboratory study of the H2 uptake behavior in accordance with the complex life cycle development of the streptomycetes. Several strains of the genus Streptomyces containing a high-affinity NiFe- hydrogenase were isolated from soil at the Harvard Forest. The presence of this hydrogenase, detected by PCR amplification of the hydrogenase large subunit, predicted H2 uptake behavior in wild-type streptomycetes and in phylogenetically different organisms containing more distantly related versions of the gene. H2 uptake depended on the streptomyces' life cycle, reaching a maximum during spore formation. These findings reveal connections between environmental conditions, organismal life cycle, and H2 uptake. With the rise of H2-based energy sources and a potential change in the tropospheric concentration of H2, understanding the sources and sinks of this trace gas is important for the future.

  14. Immobilization of hydrogenase on carbon nanotube polyelectrolytes as heterogeneous catalysts for electrocatalytic interconversion of protons and hydrogen

    Science.gov (United States)

    Liu, Jiang; Wu, Wen-Jie; Fang, Fang; Zorin, Nikolay A.; Chen, Meng; Qian, Dong-Jin

    2016-08-01

    Immobilization of active enzymes on the surfaces of electrodes and nanomaterials is important in the fields of bioscience, and biotechnology. In this study, we investigated electrocatalytic properties of the interconversion of protons and hydrogen by means of hydrogenase (H2ase)-functionalized carbon nanotube polyelectrolyte composites. Multiwalled carbon nanotube polyelectrolytes (MWNT-PEs) were synthesized through a diazonium and an addition reaction with poly(4-vinylpyridine) (P4VP), followed by another addition reaction with either methyl iodide (CH3I) or N-methyl- N'-benzyl bromide bipyridinium (VBenBr) to produce MWNT-P4VPMe or MWNT-P4VPBenV polyelectrolytes, respectively. The MWNT-PE@H2ase bio-nanocomposites were then prepared by means of MWNT-PEs as substrates to bind with H2ase. The redox current density of the MWNT-PE@H2ase-modified electrodes increased with a decrease in pH values of the Ar-saturated electrolyte solution owing to the catalytic reduction of protons (H2 production); further, it increased with the increasing pH values of the H2-saturated solution owing to the catalytic oxidation of hydrogen. The reversible color change between blue-colored and colorless viologen (catalyzed by the MWNT-PE@H2ase bio-nanocomposites) suggested that they may be developed as nano-biosensors for molecular H2. The as-synthesized bio-nanocomposites showed strong long-term stability and high bioactivity.

  15. Iron Hydride Detection and Intramolecular Hydride Transfer in a Synthetic Model of Mono-Iron Hydrogenase with a CNS Chelate.

    Science.gov (United States)

    Durgaprasad, Gummadi; Xie, Zhu-Lin; Rose, Michael J

    2016-01-19

    We report the identification and reactivity of an iron hydride species in a synthetic model complex of monoiron hydrogenase. The hydride complex is derived from a phosphine-free CNS chelate that includes a Fe-C(NH)(═O) bond (carbamoyl) as a mimic of the active site iron acyl. The reaction of [((O═)C(HN)N(py)S(Me))Fe(CO)2(Br)] (1) with NaHBEt3 generates the iron hydride intermediate [((O═)C(HN)N(py)S(Me))Fe(H)(CO)2] (2; δFe-H = -5.08 ppm). Above -40 °C, the hydride species extrudes CH3S(-) via intramolecular hydride transfer, which is stoichiometrically trapped in the structurally characterized dimer μ2-(CH3S)2-[((O═)C(HN)N(Ph))Fe(CO)2]2 (3). Alternately, when activated by base ((t)BuOK), 1 undergoes desulfurization to form a cyclometalated species, [((O═)C(NH)NC(Ph))Fe(CO)2] (5); derivatization of 5 with PPh3 affords the structurally characterized species [((O═)C(NH)NC)Fe(CO)(PPh3)2] (6), indicating complex 6 as the common intermediate along each pathway of desulfurization.

  16. Electrochemistry of metalloproteins: protein film electrochemistry for the study of E. coli [NiFe]-hydrogenase-1.

    Science.gov (United States)

    Evans, Rhiannon M; Armstrong, Fraser A

    2014-01-01

    Protein film electrochemistry is a technique which allows the direct control of redox-active enzymes, providing particularly detailed information on their catalytic properties. The enzyme is deposited onto a working electrode tip, and through control of the applied potential the enzyme activity is monitored as electrical current, allowing for direct study of inherent activity as electrons are transferred to and from the enzyme redox center(s). No mediators are used. Because the only enzyme present in the experiment is bound at the electrode surface, gaseous and liquid phase inhibitors can be introduced and removed whilst the enzyme remains in situ. Potential control means that kinetics and thermodynamics are explored simultaneously; the kinetics of a reaction can be studied as a function of potential. Steady-state catalytic rates are observed directly as current (for a given potential) and non-steady-state rates (such as interconversions between different forms of the enzyme) are observed from the change in current with time. The more active the enzyme, the higher the current and the better the signal-to-noise. In this chapter we outline the practical aspects of PFE for studying electroactive enzymes, using the Escherichia coli [NiFe]-hydrogenase 1 (Hyd-1) as an example.

  17. 可溶性阳极电刷镀纳米晶Ni-Fe合金镀层的退火再强化%Annealing hardening of nanocrystalline Ni-Fe alloy coatings synthesized by brush plating using soluble anode

    Institute of Scientific and Technical Information of China (English)

    戴品强; 陈晓文; 项忠楠; 许伟长

    2009-01-01

    Nanocrystalline Ni-Fe alloy coatings were synthesized by brush plating using soluble anode. Microhardness test and techniques of XRD, SEM and TEM were applied to characterize the microstructures and mechanical properties of Ni-Fe alloy coatings. The results show that the microhardness of Ni-Fe alloy coatings increases with increasing annealing temperature and reaches a peak value at 200 ℃, showing a significant hardening during annealing. With further increasing annealing temperature, the microhardness decreases gradually, but the microhardness of the coating after being annealed at 400 ℃ is still as high as that of the as-deposited sample. The abnormal grain growth is not observed during annealing of Ni-Fe alloy coatings, which indicates a better thermal stability.%采用可溶性Ni阳极电刷镀方法制备纳米晶Ni-Fe合金镀层,利用XRD、SEM、TEM、显微硬度计等测试方法分析低温退火对镀层结构和性能的影响.结果表明:纳米晶Ni-Fe合金镀层的硬度随退火温度的升高而提高,在200 ℃时达到最大值,存在明显的退火再强化;继续提高退火温度导致镀层硬度降低;400 ℃退火后的镀层硬度与镀态的接近;纳米晶Ni-Fe合金镀层退火过程没有出现晶粒异常长大,表现出比纯Ni镀层更高的热稳定性.

  18. Synthesis and Characterization of Spongy NiFe2O4 Clusters%海绵状NiFe2O4团簇的合成与表征

    Institute of Scientific and Technical Information of China (English)

    王晶; 杨巧珍; 董平; 张俊玲

    2011-01-01

    Spongy NiFe2O4 clusters were synthesized via solvothermal process with poly (vinylpyrrolidone) (PVP) and hexamethylenetetramine (HMTA) as additives. The samples were characterized by XRD, VSM, SEM, TEM,BET and ICP. The results show that when PVP and HMTA are added to the reaction system together, the as-synthesized spongy NiFe2O4 clusters have spinel crystal structure with small crystal size, their saturation magnetization is 57. 30emu/g and specific surface area and pore volume are 91. 57m2/g and 0. 26cm3/g, respectively.%以乙醇-水为混合溶剂,六亚甲基四胺(HMTA)与聚乙烯吡咯烷酮(PVP)为添加剂,通过溶剂热法合成海绵状的镍铁氧体( NiFe2O4)团簇,并对其进行了XRD、VSM、SEM、TEM、BET与ICP表征.结果表明,在反应体系中同时引入PVP与HMTA时,合成的海绵状NiFe2O4团簇具有尖晶石晶体结构,晶粒均匀细小,比饱和磁化强度为57.30emu/g,比表面积与孔容分别为91.57m2/g、0.26cm3/g.

  19. A hybrid photocatalytic system comprising ZnS as light harvester and an [Fe(2)S(2)] hydrogenase mimic as hydrogen evolution catalyst.

    Science.gov (United States)

    Wen, Fuyu; Wang, Xiuli; Huang, Lei; Ma, Guijun; Yang, Jinhui; Li, Can

    2012-05-01

    Photo opportunity: A highly efficient and stable hybrid artificial photosynthetic H(2) evolution system is assembled by using a semiconductor (ZnS) as light-harvester and an [Fe(2)S(2)] hydrogenase mimic ([(μ-SPh-4-NH(2) )(2) Fe(2) (CO)(6)]) as catalyst for H(2) evolution. Photocatalytic H(2) production is achieved with more than 2607 turnovers (based on [Fe(2)S(2)]) and an initial turnover frequency of 100 h(-1) through the efficient transfer of photogenerated electrons from ZnS to the [Fe(2)S(2)] complex.

  20. Immobilization of FeFe hydrogenase mimics onto carbon and gold electrodes by controlled aryldiazonium salt reduction: an electrochemical, XPS and ATR-IR study

    Energy Technology Data Exchange (ETDEWEB)

    Le Goff, Alan; Metaye, Romain; Moggia, Fabrice; Jousselme, Bruno; Palacin, Serge [CEA, IRAMIS, SPCSI, Chemistry of Surfaces and Interfaces group, F-91191 Gif sur Yvette Cedex (France); Artero, Vincent; Razavet, Mathieu; Tran, Phong D.; Fontecave, Marc [Laboratoire de Chimie et Biologie des Metaux, Universite Joseph Fourier, CNRS UMR 5249, CEA DSV/iRTSV, 17 rue des Martyrs, F-38054 Grenoble cedex 9 (France)

    2010-10-15

    A dithiolate-bridged hexacarbonyldiiron complex was synthesized from the reaction of the N-hydroxysuccinimide (NHS) ester of lipoic acid with Fe{sub 3}(CO){sub 12} in toluene. This mimic of the active site of FeFe hydrogenases could be covalently attached, using an NHS ester route, to carbon or gold electrode first decorated with amino functions. Once grafted this complex catalyzes hydrogen electro-evolution under strongly acidic conditions but is rapidly inactivated. We could evidence that the activity loss was due to the elimination of the carbonyl ligands rather than a leaching of the catalyst as a consequence of hydrolysis of the amide linkages. (author)

  1. New redox states observed in [FeFe] hydrogenases reveal redox coupling within the H-cluster.

    Science.gov (United States)

    Adamska-Venkatesh, Agnieszka; Krawietz, Danuta; Siebel, Judith; Weber, Katharina; Happe, Thomas; Reijerse, Edward; Lubitz, Wolfgang

    2014-08-13

    Active [FeFe] hydrogenases can be obtained by expressing the unmaturated enzyme in Escherichia coli followed by incubation with a synthetic precursor of the binuclear [2Fe] subcluster, namely: [NEt4]2[Fe2(adt)(CO)4(CN)2] (adt = [S-CH2-NH-CH2-S](2-)). The binuclear subsite Fe2(adt)(CO)3(CN)2 is attached through a bridging cysteine side chain to a [4Fe-4S] subcluster already present in the unmaturated enzyme thus yielding the intact native "H-cluster". We present FTIR electrochemical studies of the [FeFe] hydrogenase from Chlamydomonas reinhardtii, CrHydA1, maturated with the precursor of the native cofactor [Fe2(adt)(CO)4(CN)2](2-) as well as a non-natural variant [Fe2(pdt)(CO)4(CN)2](2-) in which the bridging amine functionality is replaced by CH2. The obtained active enzyme CrHydA1(adt) shows the same redox states in the respective potential range as observed for the native system (E(ox/red) = -400 mV, E(red/sred) = -470 mV). For the Hox → Hred transition the reducing equivalent is stored on the binuclear part, ([4Fe-4S](2+)Fe(II)Fe(I) → [4Fe-4S](2+)Fe(I)Fe(I)), while the Hred → Hsred transition is characterized by a reduction of the [4Fe-4S] part of the H-cluster ([4Fe-4S](2+)Fe(I)Fe(I) → [4Fe-4S](+)Fe(I)Fe(I)). A similar transition is reported here for the CO inhibited state of the H-cluster: ([4Fe-4S](2+)Fe(I)Fe(II)CO → [4Fe-4S](+)Fe(I)Fe(II)CO). An FTIR electrochemical study of the inactive variant with the pdt ligand, CrHydA1(pdt), identified two redox states H(pdt)-ox and H(pdt)-"red". Both EPR and FTIR spectra of H(pdt)-ox are virtually identical to those of the H(adt)-ox and the native Hox state. The H(pdt)-"red" state is also characterized by a reduced [4Fe-4S] subcluster. In contrast to CrHydA1(adt), the H(pdt)-ox state of CrHydA1(pdt) is stable up to rather high potentials (+200 mV). This study demonstrates the distinct redox coupling between the two parts of the H-cluster and confirms that the [4Fe-4S]H subsite is also redox active and as

  2. Characterization of Photochemical Processes for H2 Production by CdS Nanorod-[FeFe] Hydrogenase Complexes

    Energy Technology Data Exchange (ETDEWEB)

    Brown, K. A.; Wilker, M. B.; Boehm, M.; Dukovic, G.; King, P. W.

    2012-03-28

    We have developed complexes of CdS nanorods capped with 3-mercaptopropionic acid (MPA) and Clostridium acetobutylicum [FeFe]-hydrogenase I (CaI) that photocatalyze reduction of H{sup +} to H{sub 2} at a CaI turnover frequency of 380-900 s{sup -1} and photon conversion efficiencies of up to 20% under illumination at 405 nm. In this paper, we focus on the compositional and mechanistic aspects of CdS:CaI complexes that control the photochemical conversion of solar energy into H{sub 2}. Self-assembly of CdS with CaI was driven by electrostatics, demonstrated as the inhibition of ferredoxin-mediated H{sub 2} evolution by CaI. Production of H{sub 2} by CdS:CaI was observed only under illumination and only in the presence of a sacrificial donor. We explored the effects of the CdS:CaI molar ratio, sacrificial donor concentration, and light intensity on photocatalytic H{sub 2} production, which were interpreted on the basis of contributions to electron transfer, hole transfer, or rate of photon absorption, respectively. Each parameter was found to have pronounced effects on the CdS:CaI photocatalytic activity. Specifically, we found that under 405 nm light at an intensity equivalent to total AM 1.5 solar flux, H{sub 2} production was limited by the rate of photon absorption ({approx}1 ms{sup -1}) and not by the turnover of CaI. Complexes were capable of H{sub 2} production for up to 4 h with a total turnover number of 106 before photocatalytic activity was lost. This loss correlated with inactivation of CaI, resulting from the photo-oxidation of the CdS capping ligand MPA.

  3. Model of the iron hydrogenase active site covalently linked to a ruthenium photosensitizer: synthesis and photophysical properties.

    Science.gov (United States)

    Ott, Sascha; Borgström, Magnus; Kritikos, Mikael; Lomoth, Reiner; Bergquist, Jonas; Akermark, Björn; Hammarström, Leif; Sun, Licheng

    2004-07-26

    A model of the iron hydrogenase active site with the structure [(mu-ADT)Fe2(CO)6] (ADT = azadithiolate (S-CH2-NR-CH2-S), (2: R = 4-bromophenyl, 3: R = 4-iodophenyl)) has been assembled and covalently linked to a [Ru(terpy)2]2+ photosensitizer. This trinuclear complex 1 represents one synthetic step toward the realization of our concept of light-driven proton reduction. A rigid phenylacetylene tether has been incorporated as the linking unit in 1 in order to prolong the lifetime of the otherwise short-lived [Ru(terpy)2]2+ excited state. The success of this strategy is demonstrated by comparison of the photophysical properties of 1 and of two related ruthenium complexes bearing acetylenic terpyridine ligands, with those of [Ru(terpy)2]2+. IR and electrochemical studies reveal that the nitrogen heteroatom of the ADT bridge has a marked influence on the electronic properties of the [Fe2(CO)6] core. Using the Rehm-Weller equation, the driving force for an electron transfer from the photoexcited *[Ru(terpy)2]2+ to the diiron site in 1 was calculated to be uphill by 0.59 eV. During the construction of the trinuclear complex 1, n-propylamine has been identified as a decarbonylation agent on the [(mu-ADT)Fe2(CO)6] portion of the supermolecule. Following this procedure, the first azadithiolate-bridged dinuclear iron complex coordinated by a phosphine ligand [(mu-ADT)Fe2(CO)5PPh3] (4, R = 4-bromophenyl) was synthesized. Copyright 2004 American Chemical Society

  4. Coordination and conformational isomers in mononuclear iron complexes with pertinence to the [FeFe] hydrogenase active site.

    Science.gov (United States)

    Orthaber, Andreas; Karnahl, Michael; Tschierlei, Stefanie; Streich, Daniel; Stein, Matthias; Ott, Sascha

    2014-03-21

    A series of six mononuclear iron complexes of the type [Fe(X-bdt)(P(R)2N(Ph)2)(CO)] (P(R)2N(Ph)2 = 1,5-diaza-3,7-diphosphaoctane, bdt = benzenedithiolate with X = H, Cl2 or Me and R = Ph, Bn, Cyc or tert-Bu) was prepared. This new class of penta-coordinate iron complexes contains a free coordination site and a pendant base as essential structural features of the [FeFe]-hydrogenase active site. The bidentate nature of the P(R)2N(Ph)2 ligands was found to be crucial for the preferential formation of coordinatively unsaturated penta-coordinate complexes, which is supported by first principle calculations. IR-spectroscopic data suggest the presence of coordination isomers around the metal center, as well as multiple possible conformers of the P(R)2N(Ph)2 ligand. This finding is further corroborated by X-ray crystallographic and computational studies. (31)P{(1)H}-NMR- and IR-spectroscopic as well as electrochemical measurements show that the electronic properties of the complexes are strongly, and independently, influenced by the P-substituents at the P(R)2N(Ph)2 ligand as well as by modifications of the bdt bridge. These results illustrate the advantages of this modular platform, which allows independent and selective tuning through site specific modifications. Potential catalytic intermediates, namely singly reduced and protonated complexes, have been further investigated by spectroscopic methods and exhibit remarkable stability. Finally, their general capacity for electro-catalytic reduction of protons to molecular hydrogen was verified.

  5. Fabrication of a novel NiFe2O4/Zn-Al layered double hydroxide intercalated with EDTA composite and its adsorption behavior for Cr(VI) from aqueous solution

    Science.gov (United States)

    Deng, Lin; Shi, Zhou; Wang, Li; Zhou, Shiqing

    2017-05-01

    A novel magnetic NiFe2O4/Zn-Al layered double hydroxide intercalated with EDTA composite (NiFe2O4/ZnAl-EDTA LDH) was prepared through modified coprecipitation method and employed for adsorptive removal of Cr(VI) from aqueous solution. The adsorbents were characterized using Brunauer-Emmett-Teller (BET), scanning electron microscopy (SEM), transmission electron microscope (TEM), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), vibrating sample magnetometer (VSM), and X-ray photoelectron spectroscopy (XPS). Factors affecting the Cr(VI) adsorption, such as initial solution pH, adsorbent dosage, contact time, initial Cr(VI) concentration, temperature and coexisting ions, were studied systematically. Experiments results show that the magnetic NiFe2O4/ZnAl-EDTA LDH exhibits high adsorption efficiency within a wide pH range of 3.0-7.0 (R>80% at Cr(VI) concentration 50 mg L-1, contact time 360 min, and adsorbent dosage 2 g/L) and quick separation property. The adsorption process is fitted well with the Langmuir isotherm and pseudo-second-order kinetic model. The maximum theoretical adsorption capacity is found to be 77.22 mg g-1 at pH 6.0 and 318 K. The positive ΔH value (2.907 kJ mol-1) and negative ΔG value (-4.722 kJ mol-1) at 298-318 K reveals that the adsorption process is feasible, spontaneous and endothermic. Coexisting anions (PO43-, SO42-, CO32-, HCO3-, Cl-, and NO3-) have no significant effect on Cr(VI) removal. The mechanism study indicates that the adsorption of Cr(VI) onto NiFe2O4/ZnAl-EDTA LDH mainly involves electrostatic attraction and ion exchange interaction. It is interesting to note that a proportion of Cr(VI) adsorbed on the adsorbent surface are reduced to Cr(III) during the adsorption process. Results from this study demonstrate the potential utility of the magnetic NiFe2O4/ZnAl-EDTA LDH that could be developed into a viable technology for efficient removal of Cr(VI) from aqueous solution.

  6. Mononuclear [NiII(L)(P-(o-C6H4S)2(o-C6H4SH))]0/1- (L = thiolate, selenolate, PPh3, and Cl) complexes with intramolecular [Ni...S...H...S]/[Ni...H...S] interactions modulated by the coordinated ligand L: relevance to the [NiFe] hydrogenases.

    Science.gov (United States)

    Chen, Chien-Hong; Lee, Gene-Hsiang; Liaw, Wen-Feng

    2006-03-06

    The shift of the IR nu(S)(-)(H) frequency to lower wavenumbers for the series of complexes [Ni(II)(L)(P-(o-C6H4S)2(o-C6H4SH))]0/1- (L = PPh3 (1), Cl (6), Se-p-C6H4-Cl (5), S-C4H3S (7), SePh (4)) indicates that a trend of increasing electronic donation of the L ligands coordinated to the Ni(II) center promotes intramolecular [Ni-S...H-S] interactions. Compared to the Ni...S(H) distance, in the range of 3.609-3.802 A in complexes 1 and 4-7, the Ni...S(CH3) distances of 2.540 and 2.914 A observed in the [Ni(II)(PPh3)(P(o-C6H4S)2(o-C6H4-SCH3))] complexes (8a and 8b, two conformational isomers with the chemical shift of the thioether methyl group at delta 1.820 (-60 degrees C) and 2.109 ppm (60 degrees C) (C4D8O)) and the Ni...S(CH3) distances of 3.258 and 3.229 A found in the [Ni(II)(L)(P(o-C6H4S)2(o-C6H4-SCH3))]1- complexes (L = SPh (9), SePh (10)) also support the idea that the pendant thiol protons of the Ni(II)-thiol complexes 1/4-7 were attracted by both the sulfur of thiolate and the nickel. The increased basicity (electronic density) of the nickel center regulated by the monodentate ligand attracted the proton of the pendant thiol effectively and caused the weaker S...H bond. In addition, the pendant thiol interaction modes in the solid state (complexes 1a and 1b, Scheme 1) may be controlled by the solvent of crystallization. Compared to complex 1a, the stronger intramolecular [Ni-S...H-S] interaction (or a combination of [Ni-S...H-S]/[Ni...H-S] interactions) found in complexes 4-7 led to the weaker S-H bond strength and accelerated the oxidation (by O2) of complexes 4-7 to produce the [Ni(Y)(L)(P(o-C6H4S)3)]1- (L = Se-p-C6H4-Cl (11), SePh (12), S-C4H3S (13)) complexes.

  7. Direct Comparison of the Performance of a Bio-inspired Synthetic Nickel Catalyst and a [NiFe]-Hydrogenase, Both Covalently Attached to Electrodes

    Energy Technology Data Exchange (ETDEWEB)

    Rodriguez-Macia, Patricia; Dutta, Arnab; Lubitz, Wolfgang; Shaw, Wendy J.; Rudiger, Olaf

    2015-10-12

    The active site of hydrogenases has been a source of inspiration for the development of molecular catalysts. However, direct comparisons between molecular catalysts and enzymes have not been possible because different techniques are used to evaluate both types of catalysts, minimizing our ability to determine how far we’ve come in mimicking the impressive enzymatic performance. Here we directly compare the catalytic properties of the [Ni(PCy2NGly2)2]2+ complex with the [NiFe]-hydrogenase from Desulfobivrio vulgaris Miyazaki F (DvMF) immobilized to a functionalized electrode under identical conditions. At pH=7, the enzyme has higher performance in both activity and overpotential, and is more stable, while at low pH, the molecular catalyst outperforms the enzyme in all respects. The Ni complex also has increased tolerance to CO. This is the first direct comparison of enzymes and molecular complexes, enabling a unique understanding of the benefits and detriments of both systems, and advancing our understanding of the utilization of these bioinspired complexes in fuel cells. AD and WJS acknowledge the Office of Science Early Career Research Program through the US Department of Energy (US DOE), Office of Science, Office of Basic Energy Sciences (BES), and Pacific Northwest National Laboratory (PNNL). PNNL is operated by Battelle for the US DOE.

  8. Effects of Iron on Hydrogen-producing Capacity,Hydrogenase and NADH-fd Reductase Activities of a Fermentative Hydrogen-producing Bacterial Strain B49

    Institute of Scientific and Technical Information of China (English)

    Wang Xiangjing(王相晶); Ren Nanqi; Xiang Wensheng

    2004-01-01

    Iron plays an important role in hydrogen production, cell growth, hydrogenase and NADH-fd reductase activities of hydrogen-producing bacterial strain B49 (AF481148 in EMBL). At the end of fermentation from 10 g/L glucose, for the culture containing 10 mg/L FeSO4*7H2O the cell growth in terms of optical density (OD) at 600nm was 1.13, the ratio of ethanol amount (mg/L) to acetate amount (mg/L) was 1.55, and the accumulated hydrogen volume was 1816.3 ml H2/L culture; whereas for the culture of 80 mg/L FeSO4*7H2O OD600nm was increased to 1.34, the accumulated hydrogen volume was increased to 2360.5 ml H2/L culture, and the ratio of ethanol amount (mg/L) to acetate amount (mg/L) decreased to 1.31. Moreover, the iron addition to the medium at different fermentation time could affect hydrogen-producing ability. However, the later the addition time of FeSO4*7H2O was postponed, the less the effect on hydrogen evolution was. In the course of fermentation, the specific activities of hydrogenase and NADH-fd reductase of hydrogen-producing bacterial strain B49 decreased with the consumption of iron.

  9. Fabrication and electromagnetic properties of bio-based helical soft-core particles by way of Ni-Fe alloy electroplating

    Energy Technology Data Exchange (ETDEWEB)

    Lan Mingming, E-mail: lan_mingming@163.com [Bionic and Micro/Nano/Bio Manufacturing Technology Research Center, School of Mechanical Engineering and Automation, Beihang University, Beijing 100191 (China); Zhang Deyuan; Cai Jun; Zhang Wenqiang; Yuan Liming [Bionic and Micro/Nano/Bio Manufacturing Technology Research Center, School of Mechanical Engineering and Automation, Beihang University, Beijing 100191 (China)

    2011-12-15

    Ni-Fe alloy electroplating was used as a bio-limited forming process to fabricate bio-based helical soft-core ferromagnetic particles, and a low frequency vibration device was applied to the cathode to avoid microorganism (Spirulina platens) cells adhesion to the copper net during the course of plating. The morphologies and ingredients of the coated Spirulina cells were characterized using scanning electron microscopy and energy dispersive spectrometer. The complex permittivity and permeability of the samples containing the coated Spirulina cells before and after heat treatment were measured and investigated by a vector network analyzer. The results show that the Spirulina cells after plating keep their initial helical shape, and applying low frequency vibration to the copper net cathode in the plating process can effectively prevent agglomeration and intertwinement of the Spirulina cells. The microwave absorbing and electromagnetic properties of the samples containing the coated Spirulina cells particles with heat treatment are superior to those samples containing the coated Spirulina cells particles without heat treatment. - Highlights: > We used the microorganism cells as forming template to fabricate the bio-based helical soft-core ferromagnetic particles. > Microorganism selected as forming templates was Spirulina platens, which are of natural helical shape and have high aspect ratio. > Coated Spirulina cells were a kind lightweight ferromagnetic particle.

  10. NiFe layered double hydroxide/reduced graphene oxide nanohybrid as an efficient bifunctional electrocatalyst for oxygen evolution and reduction reactions

    Science.gov (United States)

    Zhan, Tianrong; Zhang, Yumei; Liu, Xiaolin; Lu, SiSi; Hou, Wanguo

    2016-11-01

    Highly active and low-cost bifunctional electrocatalysts for oxygen evolution and reduction reactions (OER and ORR) hold a heart position for the renewable energy technologies such as metal-air batteries and fuel cells. Here, we reported the synthesis of NiFe layered double hydroxide/reduced graphene oxide (NiFe-LDH/rGO) nanohybrid via the facile solvothermal method followed by chemical reduction. The template role of surfactant and the hybridization of rGO supplied the NiFe-LDH/rGO catalyst with a porous nanostructure and an enhanced conductivity, favoring both mass transport and charge communication of electrocatalytic reactions. The NiFe-LDH/rGO composite not only displayed highly efficient OER activity in alkaline solution with a low onset overpotential of 240 mV, but also only needed an overpotential of 250 mV to reach the 10 mA cm-2 current density. The NiFe-LDH/rGO nanohybrid also offered excellent ORR catalytic activity with onset potential at 0.796 V in alkaline media. The rotating-disk and rotating-ring-disk electrodes both revealed that the ORR on NiFe-LDH/rGO mainly involved a direct four-electron reaction pathways accompanying part of the two-electron process. The excellent bifunctional activity of the NiFe-LDH/rGO nanohybrid could be attributed to the synergistic effects of rGO and NiFe-LDH components due to the strongly coupled interactions.

  11. N-doped graphene layers encapsulated NiFe alloy nanoparticles derived from MOFs with superior electrochemical performance for oxygen evolution reaction

    Science.gov (United States)

    Feng, Yi; Yu, Xin-Yao; Paik, Ungyu

    2016-01-01

    Water splitting, an efficient approach for hydrogen production, is often hindered by unfavorable kinetics of oxygen evolution reaction (OER). In order to reduce the overpotential, noble metal oxides-based electrocatalysts like RuO2 and IrO2 are usually utilized. However, due to their scarcity, the development of cost-effective non-precious OER electrocatalysts with high efficiency and good stability is urgently required. Herein, we report a facile one-step annealing of metal-organic frameworks (MOFs) strategy to synthesize N-doped graphene layers encapsulated NiFe alloy nanoparticles (NiFe@C). Through tuning the nanoparticle size and calcination temperature, NiFe@C with an average size of around 16 nm obtained at 700 °C exhibits superior OER performance with an overpotential of only 281 mV at 10 mA cm−2 and high durability. The facile synthesis method and excellent electrochemical performance show great potential of NiFe@C in replacing the precious metal-based electrocatalysts in the OER. PMID:27658968

  12. Biochemical Characterization of HydF, a Scaffolding Enzyme, in the Synthesis of the Hydrogenase Active Site Metal Center: Implications Towards the Evolution of Biocatalysts from Mineral-based Components on Early Earth

    Science.gov (United States)

    Duffus, B. R.; Shepard, E. M.; McGlynn, S. E.; Bueling, A. L.; Winslow, M. A.; Peters, J. W.; Broderick, J. B.

    2010-04-01

    [FeFe]-hydrogenase active site biosynthesis utilizes radical chemistry on a scaffold protein whose ancestor may have been one of the earliest examples of a protein that couples the chemistry of an Fe-S peptide nest with a nucleotide binding nest.

  13. [Nitrogenase, hydrogenase and nitrate reductase activities, oxygen consumption, and ATP content in nodules formed by strains of Rhizobium leguminosarum 128C53 and 300 in symbiosis with pea plants].

    Science.gov (United States)

    Bedmar, E J; Olivares, J

    1986-10-01

    The nitrogenase activity, nitrate reductase activity and oxygen uptake as well as the hydrogen incorporation and ATP content were examined in the root nodules and bacteroids, respectively, formed by Rhizobium leguminosarum strains 128C53 (hydrogenase positive) and 300 (hydrogenase negative) in symbiosis with Pisum sativum plants grown in the presence of 2 mM KNO3. The strain 128C53 showed the greatest values for all parameters analyzed, except for the nitrate reductase activity, which was higher for the strain 300. Similarly, nodule nitrate reductase activity in strain 300 was greater than that in strain 128C53 when plants grew in the absence of combined nitrogen. In general, the highest values were obtained when determinations were made after 7 hours of plant illumination. However, the hydrogenase activity of strain 128C53 and the nitrate reductase activities of both strains increased with the light period, reaching a maximum after 14 hours of illumination. These results suggest that the benefits derived from the superior symbiotic properties and from the presence of hydrogenase activity in strain 128C53 could be counteracted by the higher rates of the nodule nitrate reductase activity in strain 300.

  14. Mechanism of H2 Production by Models for the [NiFe]-Hydrogenases: Role of Reduced Hydrides.

    Science.gov (United States)

    Ulloa, Olbelina A; Huynh, Mioy T; Richers, Casseday P; Bertke, Jeffery A; Nilges, Mark J; Hammes-Schiffer, Sharon; Rauchfuss, Thomas B

    2016-07-27

    The intermediacy of a reduced nickel-iron hydride in hydrogen evolution catalyzed by Ni-Fe complexes was verified experimentally and computationally. In addition to catalyzing hydrogen evolution, the highly basic and bulky (dppv)Ni(μ-pdt)Fe(CO)(dppv) ([1](0); dppv = cis-C2H2(PPh2)2) and its hydride derivatives have yielded to detailed characterization in terms of spectroscopy, bonding, and reactivity. The protonation of [1](0) initially produces unsym-[H1](+), which converts by a first-order pathway to sym-[H1](+). These species have C1 (unsym) and Cs (sym) symmetries, respectively, depending on the stereochemistry of the octahedral Fe site. Both experimental and computational studies show that [H1](+) protonates at sulfur. The S = 1/2 hydride [H1](0) was generated by reduction of [H1](+) with Cp*2Co. Density functional theory (DFT) calculations indicate that [H1](0) is best described as a Ni(I)-Fe(II) derivative with significant spin density on Ni and some delocalization on S and Fe. EPR spectroscopy reveals both kinetic and thermodynamic isomers of [H1](0). Whereas [H1](+) does not evolve H2 upon protonation, treatment of [H1](0) with acids gives H2. The redox state of the "remote" metal (Ni) modulates the hydridic character of the Fe(II)-H center. As supported by DFT calculations, H2 evolution proceeds either directly from [H1](0) and external acid or from protonation of the Fe-H bond in [H1](0) to give a labile dihydrogen complex. Stoichiometric tests indicate that protonation-induced hydrogen evolution from [H1](0) initially produces [1](+), which is reduced by [H1](0). Our results reconcile the required reductive activation of a metal hydride and the resistance of metal hydrides toward reduction. This dichotomy is resolved by reduction of the remote (non-hydride) metal of the bimetallic unit.

  15. Single-Amino Acid Modifications Reveal Additional Controls on the Proton Pathway of [FeFe]-Hydrogenase

    Energy Technology Data Exchange (ETDEWEB)

    Cornish, Adam J.; Ginovska, Bojana; Thelen, Adam; da Silva, Julio C. S.; Soares, Thereza A.; Raugei, Simone; Dupuis, Michel; Shaw, Wendy J.; Hegg, Eric L.

    2016-06-07

    The proton pathway of [FeFe]-hydrogenase is essential for enzymatic H2 production and oxidation and is composed of four residues and a modeled water molecule. Recently, a computational analysis of this pathway revealed that the solvent-exposed residue of the pathway (Glu282) could form hydrogen bonds to two residues outside of the pathway (Arg286 and Ser320), implicating that these residues could function in regulating proton transfer. Substituting Arg286 with leucine eliminates hydrogen bonding with Glu282 and results in a 2.5-fold enhancement in H2 production activity, suggesting that Arg286 serves an important role in controlling the rate of proton delivery. In contrast, substitution of Ser320 with alanine reduces the rate approximately 5-fold, implying that it either acts as a member of the pathway or influences Glu282 to enable proton transfer. Interestingly, QM/MM and molecular dynamics calculations indicate that Ser320 does not play an electronic or structural role. QM calculations also estimate that including Ser320 in the pathway does not significantly change the barrier to proton movement, providing further support for its role as a member of the proton pathway. While further studies are needed to quantify the role of Ser320, collectively, these data provide evidence that the enzyme scaffold plays a significant role in modulating the activity of the enzyme, demonstrating that the rate of intraprotein proton transfer can be accelerated, particularly in a non-biological context. This work was supported by the DOE Great Lakes Bioenergy Research Center (DOE BER Office of Science, DE-FC02-07ER64494). In addition, support from the DOE Office of Science Early Career Research Program through the Office of Basic Energy Sciences (WJS, BGP, SR) is gratefully acknowledged. Computational resources were provided at W. R. Wiley Environmental Molecular Science Laboratory (EMSL), a national scientific user facility sponsored by the Department of Energy’s Office of

  16. Profiling the hydA gene and hydA gene transcript levels of Clostridium butyricum during continuous, mixed-culture hydrogen fermentation

    Energy Technology Data Exchange (ETDEWEB)

    Tolvanen, Katariina E.S.; Koskinen, Perttu E.P.; Santala, Ville P.; Karp, Matti T. [Department of Chemistry and Bioengineering, Tampere University of Technology, P.O. Box 541, FI-33101 Tampere (Finland); Raussi, Hanna-Mari; Ylikoski, Alice I.; Hemmilae, Ilkka A. [PerkinElmer Life Sciences, Wallac Oy, P.O. Box 10, FI-20101 Turku (Finland)

    2008-10-15

    The purpose of the research was to investigate the hydrogenase (hydA) gene and the corresponding transcript levels of Clostridium butyricum during continuous hydrogen fermentation. A quantitative real-time PCR (qrt-PCR) method was developed to specifically target the C. butyricum hydrogenase gene and mRNA in samples collected from a continuous, mixed-culture bioreactor over a period of 30 days (operation days 114-144). The detection limit of the qrt-PCR was 3.9 x 10{sup 2}hydA copies and the linear range 3.9 x 10{sup 2}-3.9 x 10{sup 7}hydA copies. The results showed that after a re-inoculation of the bioreactor on day 120 the hydA copy numbers started to rise and stabilized after day 127. The number of hydA transcript continued to rise until day 142. The results demonstrate that this method is suitable for detecting the hydA gene and gene transcript levels of C. butyricum from bioreactor samples. The expression level of hydA gene changed during continuous operation and can, therefore, be a useful target for process performance monitoring. (author)

  17. The genome of Paenibacillus sabinae T27 provides insight into evolution, organization and functional elucidation of nif and nif-like genes.

    Science.gov (United States)

    Li, Xinxin; Deng, Zhiping; Liu, Zhanzhi; Yan, Yongliang; Wang, Tianshu; Xie, Jianbo; Lin, Min; Cheng, Qi; Chen, Sanfeng

    2014-08-27

    Most biological nitrogen fixation is catalyzed by the molybdenum nitrogenase. This enzyme is a complex which contains the MoFe protein encoded by nifDK and the Fe protein encoded by nifH. In addition to nifHDK, nifHDK-like genes were found in some Archaea and Firmicutes, but their function is unclear. We sequenced the genome of Paenibacillus sabinae T27. A total of 4,793 open reading frames were predicted from its 5.27 Mb genome. The genome of P. sabinae T27 contains fifteen nitrogen fixation (nif) genes, including three nifH, one nifD, one nifK, four nifB, two nifE, two nifN, one nifX and one nifV. Of the 15 nif genes, eight nif genes (nifB, nifH, nifD, nifK, nifE, nifN, nifX and nifV) and two non-nif genes (orf1 and hesA) form a complete nif gene cluster. In addition to the nif genes, there are nitrogenase-like genes, including two nifH-like genes and five pairs of nifDK-like genes. IS elements on the flanking regions of nif and nif-like genes imply that these genes might have been obtained by horizontal gene transfer. Phylogenies of the concatenated 8 nif gene (nifB, nifH, nifD, nifK, nifE, nifN, nifX and nifV) products suggest that P. sabinae T27 is closely related to Frankia. RT-PCR analysis showed that the complete nif gene cluster is organized as an operon. We demonstrated that the complete nif gene cluster under the control of σ70-dependent promoter enabled Escherichia coli JM109 to fix nitrogen. Also, here for the first time we demonstrated that unlike nif genes, the transcriptions of nifHDK-like genes were not regulated by ammonium and oxygen, and nifH-like or nifD-like gene could not restore the nitrogenase activity of Klebsiella pneumonia nifH- and nifD- mutant strains, respectively, suggesting that nifHDK-like genes were not involved in nitrogen fixation. Our data and analysis reveal the contents and distribution of nif and nif-like genes and contribute to the study of evolutionary history of nitrogen fixation in Paenibacillus. For the first time we

  18. Study of angular dependence of exchange bias and misalignment in uniaxial and unidirectional anisotropy in NiFe(111)/FeMn(111)/CoFeB(amorphous) stack

    Energy Technology Data Exchange (ETDEWEB)

    Singh, Braj Bhusan; Chaudhary, Sujeet, E-mail: sujeetc@physics.iitd.ac.in

    2015-07-01

    We report the investigation of the in-plane azimuthal angular dependence of the magnetization reversal in the ion beam sputtered exchanged biased NiFe(111)/FeMn(111)/CoFeB(amorphous) stack. Compared to the as-deposited case, the magnetic annealing resulted in 3 fold enhancement in exchange bias but decrease in coercivity. The observed cosine dependence of exchange biased CoFeB layer on the in-plane azimuthal angle of applied field is corroborated with Meiklejohn and Bean model. The training effect associated with the exchange bias showed unconventional increase in coercivity after first cycle of hysteresis loop, while the exchange bias decreases sharply, and for subsequent cycles the exchange bias follows the empirical relation based on the energy dissipation in the AF layer. The ferromagnetic resonance (FMR) measurements also exhibited the in-plane azimuthal angle dependence of the magnetic resonance field indicating that the uniaxial and unidirectional anisotropies are not collinear, although they lie in the same plane. However, no misalignment between the unidirectional anisotropy and the exchange bias direction is observed. The misalignment angle between the uniaxial and unidirectional anisotropy, as measured by FMR, is found to be 10° and 14° for CoFeB and NiFe, respectively. This misalignment is attributed to the interface roughness as revealed by x-ray reflectance measurements. - Highlights: • In-plane azimuthal angular dependence of the magnetization reversal in the ion beam sputtered exchanged biased NiFe(111)/FeMn(111)/ CoFeB(amorphous) stack. • The observed cosine dependence of exchange biased CoFeB layer on the in-plane azimuthal angle of applied field is corroborated with Meiklejohn and Bean model. • In-plane azimuthal angle dependence of the magnetic resonance field indicates that the uniaxial and unidirectional anisotropies are not collinear, although they lie in the same plane. • The misalignment angle between the uniaxial and

  19. From Fe3O4 /NiO bilayers to NiFe2O4 -like thin films through Ni interdiffusion

    Science.gov (United States)

    Kuschel, O.; Buß, R.; Spiess, W.; Schemme, T.; Wöllermann, J.; Balinski, K.; N'Diaye, A. T.; Kuschel, T.; Wollschläger, J.; Kuepper, K.

    2016-09-01

    Ferrites with (inverse) spinel structure display a large variety of electronic and magnetic properties, making some of them interesting for potential applications in spintronics. We investigate the thermally induced interdiffusion of Ni2 + ions out of NiO into Fe3O4 ultrathin films, resulting in off-stoichiometric nickel ferrite-like thin layers. We synthesized epitaxial Fe3O4 /NiO bilayers on Nb-doped SrTiO3(001) substrates by means of reactive molecular beam epitaxy. Subsequently, we performed an annealing cycle comprising three steps at temperatures of 400 ∘C , 600 ∘C , and 800 ∘C under an oxygen background atmosphere. We studied the changes of the chemical and electronic properties as result of each annealing step with help of hard x-ray photoelectron spectroscopy and found a rather homogeneous distribution of Ni and Fe cations throughout the entire film after the overall annealing cycle. For one sample we observed a cationic distribution close to that of the spinel ferrite NiFe2O4 . Further evidence comes from low-energy electron diffraction patterns indicating a spinel-type structure at the surface after annealing. Site- and element-specific hysteresis loops performed by x-ray magnetic circular dichroism uncovered the antiferrimagnetic alignment between the octahedral coordinated Ni2 + and Fe3 + ions and the Fe3 + ion in tetrahedral coordination. We find a quite low coercive field of 0.02 T, indicating a rather low defect concentration within the thin ferrite films.

  20. Hydrogen production at high Faradaic efficiency by a bio-electrode based on TiO2 adsorption of a new [FeFe]-hydrogenase from Clostridium perfringens.

    Science.gov (United States)

    Morra, Simone; Valetti, Francesca; Sarasso, Veronica; Castrignanò, Silvia; Sadeghi, Sheila J; Gilardi, Gianfranco

    2015-12-01

    The [FeFe]-hydrogenase CpHydA from Clostridium perfringens was immobilized by adsorption on anatase TiO2 electrodes for clean hydrogen production. The immobilized enzyme proved to perform direct electron transfer to and from the electrode surface and catalyses both H2 oxidation (H2 uptake) and H2 production (H2 evolution) with a current density for H2 evolution of about 2 mA cm(-1). The TiO2/CpHydA bioelectrode remained active for several days upon storage and when a reducing potential was set, H2 evolution occurred with a mean Faradaic efficiency of 98%. The high turnover frequency of H2 production and the tight coupling of electron transfer, resulting in a Faradaic efficiency close to 100%, support the exploitation of the novel TiO2/CpHydA stationary electrode as a powerful device for H2 production.

  1. X-ray crystallographic and EPR spectroscopic analysis of HydG, a maturase in [FeFe]-hydrogenase H-cluster assembly.

    Science.gov (United States)

    Dinis, Pedro; Suess, Daniel L M; Fox, Stephen J; Harmer, Jenny E; Driesener, Rebecca C; De La Paz, Liliana; Swartz, James R; Essex, Jonathan W; Britt, R David; Roach, Peter L

    2015-02-03

    Hydrogenases use complex metal cofactors to catalyze the reversible formation of hydrogen. In [FeFe]-hydrogenases, the H-cluster cofactor includes a diiron subcluster containing azadithiolate, three CO, and two CN(-) ligands. During the assembly of the H cluster, the radical S-adenosyl methionine (SAM) enzyme HydG lyses the substrate tyrosine to yield the diatomic ligands. These diatomic products form an enzyme-bound Fe(CO)x(CN)y synthon that serves as a precursor for eventual H-cluster assembly. To further elucidate the mechanism of this complex reaction, we report the crystal structure and EPR analysis of HydG. At one end of the HydG (βα)8 triosephosphate isomerase (TIM) barrel, a canonical [4Fe-4S] cluster binds SAM in close proximity to the proposed tyrosine binding site. At the opposite end of the active-site cavity, the structure reveals the auxiliary Fe-S cluster in two states: one monomer contains a [4Fe-5S] cluster, and the other monomer contains a [5Fe-5S] cluster consisting of a [4Fe-4S] cubane bridged by a μ2-sulfide ion to a mononuclear Fe(2+) center. This fifth iron is held in place by a single highly conserved protein-derived ligand: histidine 265. EPR analysis confirms the presence of the [5Fe-5S] cluster, which on incubation with cyanide, undergoes loss of the labile iron to yield a [4Fe-4S] cluster. We hypothesize that the labile iron of the [5Fe-5S] cluster is the site of Fe(CO)x(CN)y synthon formation and that the limited bonding between this iron and HydG may facilitate transfer of the intact synthon to its cognate acceptor for subsequent H-cluster assembly.

  2. Study of angular dependence of exchange bias and misalignment in uniaxial and unidirectional anisotropy in NiFe(111)/FeMn(111)/CoFeB(amorphous) stack

    Science.gov (United States)

    Singh, Braj Bhusan; Chaudhary, Sujeet

    2015-07-01

    We report the investigation of the in-plane azimuthal angular dependence of the magnetization reversal in the ion beam sputtered exchanged biased NiFe(111)/FeMn(111)/CoFeB(amorphous) stack. Compared to the as-deposited case, the magnetic annealing resulted in 3 fold enhancement in exchange bias but decrease in coercivity. The observed cosine dependence of exchange biased CoFeB layer on the in-plane azimuthal angle of applied field is corroborated with Meiklejohn and Bean model. The training effect associated with the exchange bias showed unconventional increase in coercivity after first cycle of hysteresis loop, while the exchange bias decreases sharply, and for subsequent cycles the exchange bias follows the empirical relation based on the energy dissipation in the AF layer. The ferromagnetic resonance (FMR) measurements also exhibited the in-plane azimuthal angle dependence of the magnetic resonance field indicating that the uniaxial and unidirectional anisotropies are not collinear, although they lie in the same plane. However, no misalignment between the unidirectional anisotropy and the exchange bias direction is observed. The misalignment angle between the uniaxial and unidirectional anisotropy, as measured by FMR, is found to be 10° and 14° for CoFeB and NiFe, respectively. This misalignment is attributed to the interface roughness as revealed by x-ray reflectance measurements.

  3. 具有条纹磁畴结构的NiFe薄膜的制备与磁各向异性研究∗%Preparation and magnetic anisotropy of NiFe film with strip e domains

    Institute of Scientific and Technical Information of China (English)

    李金财; 詹清峰; 潘民杰; 刘鲁萍; 杨华礼; 谢亚丽; 谢淑红; 李润伟

    2016-01-01

    Magnetic anisotropy is one of the most important fundamental properties of magnetic film. For the high-frequency applications, the magnetic anisotropy determines the ferromagnetic resonance frequency of magnetic film. Due to the directionality of conventional static magnetic anisotropy in magnetic film, the high-frequency device usually exhibits a remarkable angular dependent behavior. Only when the microwave magnetic field is perpendicular to the magnetic anisotropy, can the device work at the best performance. The magnetic film with a thickness beyond a critical value displays a stripe domain structure as well as an in-plane rotatable magnetic anisotropy, which can be an important strategy to solve the problem of magnetic field orientation dependent performance in high-frequency device. Thus, the fabrication, the magnetic anisotropy, the magnetic domain and the high-frequency behavior for magnetic film with stripe domain structure have received extensive attention. Previously, a lot of studies have qualitatively indicated that the different fabrication processes could change the critical thickness values of displaying stripe domains, the magnetic domains, and the magnetic anisotropies in many magnetic films. However, the quantitative investigation, especially regarding the magnetic anisotropy which determines the high-frequency behaviors of magnetic films, is less. NiFe alloys display excellent soft magnetic properties, which have been extensively applied to various spintronic devices. In addition, the stripe magnetic domain is discovered for the first time in NiFe film. In this work, we fabricate NiFe magnetic thin films by using radio frequency magnetron sputtering technique at room temperature and quantitatively study the effects of film thickness, sputtering power density and Ar pressure on the magnetic domain structure, in-plane static magnetic anisotropy, in-plane rotatable magnetic anisotropy and out-of-plane magnetic anisotropy. For NiFe films fabricated

  4. Molecular evolution of the nif gene cluster carrying nifI1 and nifI2 genes in the Gram-positive phototrophic bacterium Heliobacterium chlorum.

    Science.gov (United States)

    Enkh-Amgalan, Jigjiddorj; Kawasaki, Hiroko; Seki, Tatsuji

    2006-01-01

    A major nif cluster was detected in the strictly anaerobic, Gram-positive phototrophic bacterium Heliobacterium chlorum. The cluster consisted of 11 genes arranged within a 10 kb region in the order nifI1, nifI2, nifH, nifD, nifK, nifE, nifN, nifX, fdx, nifB and nifV. The phylogenetic position of Hbt. chlorum was the same in the NifH, NifD, NifK, NifE and NifN trees; Hbt. chlorum formed a cluster with Desulfitobacterium hafniense, the closest neighbour of heliobacteria based on the 16S rRNA phylogeny, and two species of the genus Geobacter belonging to the Deltaproteobacteria. Two nifI genes, known to occur in the nif clusters of methanogenic archaea between nifH and nifD, were found upstream of the nifH gene of Hbt. chlorum. The organization of the nif operon and the phylogeny of individual and concatenated gene products showed that the Hbt. chlorum nif operon carrying nifI genes upstream of the nifH gene was an intermediate between the nif operon with nifI downstream of nifH (group II and III of the nitrogenase classification) and the nif operon lacking nifI (group I). Thus, the phylogenetic position of Hbt. chlorum nitrogenase may reflect an evolutionary stage of a divergence of the two nitrogenase groups, with group I consisting of the aerobic diazotrophs and group II consisting of strictly anaerobic prokaryotes.

  5. Preyssler heteropolyacid supported on silica coated NiFe2O4 nanoparticles for the catalytic synthesis of bis(dihydropyrimidinone)benzene and 3,4-dihydropyrimidin-2(1H)-ones

    Institute of Scientific and Technical Information of China (English)

    Hossein Eshghi; Ali Javid; Amir Khojastehnezhad; Farid Moeinpour; Fatemeh F. Bamoharram; Mehdi Bakavoli; Masoud Mirzaei

    2015-01-01

    A novel magnetic acidic catalyst comprising Preyssler (H14[NaP5W30O110]) heteropoly acid support‐ed on silica coated nickel ferrite nanoparticles (NiFe2O4@SiO2) was prepared. The catalyst was characterized by Fourier transform infrared, scanning electron microscopy, transmission electron microscopy, X‐ray diffraction, energy dispersive spectrum, VSM and particle size neasurement. Its catalytic activity was investigated for the synthesis of bis(dihydropyrimidinone)benzene and 3,4‐dihydropyrimidin‐2(1H)‐ones derivatives by the Biginelli reaction. With the catalyst, the reac‐tions occurred in less than 1 h with good to excellent yields. More importantly, the catalyst was easily separated from the reaction mixture by an external magnet and reused at least five times without degradation in the activity.

  6. Nickel-zinc ferrite/permalloy (Ni0.5Zn0.5Fe2O4/Ni-Fe soft magnetic nanocomposites fabricated by electro-infiltration

    Directory of Open Access Journals (Sweden)

    Xiao Wen

    2016-05-01

    Full Text Available Magnetically soft NiZn ferrite (Ni0.5Zn0.5Fe2O4 nanoparticles are embedded within a permalloy (Ni-Fe matrix via an electro-infiltration process as thin films intended for use as on-chip inductor cores in the MHz frequency regime. A layer of NiZn ferrite nanoparticles is first deposited, and then permalloy is electroplated through the voids to encapsulate the particles and form three-dimensional ferrite/alloy nanocomposites. The composites are estimated to contain 37% ferrite by volume and exhibit a relative permeability of ∼320, a saturation of ∼1.15 T, and an operational bandwidth of 93 MHz. Compared to a permalloy thin film of similar thickness, the nanocomposite exhibits 39% higher electrical resistivity and 50% higher bandwidth.

  7. Preliminary testing of NiFe2O4-NiO-Ni cermet as inert anode in Na3AlF6-AlF3 melts

    Institute of Scientific and Technical Information of China (English)

    LAI Yan-qing; TIAN Zhong-liang; LI Jie; YE Shao-long; LIU Ye-xiang

    2006-01-01

    The electrical conductivity of cermet 83(90NiFe2O4-10NiO)-17Ni at different temperatures was measured in air, the operating performance of inert anode was evaluated in a laboratory electrolysis cell with various electrolyte compositions. The results indicate that the electrical resistivity of cermet studied has negative temperature coefficient, which is the characteristic of semi-conducting material. The proper addition of AlF3 in the bath can improve the corrosion resistance of cermet inert anode, but excess adding amount will cause the catastrophic corrosion. Post-examination of anodes shows that metal Ni leaches preferentially on the anode surface. Chemical dissolution, electrolyte penetration as well as electrochemical dissolution serve as major corrosion mechanisms.

  8. Systems Level Approaches to Understanding and Manipulating Heterocyst Differentiation in Nostoc Punctiforme: Sites of Hydrogenase and Nitrogenase Synthesis and Activity

    Energy Technology Data Exchange (ETDEWEB)

    Meeks, John C. [University of California, Davis, CA (United States)

    2015-04-02

    Heterocysts are specialized cells that establish a physiologically low oxygen concentration; they function as the sites of oxygen-sensitive nitrogen fixation and hydrogen metabolism in certain filamentous cyanobacteria. They are present at a frequency of less than 10% of the cells and singly in a nonrandom spacing pattern in the filaments. The extent of differential gene expression during heterocyst differentiation was defined by DNA microarray analysis in wild type and mutant cultures of Nostoc punctiforme. The results in wild-type cultures identified two groups of genes; approximately 440 that are unique to heterocyst formation and function, and 500 that respond positively and negatively to the transient stress of nitrogen starvation. Nitrogen fixation is initiated within 24 h after induction, but the cultures require another 24 h before growth is reinitiated. Microarray analyses were conducted on strains with altered expression of three genes that regulate the presence and spacing of heterocysts in the filaments; loss of function or over expression of these genes increases the heterocyst frequency 2 to 3 fold compared to the wild-type. Mutations in the genes hetR and hetF result in the inability to differentiate heterocysts, whereas over expression of each gene individually yields multiple contiguous heterocysts at sites in the filaments; they are positive regulatory elements. Mutation of the gene patN results in an increase in heterocysts frequency, but, in this case, the heterocysts are singly spaced in the filaments with a decrease in the number of vegetative cells in the interval between heterocysts; this is a negative regulatory element. However, over expression of patN resulted in the wild-type heterocyst frequency and spacing pattern. Microarray results indicated HetR and HetF influence the transcription of a common set of about 395 genes, as well as about 350 genes unique to each protein. HetR is known to be a transcriptional regulator and HetF is

  9. Molecular recognition and self-assembly special feature: Self-assembled biomimetic [2Fe2S]-hydrogenase-based photocatalyst for molecular hydrogen evolution.

    Science.gov (United States)

    Kluwer, A M; Kapre, R; Hartl, F; Lutz, M; Spek, A L; Brouwer, A M; van Leeuwen, P W N M; Reek, J N H

    2009-06-30

    The large-scale production of clean energy is one of the major challenges society is currently facing. Molecular hydrogen is envisaged as a key green fuel for the future, but it becomes a sustainable alternative for classical fuels only if it is also produced in a clean fashion. Here, we report a supramolecular biomimetic approach to form a catalyst that produces molecular hydrogen using light as the energy source. It is composed of an assembly of chromophores to a bis(thiolate)-bridged diiron ([2Fe2S]) based hydrogenase catalyst. The supramolecular building block approach introduced in this article enabled the easy formation of a series of complexes, which are all thoroughly characterized, revealing that the photoactivity of the catalyst assembly strongly depends on its nature. The active species, formed from different complexes, appears to be the [Fe(2)(micro-pdt)(CO)(4){PPh(2)(4-py)}(2)] (3) with 2 different types of porphyrins (5a and 5b) coordinated to it. The modular supramolecular approach was important in this study as with a limited number of building blocks several different complexes were generated.

  10. X-ray crystal structure of the Fe-only hydrogenase (CpI) from Clostridium pasteurianum to 1.8 angstrom resolution.

    Science.gov (United States)

    Peters, J W; Lanzilotta, W N; Lemon, B J; Seefeldt, L C

    1998-12-01

    A three-dimensional structure for the monomeric iron-containing hydrogenase (CpI) from Clostridium pasteurianum was determined to 1.8 angstrom resolution by x-ray crystallography using multiwavelength anomalous dispersion (MAD) phasing. CpI, an enzyme that catalyzes the two-electron reduction of two protons to yield dihydrogen, was found to contain 20 gram atoms of iron per mole of protein, arranged into five distinct [Fe-S] clusters. The probable active-site cluster, previously termed the H-cluster, was found to be an unexpected arrangement of six iron atoms existing as a [4Fe-4S] cubane subcluster covalently bridged by a cysteinate thiol to a [2Fe] subcluster. The iron atoms of the [2Fe] subcluster both exist with an octahedral coordination geometry and are bridged to each other by three non-protein atoms, assigned as two sulfide atoms and one carbonyl or cyanide molecule. This structure provides insights into the mechanism of biological hydrogen activation and has broader implications for [Fe-S] cluster structure and function in biological systems.

  11. Comparison of H{sub 2} accumulation by Rhodobacter sphaeroides KD131 and its uptake hydrogenase and PHB synthase deficient mutant

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Mi-Sun; Baek, Jin-Sook [Biomass Research Center, Korea Institute of Energy Research, 71-2 Jang-dong, Yuseong-gu, Daejeon 305-343 (Korea, Republic of); Lee, Jeong K. [Department of Life Science, Sogang University, Shinsu-dong, Mapo-gu, Seoul 121-742 (Korea, Republic of)

    2006-01-15

    Rhodobacter sphaeroides KD131 and its mutant strain lacking uptake hydrogenase (Hup{sup -}) and PHB synthase (Phb{sup -}) have been studied on H{sub 2} production and cell growth under different culture conditions. Both strains started producing H{sub 2} from the middle of the logarithmic growth phase and continued until the cell concentration leveled out. The rates of H{sub 2} production were 1.32 and 3.34ml H{sub 2}/mg-dcw for the wild-type and Hup{sup -}/Phb{sup -} mutant strain, respectively, at the optimum conditions. Malate and lactate were better carbon sources than starch, sucrose or glycerol. Approximately 60% of acetic acid was degraded in 48h by the wild-type strain and pH increased to 9.4. However, the Hup{sup -}/Phb{sup -} mutant strain did not grow well and degraded only 19% of acetic acid. The pH ranges of 7.0 were the optimum for the cell growth and pH 7.5 for the H{sub 2} production. Both strains grew and produced hydrogen under the irradiance of 12-120W/m{sup 2}, but cell growth was inhibited over 400W/m{sup 2}. (author)

  12. Secondary coordination sphere accelerates hole transfer for enhanced hydrogen photogeneration from [FeFe]-hydrogenase mimic and CdSe QDs in water

    Science.gov (United States)

    Wen, Min; Li, Xu-Bing; Jian, Jing-Xin; Wang, Xu-Zhe; Wu, Hao-Lin; Chen, Bin; Tung, Chen-Ho; Wu, Li-Zhu

    2016-07-01

    Achieving highly efficient hydrogen (H2) evolution via artificial photosynthesis is a great ambition pursued by scientists in recent decades because H2 has high specific enthalpy of combustion and benign combustion product. [FeFe]-Hydrogenase ([FeFe]-H2ase) mimics have been demonstrated to be promising catalysts for H2 photoproduction. However, the efficient photocatalytic H2 generation system, consisting of PAA-g-Fe2S2, CdSe QDs and H2A, suffered from low stability, probably due to the hole accumulation induced photooxidation of CdSe QDs and the subsequent crash of [FeFe]-H2ase mimics. In this work, we take advantage of supramolecular interaction for the first time to construct the secondary coordination sphere of electron donors (HA‑) to CdSe QDs. The generated secondary coordination sphere helps realize much faster hole removal with a ~30-fold increase, thus leading to higher stability and activity for H2 evolution. The unique photocatalytic H2 evolution system features a great increase of turnover number to 83600, which is the highest one obtained so far for photocatalytic H2 production by using [FeFe]-H2ase mimics as catalysts.

  13. Activation Thermodynamics and H/D Kinetic Isotope Effect of the Hox to HredH+ Transition in [FeFe] Hydrogenase

    Energy Technology Data Exchange (ETDEWEB)

    King, Paul W [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Ratzloff, Michael W [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Mulder, David W [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Lubner, Carolyn E [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Brown, Katherine A [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Wilker, Molly B. [University of Colorado; Hamby, Hayden [University of Colorado; Dukovic, Gordana [University of Colorado

    2017-08-29

    Molecular complexes between CdSe nanocrystals and Clostridium acetobutylicum [FeFe] hydrogenase I (CaI) enabled light-driven control of electron transfer for spectroscopic detection of redox intermediates during catalytic proton reduction. Here we address the route of electron transfer from CdSe->CaI and activation thermodynamics of the initial step of proton reduction in CaI. The electron paramagnetic spectroscopy of illuminated CdSe:CaI showed how the CaI accessory FeS cluster chain (F-clusters) functions in electron transfer with CdSe. The Hox->HredH+ reduction step measured by Fourier-transform infrared spectroscopy showed an enthalpy of activation of 19 kJ mol-1 and a ~2.5-fold kinetic isotope effect. Overall these results support electron injection from CdSe into CaI involving F-clusters, and that the Hox->HredH+ step of catalytic proton reduction in CaI proceeds by a proton-dependent process.

  14. Investigating the Role of the Outer-Coordination Sphere in [Ni(PPh2NPh-R2)2]2+ Hydrogenase Mimics

    Energy Technology Data Exchange (ETDEWEB)

    Jain, Avijita; Reback, Matthew L.; Lindstrom, Mary L.; Thogerson, Colleen E.; Helm, Monte L.; Appel, Aaron M.; Shaw, Wendy J.

    2012-06-18

    A series of dipeptide nickel complexes with the general formula, [Ni(PPh2NNNA-amino acid/ester2)2](BF4)2, have been synthesized and characterized (P2N2= 1,5-diaza-3,7-diphosphacyclooctane, amino acid/esters = glutamic acid, alanine, lysine, and aspartic acid). Each of these complexes is an efficient electrocatalyst for H2 production. The contribution of the outer-coordination sphere, specifically the impact of sterics, the ability to protonate and the pKa of amino acid side chain on the hydrogen production activity of these complexes, was investigated. The rates of all of the catalysts ranged over an order of magnitude. The amino acid containing complexes display 2-3 times higher rates of hydrogen production than the corresponding ester complexes, suggesting the significance of protonated species (side chains/backbone of amino acids) in the outer-coordination sphere. The largest had the fastest rates suggesting that catalytic activity is not hindered by sterics. However, the shapes of catalytic waves are indicative of hindered electron transfer and may suggest a competing mechanism for catalysis than that observed for the unsubstituted parent complex. These studies demonstrate the significant contribution that the outer-coordination sphere can have in tuning the catalytic activity of small molecule hydrogenase mimics.

  15. A new cumulene diiron complex related to the active site of Fe-only hydrogenases and its phosphine substituted derivatives: synthesis, electrochemistry and structural characterization.

    Science.gov (United States)

    Wen, Na; Xu, Fenfen; Feng, Yanan; Du, Shaowu

    2011-09-01

    A new cumulene diiron complex related to the Fe-only hydrogenase active site [(μ-SCH(2)C(S)CCH(2))Fe(2)(CO)(6)] (1) was obtained by treatment of (μ-LiS)(2)Fe(2)(CO)(6) with excess 1,4-dichloro-2-butyne. By controllable CO displacement of 1 with PPh(3) and bis(diphenylphosphino)methane (dppm), mono- and di-substituted complexes, namely [(μ-SCH(2)C(S)CCH(2))Fe(2)(CO)(5)L] (2: L=PPh(3); 3: L=dppm) and [(μ-SCH(2)C(S)CCH(2))Fe(2)(CO)(4)L(2)] (4: L=PPh(3); 5: L=dppm) could be prepared in moderate yields. Treatment of 1 with bis(diphenylphosphino)ethane (dppe) afforded a double butterfly complex [(μ-SCH(2)C(S)CCH(2))Fe(2)(CO)(5)](2)(μ-dppe) (7). With dppm in refluxing toluene, a dppm-bridged complex [(μ-SCH(2)C(S)CCH(2))Fe(2)(CO)(4)(μ-dppm)] (6) was obtained. These model complexes were characterized by IR, (1)H, (31)P NMR spectra and the molecular structures of 1, 2 and 5-7 were determined by single crystal X-ray analyses. The electrochemistry of 1-3 was studied and the electrocatalytic property of 1 was investigated for proton reduction in the presence of HOAc. Copyright © 2011 Elsevier Inc. All rights reserved.

  16. Reduction Potentials of [FeFe]-Hydrogenase Accessory Iron–Sulfur Clusters Provide Insights into the Energetics of Proton Reduction Catalysis

    Energy Technology Data Exchange (ETDEWEB)

    Artz, Jacob H. [Institute; Mulder, David W. [Biosciences; Ratzloff, Michael W. [Biosciences; Lubner, Carolyn E. [Biosciences; Zadvornyy, Oleg A. [Institute; LeVan, Axl X. [Department; Williams, S. Garrett [School; Adams, Michael W. W. [B216B; Jones, Anne K. [School; King, Paul W. [Biosciences; Peters, John W. [Institute

    2017-07-06

    An [FeFe]-hydrogenase from Clostridium pasteurianum, CpI, is a model system for biological H2 activation. In addition to the catalytic H-cluster, CpI contains four accessory iron-sulfur [FeS] clusters in a branched series that transfer electrons to and from the active site. In this work, potentiometric titrations have been employed in combination with electron paramagnetic resonance (EPR) spectroscopy at defined electrochemical potentials to gain insights into the role of the accessory clusters in catalysis. EPR spectra collected over a range of potentials were deconvoluted into individual components attributable to the accessory [FeS] clusters and the active site H-cluster, and reduction potentials for each cluster were determined. The data suggest a large degree of magnetic coupling between the clusters. The distal [4Fe-4S] cluster is shown to have a lower reduction potential (~<-450 mV) than the other clusters, and molecular docking experiments indicate that the physiological electron donor, ferredoxin (Fd), most favorably interacts with this cluster. The low reduction potential of the distal [4Fe-4S] cluster thermodynamically restricts the Fdox/Fdred ratio at which CpI can operate, consistent with the role of CpI in recycling Fdred that accumulates during fermentation. Subsequent electron transfer through the additional accessory [FeS] clusters to the H-cluster is thermodynamically favorable.

  17. Synthesis, structures and electrochemical properties of nitro- and amino-functionalized diiron azadithiolates as active site models of Fe-only hydrogenases.

    Science.gov (United States)

    Liu, Tianbiao; Wang, Mei; Shi, Zhan; Cui, Hongguang; Dong, Weibing; Chen, Jiesheng; Akermark, Björn; Sun, Licheng

    2004-09-20

    Complex [[(mu-SCH2)2N(4-NO2C6H4)]Fe2(CO)6] (4) was prepared by the reaction of the dianionic intermediate [(mu-S)2Fe2(CO)6](2-) and N,N-bis(chloromethyl)-4-nitroaniline as a biomimetic model of the active site of Fe-only hydrogenase. The reduction of 4 by Pd-C/H2 under a neutral condition afforded complex [[(mu-SCH2)2N(4-NH2C6H4)]Fe2(CO)6] (5) in 67 % yield. Both complexes were characterized by IR, 1H and 13C NMR spectroscopy and MS spectrometry. The molecular structure of 4, as determined by X-ray analysis, has a butterfly 2Fe2S core and the aryl group on the bridged-N atom slants to the Fe(2) site. Cyclic voltammograms of 4 and 5 were studied to evaluate their redox properties. It was found that complex 4 catalyzed electrochemical proton reduction in the presence of acetic acid. A plausible mechanism of the electrocatalytic proton reduction is discussed.

  18. Super magnetic nanoparticles NiFe2O4, coated with aluminum-nickel oxide sol-gel lattices to safe, sensitive and selective purification of his-tagged proteins.

    Science.gov (United States)

    Mirahmadi-Zare, Seyede Zohreh; Allafchian, Alireza; Aboutalebi, Fatemeh; Shojaei, Pendar; Khazaie, Yahya; Dormiani, Kianoush; Lachinani, Liana; Nasr-Esfahani, Mohammad-Hossein

    2016-05-01

    Super magnetic nanoparticle NiFe2O4 with high magnetization, physical and chemical stability was introduced as a core particle which exhibits high thermal stability (>97%) during the harsh coating process. Instead of multi-stage process for coating, the magnetic nanoparticles was mineralized via one step coating by a cheap, safe, stable and recyclable alumina sol-gel lattice (from bohemite source) saturated by nickel ions. The TEM, SEM, VSM and XRD imaging and BET analysis confirmed the structural potential of NiFe2O4@NiAl2O4 core-shell magnetic nanoparticles for selective and sensitive purification of His-tagged protein, in one step. The functionality and validity of the nickel magnetic nanoparticles were attested by purification of three different bioactive His-tagged recombinant fusion proteins including hIGF-1, GM-CSF and bFGF. The bonding capacity of the nickel magnetics nanoparticles was studied by Bradford assay and was equal to 250 ± 84 μg Protein/mg MNP base on protein size. Since the metal ion leakage is the most toxicity source for purification by nickel magnetic nanoparticles, therefor the nickel leakage in purified final protein was determined by atomic absorption spectroscopy and biological activity of final purified protein was confirmed in comparison with reference. Also, in vitro cytotoxicity of nickel magnetic nanoparticles and trace metal ions were investigated by MTS assay analysis. The results confirmed that the synthesized nickel magnetic nanoparticles did not show metal ion toxicity and not affected on protein folding.

  19. Production of Ni-Fe alloy from nickeliferous laterite ore by prereduction-electric furnace smelting separation%红土镍矿转底炉预还原-电炉熔分制取镍铁合金

    Institute of Scientific and Technical Information of China (English)

    范兴祥; 董海刚; 汪云华; 吴跃东; 李柏榆; 李博捷; 赵家春; 暂林寒

    2012-01-01

    对某红土镍矿采用转底炉预还原-电炉熔分工艺制取镍铁合金进行研究.实验结果表明:根据矿石性质,选择合适的渣型为SiO2-MgO-CaO-FeO四元渣系,在熔剂石灰配比为25%,还原剂配比为3.5%,预还原温度为1 150℃,预还原时间为30 min,电炉熔分温度为1 450℃,熔分时间为15 min的条件下,经转底炉预还原-电炉熔分后,获得镍质量分数为8.68%、镍回收率97.62%、铁质量分数为86.23%的镍铁合金,该合金可用作不锈钢生产原料.%The production of Ni-Fe alloy from nickeliferous laterite ore was studied by rotary hearth furnace prereduction-electric furnace smelting separation. The results show that a SiOi-MgO-CaO-FeO slag style is chosen according to the characteristic of the nickeliferous laterite ore and Ni-Fe alloy with 8.68% Ni (mass fraction), Ni 97.62% recovery and 86.23% Fe is obtained under the conditions of lime dosage 25%, reductant coke powder dosage 3.5%, prereduction temperature 1 150 ℃, prereduction time 30 min, smelting temperature 1 450 ℃, smelting time 15 rain, respectively, which can be used as the raw material of production of stainless steel.

  20. Hydrogen from Water in a Novel Recombinant Cyanobacterial System

    Energy Technology Data Exchange (ETDEWEB)

    Weyman, Philip D [J. Craig Venter Institute; Smith, Hamillton O.

    2014-12-03

    Photobiological processes are attractive routes to renewable H2 production. With the input of solar energy, photosynthetic microbes such as cyanobacteria and green algae carry out oxygenic photosynthesis, using sunlight energy to extract protons and high energy electrons from water. These protons and high energy electrons can be fed to a hydrogenase system yielding H2. However, most hydrogen-evolving hydrogenases are inhibited by O2, which is an inherent byproduct of oxygenic photosynthesis. The rate of H2 production is thus limited. Certain photosynthetic bacteria are reported to have an O2-tolerant evolving hydrogenase, yet these microbes do not split water, and require other more expensive feedstocks. To overcome these difficulties, the goal of this work has been to construct novel microbial hybrids by genetically transferring O2-tolerant hydrogenases from other bacteria into a class of photosynthetic bacteria called cyanobacteria. These hybrid organisms will use the photosynthetic machinery of the cyanobacterial hosts to perform the water-oxidation reaction with the input of solar energy, and couple the resulting protons and high energy electrons to the O2-tolerant bacterial hydrogenase, all within the same microbe (Fig. 1). The ultimate goal of this work has been to overcome the sensitivity of the hydrogenase enzyme to O2 and address one of the key technological hurdles to cost-effective photobiological H2 production which currently limits the production of hydrogen in algal systems. In pursuit of this goal, work on this project has successfully completed many subtasks leading to a greatly increased understanding of the complicated [NiFe]-hydrogenase enzymes. At the beginning of this project, [NiFe] hydrogenases had never been successfully moved across wide species barriers and had never been heterologously expressed in cyanobacteria. Furthermore, the idea that whole, functional genes could be extracted from complicated, mixed-sequence meta-genomes was not

  1. Detection of the gene encoding the small subunit of the CO dehydrogenase enzyme in the H{sub 2}-evolving bacterium Rubrivivax gelatinosus CBS

    Energy Technology Data Exchange (ETDEWEB)

    Kish, A.; Levin, D. [Victoria Univ., BC (Canada)]|[Victoria Univ., BC (Canada)

    2001-06-01

    A purple non-sulfur bacterium, Rubrivivax gelatinosus CBS presents great opportunities, on a commercial scale, for the biological hydrogen production. A water-gas shift reaction is catalyzed when the bacterium is cultured in the presence of carbon oxide in the dark. The result is carbon monoxide (and water) being shifted into hydrogen (H{sub 2}) and carbon dioxide in near stoichiometric quantities. The production of hydrogen as a clean alternative fuel could be accomplished by using carbon monoxide generated from gasified waste biomass, using the bacterial water-gas shift reaction for that purpose. The characterization of three key enzymes and the genes encoding them was performed in a closely related purple non-sulfur bacterium called Rhodospirillum rubrum. They were: (1) a carbon monoxide dehydrogenase (CODH), (2) the ferredoxin-like electron-carrier small subunit of the CODH enzyme, and (3) an hydrogen-evolving hydrogenase. A transcriptional unit separate from the genes encoding the CODH and its ferredoxin-like small subunit encode the genes for the hydrogenase. A fragment of the Rhodospirillum rubrum ferredoxin-like subunit gene was amplified through the use of a polymerase chain reaction. Southern blots of restriction endonuclease digested genomic deoxyribonucleic acid (DNA) extracted from Rubrivivax gelatinosus CBS was probed with the fragment of the Rhodospirillum rubrum previously amplified using the polymerase chain reaction. Confirmation of the identification is being confirmed, while the gene is sequenced. 25 refs., 2 figs.

  2. Further analysis of nitrogen fixation (nif) genes in Azotobacter chroococcum: identification and expression in Klebsiella pneumoniae of nifS, nifV, nifM, and nifB genes and localization of nifE/N-, nifU-, nifA- and fixABC-like genes.

    Science.gov (United States)

    Evans, D; Jones, R; Woodley, P; Robson, R

    1988-04-01

    The results presented extend previous investigations on the genetics of nitrogen fixation in Azotobacter chroococcum and indicate that nif- and fix-like DNA is located in at least five different regions of the genome. Region I contains functional copies of nifS,V and M, as well as nifH, D and K, all of which complemented mutants of Klebsiella pneumoniae. In addition, nifE- and/or nifN-like and nifU-like DNA is located in this region. The organization of the nif cluster in region I closely resembles that of K. pneumoniae. though spread over 22 kb as compared with 14 kb. Region II contains a functional nifB gene, which complemented a K. pneumoniae nifB mutant, and seems to be adjacent to ap nifA-like gene. Region III harbours nifH*, encoding a second nitrogenase Fe-protein. Region IV contains a reiteration of nifE- on and/or nifN-like sequences, and DNA homologous to Rhizobium meliloti fixABC is present in region V. The apparent complexity of nifDNA in A. chroococcum is probably related to the two systems for N2-fixation pr present in this organism.

  3. STUDIES ON TRANSFERENCE OF HYDROGENASE GENES OF RHIZOBIUM ARACHIS%根瘤菌吸氢酶基因转移的研究

    Institute of Scientific and Technical Information of China (English)

    王振宇; 龙敏南; 刘月英; 张风章; 许良树; 朱天赐; 蒋水柳

    2001-01-01

    通过三亲本杂交将含有花生根瘤菌吸氢基因的质粒pZ 55(Tcr)转入不吸氢的花生根瘤菌Ra 34等菌株(Hup-,Nif+,Ap.)中,筛选到既具有吸氢又具有固氮能力的花生根瘤菌结合株Rz34-2.在自生和共生条件下,结合株均可表达高吸氢和高固氮活性.以结合株Rz34-2接种的花生植株叶片的干重比不接种的、接种受体株Ra34(Hup-)和接种对照菌株L8-3(Hup+,Nif+)的分别高6.2%、7.6%和6.3%;种子的含氮量分别高8.9%、10.0%和6.0%;产量分别高18.8%、10.5%和10.7%.研究结果表明,以含吸氢基因的结合株接种花生能提高根瘤菌与花生的共生固氮效率,增加作物的产量.

  4. Carbon-dependent control of electron transfer and central carbon pathway genes for methane biosynthesis in the Archaean, Methanosarcina acetivorans strain C2A

    Directory of Open Access Journals (Sweden)

    Gunsalus Robert P

    2010-02-01

    Full Text Available Abstract Background The archaeon, Methanosarcina acetivorans strain C2A forms methane, a potent greenhouse gas, from a variety of one-carbon substrates and acetate. Whereas the biochemical pathways leading to methane formation are well understood, little is known about the expression of the many of the genes that encode proteins needed for carbon flow, electron transfer and/or energy conservation. Quantitative transcript analysis was performed on twenty gene clusters encompassing over one hundred genes in M. acetivorans that encode enzymes/proteins with known or potential roles in substrate conversion to methane. Results The expression of many seemingly "redundant" genes/gene clusters establish substrate dependent control of approximately seventy genes for methane production by the pathways for methanol and acetate utilization. These include genes for soluble-type and membrane-type heterodisulfide reductases (hdr, hydrogenases including genes for a vht-type F420 non-reducing hydrogenase, molybdenum-type (fmd as well as tungsten-type (fwd formylmethanofuran dehydrogenases, genes for rnf and mrp-type electron transfer complexes, for acetate uptake, plus multiple genes for aha- and atp-type ATP synthesis complexes. Analysis of promoters for seven gene clusters reveal UTR leaders of 51-137 nucleotides in length, raising the possibility of both transcriptional and translational levels of control. Conclusions The above findings establish the differential and coordinated expression of two major gene families in M. acetivorans in response to carbon/energy supply. Furthermore, the quantitative mRNA measurements demonstrate the dynamic range for modulating transcript abundance. Since many of these gene clusters in M. acetivorans are also present in other Methanosarcina species including M. mazei, and in M. barkeri, these findings provide a basis for predicting related control in these environmentally significant methanogens.

  5. Characterization of the Fe site in iron-sulfur cluster-free hydrogenase (Hmd) and of a model compound via nuclear resonance vibrational spectroscopy (NRVS).

    Science.gov (United States)

    Guo, Yisong; Wang, Hongxin; Xiao, Yuming; Vogt, Sonja; Thauer, Rudolf K; Shima, Seigo; Volkers, Phillip I; Rauchfuss, Thomas B; Pelmenschikov, Vladimir; Case, David A; Alp, Ercan E; Sturhahn, Wolfgang; Yoda, Yoshitaka; Cramer, Stephen P

    2008-05-19

    We have used (57)Fe nuclear resonance vibrational spectroscopy (NRVS) to study the iron site in the iron-sulfur cluster-free hydrogenase Hmd from the methanogenic archaeon Methanothermobacter marburgensis. The spectra have been interpreted by comparison with a cis-(CO)2-ligated Fe model compound, Fe(S2C2H4)(CO)2(PMe3)2, as well as by normal mode simulations of plausible active site structures. For this model complex, normal mode analyses both from an optimized Urey-Bradley force field and from complementary density functional theory (DFT) calculations produced consistent results. For Hmd, previous IR spectroscopic studies found strong CO stretching modes at 1944 and 2011 cm(-1), interpreted as evidence for cis-Fe(CO)2 ligation. The NRVS data provide further insight into the dynamics of the Fe site, revealing Fe-CO stretch and Fe-CO bend modes at 494, 562, 590, and 648 cm(-1), consistent with the proposed cis-Fe(CO)2 ligation. The NRVS also reveals a band assigned to Fe-S stretching motion at approximately 311 cm(-1) and another reproducible feature at approximately 380 cm(-1). The (57)Fe partial vibrational densities of states (PVDOS) for Hmd can be reasonably well simulated by a normal mode analysis based on a Urey-Bradley force field for a five-coordinate cis-(CO)2-ligated Fe site with additional cysteine, water, and pyridone cofactor ligands. A "truncated" model without a water ligand can also be used to match the NRVS data. A final interpretation of the Hmd NRVS data, including DFT analysis, awaits a three-dimensional structure for the active site.

  6. Pd(II)-Directed Encapsulation of Hydrogenase within the Layer-by-Layer Multilayers of Carbon Nanotube Polyelectrolyte Used as a Heterogeneous Catalyst for Oxidation of Hydrogen.

    Science.gov (United States)

    Liu, Jiang; Zorin, Nikolay A; Chen, Meng; Qian, Dong-Jin

    2015-06-16

    A metal-directed assembling approach has been developed to encapsulate hydrogenase (H2ase) within a layer-by-layer (LBL) multilayer of carbon nanotube polyelectrolyte (MWNT-PVPMe), which showed efficient biocatalytic oxidation of H2 gas. The MWNT-PVPMe was prepared via a diazonium process and addition reactions with poly(4-vinylpyridine) (PVP) and methyl iodide (MeI). The covalently attached polymers and organic substituents in the polyelectrolyte comprised 60-70% of the total weight. The polyelectrolyte was then used as a substrate for H2ase binding to produce MWNT-PVPMe@H2ase bionanocomposites. X-ray photoelectron spectra revealed that the bionanocomposites included the elements of Br, S, C, N, O, I, Fe, and Ni, which confirmed that they were composed of MWNT-PVPMe and H2ase. Field emission transmission electron microscope images revealed that the H2ase was adsorbed on the surface of MWNT-PVPMe with the domains ranging from 20 to 40 nm. Further, with the use of the bionanocomposites as nanolinkers and Na2PdCl4 as connectors, the (Pd/MWNT-PVPMe@H2ase)n multilayers were constructed on the quartz and gold substrate surfaces by the Pd(II)-directed LBL assembling technique. Finally, the as-prepared LBL multilayers were used as heterogeneous catalysts for hydrogen oxidation with methyl viologen (MV(2+)) as an electron carrier. The dynamic processes for the reversible color change between blue-colored MV(+) and colorless MV(2+) (catalyzed by the LBL multilayers) were video recorded, which confirmed that the H2ase encapsulated within the present LBL multilayers was of much stronger stability and higher biocatalytic activity of H2 oxidation resulting in potential applications for the development of H2 biosensors and fuel cells.

  7. Synthesis, structural characterization, and some properties of 2-acylmethyl-6-ester group-difunctionalized pyridine-containing iron complexes related to the active site of [Fe]-hydrogenase.

    Science.gov (United States)

    Song, Li-Cheng; Hu, Fu-Qiang; Wang, Miao-Miao; Xie, Zhao-Jun; Xu, Kai-Kai; Song, Hai-Bin

    2014-06-07

    As biomimetic models for [Fe]-hydrogenase, the 2-acylmethyl-6-ester group-difunctionalized pyridine-containing iron(II) complexes 1-4 have been successfully prepared via the following three separate steps. In the first step, the acylation or esterification of difunctionalized pyridine 2-(p-MeC6H4SO3CH2)-6-HOCH2C5H3N with acetyl chloride or benzoic acid gives the corresponding pyridine derivatives 2-(p-MeC6H4SO3CH2)-6-RCO2CH2C5H3N (A, R = Me; B, R = Ph). The second step involves reaction of A or B with Na2Fe(CO)4 followed by treatment of the intermediate Fe(0) complexes [Na(2-CH2-6-RCO2CH2C5H3N)Fe(CO)4] (M1, R = Me; M2, R = Ph) with iodine to afford 2-acylmethyl-6-acetoxymethyl or 6-benzoyloxymethyl-difunctionalized pyridine-containing Fe(II) iodide complexes [2-C(O)CH2-6-RCO2CH2C5H3N]Fe(CO)2I (1, R = Me; 3, R = Ph). Finally, when 1 or 3 is treated with sodium 2-mercaptopyridinate, the corresponding difunctionalized pyridine-containing Fe(ii) mercaptopyridinate complexes [2-C(O)CH2-6-RCO2C5H3N]Fe(CO)2(2-SC5H4N) (2, R = Me; 4, R = Ph) are produced. While the structures of model complexes 1-4 are confirmed by X-ray crystallography, the electrochemical properties of 2 and 4 are compared with those of the two previously reported models. In addition, complexes 2 and 4 have been found to be catalysts for H2 production in the presence of TFA under CV conditions.

  8. GenBank blastx search result: AK119179 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK119179 001-037-G09 M55230.1 Ralstonia eutropha NAD-reducing hydrogenase (hoxF), NAD-reducing... hydrogenase (hoxU), NAD-reducing hydrogenase (hoxY), and NAD-reducing hydrogenase (hoxH) genes, complete cds.|BCT BCT 1e-38 +3 ...

  9. The nif Gene Operon of the Methanogenic Archaeon Methanococcus maripaludis

    Science.gov (United States)

    Kessler, Peter S.; Blank, Carrine; Leigh, John A.

    1998-01-01

    Nitrogen fixation occurs in two domains, Archaea and Bacteria. We have characterized a nif (nitrogen fixation) gene cluster in the methanogenic archaeon Methanococcus maripaludis. Sequence analysis revealed eight genes, six with sequence similarity to known nif genes and two with sequence similarity to glnB. The gene order, nifH, ORF105 (similar to glnB), ORF121 (similar to glnB), nifD, nifK, nifE, nifN, and nifX, was the same as that found in part in other diazotrophic methanogens and except for the presence of the glnB-like genes, also resembled the order found in many members of the Bacteria. Using transposon insertion mutagenesis, we determined that an 8-kb region required for nitrogen fixation corresponded to the nif gene cluster. Northern analysis revealed the presence of either a single 7.6-kb nif mRNA transcript or 10 smaller mRNA species containing portions of the large transcript. Polar effects of transposon insertions demonstrated that all of these mRNAs arose from a single promoter region, where transcription initiated 80 bp 5′ to nifH. Distinctive features of the nif gene cluster include the presence of the six primary nif genes in a single operon, the placement of the two glnB-like genes within the cluster, the apparent physical separation of the cluster from any other nif genes that might be in the genome, the fragmentation pattern of the mRNA, and the regulation of expression by a repression mechanism described previously. Our study and others with methanogenic archaea reporting multiple mRNAs arising from gene clusters with only a single putative promoter sequence suggest that mRNA processing following transcription may be a common occurrence in methanogens. PMID:9515920

  10. Onsite magnetic moment through cation distribution and magnetocrystalline anisotropy studies in NiFe2-xRxO4 (R = Y and Lu; x = 0, 0.05, and 0.075)

    Science.gov (United States)

    Kodam, Ugendar; Kamala Bharathi, K.; Raghavendra Reddy, V.; Rayaprol, Sudhindra; Siruguri, Vasudeva; Garimalle, Markandeyulu

    2017-02-01

    Onsite magnetic moments through cation distribution and magnetocrystalline anisotropy studies of NiFe2-xRxO4 (R = Y and Lu; x = 0, 0.05, and 0.075) compounds were investigated, and the results are discussed and presented in this paper. All the compounds were prepared by solid state reaction, and the compounds formed in the cubic inverse spinel phase with the space group Fd 3 ¯ m . The cation distribution, bond lengths, u-parameter, etc. were estimated through the Rietveld refinement of XRD patterns. Increment in the lattice constant was observed upon partial substitution of Fe3+ by Y3+/Lu3+. The presence of all elements and their ionic states were confirmed from X-ray photoelectron spectroscopy studies. Analyses of Mössbauer spectra revealed that the hyperfine fields and the magnetic moments at the B-site (and hence net moment) decreased with increasing Y3+/Lu3+ occupancy and that the compounds exhibited a Néel-type, collinear ferrimagnetic structure. Magnetization measurements revealed that the magnetic moment decreased with Y3+/Lu3+ substitution. The high field regimes of the magnetization curves were modeled using the law of approach to the saturation magnetization equation, and the first order cubic anisotropy constants (K1) were calculated. The temperature variation of K1 and effects of Y3+/Lu3+ substitution are explained.

  11. Relationship of proton motive force and the F(0)F (1)-ATPase with bio-hydrogen production activity of Rhodobacter sphaeroides: effects of diphenylene iodonium, hydrogenase inhibitor, and its solvent dimethylsulphoxide.

    Science.gov (United States)

    Hakobyan, Lilit; Gabrielyan, Lilit; Trchounian, Armen

    2012-08-01

    Rhodobacter sphaeroides MDC 6521 was able to produce bio-hydrogen (H(2)) in anaerobic conditions under illumination. In this study the effects of the hydrogenase inhibitor-diphenylene iodonium (Ph(2)I) and its solvent dimethylsulphoxide (DMSO) on growth characteristics and H(2) production by R. sphaeroides were investigated. The results point out the concentration dependent DMSO effect: in the presence of 10 mM DMSO H(2) yield was ~6 fold lower than that of the control. The bacterium was unable to produce H(2) in the presence of Ph(2)I. In order to examine the mediatory role of proton motive force (∆p) or the F(0)F(1)-ATPase in H(2) production by R. sphaeroides, the effects of Ph(2)I and DMSO on ∆p and its components (membrane potential (∆ψ) and transmembrane pH gradient), and ATPase activity were determined. In these conditions ∆ψ was of -98 mV and the reversed ∆pH was +30 mV, resulting in ∆p of -68 mV. Ph(2)I decreased ∆ψ in concentrations of 20 μM and higher; lower concentrations of Ph(2)I as DMSO had no valuable effect on ∆ψ. The R. sphaeroides membrane vesicles demonstrated significant ATPase activity sensitive to N,N'-dicyclohexylcarbodiimide. The 10-20 μM Ph(2)I did not affect the ATPase activity, whereas 40 μM Ph(2)I caused a marked inhibition (~2 fold) in ATPase activity. The obtained results provide novel evidence on the involvement of hydrogenase and the F(0)F(1)-ATPase in H(2) production by R. sphaeroides. Moreover, these data indicate the role of hydrogenase and the F(0)F(1)-ATPase in ∆p generation. In addition, DMSO might increase an interaction of nitrogenase with CO(2), decreasing nitrogenase activity and affecting H(2) production.

  12. Genes and Gene Therapy

    Science.gov (United States)

    ... correctly, a child can have a genetic disorder. Gene therapy is an experimental technique that uses genes to ... or prevent disease. The most common form of gene therapy involves inserting a normal gene to replace an ...

  13. Requirements for functional models of the iron hydrogenase active site: D2/H2O exchange activity in ((mu-SMe)(mu-pdt)[Fe(CO)2(PMe3)]2+)[BF4-].

    Science.gov (United States)

    Georgakaki, Irene P; Miller, Matthew L; Darensbourg, Marcetta Y

    2003-04-21

    Hydrogen uptake in hydrogenase enzymes can be assayed by H/D exchange reactivity in H(2)/D(2)O or H(2)/D(2)/H(2)O mixtures. Diiron(I) complexes that serve as structural models for the active site of iron hydrogenase are not active in such isotope scrambling but serve as precursors to Fe(II)Fe(II) complexes that are functional models of [Fe]H(2)ase. Using the same experimental protocol as used previously for ((mu-H)(mu-pdt)[Fe(CO)(2)(PMe(3))](2)(+)), 1-H(+) (Zhao et al. J. Am. Chem. Soc. 2001, 123, 9710), we now report the results of studies of ((mu-SMe)(mu-pdt)[Fe(CO)(2)(PMe(3))](2)(+)), 1-SMe(+), toward H/D exchange. The 1-SMe(+) complex can take up H(2) and catalyze the H/D exchange reaction in D(2)/H(2)O mixtures under photolytic, CO-loss conditions. Unlike 1-H(+), it does not catalyze H(2)/D(2) scrambling under anhydrous conditions. The molecular structure of 1-SMe(+) involves an elongated Fe.Fe separation, 3.11 A, relative to 2.58 A in 1-H(+). It is proposed that the strong SMe(-) bridging ligand results in catalytic activity localized on a single Fe(II) center, a scenario that is also a prominent possibility for the enzyme active site. The single requirement is an open site on Fe(II) available for binding of D(2) (or H(2)), followed by deprotonation by the external base H(2)O (or D(2)O).

  14. Combined genomics and experimental analyses of respiratory characteristics of Shewanella putrefaciens W3-18-1.

    Science.gov (United States)

    Qiu, Dongru; Wei, Hehong; Tu, Qichao; Yang, Yunfeng; Xie, Ming; Chen, Jingrong; Pinkerton, Mark H; Liang, Yili; He, Zhili; Zhou, Jizhong

    2013-09-01

    It has previously been shown that the Shewanella putrefaciens W3-18-1 strain produces remarkably high current in microbial fuel cells (MFCs) and can form magnetite at 0°C. To explore the underlying mechanisms, we developed a genetic manipulation method by deleting the restriction-modification system genes of the SGI1 (Salmonella genome island 1)-like prophage and analyzed the key genes involved in bacterial respiration. W3-18-1 has less respiratory flexibility than the well-characterized S. oneidensis MR-1 strain, as it possesses fewer cytochrome c genes and lacks the ability to oxidize sulfite or reduce dimethyl sulfoxide (DMSO) and timethylamine oxide (TMAO). W3-18-1 lacks the hydrogen-producing Fe-only hydrogenase, and the hydrogen-oxidizing Ni-Fe hydrogenase genes were split into two separate clusters. Two periplasmic nitrate reductases (NapDAGHB and NapDABC) were functionally redundant in anaerobic growth of W3-18-1 with nitrate as the electron acceptor, though napDABC was not regulated by Crp. Moreover, nitrate respiration started earlier in W3-18-1 than in MR-1 (with NapDAGHB only) under microoxic conditions. These results indicate that Shewanella putrefaciens W3-18-1 is well adapted to habitats with higher oxygen levels. Taken together, the results of this study provide valuable insights into bacterial genome evolution.

  15. Montmorillonite supported Ni-Fe catalysts for hydrogen production from steam reforming of ethanol%Ni-Fe/蒙脱土催化剂催化乙醇水蒸气重整制氢的研究

    Institute of Scientific and Technical Information of China (English)

    李宝茹; 殷雪梅; 吴旭; 安霞; 谢鲜梅

    2016-01-01

    采用浸渍法制备了一系列Ni-Fe/蒙脱土( MMT)催化剂,并应用于乙醇水蒸气重整制氢反应( ESR)。采用X射线衍射( XRD)、N2吸附脱附分析和H2-程序升温还原( H2-TPR)表征手段对催化剂的物理化学性质、还原性能、碳沉积等进行了研究。结果表明,Ni-Fe/MMT催化剂中,Ni、Fe高度分散在载体MMT层间及表面,而且Fe的加入降低了Ni颗粒的粒径,增强了Ni2+与载体的相互作用力。以10Ni5Fe/MMT为催化剂,在反应温度为500℃、水醇比为3:1、空速为12 h-1,反应进行30 h后,乙醇转化率为100%,氢气选择性仍保持72%,副产物CO和CH4含量明显降低。这是因为催化助剂Fe的引入,一方面,提高了Ni的分散度,使得ESR低温活性较好;另一方面,减小了Ni颗粒粒径,小颗粒的Ni有利于抑制甲烷的生成,并且Fe的加入加强了甲烷重整和水煤气变换反应,提高产物中氢气的选择性。%Ni-Fe/montmorillonite ( MMT ) catalysts were prepared by impregnation method for hydrogen production via ethanol steam reforming. The catalysts were characterized by XRD, H2-TPR, and N2 adsorption-desorption . It was found that Ni-Fe bimetallic catalysts exhibited higher activities and stability than single metallic catalysts due to the well dispersed Ni-Fe, small nickel crystallites and stronger interaction between Ni2+ and carrier. The conversion and selectivity were affected by the ratio of Ni to Fe. The 10Ni5Fe/MMT catalyst showed the optimum catalytic performance, its ethanol conversion was 100%, the selectivity of hydrogen gas remained at 72%, and selectivity of CO and CH4 were significantly decreased at 500℃ during 30 h testing. This could be attributed to the promoter Fe, which improves the dispersion of Ni and results in a good ESR activity at low reaction temperature. Small Ni particles can suppress methane formation and Fe addition can enhance the methane reforming with water and water gas shift reaction, resulting in higher

  16. 聚丙烯酰胺凝胶法制备NiFe2O4纳米颗粒及性能%Preparation and Properties of NiFe2O4 Nanoparticles by a Polyacrylamide Gel Route

    Institute of Scientific and Technical Information of China (English)

    赵东方; 杨华; 县涛; 王伟鹏; 魏智强; 李瑞山; 冯旺军; 姜金龙

    2013-01-01

    采用聚丙烯酰胺凝胶法制备了NiFe2O4纳米颗粒,利用XRD、SEM、紫外-可见漫反射光谱、FTIR、XPS、VSM等对样品进行了表征分析.结果表明:分别以EDTA和乙酸作络合剂时,在600℃烧结温度下可制得单相NiFe2O4纳米颗粒;两个样品的颗粒形貌较为规整,主要以类球形为主,粒度分布较为均匀,平均粒径分别为55 nm和75nm.根据紫外-可见漫反射光谱求得样品的带隙为1.85 eV.Fe2p3/2和Ni2p3/2的XPS谱分析表明:样品为反尖晶石型结构,其化学通式可表示为(Fe3+)[Ni2+ Fe3+]O4.磁滞回线测量结果表明:样品具有良好的软磁特性,颗粒尺寸较小的样品其饱和磁化强度、剩余磁化强度和矫顽力相对较小.%A polyacrylamide gel route was used to prepare NiFe2O4 nanoparticles. The prepared samples were characterized by X-ray diffraction (XRD) , scanning electron microscope (SEM) , ultraviolet-visible diffuse reflectance spectroscopy, fourier transform infrared spectroscopy (FTIR) , X-ray photoelectron spectroscopy (XPS) , and vibrating sample magnetometer (VSM). It is demonstrated that single-phase NiFe2O4 nanoparticles can be prepared separately using ethylenediamine-tetraacetic acid (EDTA) and acetic acid as the chelating agent at a calcining temperature of 600 ℃. The two as-prepared samples have regular spherical particle shapes and uniform particle sizes with an average diameter of 55 run and 75 nm, respectively. The energy bandgap of the samples is estimated to be 1.85 eV by ultraviolet-visible diffuse reflectance spectroscopy. Fe 2p3/2 and Ni 2p3/2 XPS spectra reveal that the samples have an inverse spinel-type structure with the chemical formula (Fe3+ ) [Ni2+Fe3+ ]O4. Magnetic hysteresis loop measurements indicate that the samples exhibit good soft magnetic properties, and their saturation magnetization, remanent magnetization and coercivity have a decrease with decrease in particle size.

  17. The nifU, nifS and nifV gene products are required for activity of all three nitrogenases of Azotobacter vinelandii.

    Science.gov (United States)

    Kennedy, C; Dean, D

    1992-02-01

    Strains with mutations in 23 of the 30 genes and open reading frames in the major nif gene cluster of A. vinelandii were tested for ability to grow on N-free medium with molybdenum (Nif phenotype), with vanadium (Vnf phenotype), or with neither metal present (Anf phenotype). As reported previously, nifE, nifN, nifU, nifS and nifV mutants were Nif- (failed to grow on molybdenum) while nifM mutants were Nif-, Vnf- and Anf-. nifV, nifS, and nifU mutants were found to be unable to grow on medium with or without vanadium, i.e. were Vnf- Anf-. Therefore neither vnf nor anf analogoues of nifU, nifS, nifV or nifM are expected to be present in A. vinelandii.

  18. Kinetics and Mechanism of Carbon Tetrachloride Rapid Reduction by Nanoscale Ni-Fe Particles Using Scanning Electron Microscope%利用扫描电镜技术研究纳米Ni-Fe 颗粒对四氯化碳快速脱氯的机理

    Institute of Scientific and Technical Information of China (English)

    黄园英; 王倩; 韩子金; 刘菲

    2015-01-01

    纳米铁具有高的比表面积和高反应活性,能快速将氯代烯烃还原成无毒氯离子、乙烯和乙烷,但对于氯代烷烃的脱氯仍能产生大量的氯代中间或最终产物,可以通过合成制得纳米双金属提高脱氯速率和减少氯代中间产物。本文利用扫描电镜测得实验室制备的纳米 Ni -Fe(2%,质量分数)颗粒直径为20~60 nm,通过批实验方式对纳米 Ni -Fe 降解四氯化碳的反应动力学性质、产物、持久性能和反应机理进行了探讨。结果表明,纳米 Ni -Fe 体系主要最终产物为42% CH4和17% CH2 Cl2。与铸铁屑和纳米铁相比,纳米Ni -Fe由于催化脱氯加氢,显著提高了氯代烃脱氯速率,同时降低了有毒氯代产物的产量,且 Ni 作为催化剂不会进入水体引起二次污染。纳米 Ni -Fe 颗粒在空气中具有很好的稳定性,虽然降解四氯化碳的最终产物CH4与纳米 Pd -Fe 相比少13%,但由于价格便宜,有望在工程上应用于氯代有机化合物水土污染治理。%The nanoscale iron particles have high specific surface area and high reactivity,can be used to rapidly reduce chlorinated alkene to the non-toxic chloride ions,chlorine-free end products ethene and ethane.But nanoscale iron particle degrades chlorinated alkanes with much toxic intermediate or end products.A small amount of a second metal deposited on the iron surface has proved to enhance the reactivity of metal particles.In this paper,laboratory-synthesized nanoscale Ni-Fe (2% by weight)particles have diameters on the order of 20 -60 nm using Scanning Electron Microscope.In batch experiments,the kinetics,products,stability of performance,and mechanism of carbon tetrachloride (CT)by Ni-Fe nanoparticles were investigated.CH4 (~42%)and CH2 Cl2 (~17%)in nanoscale Ni-Fe system were the major end products.Compared to nanoscale iron and the cast iron scarp,a major benefit of the nanoscale Ni-Fe particles

  19. Use of algae technology for production of biohydrogen from green microalgae. Possibilities for a practical sustainable process and diversity at both species selection, culturing and gene transcript levels

    Energy Technology Data Exchange (ETDEWEB)

    Skjaanes, Kari

    2011-01-15

    species of green algae under controlled conditions, and Chlamydomonas noctigama shows the ability to produce hydrogen with efficiency comparable to the model organism Chlamydomonas reinhardtii. The ability to produce hydrogen under sulfur deprivation is also explored in relation to the different species' ability to show heterotrophic or mixotrophic growth on acetate. A photo bioreactor specifically designed for algae hydrogen production is described for lab scale research purposes, including considerations for measurement devices and materials choice. Hydrogen production by the algae C. noctigama is further explored at molecular level. By using RT-PCR followed by PCR with degenerate primers, mRNA with homology towards green algal hydrogenases was identified. The cDNA sequences were translated to putative amino acid sequences, and analyzed in respect to amino acids characteristic for green algal hydrogenases and amino acids which share characteristics with both hydrogenases and narf-like proteins. These results were used to evaluate the identification of the mRNA sequences found in C. noctigama. While other green algae have been shown to contain two different hydrogenases, it is here demonstrated that C. noctigama is able to transcribe three distinct genes which share essential characteristics with hydrogenases. The combination of these results provides valuable insights at several levels of a combined process for production of bio hydrogen and other valuable products. Further studies of these topics may result in a sustainable process where solar energy can be converted into hydrogen in an integrated manner, where production efficiencies are sufficient for an economic exploitation of algal technology using algal stress reactions. (Author)

  20. Phylogeny and phylogeography of functional genes shared among seven terrestrial subsurface metagenomes reveal N-cycling and microbial evolutionary relationships

    Directory of Open Access Journals (Sweden)

    Maggie CY Lau

    2014-10-01

    Full Text Available Comparative studies on community phylogenetics and phylogeography of microorganisms living in extreme environments are rare. Terrestrial subsurface habitats are valuable for studying microbial biogeographical patterns due to their isolation and the restricted dispersal mechanisms. Since the taxonomic identity of a microorganism does not always correspond well with its functional role in a particular community, the use of taxonomic assignments or patterns may give limited inference on how microbial functions are affected by historical, geographical and environmental factors. With seven metagenomic libraries generated from fracture water samples collected from five South African mines, this study was carried out to (1 screen for ubiquitous functions or pathways of biogeochemical cycling of CH4, S and N; (2 to characterize the biodiversity represented by the common functional genes; (3 to investigate the subsurface biogeography as revealed by this subset of genes; and (4 to explore the possibility of using metagenomic data for evolutionary study. The ubiquitous functional genes are NarV, NPD, PAP reductase, NifH, NifD, NifK, NifE and NifN genes. Although these 8 common functional genes were taxonomically and phylogenetically diverse and distinct from each other, the dissimilarity between samples did not correlate strongly with either geographical, environmental or residence time of the water. Por genes homologous to those of Thermodesulfovibrio yellowstonii detected in all metagenomes were deep lineages of Nitrospirae, suggesting that subsurface habitats have preserved ancestral genetic signatures that inform the study of the origin and evolution of prokaryotes.

  1. Phylogeny and phylogeography of functional genes shared among seven terrestrial subsurface metagenomes reveal N-cycling and microbial evolutionary relationships.

    Science.gov (United States)

    Lau, Maggie C Y; Cameron, Connor; Magnabosco, Cara; Brown, C Titus; Schilkey, Faye; Grim, Sharon; Hendrickson, Sarah; Pullin, Michael; Sherwood Lollar, Barbara; van Heerden, Esta; Kieft, Thomas L; Onstott, Tullis C

    2014-01-01

    Comparative studies on community phylogenetics and phylogeography of microorganisms living in extreme environments are rare. Terrestrial subsurface habitats are valuable for studying microbial biogeographical patterns due to their isolation and the restricted dispersal mechanisms. Since the taxonomic identity of a microorganism does not always correspond well with its functional role in a particular community, the use of taxonomic assignments or patterns may give limited inference on how microbial functions are affected by historical, geographical and environmental factors. With seven metagenomic libraries generated from fracture water samples collected from five South African mines, this study was carried out to (1) screen for ubiquitous functions or pathways of biogeochemical cycling of CH4, S, and N; (2) to characterize the biodiversity represented by the common functional genes; (3) to investigate the subsurface biogeography as revealed by this subset of genes; and (4) to explore the possibility of using metagenomic data for evolutionary study. The ubiquitous functional genes are NarV, NPD, PAPS reductase, NifH, NifD, NifK, NifE, and NifN genes. Although these eight common functional genes were taxonomically and phylogenetically diverse and distinct from each other, the dissimilarity between samples did not correlate strongly with geographical or environmental parameters or residence time of the water. Por genes homologous to those of Thermodesulfovibrio yellowstonii detected in all metagenomes were deep lineages of Nitrospirae, suggesting that subsurface habitats have preserved ancestral genetic signatures that inform the study of the origin and evolution of prokaryotes.

  2. Physical and genetic map of the major nif gene cluster from Azotobacter vinelandii.

    Science.gov (United States)

    Jacobson, M R; Brigle, K E; Bennett, L T; Setterquist, R A; Wilson, M S; Cash, V L; Beynon, J; Newton, W E; Dean, D R

    1989-02-01

    Determination of a 28,793-base-pair DNA sequence of a region from the Azotobacter vinelandii genome that includes and flanks the nitrogenase structural gene region was completed. This information was used to revise the previously proposed organization of the major nif cluster. The major nif cluster from A. vinelandii encodes 15 nif-specific genes whose products bear significant structural identity to the corresponding nif-specific gene products from Klebsiella pneumoniae. These genes include nifH, nifD, nifK, nifT, nifY, nifE, nifN, nifX, nifU, nifS, nifV, nifW, nifZ, nifM, and nifF. Although there are significant spatial differences, the identified A. vinelandii nif-specific genes have the same sequential arrangement as the corresponding nif-specific genes from K. pneumoniae. Twelve other potential genes whose expression could be subject to nif-specific regulation were also found interspersed among the identified nif-specific genes. These potential genes do not encode products that are structurally related to the identified nif-specific gene products. Eleven potential nif-specific promoters were identified within the major nif cluster, and nine of these are preceded by an appropriate upstream activator sequence. A + T-rich regions were identified between 8 of the 11 proposed nif promoter sequences and their upstream activator sequences. Site-directed deletion-and-insertion mutagenesis was used to establish a genetic map of the major nif cluster.

  3. Relationship among growth parameters for Clostridium butyricum, hydA gene expression, and biohydrogen production in a sucrose-supplemented batch reactor.

    Science.gov (United States)

    Wang, Mei-Yun; Olson, Betty H; Chang, Jo-Shu

    2008-03-01

    This study was undertaken to identify the relationship between the performance of dark H2 fermentation and expression of the key functional gene (i.e., hydrogenase gene) involved in the bioH2 production process. Clostridium butyricum CGS5 isolated from anaerobic sewage sludge was used as the model strain for this study. Copy number of the hydrogenase gene (hydA) and mRNA transcripts (cDNA hydA) (after amplification) and the total DNA and RNA (before amplification) were measured over the course of the growth of strain CGS5. Cell concentration was also determined by optical density and converted to dry weight. After amplification, the hydA gene increased 1,500-fold during late exponential growth phase after normalization to the copy number at time 0, and cDNA from mRNA transcripts of hydA also increased 500-fold after normalization. mRNA transcripts of hydA lagged behind the increase of total DNA and RNA, and increases in hydA more closely mimicked those of total DNA. Increases in both of these parameters corresponded with hydrogen production. Transcripts of 16s ribosomal RNA reached a maximum value earlier (38 h) than did those of hydA (47 h). All molecular characteristics matched those for sucrose utilization, growth, and hydrogen production. These experiments indicated that transcription as measured by cDNA can be related to hydrogen production and possesses the potential to be used as tool for process control.

  4. Direct purification of his-tagged recombinant protein by NiFe2 O4 magnetic nanoparticles%铁镍磁性微球直接纯化组氨酸标签蛋白

    Institute of Scientific and Technical Information of China (English)

    刘宝全; 张停停; 鲍丽; 吕晓菊; 王佳宁; 张艳梅; 王剑锋; 范圣第

    2015-01-01

    The iron-nickel magnetic nanoparticle was synthesized with thermal decomposition method. Ferric chloride,nickel sulfate and sodium acetate were stirred in ethylene glycol at 80℃ for 30 minutes,and then stirred at 180℃ under the protection of nitrogen for 6 hours after the addition of ethanol amine,the iron-nickel magnetic nanoparticles were obtained with one pot method and were washed with water and ethanol. The nanoparticles could be dispersed in water and gathered in magnetic field. According to the XRD spectrum and the standard database,these results were obtained:the magnetic particle was cube,the crystal type was NiFe2 O4 ,and the particle size was 6. 75nm calculated with Scherrer formal. The His-tagged green fluorescent protein( GFP) was expressed in E-. coli under the induction of IPTG. The supernatant from the ultrasonic lysates was associated with the magnetic nanoparticles to pu-rify the tagged GFP. The GFP was obtained with different elution buffers from the magnetic nanoparticles or Ni-NTA agarose and de-tected with SDS-PAGE. The results showed that the protein yield of Ni-NTA agarose was five times than the magnetic nanoparti-cles. The low-cost nickel-iron magnetic nanoparticles were obtained by the hydrothermal synthesis method,and could replace the Ni-NTA agarose to purify the tagged green fluorescent protein directly.%以氯化铁与硫酸镍为主要原料基于热分解法原理一锅法合成铁镍磁性微球。氯化铁、硫酸镍和乙酸钠在(80℃)乙二醇中搅拌30 min,在氮气保护下,加入乙醇胺后180℃搅拌6h,经过水清洗与乙醇清洗后获得有磁性的含镍微球,XRD检测结果证实晶型为NiFe2 O4,晶体的平均粒径为6.75 nm。磁性微球不经任何修饰直接与组氨酸标签蛋白结合,完成带有组氨酸标签的绿色荧光蛋白纯化,SDS-PAGE检测结果表明:使用5倍于镍配位凝胶质量的铁镍磁性微球可达到与镍配位凝胶相近的蛋白质吸附量。低成本的铁

  5. RNA-seq profiling reveals novel target genes of LexA in the cyanobacterium Synechocystis sp. PCC 6803

    Directory of Open Access Journals (Sweden)

    Ayumi eKizawa

    2016-02-01

    Full Text Available LexA is a well-established transcriptional repressor of SOS genes induced by DNA damage in E. coli and other bacterial species. However, LexA in the cyanobacterium Synechocystis sp. PCC 6803 has been suggested not to be involved in SOS response. In this study, we performed RNA-seq analysis of the wild-type strain and the lexA-disrupted mutant to obtain the comprehensive view of LexA-regulated genes in Synechocystis. Disruption of lexA positively or negatively affected expression of genes related to various cellular functions such as phototactic motility, accumulation of the major compatible solute glucosylglycerol and subunits of bidirectional hydrogenase, photosystem I and phycobilisome complexes. We also observed increase in the expression level of genes related to iron and manganese uptake in the mutant at the later stage of cultivation. However, none of the genes related to DNA metabolism were affected by disruption of lexA. DNA gel mobility shift assay using the recombinant LexA protein suggested that LexA binds to the upstream region of pilA7, pilA9, ggpS and slr1670 to directly regulate their expression, but changes in the expression level of photosystem I genes by disruption of lexA is likely a secondary effect.

  6. RNA-seq Profiling Reveals Novel Target Genes of LexA in the Cyanobacterium Synechocystis sp. PCC 6803

    Science.gov (United States)

    Kizawa, Ayumi; Kawahara, Akihito; Takimura, Yasushi; Nishiyama, Yoshitaka; Hihara, Yukako

    2016-01-01

    LexA is a well-established transcriptional repressor of SOS genes induced by DNA damage in Escherichia coli and other bacterial species. However, LexA in the cyanobacterium Synechocystis sp. PCC 6803 has been suggested not to be involved in SOS response. In this study, we performed RNA-seq analysis of the wild-type strain and the lexA-disrupted mutant to obtain the comprehensive view of LexA-regulated genes in Synechocystis. Disruption of lexA positively or negatively affected expression of genes related to various cellular functions such as phototactic motility, accumulation of the major compatible solute glucosylglycerol and subunits of bidirectional hydrogenase, photosystem I, and phycobilisome complexes. We also observed increase in the expression level of genes related to iron and manganese uptake in the mutant at the later stage of cultivation. However, none of the genes related to DNA metabolism were affected by disruption of lexA. DNA gel mobility shift assay using the recombinant LexA protein suggested that LexA binds to the upstream region of pilA7, pilA9, ggpS, and slr1670 to directly regulate their expression, but changes in the expression level of photosystem I genes by disruption of lexA is likely a secondary effect. PMID:26925056

  7. Studying Genes

    Science.gov (United States)

    ... NIGMS NIGMS Home > Science Education > Studying Genes Studying Genes Tagline (Optional) Middle/Main Content Area Other Fact Sheets What are genes? Genes are segments of DNA that contain instructions ...

  8. Effect of a C298D Mutation in CaHydA [FeFe]-Hydrogenase: Insights into the Protein-Metal Cluster Interaction by EPR and FTIR Spectroscopic Investigation

    Energy Technology Data Exchange (ETDEWEB)

    Morra, Simone; Maurelli, Sara; Chiesa, Mario; Mulder, David W.; Ratzloff, Michael W.; Giamello, Elio; King, Paul W.; Gilardi, Gianfranco; Valettia, Francesca

    2016-01-01

    A conserved cysteine located in the signature motif of the catalytic center (H-cluster) of [FeFe]-hydrogenases functions in proton transfer. This residue corresponds to C298 in Clostridium acetobutylicum CaHydA. Despite the chemical and structural difference, the mutant C298D retains fast catalytic activity, while replacement with any other amino acid caused significant activity loss. Given the proximity of C298 to the H-cluster, the effect of the C298D mutation on the catalytic center was studied by continuous wave (CW) and pulse electron paramagnetic resonance (EPR) and by Fourier transform infrared (FTIR) spectroscopies. Comparison of the C298D mutant with the wild type CaHydA by CW and pulse EPR showed that the electronic structure of the center is not altered. FTIR spectroscopy confirmed that absorption peak values observed in the mutant are virtually identical to those observed in the wild type, indicating that the H-cluster is not generally affected by the mutation. Significant differences were observed only in the inhibited state Hox-CO: the vibrational modes assigned to the COexo and Fed-CO in this state are shifted to lower values in C298D, suggesting different interaction of these ligands with the protein moiety when C298 is changed to D298. More relevant to the catalytic cycle, the redox equilibrium between the Hox and Hred states is modified by the mutation, causing a prevalence of the oxidized state. This work highlights how the interactions between the protein environment and the H-cluster, a dynamic closely interconnected system, can be engineered and studied in the perspective of designing bio-inspired catalysts and mimics.

  9. Favorable Protonation of the (μ-edt)[Fe(2)(PMe(3))(4)(CO)(2)(H-terminal)](+) Hydrogenase Model Complex Over Its Bridging μ-H Counterpart: A Spectroscopic and DFT Study.

    Science.gov (United States)

    Galinato, Mary Grace I; Whaley, C Matthew; Roberts, Dean; Wang, Peng; Lehnert, Nicolai

    2011-03-01

    The mechanism of hydrogen production in [FeFe] hydrogenase remains elusive. However, a species featuring a terminal hydride bound to the distal Fe is thought to be the key intermediate leading to hydrogen production. In this study, density functional theory (DFT) calculations on the terminal (H-term) and bridging (μ-H) hydride isomers of (μ-edt)-[Fe(2)(PMe(3))(4)(CO)(2)H](+) are presented in order to understand the factors affecting their propensity for protonation. Relative to H-term, μ-H is 12.7 kcal/mol more stable, which contributes to its decreased reactivity towards an acid. Potential energy surface (PES) calculations for the reaction of the H-term isomer with 4-nitropyridinium, a proton source, further reveal a lower activation energy barrier (14.5 kcal/mol) for H-term than for μ-H (29 kcal/mol). Besides these energetic considerations, the H-term isomer displays a key molecular orbital (MO ) that has a relatively strong hydride (1s) contribution (23%), which is not present in the μ-H isomer. This indicates a potential orbital control of the reaction of the hydride complexes with acid. The lower activation energy barrier and this key MO together control the overall catalytic activity of (μ-edt)[Fe(2)(PMe(3))(4)(CO)(2)(H-term)](+). Lastly, Raman and IR spectroscopy were performed in order to probe the ν(Fe-H) stretching mode of the two isomers and their deuterated counterparts. A ν(Fe-H) stretching mode was observed for the μ-H complex at 1220 cm(-1). However, the corresponding mode is not observed for the less stable H-term isomer.

  10. Challenges to develop nitrogen-fixing cereals by direct nif-gene transfer.

    Science.gov (United States)

    Curatti, Leonardo; Rubio, Luis M

    2014-08-01

    Some regions of the developing world suffer low cereal production yields due to low fertilizer inputs, among other factors. Biological N2 fixation, catalyzed by the prokaryotic enzyme nitrogenase, is an alternative to the use of synthetic N fertilizers. The molybdenum nitrogenase is an O2-labile metalloenzyme composed of the NifDK and NifH proteins, which biosyntheses require a number of nif gene products. A challenging strategy to increase cereal crop productivity in a scenario of low N fertilization is the direct transfer of nif genes into cereals. The sensitivity of nitrogenase to O2 and the apparent complexity of nitrogenase biosynthesis are the main barriers identified so far. Expression of active NifH requires the products of nifM, nifH, and possibly nifU and nifS, whereas active NifDK requires the products of nifH, nifD, nifK, nifB, nifE, nifN, and possibly nifU, nifS, nifQ, nifV, nafY, nifW and nifZ. Plastids and mitochondria are potential subcellular locations for nitrogenase. Both could provide the ATP and electrons required for nitrogenase to function but they differ in their internal O2 levels and their ability to incorporate ammonium into amino acids.

  11. Immunoglobulin genes

    Energy Technology Data Exchange (ETDEWEB)

    Honjo, T. (Kyoto Univ. (Japan)); Alt, F.W. (Columbia Univ., Dobbs Ferry, NY (USA). Hudson Labs.); Rabbitts, T.H. (Medical Research Council, Cambridge (UK))

    1989-01-01

    This book reports on the structure, function, and expression of the genes encoding antibodies in normal and neoplastic cells. Topics covered are: B Cells; Organization and rearrangement of immunoglobin genes; Immunoglobin genes in disease; Immunoglobin gene expression; and Immunoglobin-related genes.

  12. Identification of the nik Gene Cluster of Brucella suis: Regulation and Contribution to Urease Activity

    Science.gov (United States)

    Jubier-Maurin, Véronique; Rodrigue, Agnès; Ouahrani-Bettache, Safia; Layssac, Marion; Mandrand-Berthelot, Marie-Andrée; Köhler, Stephan; Liautard, Jean-Pierre

    2001-01-01

    Analysis of a Brucella suis 1330 gene fused to a gfp reporter, and identified as being induced in J774 murine macrophage-like cells, allowed the isolation of a gene homologous to nikA, the first gene of the Escherichia coli operon encoding the specific transport system for nickel. DNA sequence analysis of the corresponding B. suis nik locus showed that it was highly similar to that of E. coli except for localization of the nikR regulatory gene, which lies upstream from the structural nikABCDE genes and in the opposite orientation. Protein sequence comparisons suggested that the deduced nikABCDE gene products belong to a periplasmic binding protein-dependent transport system. The nikA promoter-gfp fusion was activated in vitro by low oxygen tension and metal ion deficiency and was repressed by NiCl2 excess. Insertional inactivation of nikA strongly reduced the activity of the nickel metalloenzyme urease, which was restored by addition of a nickel excess. Moreover, the nikA mutant of B. suis was functionally complemented with the E. coli nik gene cluster, leading to the recovery of urease activity. Reciprocally, an E. coli strain harboring a deleted nik operon recovered hydrogenase activity by heterologous complementation with the B. suis nik locus. Taking into account these results, we propose that the nik locus of B. suis encodes a nickel transport system. The results further suggest that nickel could enter B. suis via other transport systems. Intracellular growth rates of the B. suis wild-type and nikA mutant strains in human monocytes were similar, indicating that nikA was not essential for this step of infection. We discuss a possible role of nickel transport in maintaining enzymatic activities which could be crucial for survival of the bacteria under the environmental conditions encountered within the host. PMID:11133934

  13. A minimal nitrogen fixation gene cluster from Paenibacillus sp. WLY78 enables expression of active nitrogenase in Escherichia coli.

    Science.gov (United States)

    Wang, Liying; Zhang, Lihong; Liu, Zhanzhi; Liu, Zhangzhi; Zhao, Dehua; Liu, Xiaomeng; Zhang, Bo; Xie, Jianbo; Hong, Yuanyuan; Li, Pengfei; Chen, Sanfeng; Dixon, Ray; Li, Jilun

    2013-01-01

    Most biological nitrogen fixation is catalyzed by molybdenum-dependent nitrogenase, an enzyme complex comprising two component proteins that contains three different metalloclusters. Diazotrophs contain a common core of nitrogen fixation nif genes that encode the structural subunits of the enzyme and components required to synthesize the metalloclusters. However, the complement of nif genes required to enable diazotrophic growth varies significantly amongst nitrogen fixing bacteria and archaea. In this study, we identified a minimal nif gene cluster consisting of nine nif genes in the genome of Paenibacillus sp. WLY78, a gram-positive, facultative anaerobe isolated from the rhizosphere of bamboo. We demonstrate that the nif genes in this organism are organized as an operon comprising nifB, nifH, nifD, nifK, nifE, nifN, nifX, hesA and nifV and that the nif cluster is under the control of a σ(70) (σ(A))-dependent promoter located upstream of nifB. To investigate genetic requirements for diazotrophy, we transferred the Paenibacillus nif cluster to Escherichia coli. The minimal nif gene cluster enables synthesis of catalytically active nitrogenase in this host, when expressed either from the native nifB promoter or from the T7 promoter. Deletion analysis indicates that in addition to the core nif genes, hesA plays an important role in nitrogen fixation and is responsive to the availability of molybdenum. Whereas nif transcription in Paenibacillus is regulated in response to nitrogen availability and by the external oxygen concentration, transcription from the nifB promoter is constitutive in E. coli, indicating that negative regulation of nif transcription is bypassed in the heterologous host. This study demonstrates the potential for engineering nitrogen fixation in a non-nitrogen fixing organism with a minimum set of nine nif genes.

  14. A minimal nitrogen fixation gene cluster from Paenibacillus sp. WLY78 enables expression of active nitrogenase in Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Liying Wang

    Full Text Available Most biological nitrogen fixation is catalyzed by molybdenum-dependent nitrogenase, an enzyme complex comprising two component proteins that contains three different metalloclusters. Diazotrophs contain a common core of nitrogen fixation nif genes that encode the structural subunits of the enzyme and components required to synthesize the metalloclusters. However, the complement of nif genes required to enable diazotrophic growth varies significantly amongst nitrogen fixing bacteria and archaea. In this study, we identified a minimal nif gene cluster consisting of nine nif genes in the genome of Paenibacillus sp. WLY78, a gram-positive, facultative anaerobe isolated from the rhizosphere of bamboo. We demonstrate that the nif genes in this organism are organized as an operon comprising nifB, nifH, nifD, nifK, nifE, nifN, nifX, hesA and nifV and that the nif cluster is under the control of a σ(70 (σ(A-dependent promoter located upstream of nifB. To investigate genetic requirements for diazotrophy, we transferred the Paenibacillus nif cluster to Escherichia coli. The minimal nif gene cluster enables synthesis of catalytically active nitrogenase in this host, when expressed either from the native nifB promoter or from the T7 promoter. Deletion analysis indicates that in addition to the core nif genes, hesA plays an important role in nitrogen fixation and is responsive to the availability of molybdenum. Whereas nif transcription in Paenibacillus is regulated in response to nitrogen availability and by the external oxygen concentration, transcription from the nifB promoter is constitutive in E. coli, indicating that negative regulation of nif transcription is bypassed in the heterologous host. This study demonstrates the potential for engineering nitrogen fixation in a non-nitrogen fixing organism with a minimum set of nine nif genes.

  15. A minimal nitrogen fixation gene cluster from Paenibacillus sp. WLY78 enables expression of active nitrogenase in Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Liying Wang

    Full Text Available Most biological nitrogen fixation is catalyzed by molybdenum-dependent nitrogenase, an enzyme complex comprising two component proteins that contains three different metalloclusters. Diazotrophs contain a common core of nitrogen fixation nif genes that encode the structural subunits of the enzyme and components required to synthesize the metalloclusters. However, the complement of nif genes required to enable diazotrophic growth varies significantly amongst nitrogen fixing bacteria and archaea. In this study, we identified a minimal nif gene cluster consisting of nine nif genes in the genome of Paenibacillus sp. WLY78, a gram-positive, facultative anaerobe isolated from the rhizosphere of bamboo. We demonstrate that the nif genes in this organism are organized as an operon comprising nifB, nifH, nifD, nifK, nifE, nifN, nifX, hesA and nifV and that the nif cluster is under the control of a σ(70 (σ(A-dependent promoter located upstream of nifB. To investigate genetic requirements for diazotrophy, we transferred the Paenibacillus nif cluster to Escherichia coli. The minimal nif gene cluster enables synthesis of catalytically active nitrogenase in this host, when expressed either from the native nifB promoter or from the T7 promoter. Deletion analysis indicates that in addition to the core nif genes, hesA plays an important role in nitrogen fixation and is responsive to the availability of molybdenum. Whereas nif transcription in Paenibacillus is regulated in response to nitrogen availability and by the external oxygen concentration, transcription from the nifB promoter is constitutive in E. coli, indicating that negative regulation of nif transcription is bypassed in the heterologous host. This study demonstrates the potential for engineering nitrogen fixation in a non-nitrogen fixing organism with a minimum set of nine nif genes.

  16. A Minimal Nitrogen Fixation Gene Cluster from Paenibacillus sp. WLY78 Enables Expression of Active Nitrogenase in Escherichia coli

    Science.gov (United States)

    Zhao, Dehua; Liu, Xiaomeng; Zhang, Bo; Xie, Jianbo; Hong, Yuanyuan; Li, Pengfei; Chen, Sanfeng; Dixon, Ray; Li, Jilun

    2013-01-01

    Most biological nitrogen fixation is catalyzed by molybdenum-dependent nitrogenase, an enzyme complex comprising two component proteins that contains three different metalloclusters. Diazotrophs contain a common core of nitrogen fixation nif genes that encode the structural subunits of the enzyme and components required to synthesize the metalloclusters. However, the complement of nif genes required to enable diazotrophic growth varies significantly amongst nitrogen fixing bacteria and archaea. In this study, we identified a minimal nif gene cluster consisting of nine nif genes in the genome of Paenibacillus sp. WLY78, a gram-positive, facultative anaerobe isolated from the rhizosphere of bamboo. We demonstrate that the nif genes in this organism are organized as an operon comprising nifB, nifH, nifD, nifK, nifE, nifN, nifX, hesA and nifV and that the nif cluster is under the control of a σ70 (σA)-dependent promoter located upstream of nifB. To investigate genetic requirements for diazotrophy, we transferred the Paenibacillus nif cluster to Escherichia coli. The minimal nif gene cluster enables synthesis of catalytically active nitrogenase in this host, when expressed either from the native nifB promoter or from the T7 promoter. Deletion analysis indicates that in addition to the core nif genes, hesA plays an important role in nitrogen fixation and is responsive to the availability of molybdenum. Whereas nif transcription in Paenibacillus is regulated in response to nitrogen availability and by the external oxygen concentration, transcription from the nifB promoter is constitutive in E. coli, indicating that negative regulation of nif transcription is bypassed in the heterologous host. This study demonstrates the potential for engineering nitrogen fixation in a non-nitrogen fixing organism with a minimum set of nine nif genes. PMID:24146630

  17. Open reading frame 5 (ORF5), encoding a ferredoxinlike protein, and nifQ are cotranscribed with nifE, nifN, nifX, and ORF4 in Rhodobacter capsulatus.

    OpenAIRE

    Moreno-Vivian, C; Hennecke, S; Pühler, A.; Klipp, W

    1989-01-01

    DNA sequence analysis of a 1,600-base-pair fragment located downstream of nifENX in nif region A of Rhodobacter capsulatus revealed two additional open reading frames (ORFs): ORF5, encoding a ferredoxinlike protein, and nifQ. The ferredoxinlike gene product contained two cysteine motifs, typical of ferredoxins coordinating two 4Fe-4S clusters, but the distance between these two motifs was unusual for low-molecular-weight ferredoxins. The R. capsulatus nifQ gene product shared a high degree of...

  18. Metatranscriptomic insights on gene expression and regulatory controls in Candidatus Accumulibacter phosphatis

    Science.gov (United States)

    Oyserman, Ben O; Noguera, Daniel R; del Rio, Tijana Glavina; Tringe, Susannah G; McMahon, Katherine D

    2016-01-01

    Previous studies on enhanced biological phosphorus removal (EBPR) have focused on reconstructing genomic blueprints for the model polyphosphate-accumulating organism Candidatus Accumulibacter phosphatis. Here, a time series metatranscriptome generated from enrichment cultures of Accumulibacter was used to gain insight into anerobic/aerobic metabolism and regulatory mechanisms within an EBPR cycle. Co-expressed gene clusters were identified displaying ecologically relevant trends consistent with batch cycle phases. Transcripts displaying increased abundance during anerobic acetate contact were functionally enriched in energy production and conversion, including upregulation of both cytoplasmic and membrane-bound hydrogenases demonstrating the importance of transcriptional regulation to manage energy and electron flux during anerobic acetate contact. We hypothesized and demonstrated hydrogen production after anerobic acetate contact, a previously unknown strategy for Accumulibacter to maintain redox balance. Genes involved in anerobic glycine utilization were identified and phosphorus release after anerobic glycine contact demonstrated, suggesting that Accumulibacter routes diverse carbon sources to acetyl-CoA formation via previously unrecognized pathways. A comparative genomics analysis of sequences upstream of co-expressed genes identified two statistically significant putative regulatory motifs. One palindromic motif was identified upstream of genes involved in PHA synthesis and acetate activation and is hypothesized to be a phaR binding site, hence representing a hypothetical PHA modulon. A second motif was identified ~35 base pairs (bp) upstream of a large and diverse array of genes and hence may represent a sigma factor binding site. This analysis provides a basis and framework for further investigations into Accumulibacter metabolism and the reconstruction of regulatory networks in uncultured organisms. PMID:26555245

  19. Models of the iron-only hydrogenase: a comparison of chelate and bridge isomers of Fe2(CO)4{Ph2PN(R)PPh2}(μ-pdt) as proton-reduction catalysts.

    Science.gov (United States)

    Ghosh, Shishir; Hogarth, Graeme; Hollingsworth, Nathan; Holt, Katherine B; Richards, Idris; Richmond, Michael G; Sanchez, Ben E; Unwin, David

    2013-05-21

    the two metal centres based on the DFT calculations. The LUMO in the isomeric model compounds is similar in nature and is best described as an antibonding Fe-Fe interaction that contains differing amounts of aryl π* contributions from the ancillary PNP ligand. The proton reduction catalysis observed under electrochemical conditions at ca. -1.55 V is discussed as a function of the initial isomer and a mechanism that involves an initial protonation step involving the iron-iron bond. The measured CV currents were higher at this potential for the chelating complex, indicating faster turnover. Digital simulations showed that the faster rate of catalysis of the chelating complex can be traced to its greater propensity for protonation. This supports the theory that asymmetric distribution of electron density along the iron-iron bond leads to faster catalysis for models of the Fe-Fe hydrogenase active site.

  20. Ecological and genomic profiling of anaerobic methane-oxidizing archaea in a deep granitic environment.

    Science.gov (United States)

    Ino, Kohei; Hernsdorf, Alex W; Konno, Uta; Kouduka, Mariko; Yanagawa, Katsunori; Kato, Shingo; Sunamura, Michinari; Hirota, Akinari; Togo, Yoko S; Ito, Kazumasa; Fukuda, Akari; Iwatsuki, Teruki; Mizuno, Takashi; Komatsu, Daisuke D; Tsunogai, Urumu; Ishimura, Toyoho; Amano, Yuki; Thomas, Brian C; Banfield, Jillian F; Suzuki, Yohey

    2017-09-08

    Recent single-gene-based surveys of deep continental aquifers demonstrated the widespread occurrence of archaea related to Candidatus Methanoperedens nitroreducens (ANME-2d) known to mediate anaerobic oxidation of methane (AOM). However, it is unclear whether ANME-2d mediates AOM in the deep continental biosphere. In this study, we found the dominance of ANME-2d in groundwater enriched in sulfate and methane from a 300-m deep underground borehole in granitic rock. A near-complete genome of one representative species of the ANME-2d obtained from the underground borehole has most of functional genes required for AOM and assimilatory sulfate reduction. The genome of the subsurface ANME-2d is different from those of other members of ANME-2d by lacking functional genes encoding nitrate and nitrite reductases and multiheme cytochromes. In addition, the subsurface ANME-2d genome contains a membrane-bound NiFe hydrogenase gene putatively involved in respiratory H2 oxidation, which is different from those of other methanotrophic archaea. Short-term incubation of microbial cells collected from the granitic groundwater with (13)C-labeled methane also demonstrates that AOM is linked to microbial sulfate reduction. Given the prominence of granitic continental crust and sulfate and methane in terrestrial subsurface fluids, we conclude that AOM may be widespread in the deep continental biosphere.The ISME Journal advance online publication, 8 September 2017; doi:10.1038/ismej.2017.140.

  1. Genomic Analysis of Caldithrix abyssi, the Thermophilic Anaerobic Bacterium of the Novel Bacterial Phylum Calditrichaeota

    Science.gov (United States)

    Kublanov, Ilya V.; Sigalova, Olga M.; Gavrilov, Sergey N.; Lebedinsky, Alexander V.; Rinke, Christian; Kovaleva, Olga; Chernyh, Nikolai A.; Ivanova, Natalia; Daum, Chris; Reddy, T.B.K.; Klenk, Hans-Peter; Spring, Stefan; Göker, Markus; Reva, Oleg N.; Miroshnichenko, Margarita L.; Kyrpides, Nikos C.; Woyke, Tanja; Gelfand, Mikhail S.; Bonch-Osmolovskaya, Elizaveta A.

    2017-01-01

    The genome of Caldithrix abyssi, the first cultivated representative of a phylum-level bacterial lineage, was sequenced within the framework of Genomic Encyclopedia of Bacteria and Archaea (GEBA) project. The genomic analysis revealed mechanisms allowing this anaerobic bacterium to ferment peptides or to implement nitrate reduction with acetate or molecular hydrogen as electron donors. The genome encoded five different [NiFe]- and [FeFe]-hydrogenases, one of which, group 1 [NiFe]-hydrogenase, is presumably involved in lithoheterotrophic growth, three other produce H2 during fermentation, and one is apparently bidirectional. The ability to reduce nitrate is determined by a nitrate reductase of the Nap family, while nitrite reduction to ammonia is presumably catalyzed by an octaheme cytochrome c nitrite reductase εHao. The genome contained genes of respiratory polysulfide/thiosulfate reductase, however, elemental sulfur and thiosulfate were not used as the electron acceptors for anaerobic respiration with acetate or H2, probably due to the lack of the gene of the maturation protein. Nevertheless, elemental sulfur and thiosulfate stimulated growth on fermentable substrates (peptides), being reduced to sulfide, most probably through the action of the cytoplasmic sulfide dehydrogenase and/or NAD(P)-dependent [NiFe]-hydrogenase (sulfhydrogenase) encoded by the genome. Surprisingly, the genome of this anaerobic microorganism encoded all genes for cytochrome c oxidase, however, its maturation machinery seems to be non-operational due to genomic rearrangements of supplementary genes. Despite the fact that sugars were not among the substrates reported when C. abyssi was first described, our genomic analysis revealed multiple genes of glycoside hydrolases, and some of them were predicted to be secreted. This finding aided in bringing out four carbohydrates that supported the growth of C. abyssi: starch, cellobiose, glucomannan and xyloglucan. The genomic analysis

  2. AlCoCrCu0.5 NiFe高熵合金氧化物薄膜光学特性的研究%Optical Properties of Sputtered Oxide Films of AlCoCrCu0. 5 NiFe High-entropy Alloy

    Institute of Scientific and Technical Information of China (English)

    黄元盛; 蔡铭洪; 叶均蔚

    2016-01-01

    目的:制备AlCoCrCu0.5 NiFe高熵合金氧化物薄膜,并对其光学性能进行表征。方法使用磁控溅射设备在单晶硅片和玻璃上制备AlCoCrCu0.5 NiFe高熵合金氧化物薄膜,并对膜进行退火处理。使用椭圆偏振光谱仪对薄膜的光学特性进行分析。结果随着氧含量的增加,折射系数减小。当光波长为633 nm时,折射系数为1.69~2.40。当氧分压为10%,折射率色散曲线在475 nm和600 nm处出现拐点,在600 nm之后折射率随着波长的增大而逐渐减小。当氧分压为30%时,折射率曲线在500 nm和600 nm处出现拐点,在600 nm后折射率趋于稳定。当氧分压为50%时,折射率曲线在525 nm处出现拐点,之后折射率随波长的增大而逐渐增大。在450~550 nm波段内,AlCoCrCu0.5 FeNi氧化物薄膜的吸收系数随氧分压的增加而增加。在550~850 nm波段内,薄膜的吸收系数随工作气压的变化趋势不明显。随着氧分压的增加膜的颜色逐渐变深。经过退火处理后,膜的颜色进一步加深。在相同工艺参数的情况下,氧的分压增加,膜厚减小。结论适当减小氧分压,能获得具有高折射率的AlCoCrCu0.5 FeNi氧化物薄膜。不同的分压下,AlCoCrCu0.5 FeNi氧化物薄膜的吸收系数随波长的增加均存在一个拐点,并且随氧分压的增加,拐点的波长减小。氧含量增加导致氧化物薄膜厚度减小,颜色加深。%Objective To synthesize the oxide films of AlCoCrCu0. 5 NiFe high-entropy alloy and characterize their optical proper-ties. Methods The sputtered oxide films of AlCoCrCu0. 5 NiFe high-entropy alloy were deposited on the silicon wafer and glass using radio frequency sputter system, and were subsequently annealed. Ellipsometer was employed to analyze thickness, refractive index ( n) and absorption index ( k) . Results The refractive index n decreased with the oxygen concentration. When the wavelength was 633 nm, n varied between 1. 69 and 2. 40. At the oxygen

  3. Gene therapy

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    2005147 CNHK200-hA-a gene-viral therapeutic system and its antitumor effect on lung cancer. WANG Wei-guo(王伟国),et al. Viral & Gene Ther Center, Eastern Hepatobilli Surg Instit 2nd Milit Univ, Shanghai 200438. Chin J Oncol,2005:27(2):69-72. Objective: To develop a novel vector system, which combines the advantages of the gene therapy,

  4. Inferring Gene Networks for Strains of Dehalococcoides Highlights Conserved Relationships between Genes Encoding Core Catabolic and Cell-Wall Structural Proteins.

    Science.gov (United States)

    Mansfeldt, Cresten B; Heavner, Gretchen W; Rowe, Annette R; Hayete, Boris; Church, Bruce W; Richardson, Ruth E

    2016-01-01

    The interpretation of high-throughput gene expression data for non-model microorganisms remains obscured because of the high fraction of hypothetical genes and the limited number of methods for the robust inference of gene networks. Therefore, to elucidate gene-gene and gene-condition linkages in the bioremediation-important genus Dehalococcoides, we applied a Bayesian inference strategy called Reverse Engineering/Forward Simulation (REFS™) on transcriptomic data collected from two organohalide-respiring communities containing different Dehalococcoides mccartyi strains: the Cornell University mixed community D2 and the commercially available KB-1® bioaugmentation culture. In total, 49 and 24 microarray datasets were included in the REFS™ analysis to generate an ensemble of 1,000 networks for the Dehalococcoides population in the Cornell D2 and KB-1® culture, respectively. Considering only linkages that appeared in the consensus network for each culture (exceeding the determined frequency cutoff of ≥ 60%), the resulting Cornell D2 and KB-1® consensus networks maintained 1,105 nodes (genes or conditions) with 974 edges and 1,714 nodes with 1,455 edges, respectively. These consensus networks captured multiple strong and biologically informative relationships. One of the main highlighted relationships shared between these two cultures was a direct edge between the transcript encoding for the major reductive dehalogenase (tceA (D2) or vcrA (KB-1®)) and the transcript for the putative S-layer cell wall protein (DET1407 (D2) or KB1_1396 (KB-1®)). Additionally, transcripts for two key oxidoreductases (a [Ni Fe] hydrogenase, Hup, and a protein with similarity to a formate dehydrogenase, "Fdh") were strongly linked, generalizing a strong relationship noted previously for Dehalococcoides mccartyi strain 195 to multiple strains of Dehalococcoides. Notably, the pangenome array utilized when monitoring the KB-1® culture was capable of resolving signals from multiple

  5. Inferring Gene Networks for Strains of Dehalococcoides Highlights Conserved Relationships between Genes Encoding Core Catabolic and Cell-Wall Structural Proteins

    Science.gov (United States)

    Mansfeldt, Cresten B.; Heavner, Gretchen W.; Rowe, Annette R.; Hayete, Boris; Church, Bruce W.; Richardson, Ruth E.

    2016-01-01

    The interpretation of high-throughput gene expression data for non-model microorganisms remains obscured because of the high fraction of hypothetical genes and the limited number of methods for the robust inference of gene networks. Therefore, to elucidate gene-gene and gene-condition linkages in the bioremediation-important genus Dehalococcoides, we applied a Bayesian inference strategy called Reverse Engineering/Forward Simulation (REFS™) on transcriptomic data collected from two organohalide-respiring communities containing different Dehalococcoides mccartyi strains: the Cornell University mixed community D2 and the commercially available KB-1® bioaugmentation culture. In total, 49 and 24 microarray datasets were included in the REFS™ analysis to generate an ensemble of 1,000 networks for the Dehalococcoides population in the Cornell D2 and KB-1® culture, respectively. Considering only linkages that appeared in the consensus network for each culture (exceeding the determined frequency cutoff of ≥ 60%), the resulting Cornell D2 and KB-1® consensus networks maintained 1,105 nodes (genes or conditions) with 974 edges and 1,714 nodes with 1,455 edges, respectively. These consensus networks captured multiple strong and biologically informative relationships. One of the main highlighted relationships shared between these two cultures was a direct edge between the transcript encoding for the major reductive dehalogenase (tceA (D2) or vcrA (KB-1®)) and the transcript for the putative S-layer cell wall protein (DET1407 (D2) or KB1_1396 (KB-1®)). Additionally, transcripts for two key oxidoreductases (a [Ni Fe] hydrogenase, Hup, and a protein with similarity to a formate dehydrogenase, “Fdh”) were strongly linked, generalizing a strong relationship noted previously for Dehalococcoides mccartyi strain 195 to multiple strains of Dehalococcoides. Notably, the pangenome array utilized when monitoring the KB-1® culture was capable of resolving signals from

  6. Trichoderma genes

    Science.gov (United States)

    Foreman, Pamela [Los Altos, CA; Goedegebuur, Frits [Vlaardingen, NL; Van Solingen, Pieter [Naaldwijk, NL; Ward, Michael [San Francisco, CA

    2012-06-19

    Described herein are novel gene sequences isolated from Trichoderma reesei. Two genes encoding proteins comprising a cellulose binding domain, one encoding an arabionfuranosidase and one encoding an acetylxylanesterase are described. The sequences, CIP1 and CIP2, contain a cellulose binding domain. These proteins are especially useful in the textile and detergent industry and in pulp and paper industry.

  7. Gene Therapy.

    Science.gov (United States)

    Thorne, Barb; Takeya, Ryan; Vitelli, Francesca; Swanson, Xin

    2017-03-14

    Gene therapy refers to a rapidly growing field of medicine in which genes are introduced into the body to treat or prevent diseases. Although a variety of methods can be used to deliver the genetic materials into the target cells and tissues, modified viral vectors represent one of the more common delivery routes because of its transduction efficiency for therapeutic genes. Since the introduction of gene therapy concept in the 1970s, the field has advanced considerably with notable clinical successes being demonstrated in many clinical indications in which no standard treatment options are currently available. It is anticipated that the clinical success the field observed in recent years can drive requirements for more scalable, robust, cost effective, and regulatory-compliant manufacturing processes. This review provides a brief overview of the current manufacturing technologies for viral vectors production, drawing attention to the common upstream and downstream production process platform that is applicable across various classes of viral vectors and their unique manufacturing challenges as compared to other biologics. In addition, a case study of an industry-scale cGMP production of an AAV-based gene therapy product performed at 2,000 L-scale is presented. The experience and lessons learned from this largest viral gene therapy vector production run conducted to date as discussed and highlighted in this review should contribute to future development of commercial viable scalable processes for vial gene therapies.

  8. A comparative genomic analysis of energy metabolism in sulfate reducing bacteria and archaea

    Directory of Open Access Journals (Sweden)

    Inês A. C. ePereira

    2011-04-01

    Full Text Available The number of sequenced genomes of sulfate-reducing organisms (SRO has increased significantly in the recent years, providing an opportunity for a broader perspective into the energy metabolism of such organisms. In this work we carried out a comparative survey of energy metabolism genes found in twenty-five available genomes of SRO. This analysis revealed a higher diversity of possible energy conserving pathways than classically considered to be present in these organisms, and permitted the identification of new proteins not known to be present in this group. The Deltaproteobacteria (and Thermodesulfovibrio yellowstonii are characterized by a large number of cytochromes c and cytochrome c-associated membrane redox complexes, indicating that periplasmic electron transfer pathways are important in these bacteria. The Archaea and Clostridia groups contain practically no cytochromes c or associated membrane complexes. However, despite the absence of a periplasmic space, a few extracytoplasmic membrane redox proteins were detected in the Gram-positive bacteria. Several ion-translocating complexes were detected in SRO including H+-pyrophosphatases, complex I homologues, Rnf and Ech/Coo hydrogenases. Furthermore, we found evidence that cytoplasmic electron bifurcating mechanisms, recently described for other anaerobes, are also likely to play an important role in energy metabolism of SRO. A number of cytoplasmic [NiFe] and [FeFe] hydrogenases, formate dehydrogenases and heterodisulfide reductase-related proteins are likely candidates to be involved in energy coupling through electron bifurcation, from diverse electron donors such as H2, formate, pyruvate, NAD(PH, β-oxidation and others. In conclusion, this analysis indicates that energy metabolism of SRO is far more versatile than previously considered, and that both chemiosmotic and flavin-based electron bifurcating mechanisms provide alternative strategies for energy conservation.

  9. Gene Therapy

    Science.gov (United States)

    ... or improve your body's ability to fight disease. Gene therapy holds promise for treating a wide range of diseases, such as cancer, cystic fibrosis, heart disease, diabetes, hemophilia and AIDS. Researchers are still studying how and ...

  10. Genes V.

    Energy Technology Data Exchange (ETDEWEB)

    Lewin, B.

    1994-12-31

    This fifth edition book encompasses a wide range of topics covering 1,272 pages. The book is arranged into nine parts with a total of 36 chapters. These nine parts include Introduction; DNA as a Store of Information; Translation; Constructing Cells; Control of Prokaryotypic Gene Expression; Perpetuation of DNA; Organization of the Eukaryotypic Genome; Eukaryotypic Transcription and RNA Processing; The Dynamic Genome; and Genes in Development.

  11. NiFe alloy particles doping effect of Gd-Ba-Cu-O bulks processed by a new cold-seeding technology%钆钡铜氧超导单晶块材的冷籽晶诱导生长及铁镍合金的掺杂作用

    Institute of Scientific and Technical Information of China (English)

    周迪帆; 徐坤; Shogo HARA; 李备战; Mitsuru IZUMI

    2013-01-01

      介绍了利用在MgO衬底上生长NdBa2Cu3Oy(Nd123)薄膜从而诱导生长GdBa2Cu3Oy(Gd123)超导单畴块材的工艺,并在此基础上研究了使用MgO缓冲层诱导生长钆钡铜氧单畴块材的工艺。通过调整热处理参数和缓冲层中Gd2BaCuO5(Gd211)相的含量,克服了MgO与Gd123母体的晶格失配和低反应率的问题。成功制备了Gd123单畴块材,研究了铁镍软磁合金粒子在超导块材中的掺杂作用。结果表明,0.4%(摩尔分数)为最优掺杂比例,超导临界电流在低场和中场下得到了很大的提升,对工业应用有重要意义。超导电流的提升主要源于铁、镍离子对铜位的替代,并提供了额外的磁通钉扎。%The process of cold seeding melt growth of GdBa2Cu3Oy (Gd123) bulk superconductors using NdBa2Cu3Oy (Nd123) thin films was reported. In addition, a novel cold seeding concept of combining MgO crystal and buffer pellet was also introduced. The misorientation caused by the lattice mismatch between MgO and Gd123 melt was overcome by choosing suitable heat treatment program and Gd2BaCuO5 (Gd211) content of the buffer pellet. The doping effect of soft ferromagnetic NiFe alloy particles was also reported. The bulk sample with 0.4%(mole fraction) doping amount shows the best performance on the flux trapping. The critical current density is largely enhanced under the external field of 1−2 T, which is promising for large-scale applications. This effect is originated from the substitution of Fe and Ni ions for the Cu sites contributing to magnetic flux pinning.

  12. DREB genes

    African Journals Online (AJOL)

    Unipar

    2015-03-12

    Mar 12, 2015 ... to AP2/ERF family, dehydration-responsive element-binding protein (DREB) genes, (CitsERF01 to ... Protein sequences of DREB subfamilies belonging to group I, .... position 37, and it was present in consensus in all protein.

  13. A proteomic view of Desulfovibrio vulgaris metabolism as determined by liquid chromatography coupled with tandem mass spectrometry.

    Science.gov (United States)

    Zhang, Weiwen; Gritsenko, Marina A; Moore, Ronald J; Culley, David E; Nie, Lei; Petritis, Konstantinos; Strittmatter, Eric F; Camp, David G; Smith, Richard D; Brockman, Fred J

    2006-08-01

    Direct LC-MS/MS was used to examine the proteins extracted from exponential or stationary phase Desulfovibrio vulgaris cells that had been grown on a minimal medium containing either lactate or formate as the primary carbon source. Across all four growth conditions, 976 gene products were identified with high confidence, which is equal to approximately 28% of all predicted proteins in the D. vulgaris genome. Bioinformatic analysis showed that the proteins identified were distributed among almost all functional classes, with the energy metabolism category containing the greatest number of identified proteins. At least 154 ORFs originally annotated as hypothetical proteins were found to encode the expressed proteins, which provided verification for the authenticity of these hypothetical proteins. Proteomic analysis showed that proteins potentially involved in ATP biosynthesis using the proton gradient across membrane, such as ATPase, alcohol dehydrogenases, heterodisulfide reductases, and [NiFe] hydrogenase (HynAB-1) of the hydrogen cycling were highly expressed in all four growth conditions, suggesting they may be the primary pathways for ATP synthesis in D. vulgaris. Most of the enzymes involved in substrate-level phosphorylation were also detected in all tested conditions. However, no enzyme involved in CO cycling or formate cycling was detected, suggesting that they are not the primary ATP-biosynthesis pathways under the tested conditions. This study provides the first proteomic overview of the cellular metabolism of D. vulgaris. The complete list of proteins identified in this study and their abundances (peptide hits) is provided in Supplementary Table 1.

  14. Engineering of Corynebacterium glutamicum to Enhance L-ornithine Production by Gene Knockout and Comparative Proteomic Analysis

    Institute of Scientific and Technical Information of China (English)

    卢冬梅; 刘建忠; 毛宗万

    2012-01-01

    Engineered Corynebacterium glutamicum was constructed for L-ornithine production by disrupting genes of argF and proB to prevent the flux away from L-ornithine.Effect of the inactivation of 2-oxoglutarate de-hydrogenase complex(ODHC) on L-ornithine production was also investigated.It was found that the inactivation of ODHC by knockout of the kgd gene enhanced L-ornithine production.The engineered C.glutamicum ATCC13032(ΔargFΔproBΔkgd) produced L-ornithine up to 4.78 g·L-1 from 0.24 g·L-1 of the wild-type strain.In order to understand the mechanism of L-ornithine production in C.glutamicum ATCC13032(ΔargFΔproBΔkgd) and find out new strategies for further enhancing L-ornithine production,the comparative proteome between the wild-type and the engineered strain was analyzed.L-Ornithine overproduction in the engineered strain was related to the up-regulation of the expression levels of enzymes involved in L-ornithine biosynthesis pathway and down-regulation of the expression levels of proteins involved in pentose phosphate pathway.The overexpression of genes in the upstream pathway of glutamate to increase the availability of endogenous glutamate may further in-crease ornithine production in the engineered C.glutamicum and the ornithine synthesis enzymes(ArgCJBD) may not be the limiting enzymes in the engineered C.glutamicum.

  15. Endothelial Genes

    Science.gov (United States)

    2005-06-01

    8217Department of Surgery, Division of Oncology , and 2Department of BRCA-l and BRCA-2 (breast cancer susceptibility genes), Pathology, University of...Suppression subtractive hybridization re- Cancer: principles and practice of oncology . Philadelphia: Lippincott- vealed an RNA sequence (GenBank accession...Lippman ME. Cancer of the breast: molecular biology angiogenesis in sarcomas and carcinomas. Clin Cancer Res 1999;5: of breast cancer. In: DeVita VT

  16. Gene Ontology

    Directory of Open Access Journals (Sweden)

    Gaston K. Mazandu

    2012-01-01

    Full Text Available The wide coverage and biological relevance of the Gene Ontology (GO, confirmed through its successful use in protein function prediction, have led to the growth in its popularity. In order to exploit the extent of biological knowledge that GO offers in describing genes or groups of genes, there is a need for an efficient, scalable similarity measure for GO terms and GO-annotated proteins. While several GO similarity measures exist, none adequately addresses all issues surrounding the design and usage of the ontology. We introduce a new metric for measuring the distance between two GO terms using the intrinsic topology of the GO-DAG, thus enabling the measurement of functional similarities between proteins based on their GO annotations. We assess the performance of this metric using a ROC analysis on human protein-protein interaction datasets and correlation coefficient analysis on the selected set of protein pairs from the CESSM online tool. This metric achieves good performance compared to the existing annotation-based GO measures. We used this new metric to assess functional similarity between orthologues, and show that it is effective at determining whether orthologues are annotated with similar functions and identifying cases where annotation is inconsistent between orthologues.

  17. Gene doping: gene delivery for olympic victory

    OpenAIRE

    2012-01-01

    With one recently recommended gene therapy in Europe and a number of other gene therapy treatments now proving effective in clinical trials it is feasible that the same technologies will soon be adopted in the world of sport by unscrupulous athletes and their trainers in so called ‘gene doping’. In this article an overview of the successful gene therapy clinical trials is provided and the potential targets for gene doping are highlighted. Depending on whether a doping gene product is secreted...

  18. GeneEd -- A Genetics Educational Resource

    Science.gov (United States)

    ... Javascript on. Feature: Genetics 101 GeneEd — A Genetics Educational Resource Past Issues / Summer 2013 Table of Contents Science ... The Hereditary Material of Life / GeneEd — A Genetics Educational Resource / Using The Genetics Home Reference Website / Understanding the ...

  19. Energy and Carbon Flow: Comparing ultramafic- and basalt-hosted vents

    Science.gov (United States)

    Perner, M.; Bach, W.; Seifert, R.; Strauss, H.; Laroche, J.

    2010-12-01

    In deep-sea vent habitats hydrothermal fluids provide the grounds for life by supplying reduced inorganic compounds (e.g. H2, sulfide). Chemolithoautotrophs can oxidize these substrates hereby yielding energy, which can then be used to fuel autotrophic CO2 fixation. Depending on the type of host rocks (and the degree of admixed ambient seawater) the availability of inorganic electron donors can vary considerably. While in ultramafic-hosted vents H2 levels are high and H2-oxidizing metabolisms are thought to dominate, in basalt-hosted vents, H2 is much lower and microbial sulfide oxidation is considered to prevail [1, 2]. We have investigated the effect of H2 and sulfide availability on the microbial community of distinct H2-rich and H2-poor vent sites along the Mid-Atlantic Ridge. Hydrothermally influenced samples were collected from the H2-rich ultramafic-hosted Logatchev field (15°N) and the comparatively H2-poor basalt-hosted vents from 5°S and 9°S. We conducted catabolic energy calculations to estimate the potential of various electron donors to function as microbial energy sources. We performed incubation experiments with hydrothermal fluids amended with H2 or sulfide and radioactively labelled bicarbonate and determined H2 and sulfide consumption and carbon incorporation rates. We constructed metagenomic libraries for sequence-based screening of genes encoding key enzymes for H2 uptake (NiFe uptake hydrogenases, group 1), sulfide oxidation (sulfide quinone oxidoreductase, sqr) and CO2 fixation pathways (RubisCOs of the Calvin cycle [CBB] and beta-subunit of the ATP citrate lyase of the reductive tricarboxylic acid cycle [rTCA]). We evaluated parts of the metagenomes from basalt-hosted sites by pyrosequencing. Based on our incubation experiments - under the conditions applied - we could not confirm that generally H2 consumption rates and biomass syntheses in fluids derived from ultramafic-hosted locations are significantly enhanced over those from basalt

  20. Comparative genomic analysis of N2-fixing and non-N2-fixing Paenibacillus spp.: organization, evolution and expression of the nitrogen fixation genes.

    Directory of Open Access Journals (Sweden)

    Jian-Bo Xie

    2014-03-01

    Full Text Available We provide here a comparative genome analysis of 31 strains within the genus Paenibacillus including 11 new genomic sequences of N2-fixing strains. The heterogeneity of the 31 genomes (15 N2-fixing and 16 non-N2-fixing Paenibacillus strains was reflected in the large size of the shell genome, which makes up approximately 65.2% of the genes in pan genome. Large numbers of transposable elements might be related to the heterogeneity. We discovered that a minimal and compact nif cluster comprising nine genes nifB, nifH, nifD, nifK, nifE, nifN, nifX, hesA and nifV encoding Mo-nitrogenase is conserved in the 15 N2-fixing strains. The nif cluster is under control of a σ(70-depedent promoter and possesses a GlnR/TnrA-binding site in the promoter. Suf system encoding [Fe-S] cluster is highly conserved in N2-fixing and non-N2-fixing strains. Furthermore, we demonstrate that the nif cluster enabled Escherichia coli JM109 to fix nitrogen. Phylogeny of the concatenated NifHDK sequences indicates that Paenibacillus and Frankia are sister groups. Phylogeny of the concatenated 275 single-copy core genes suggests that the ancestral Paenibacillus did not fix nitrogen. The N2-fixing Paenibacillus strains were generated by acquiring the nif cluster via horizontal gene transfer (HGT from a source related to Frankia. During the history of evolution, the nif cluster was lost, producing some non-N2-fixing strains, and vnf encoding V-nitrogenase or anf encoding Fe-nitrogenase was acquired, causing further diversification of some strains. In addition, some N2-fixing strains have additional nif and nif-like genes which may result from gene duplications. The evolution of nitrogen fixation in Paenibacillus involves a mix of gain, loss, HGT and duplication of nif/anf/vnf genes. This study not only reveals the organization and distribution of nitrogen fixation genes in Paenibacillus, but also provides insight into the complex evolutionary history of nitrogen fixation.

  1. Comparative genomic analysis of N2-fixing and non-N2-fixing Paenibacillus spp.: organization, evolution and expression of the nitrogen fixation genes.

    Science.gov (United States)

    Xie, Jian-Bo; Du, Zhenglin; Bai, Lanqing; Tian, Changfu; Zhang, Yunzhi; Xie, Jiu-Yan; Wang, Tianshu; Liu, Xiaomeng; Chen, Xi; Cheng, Qi; Chen, Sanfeng; Li, Jilun

    2014-03-01

    We provide here a comparative genome analysis of 31 strains within the genus Paenibacillus including 11 new genomic sequences of N2-fixing strains. The heterogeneity of the 31 genomes (15 N2-fixing and 16 non-N2-fixing Paenibacillus strains) was reflected in the large size of the shell genome, which makes up approximately 65.2% of the genes in pan genome. Large numbers of transposable elements might be related to the heterogeneity. We discovered that a minimal and compact nif cluster comprising nine genes nifB, nifH, nifD, nifK, nifE, nifN, nifX, hesA and nifV encoding Mo-nitrogenase is conserved in the 15 N2-fixing strains. The nif cluster is under control of a σ(70)-depedent promoter and possesses a GlnR/TnrA-binding site in the promoter. Suf system encoding [Fe-S] cluster is highly conserved in N2-fixing and non-N2-fixing strains. Furthermore, we demonstrate that the nif cluster enabled Escherichia coli JM109 to fix nitrogen. Phylogeny of the concatenated NifHDK sequences indicates that Paenibacillus and Frankia are sister groups. Phylogeny of the concatenated 275 single-copy core genes suggests that the ancestral Paenibacillus did not fix nitrogen. The N2-fixing Paenibacillus strains were generated by acquiring the nif cluster via horizontal gene transfer (HGT) from a source related to Frankia. During the history of evolution, the nif cluster was lost, producing some non-N2-fixing strains, and vnf encoding V-nitrogenase or anf encoding Fe-nitrogenase was acquired, causing further diversification of some strains. In addition, some N2-fixing strains have additional nif and nif-like genes which may result from gene duplications. The evolution of nitrogen fixation in Paenibacillus involves a mix of gain, loss, HGT and duplication of nif/anf/vnf genes. This study not only reveals the organization and distribution of nitrogen fixation genes in Paenibacillus, but also provides insight into the complex evolutionary history of nitrogen fixation.

  2. Comparative genomic analysis of N2-fixing and non-N2-fixing Paenibacillus spp.: organization, evolution and expression of the nitrogen fixation genes.

    Directory of Open Access Journals (Sweden)

    Jian-Bo Xie

    2014-03-01

    Full Text Available We provide here a comparative genome analysis of 31 strains within the genus Paenibacillus including 11 new genomic sequences of N2-fixing strains. The heterogeneity of the 31 genomes (15 N2-fixing and 16 non-N2-fixing Paenibacillus strains was reflected in the large size of the shell genome, which makes up approximately 65.2% of the genes in pan genome. Large numbers of transposable elements might be related to the heterogeneity. We discovered that a minimal and compact nif cluster comprising nine genes nifB, nifH, nifD, nifK, nifE, nifN, nifX, hesA and nifV encoding Mo-nitrogenase is conserved in the 15 N2-fixing strains. The nif cluster is under control of a σ(70-depedent promoter and possesses a GlnR/TnrA-binding site in the promoter. Suf system encoding [Fe-S] cluster is highly conserved in N2-fixing and non-N2-fixing strains. Furthermore, we demonstrate that the nif cluster enabled Escherichia coli JM109 to fix nitrogen. Phylogeny of the concatenated NifHDK sequences indicates that Paenibacillus and Frankia are sister groups. Phylogeny of the concatenated 275 single-copy core genes suggests that the ancestral Paenibacillus did not fix nitrogen. The N2-fixing Paenibacillus strains were generated by acquiring the nif cluster via horizontal gene transfer (HGT from a source related to Frankia. During the history of evolution, the nif cluster was lost, producing some non-N2-fixing strains, and vnf encoding V-nitrogenase or anf encoding Fe-nitrogenase was acquired, causing further diversification of some strains. In addition, some N2-fixing strains have additional nif and nif-like genes which may result from gene duplications. The evolution of nitrogen fixation in Paenibacillus involves a mix of gain, loss, HGT and duplication of nif/anf/vnf genes. This study not only reveals the organization and distribution of nitrogen fixation genes in Paenibacillus, but also provides insight into the complex evolutionary history of nitrogen fixation.

  3. Functional and Expression Analysis of the Metal-Inducible dmeRF System from Rhizobium leguminosarum bv. viciae

    Science.gov (United States)

    Rubio-Sanz, L.; Prieto, R. I.; Imperial, J.; Brito, B.

    2013-01-01

    A gene encoding a homolog to the cation diffusion facilitator protein DmeF from Cupriavidus metallidurans has been identified in the genome of Rhizobium leguminosarum UPM791. The R. leguminosarum dmeF gene is located downstream of an open reading frame (designated dmeR) encoding a protein homologous to the nickel- and cobalt-responsive transcriptional regulator RcnR from Escherichia coli. Analysis of gene expression showed that the R. leguminosarum dmeRF genes are organized as a transcriptional unit whose expression is strongly induced by nickel and cobalt ions, likely by alleviating the repressor activity of DmeR on dmeRF transcription. An R. leguminosarum dmeRF mutant strain displayed increased sensitivity to Co(II) and Ni(II), whereas no alterations of its resistance to Cd(II), Cu(II), or Zn(II) were observed. A decrease of symbiotic performance was observed when pea plants inoculated with an R. leguminosarum dmeRF deletion mutant strain were grown in the presence of high concentrations of nickel and cobalt. The same mutant induced significantly lower activity levels of NiFe hydrogenase in microaerobic cultures. These results indicate that the R. leguminosarum DmeRF system is a metal-responsive efflux mechanism acting as a key element for metal homeostasis in R. leguminosarum under free-living and symbiotic conditions. The presence of similar dmeRF gene clusters in other Rhizobiaceae suggests that the dmeRF system is a conserved mechanism for metal tolerance in legume endosymbiotic bacteria. PMID:23934501

  4. Dehalococcoides' gene transcripts as quantitative bioindicators of tetrachloroethene, trichloroethene, and cis-1,2-dichloroethene dehalorespiration rates.

    Science.gov (United States)

    Rahm, Brian G; Richardson, Ruth E

    2008-07-15

    Gene transcripts corresponding to 16S rRNA, the hydrogenase (H2ase) Hup, a sequence annotated asformate dehydrogenase (Fdh) and reductive dehalogenases (RDases) TceA, PceA, DET1559, and DET1545 in Dehalococcoides ethenogenes strain 195 (DET) hold promise as potential bioindicators of the dehalorespiration of chlorinated ethenes. Here, we present quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) data taken from DET-containing mixed culture microcosms (4.4 x 10(8) DET 16S rRNA gene copies/mL) operated under continuous-feed conditions, with the aim of clarifying the relationships between pseudosteady-state abundances of bioindicator transcripts and respiration rate of various substrates: tetrachloroethene (PCE), trichloroethene (TCE), and cis-1,2-dichloroethene (cDCE). Results from PCE-fed microcosms suggested an induction threshold for transcription of some bioindicator genes between chloroethene respiration rates of 2.1 and 9.5 microeeq/L/hr. Putative RDase genes DET1559 and DET1545, however, were up-regulated at low PCE respiration rates and may be functionally significant when substrate levels are low. Data from PCE-fed microcosms operated at saturation kinetics indicated that a high respiration rate was not necessarily associated with a correspondingly high bioindicator transcript abundance. From these microcosms we calculated an approximate yield value of 1.6 x 10(8) 16S rRNA gene copies (cells) per micromol Cl- released and estimated a kmax of PCE respiration of 3 x 10(-9) micromol Cl- per 16S rRNA gene copy per day. TCE- and cDCE-fed microcosm studies indicated that Fdh, Hup, and TceA were the most abundant transcripts and could make suitable choices as bioindicators of activity for these substrates. Hup transcripts could be positively correlated to respiration rate (between approximately 8 and 45 microeeq/L/hr) regardless of chloroethene substrate, with transcript levels predicted to increase by 1.8 x 10(9) copies/mL culture for every

  5. Influence of setup and carbon source on the bacterial community of biocathodes in microbial electrolysis cells.

    Science.gov (United States)

    Croese, Elsemiek; Jeremiasse, Adriaan W; Marshall, Ian P G; Spormann, Alfred M; Euverink, Gert-Jan W; Geelhoed, Jeanine S; Stams, Alfons J M; Plugge, Caroline M

    2014-01-01

    The microbial electrolysis cell (MEC) biocathode has shown great potential as alternative for expensive metals as catalyst for H2 synthesis. Here, the bacterial communities at the biocathode of five hydrogen producing MECs using molecular techniques were characterized. The setups differed in design (large versus small) including electrode material and flow path and in carbon source provided at the cathode (bicarbonate or acetate). A hydrogenase gene-based DNA microarray (Hydrogenase Chip) was used to analyze hydrogenase genes present in the three large setups. The small setups showed dominant groups of Firmicutes and two of the large setups showed dominant groups of Proteobacteria and Bacteroidetes. The third large setup received acetate but no sulfate (no sulfur source). In this setup an almost pure culture of a Promicromonospora sp. developed. Most of the hydrogenase genes detected were coding for bidirectional Hox-type hydrogenases, which have shown to be involved in cytoplasmatic H2 production.

  6. Special Issue: Gene Conversion in Duplicated Genes

    Directory of Open Access Journals (Sweden)

    Hideki Innan

    2011-06-01

    Full Text Available Gene conversion is an outcome of recombination, causing non-reciprocal