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Sample records for hyperthermophile thermotoga maritima

  1. Crystallization and preliminary crystallographic analysis of an esterase with a novel domain from the hyperthermophile Thermotoga maritima

    NARCIS (Netherlands)

    Sun, Lei; Levisson, Mark; Hendriks, Sjon; Akveld, Twan; Kengen, Serve W. M.; Dijkstra, Bauke W.; van der Oost, John

    A predicted esterase ( EstA) with an unusual new domain from the hyperthermophilic bacterium Thermotoga maritima has been cloned and overexpressed in Escherichia coli. The purified protein was crystallized by the hanging-drop vapour-diffusion technique in the presence of lithium sulfate and

  2. Crystallization and preliminary crystallographic analysis of an esterase with a novel domain from the hyperthermophile Thermotoga maritima

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Lei [Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, Dreijenplein 10, 6703 HB Wageningen (Netherlands); Laboratory of Biophysical Chemistry, University of Groningen, Nijenborgh 4, 9747 AG Groningen (Netherlands); Levisson, Mark; Hendriks, Sjon; Akveld, Twan; Kengen, Servé W. M. [Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, Dreijenplein 10, 6703 HB Wageningen (Netherlands); Dijkstra, Bauke W. [Laboratory of Biophysical Chemistry, University of Groningen, Nijenborgh 4, 9747 AG Groningen (Netherlands); Oost, John van der, E-mail: john.vanderoost@wur.nl [Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, Dreijenplein 10, 6703 HB Wageningen (Netherlands)

    2007-09-01

    A thermostable esterase (EstA) from Thermotoga maritima was cloned and purified. Crystals of EstA and its selenomethionine derivative were grown and diffract to beyond 2.6 Å resolution at 100 K using synchrotron radiation. A predicted esterase (EstA) with an unusual new domain from the hyperthermophilic bacterium Thermotoga maritima has been cloned and overexpressed in Escherichia coli. The purified protein was crystallized by the hanging-drop vapour-diffusion technique in the presence of lithium sulfate and polyethylene glycol 8000. Selenomethionine-substituted EstA crystals were obtained under the same conditions and three different-wavelength data sets were collected to 2.6 Å resolution. The crystal belongs to space group H32, with unit-cell parameters a = b = 130.2, c = 306.2 Å. There are two molecules in the asymmetric unit, with a V{sub M} of 2.9 Å{sup 3} Da{sup −1} and 58% solvent content.

  3. Hydrogen production by the hyperthermophilic bacterium Thermotoga maritima Part II: modeling and experimental approaches for hydrogen production.

    Science.gov (United States)

    Auria, Richard; Boileau, Céline; Davidson, Sylvain; Casalot, Laurence; Christen, Pierre; Liebgott, Pierre Pol; Combet-Blanc, Yannick

    2016-01-01

    Thermotoga maritima is a hyperthermophilic bacterium known to produce hydrogen from a large variety of substrates. The aim of the present study is to propose a mathematical model incorporating kinetics of growth, consumption of substrates, product formations, and inhibition by hydrogen in order to predict hydrogen production depending on defined culture conditions. Our mathematical model, incorporating data concerning growth, substrates, and products, was developed to predict hydrogen production from batch fermentations of the hyperthermophilic bacterium, T. maritima . It includes the inhibition by hydrogen and the liquid-to-gas mass transfer of H 2 , CO 2 , and H 2 S. Most kinetic parameters of the model were obtained from batch experiments without any fitting. The mathematical model is adequate for glucose, yeast extract, and thiosulfate concentrations ranging from 2.5 to 20 mmol/L, 0.2-0.5 g/L, or 0.01-0.06 mmol/L, respectively, corresponding to one of these compounds being the growth-limiting factor of T. maritima . When glucose, yeast extract, and thiosulfate concentrations are all higher than these ranges, the model overestimates all the variables. In the window of the model validity, predictions of the model show that the combination of both variables (increase in limiting factor concentration and in inlet gas stream) leads up to a twofold increase of the maximum H 2 -specific productivity with the lowest inhibition. A mathematical model predicting H 2 production in T. maritima was successfully designed and confirmed in this study. However, it shows the limit of validity of such mathematical models. Their limit of applicability must take into account the range of validity in which the parameters were established.

  4. Structure of a d-tagatose 3-epimerase-related protein from the hyperthermophilic bacterium Thermotoga maritima

    International Nuclear Information System (INIS)

    Sakuraba, Haruhiko; Yoneda, Kazunari; Satomura, Takenori; Kawakami, Ryushi; Ohshima, Toshihisa

    2009-01-01

    The crystal structure of a hyperthermophilic d-tagatose 3-epimerase-related protein with a unique active-site architecture was determined. The crystal structure of a d-tagatose 3-epimerase-related protein (TM0416p) encoded by the hypothetical open reading frame TM0416 in the genome of the hyperthermophilic bacterium Thermotoga maritima was determined at a resolution of 2.2 Å. The asymmetric unit contained two homologous subunits and a dimer was generated by twofold symmetry. The main-chain coordinates of the enzyme monomer proved to be similar to those of d-tagatose 3-epimerase from Pseudomonas cichorii and d-psicose 3-epimerase from Agrobacterium tumefaciens; however, TM0416p exhibited a unique solvent-accessible substrate-binding pocket that reflected the absence of an α-helix that covers the active-site cleft in the two aforementioned ketohexose 3-epimerases. In addition, the residues responsible for creating a hydrophobic environment around the substrate in TM0416p differ entirely from those in the other two enzymes. Collectively, these findings suggest that the substrate specificity of TM0416p is likely to differ substantially from those of other d-tagatose 3-epimerase family enzymes

  5. Structure of a D-tagatose 3-epimerase-related protein from the hyperthermophilic bacterium Thermotoga maritima.

    Science.gov (United States)

    Sakuraba, Haruhiko; Yoneda, Kazunari; Satomura, Takenori; Kawakami, Ryushi; Ohshima, Toshihisa

    2009-03-01

    The crystal structure of a D-tagatose 3-epimerase-related protein (TM0416p) encoded by the hypothetical open reading frame TM0416 in the genome of the hyperthermophilic bacterium Thermotoga maritima was determined at a resolution of 2.2 A. The asymmetric unit contained two homologous subunits and a dimer was generated by twofold symmetry. The main-chain coordinates of the enzyme monomer proved to be similar to those of D-tagatose 3-epimerase from Pseudomonas cichorii and D-psicose 3-epimerase from Agrobacterium tumefaciens; however, TM0416p exhibited a unique solvent-accessible substrate-binding pocket that reflected the absence of an alpha-helix that covers the active-site cleft in the two aforementioned ketohexose 3-epimerases. In addition, the residues responsible for creating a hydrophobic environment around the substrate in TM0416p differ entirely from those in the other two enzymes. Collectively, these findings suggest that the substrate specificity of TM0416p is likely to differ substantially from those of other D-tagatose 3-epimerase family enzymes.

  6. Site-Directed Mutagenesis of a Hyperthermophilic Endoglucanase Cel12B from Thermotoga maritima Based on Rational Design.

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    Jinfeng Zhang

    Full Text Available To meet the demand for the application of high activity and thermostable cellulases in the production of new-generation bioethanol from nongrain-cellulose sources, a hyperthermostable β-1,4-endoglucase Cel12B from Thermotoga maritima was selected for further modification by gene site-directed mutagenesis method in the present study, based on homology modeling and rational design. As a result, two recombinant enzymes showed significant improvement in enzyme activity by 77% and 87%, respectively, higher than the parental enzyme TmCel12B. Furthermore, the two mutants could retain 80% and 90.5% of their initial activity after incubation at 80°C for 8 h, while only 45% for 5 h to TmCel12B. The Km and Vmax of the two recombinant enzymes were 1.97±0.05 mM, 4.23±0.15 μmol·mg(-1·min(-1 of TmCel12B-E225H-K207G-D37V, and 2.97±0.12 mM, 3.15±0.21 μmol·mg(-1·min(-1 of TmCel12B-E225H-K207G, respectively, when using CMC-Na as the substrate. The roles of the mutation sites were also analyzed and evaluated in terms of electron density, hydrophobicity of the modeled protein structures. The recombinant enzymes may be used in the hydrolysis of cellulose at higher temperature in the future. It was concluded that the gene mutagenesis approach of a certain active residues may effectively improve the performance of cellulases for the industrial applications and contribute to the study the thermostable mechanism of thermophilic enzymes.

  7. Transcriptional regulation of the carbohydrate utilization network in Thermotoga maritima

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    Dmitry A Rodionov

    2013-08-01

    Full Text Available Hyperthermophilic bacteria from the Thermotogales lineage can produce hydrogen by fermenting a wide range of carbohydrates. Previous experimental studies identified a large fraction of genes committed to carbohydrate degradation and utilization in the model bacterium Thermotoga maritima. Knowledge of these genes enabled comprehensive reconstruction of biochemical pathways comprising the carbohydrate utilization network. However, transcriptional factors (TFs and regulatory mechanisms driving this network remained largely unknown. Here, we used an integrated approach based on comparative analysis of genomic and transcriptomic data for the reconstruction of the carbohydrate utilization regulatory networks in 11 Thermotogales genomes. We identified DNA-binding motifs and regulons for 19 orthologous TFs in the Thermotogales. The inferred regulatory network in T. maritima contains 181 genes encoding TFs, sugar catabolic enzymes and ABC-family transporters. In contrast to many previously described bacteria, a transcriptional regulation strategy of Thermotoga does not employ global regulatory factors. The reconstructed regulatory network in T. maritima was validated by gene expression profiling on a panel of mono- and disaccharides and by in vitro DNA-binding assays. The observed upregulation of genes involved in catabolism of pectin, trehalose, cellobiose, arabinose, rhamnose, xylose, glucose, galactose, and ribose showed a strong correlation with the UxaR, TreR, BglR, CelR, AraR, RhaR, XylR, GluR, GalR, and RbsR regulons. Ultimately, this study elucidated the transcriptional regulatory network and mechanisms controlling expression of carbohydrate utilization genes in T. maritima. In addition to improving the functional annotations of associated transporters and catabolic enzymes, this research provides novel insights into the evolution of regulatory networks in Thermotogales.

  8. Optimization of expression and properties of the recombinant acetohydroxyacid synthase of Thermotoga maritima

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    Mohammad S. Eram

    2015-12-01

    Full Text Available The data provide additional support of the characterization of the biophysical and biochemical properties of the enzyme acetohydroxyacid synthase from the hyperthermophilic bacterium Thermotoga maritima (Eram et al., 2015 [1]. The genes encoding the enzyme subunits have been cloned and expressed in the mesophilic host Escherichia coli. Detailed data include information about the optimization of the expression conditions, biophysical properties of the enzyme and reconstitution of the holoenzyme from individually expressed and purified subunits.

  9. Functional and structural characterization of a thermostable acetyl esterase from Thermotoga maritima

    NARCIS (Netherlands)

    Levisson, M.; Han, G.W.; Deller, M.C.; Hendriks, S.N.A.; Oost, van der J.; Kengen, S.W.M.

    2012-01-01

    TM0077 from Thermotoga maritima is a member of the carbohydrate esterase family 7 and is active on a variety of acetylated compounds, including cephalosporin C. TM0077 esterase activity is confined to short-chain acyl esters (C2-C3), and is optimal around 100°C and pH 7.5. The positional specificity

  10. Structure of a NAD kinase from Thermotoga maritima at 2.3 Å resolution

    International Nuclear Information System (INIS)

    Oganesyan, Vaheh; Huang, Candice; Adams, Paul D.; Jancarik, Jaru; Yokota, Hisao A.; Kim, Rosalind; Kim, Sung-Hou

    2005-01-01

    The expression, purification, crystallization, and structure determination of NAD-kinase from T. maritima are reported. Similarity to other NAD-kinases as well as homo-oligomrization state of the enzyme from T. maritima are discussed. NAD kinase is the only known enzyme that catalyzes the formation of NADP, a coenzyme involved in most anabolic reactions and in the antioxidant defense system. Despite its importance, very little is known regarding the mechanism of catalysis and only recently have several NAD kinase structures been deposited in the PDB. Here, an independent investigation of the crystal structure of inorganic polyphosphate/ATP-NAD kinase, PPNK-THEMA, a protein from Thermotoga maritima, is reported at a resolution of 2.3 Å. The crystal structure was solved using single-wavelength anomalous diffraction (SAD) data collected at the Se absorption-peak wavelength in a state in which no cofactors or substrates were bound. It revealed that the 258-amino-acid protein is folded into two distinct domains, similar to recently reported NAD kinases. The N-terminal α/β-domain spans the first 100 amino acids and the last 30 amino acids of the polypeptide and has several topological matches in the PDB, whereas the other domain, which spans the middle 130 residues, adopts a unique β-sandwich architecture and only appreciably matches the recently deposited PDB structures of NAD kinases

  11. Purification, crystallization and preliminary crystallographic analysis of a thermostable endonuclease IV from Thermotoga maritima

    International Nuclear Information System (INIS)

    Hughes, Ronny C.; Tomanicek, Stephen J.; Ng, Joseph D.; Coates, Leighton

    2009-01-01

    The overexpression, purification and crystallization of endonuclease IV from T. maritima are reported. The crystals belonged to the hexagonal space group P6 1 and diffracted to 2.36 Å resolution. The DNA-repair enzyme endonuclease IV from the thermophilic bacterium Thermotoga maritima MSB8 (reference sequence NC-000853) has been expressed in Escherichia coli and crystallized for X-ray analysis. T. maritima endonuclease IV is a 287-amino-acid protein with 32% sequence identity to E. coli endonuclease IV. The protein was purified to homogeneity and was crystallized using the sitting-drop vapor-diffusion method. The protein crystallized in space group P6 1 , with one biological molecule in the asymmetric unit, corresponding to a Matthews coefficient of 2.39 Å 3 Da −1 and 47% solvent content. The unit-cell parameters of the crystals were a = b = 123.2, c = 35.6 Å. Microseeding and further optimization yielded crystals with an X-ray diffraction limit of 2.36 Å. A single 70° data set was collected and processed, resulting in an overall R merge and a completeness of 9.5% and 99.3%, respectively

  12. Genome Sequence of Thermotoga sp. Strain RQ2, a Hyperthermophilic Bacterium Isolated from a Geothermally Heated Region of the Seafloor near Ribeira Quente, the Azores

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    Swithers, Kristen S.; DiPippo, Jonathan L.; Bruce, David C.; Detter, Christopher; Tapia, Roxanne; Han, Shunsheng; Saunders, Elizabeth; Goodwin, Lynne A.; Han, James; Woyke, Tanja; Pitluck, Sam; Pennacchio, Len; Nolan, Matthew; Mikhailova, Natalia; Lykidis, Athanasios; Land, Miriam L.; Brettin, Thomas; Stetter, Karl O.; Nelson, Karen E.; Gogarten, J. Peter; Noll, Kenneth M.

    2011-01-01

    Thermotoga sp. strain RQ2 is probably a strain of Thermotoga maritima. Its complete genome sequence allows for an examination of the extent and consequences of gene flow within Thermotoga species and strains. Thermotoga sp. RQ2 differs from T. maritima in its genes involved in myo-inositol metabolism. Its genome also encodes an apparent fructose phosphotransferase system (PTS) sugar transporter. This operon is also found in Thermotoga naphthophila strain RKU-10 but no other Thermotogales. These are the first reported PTS transporters in the Thermotogales. PMID:21952543

  13. Structure of a periplasmic glucose-binding protein from Thermotoga maritima

    International Nuclear Information System (INIS)

    Palani, Kandavelu; Kumaran, Desigan; Burley, Stephen K.; Swaminathan, Subramanyam

    2012-01-01

    The periplasmic glucose-binding protein from T. maritima consists of two domains with the ligand β-d-glucose buried between them. The two domains adopt a closed conformation. ABC transport systems have been characterized in organisms ranging from bacteria to humans. In most bacterial systems, the periplasmic component is the primary determinant of specificity of the transport complex as a whole. Here, the X-ray crystal structure of a periplasmic glucose-binding protein (GBP) from Thermotoga maritima determined at 2.4 Å resolution is reported. The molecule consists of two similar α/β domains connected by a three-stranded hinge region. In the current structure, a ligand (β-d-glucose) is buried between the two domains, which have adopted a closed conformation. Details of the substrate-binding sites revealed features that determine substrate specificity. In toto, ten residues from both domains form eight hydrogen bonds to the bound sugar and four aromatic residues (two from each domain) stabilize the substrate through stacking interactions

  14. Structure of an essential bacterial protein YeaZ (TM0874) from Thermotoga maritima at 2.5 Å resolution

    International Nuclear Information System (INIS)

    Xu, Qingping; McMullan, Daniel; Jaroszewski, Lukasz; Krishna, S. Sri; Elsliger, Marc-André; Yeh, Andrew P.; Abdubek, Polat; Astakhova, Tamara; Axelrod, Herbert L.; Carlton, Dennis; Chiu, Hsiu-Ju; Clayton, Thomas; Duan, Lian; Feuerhelm, Julie; Grant, Joanna; Han, Gye Won; Jin, Kevin K.; Klock, Heath E.; Knuth, Mark W.; Miller, Mitchell D.; Morse, Andrew T.; Nigoghossian, Edward; Okach, Linda; Oommachen, Silvya; Paulsen, Jessica; Reyes, Ron; Rife, Christopher L.; Bedem, Henry van den; Hodgson, Keith O.; Wooley, John; Deacon, Ashley M.; Godzik, Adam; Lesley, Scott A.; Wilson, Ian A.

    2009-01-01

    The crystal structure of an essential bacterial protein, YeaZ, from T. maritima identifies an interface that potentially mediates protein–protein interaction. YeaZ is involved in a protein network that is essential for bacteria. The crystal structure of YeaZ from Thermotoga maritima was determined to 2.5 Å resolution. Although this protein belongs to a family of ancient actin-like ATPases, it appears that it has lost the ability to bind ATP since it lacks some key structural features that are important for interaction with ATP. A conserved surface was identified, supporting its role in the formation of protein complexes

  15. Polymerization properties of the Thermotoga maritima actin MreB: roles of temperature, nucleotides, and ions.

    Science.gov (United States)

    Bean, Greg J; Amann, Kurt J

    2008-01-15

    MreB is a bacterial orthologue of actin that affects cell shape, polarity, and chromosome segregation. Although a significant body of work has explored its cellular functions, we know very little about the biochemical behavior of MreB. We have cloned, overexpressed in Escherichia coli, and purified untagged MreB1 from Thermotoga maritima. We have characterized the conditions that regulate its monomer-to-polymer assembly reaction, the critical concentrations of that reaction, the manner in which MreB uses nucleotides, its stability, and the structure of the assembled polymer. MreB requires a bound purine nucleotide for polymerization and rapidly hydrolyzes it following assembly. MreB assembly contains two distinct components, one that does not require divalent cations and one that does, which may comprise the nucleation and elongation phases of assembly, respectively. MreB assembly is strongly favored by increasing temperature or protein concentration but inhibited differentially by high concentrations of monovalent salts. The polymerization rate increases and the bulk critical concentration decreases with increasing temperature, but in contrast to previous reports, MreB is capable of polymerizing across a broad range of temperatures. MreB polymers are shorter and stiffer and scatter more light than eukaryotic actin filaments. Due to rapid ATP hydrolysis and phosphate release, we suggest that most assembled MreB in cells is in the ADP-bound state. Because of only moderate differences between the ATP and ADP critical concentrations, treadmilling may occur, but we do not predict dynamic instability in cells. Because of the relatively low cellular concentration of MreB and the observed structural properties of the polymer, a single MreB assembly may exist in cells.

  16. Structural Insight inot the low Affinity Between Thermotoga maritima CheA and CheB Compared to their Escherichia coli/Salmonella typhimurium Counterparts

    Energy Technology Data Exchange (ETDEWEB)

    S Park; B Crane

    2011-12-31

    CheA-mediated CheB phosphorylation and the subsequent CheB-mediated demethylation of the chemoreceptors are important steps required for the bacterial chemotactic adaptation response. Although Escherichia coli CheB has been reported to interact with CheA competitively against CheY, we have observed that Thermotoga maritima CheB has no detectable CheA-binding. By determining the CheY-like domain crystal structure of T. maritima CheB, and comparing against the T. maritima CheY and Salmonella typhimurium CheB structures, we propose that the two consecutive glutamates in the {beta}4/{alpha}4 loop of T. maritima CheB that is absent in T. maritima CheY and in E. coli/S. typhimurium CheB may be one factor contributing to the low CheA affinity.

  17. Structural and biochemical characterization of the β-N-acetylglucosaminidase from Thermotoga maritima: toward rationalization of mechanistic knowledge in the GH73 family.

    Science.gov (United States)

    Lipski, Alexandra; Hervé, Mireille; Lombard, Vincent; Nurizzo, Didier; Mengin-Lecreulx, Dominique; Bourne, Yves; Vincent, Florence

    2015-03-01

    Members of the GH73 glycosidase family cleave the β-1,4-glycosidic bond between the N-acetylglucosaminyl (GlcNAc) and N-acetylmuramyl (MurNAc) moieties in bacterial peptidoglycan. A catalytic mechanism has been proposed for members FlgJ, Auto, AcmA and Atl(WM) and the structural analysis of FlgJ and Auto revealed a conserved α/β fold reminiscent of the distantly related GH23 lysozyme. Comparison of the active site residues reveals variability in the nature of the catalytic general base suggesting two distinct catalytic mechanisms: an inverting mechanism involving two distant glutamate residues and a substrate-assisted mechanism involving anchimeric assistance by the C2-acetamido group of the GlcNAc moiety. Herein, we present the biochemical characterization and crystal structure of TM0633 from the hyperthermophilic bacterium Thermotoga maritima. TM0633 adopts the α/β fold of the family and displays β-N-acetylglucosaminidase activity on intact peptidoglycan sacculi. Site-directed mutagenesis identifies Glu34, Glu65 and Tyr118 as important residues for catalysis. A thorough bioinformatic analysis of the GH73 sequences identified five phylogenetic clusters. TM0633, FlgJ and Auto belong to a group of three clusters that conserve two carboxylate residues involved in a classical inverting acid-base mechanism. Members of the other two clusters lack a conserved catalytic general base supporting a substrate-assisted mechanism. Molecular modeling of representative members from each cluster suggests that variability in length of the β-hairpin region above the active site confers ligand-binding specificity and modulates the catalytic mechanisms within the GH73 family. © The Author 2014. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  18. Impact of orientation of carbohydrate binding modules family 22 and 6 on the catalytic activity of Thermotoga maritima xylanase XynB.

    Science.gov (United States)

    Tajwar, Razia; Shahid, Saher; Zafar, Rehan; Akhtar, Muhammad Waheed

    2017-11-01

    Xylanase XynB of the hyperthermophile Thermotoga maritima, which belongs to glycoside hydrolase family 10 (GH10), does not have an associated carbohydrate binding module (CBM) in the native state. CBM6 and CBM22 from a thermophile Clostridium thermocellum were fused to the catalytic domain of XynB (XynB-C) to determine the effects on activity and other properties. XynB-B22C and XynB-CB22, produced by fusing CBM22 to the N- and C-terminal of XynB-C, showed 1.7- and 3.24-fold increase in activity against the insoluble birchwood xylan, respectively. Similarly, CBM6 when attached to the C-terminal of XynB-C resulted in 2.0-fold increase in activity, whereas its attachment to the N-terminal did not show any increase of activity. XynB-B22C and XynB-CB22 retained all the activity, whereas XynB-B6C and XynB-CB6 lost 17 and 11% of activity, respectively, at 60°C for 4h. Thermostability data and the secondary structure contents obtained by molecular modelling are in agreement with the data from circular dichroism analysis. Molecular modelling analysis showed that the active site residues of the catalytic domain and the binding residues of CBM6 and CBM22 were located on the surface of molecule, except XynB-B6C, where the binding residues were found somewhat buried. In the case of XynB-CB22, the catalytic and the binding residues seem to be located favorably adjacent to each other, thus showing higher increase in activity. This study shows that the active site residues of the catalytic domain and the binding residues of the CBM are arranged in a unique fashion, not reported before. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. Enzymatic synthesis of rare sugars with L-rhamnulose-1-phosphate aldolase from Thermotoga maritima MSB8.

    Science.gov (United States)

    Li, Zijie; Wu, Xiaoru; Cai, Li; Duan, Shenglin; Liu, Jia; Yuan, Peng; Nakanishi, Hideki; Gao, Xiao-Dong

    2015-09-15

    L-Rhamnulose-1-phosphate aldolase from a thermophilic source (Thermotoga maritima MSB8) (RhaDT.mari) was heterologously overexpressed in Escherichia coli and the stereoselectivity of this enzyme with or without Nus tag was investigated. We also applied this enzyme to the synthesis of rare sugars D-psicose, D-sorbose, L-tagatose and L-fructose using our one-pot four-enzyme system. To the best of our knowledge, this is the first use of RhaD from a thermophilic source for rare sugar synthesis and the temperature tolerance of this enzyme paves the path for large scale fermentation. Copyright © 2015 Elsevier Ltd. All rights reserved.

  20. Cytoplasmic expression of a thermostable invertase from Thermotoga maritima in Lactococcus lactis.

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    Pek, Han Bin; Lim, Pei Yu; Liu, Chengcheng; Lee, Dong-Yup; Bi, Xuezhi; Wong, Fong Tian; Ow, Dave Siak-Wei

    2017-05-01

    To evaluate the secretory and cytoplasmic expression of a thermostable Thermogata maritima invertase in Lactococcus lactis. The thermostable invertase from T. maritima was cloned with and without the USP45 secretory peptide into the pNZ8148 vector for nisin-inducible expression in L. lactis. The introduction of an USP45 secretion peptide at the N-terminal of the enzyme led to a loss of protein solubility. Computational homology modeling and hydrophobicity studies indicated that the USP45 peptide exposes a stretch of hydrophobic amino acids on the protein surface resulting in lower solubility. Removal of the USP45 secretion peptide allowed a soluble and functional invertase to be expressed intracellularly in L. lactis. Immobilized metal affinity chromatography purification of the cell lysate with nickel-NTA gave a single protein band on SDS-PAGE, while E. coli-expressed invertase consistently co-purified with an additional band. The yields of the purified invertase from E. coli and L. lactis were 14.1 and 6.3 mg/l respectively. Invertase can be expressed in L. lactis and purified in a functional form. L. lactis is a suitable host for the production of food-grade invertase for use in the food and biotechnology industries.

  1. Crystallization and preliminary X-ray crystallographic analysis of CheW from Thermotoga maritima: a coupling protein of CheA and the chemotaxis receptor

    International Nuclear Information System (INIS)

    Park, SangYoun; Crane, Brian R.

    2011-01-01

    CheW from T. maritima has been crystallized (space group P6 3 , unit-cell parameters a = b = 61.265, c = 361.045 Å). Diffraction data have been collected to 3.1 Å resolution using synchrotron X-ray radiation. The CheW protein plays a key role in bacterial chemotaxis signal transduction by coupling CheA to chemotaxis receptors. CheW from Thermotoga maritima has been overexpressed in Escherichia coli and crystallized at 298 K using ammonium sulfate as a salt precipitant. X-ray diffraction data have been collected to 3.10 Å resolution at 100 K using synchrotron radiation. The crystal belonged to space group P6 3 , with unit-cell parameters a = b = 61.265, c = 361.045 Å. The asymmetric unit may contain four to six CheW molecules

  2. Improved Activity of a Thermophilic Cellulase, Cel5A, from Thermotoga maritima on Ionic Liquid Pretreated Switchgrass

    Science.gov (United States)

    Chen, Zhiwei; Pereira, Jose H.; Liu, Hanbin; Tran, Huu M.; Hsu, Nathan S. Y.; Dibble, Dean; Singh, Seema; Adams, Paul D.; Sapra, Rajat; Hadi, Masood Z.; Simmons, Blake A.; Sale, Kenneth L.

    2013-01-01

    Ionic liquid pretreatment of biomass has been shown to greatly reduce the recalcitrance of lignocellulosic biomass, resulting in improved sugar yields after enzymatic saccharification. However, even under these improved saccharification conditions the cost of enzymes still represents a significant proportion of the total cost of producing sugars and ultimately fuels from lignocellulosic biomass. Much of the high cost of enzymes is due to the low catalytic efficiency and stability of lignocellulolytic enzymes, especially cellulases, under conditions that include high temperatures and the presence of residual pretreatment chemicals, such as acids, organic solvents, bases, or ionic liquids. Improving the efficiency of the saccharification process on ionic liquid pretreated biomass will facilitate reduced enzyme loading and cost. Thermophilic cellulases have been shown to be stable and active in ionic liquids but their activity is typically at lower levels. Cel5A_Tma, a thermophilic endoglucanase from Thermotoga maritima, is highly active on cellulosic substrates and is stable in ionic liquid environments. Here, our motivation was to engineer mutants of Cel5A_Tma with higher activity on 1-ethyl-3-methylimidazolium acetate ([C2mim][OAc]) pretreated biomass. We developed a robotic platform to screen a random mutagenesis library of Cel5A_Tma. Twelve mutants with 25–42% improvement in specific activity on carboxymethyl cellulose and up to 30% improvement on ionic-liquid pretreated switchgrass were successfully isolated and characterized from a library of twenty thousand variants. Interestingly, most of the mutations in the improved variants are located distally to the active site on the protein surface and are not directly involved with substrate binding. PMID:24244549

  3. Improved activity of a thermophilic cellulase, Cel5A, from Thermotoga maritima on ionic liquid pretreated switchgrass.

    Directory of Open Access Journals (Sweden)

    Zhiwei Chen

    Full Text Available Ionic liquid pretreatment of biomass has been shown to greatly reduce the recalcitrance of lignocellulosic biomass, resulting in improved sugar yields after enzymatic saccharification. However, even under these improved saccharification conditions the cost of enzymes still represents a significant proportion of the total cost of producing sugars and ultimately fuels from lignocellulosic biomass. Much of the high cost of enzymes is due to the low catalytic efficiency and stability of lignocellulolytic enzymes, especially cellulases, under conditions that include high temperatures and the presence of residual pretreatment chemicals, such as acids, organic solvents, bases, or ionic liquids. Improving the efficiency of the saccharification process on ionic liquid pretreated biomass will facilitate reduced enzyme loading and cost. Thermophilic cellulases have been shown to be stable and active in ionic liquids but their activity is typically at lower levels. Cel5A_Tma, a thermophilic endoglucanase from Thermotoga maritima, is highly active on cellulosic substrates and is stable in ionic liquid environments. Here, our motivation was to engineer mutants of Cel5A_Tma with higher activity on 1-ethyl-3-methylimidazolium acetate ([C2mim][OAc] pretreated biomass. We developed a robotic platform to screen a random mutagenesis library of Cel5A_Tma. Twelve mutants with 25-42% improvement in specific activity on carboxymethyl cellulose and up to 30% improvement on ionic-liquid pretreated switchgrass were successfully isolated and characterized from a library of twenty thousand variants. Interestingly, most of the mutations in the improved variants are located distally to the active site on the protein surface and are not directly involved with substrate binding.

  4. Proteolysis in hyperthermophilic microorganisms

    Directory of Open Access Journals (Sweden)

    Donald E. Ward

    2002-01-01

    Full Text Available Proteases are found in every cell, where they recognize and break down unneeded or abnormal polypeptides or peptide-based nutrients within or outside the cell. Genome sequence data can be used to compare proteolytic enzyme inventories of different organisms as they relate to physiological needs for protein modification and hydrolysis. In this review, we exploit genome sequence data to compare hyperthermophilic microorganisms from the euryarchaeotal genus Pyrococcus, the crenarchaeote Sulfolobus solfataricus, and the bacterium Thermotoga maritima. An overview of the proteases in these organisms is given based on those proteases that have been characterized and on putative proteases that have been identified from genomic sequences, but have yet to be characterized. The analysis revealed both similarities and differences in the mechanisms utilized for proteolysis by each of these hyperthermophiles and indicated how these mechanisms relate to proteolysis in less thermophilic cells and organisms.

  5. The structure and dynamic properties of the complete histidine phosphotransfer domain of the chemotaxis specific histidine autokinase CheA from Thermotoga maritima

    International Nuclear Information System (INIS)

    Vu, Anh; Hamel, Damon J.; Zhou Hongjun; Dahlquist, Frederick W.

    2011-01-01

    The bacterial histidine autokinase CheA contains a histidine phosphotransfer (Hpt) domain that accepts a phosphate from the catalytic domain and donates the phosphate to either target response regulator protein, CheY or CheB. The Hpt domain forms a helix-bundle structure with a conserved four-helix bundle motif and a variable fifth helix. Observation of two nearly equally populated conformations in the crystal structure of a Hpt domain fragment of CheA from Thermotoga maritima containing only the first four helices suggests more mobility in a tightly packed helix bundle structure than previously thought. In order to examine how the structures of Hpt domain homologs may differ from each other particularly in the conformation of the last helix, and whether an alternative conformation exists in the intact Hpt domain in solution, we have solved a high-resolution, solution structure of the CheA Hpt from T. maritima and characterized the backbone dynamics of this protein. The structure contains a four-helix bundle characteristic of histidine phosphotransfer domains. The position and orientation of the fifth helix resembles those in known Hpt domain crystal and solution structures in other histidine kinases. The alternative conformation that was reported in the crystal structure of the CheA Hpt from T. maritima missing the fifth helix is not detected in the solution structure, suggesting a role for the fifth helix in providing stabilizing forces to the overall structure.

  6. Structure of Thermotoga maritima TM0439: implications for the mechanism of bacterial GntR transcription regulators with Zn2+-binding FCD domains

    International Nuclear Information System (INIS)

    Zheng, Meiying; Cooper, David R.; Grossoehme, Nickolas E.; Yu, Minmin; Hung, Li-Wei; Cieslik, Marcin; Derewenda, Urszula; Lesley, Scott A.; Wilson, Ian A.; Giedroc, David P.; Derewenda, Zygmunt S.

    2009-01-01

    Here, the crystal structure of TM0439, a GntR regulator with an FCD domain found in the Thermotoga maritima genome, is described. The GntR superfamily of dimeric transcription factors, with more than 6200 members encoded in bacterial genomes, are characterized by N-terminal winged-helix DNA-binding domains and diverse C-terminal regulatory domains which provide a basis for the classification of the constituent families. The largest of these families, FadR, contains nearly 3000 proteins with all-α-helical regulatory domains classified into two related Pfam families: FadR-C and FCD. Only two crystal structures of FadR-family members, those of Escherichia coli FadR protein and LldR from Corynebacterium glutamicum, have been described to date in the literature. Here, the crystal structure of TM0439, a GntR regulator with an FCD domain found in the Thermotoga maritima genome, is described. The FCD domain is similar to that of the LldR regulator and contains a buried metal-binding site. Using atomic absorption spectroscopy and Trp fluorescence, it is shown that the recombinant protein contains bound Ni 2+ ions but that it is able to bind Zn 2+ with K d < 70 nM. It is concluded that Zn 2+ is the likely physiological metal and that it may perform either structural or regulatory roles or both. Finally, the TM0439 structure is compared with two other FadR-family structures recently deposited by structural genomics consortia. The results call for a revision in the classification of the FadR family of transcription factors

  7. Crystallization and preliminary X-ray crystallographic analysis of Thermotoga maritima CheA P3-P4-P5 domains in complex with CheW

    International Nuclear Information System (INIS)

    Park, SangYoun; Kim, Keon Young; Kim, Sunmin; Crane, Brian R.

    2012-01-01

    T. maritima CheA P3-P4-P5 domains were crystallized in complex with CheW. Low-resolution diffraction data were collected to ∼8 Å using synchrotron X-ray radiation. The CheA–CheW complex plays a key role in bacterial chemotaxis signal transduction by initiating phosphotransfer to response regulators via coupling to the chemoreceptors. CheA (P3-P4-P5 domains) and CheW from Thermotoga maritima were overexpressed in Escherichia coli and crystallized as a complex at 298 K using ammonium dihydrogen phosphate as a precipitant. X-ray diffraction data were collected to ∼8 Å resolution at 100 K using synchrotron radiation. The crystal belonged to space group I222 or I2 1 2 1 2 1 , with unit-cell parameters a = 184.2, b = 286.4, c = 327.7 Å. The asymmetric unit may contain six to ten CheA–CheW molecules

  8. Purification, crystallization and preliminary X-ray diffraction analysis of the putative ABC transporter ATP-binding protein from Thermotoga maritima

    International Nuclear Information System (INIS)

    Ethayathulla, Abdul S.; Bessho, Yoshitaka; Shinkai, Akeo; Padmanabhan, Balasundaram; Singh, Tej P.; Kaur, Punit; Yokoyama, Shigeyuki

    2008-01-01

    The putative ABC transporter ATP-binding protein TM0222 from T. maritima was cloned, overproduced, purified and crystallized. A complete MAD diffraction data set has been collected to 2.3 Å resolution. Adenosine triphosphate (ATP) binding cassette transporters (ABC transporters) are ATP hydrolysis-dependent transmembrane transporters. Here, the overproduction, purification and crystallization of the putative ABC transporter ATP-binding protein TM0222 from Thermotoga maritima are reported. The protein was crystallized in the hexagonal space group P6 4 22, with unit-cell parameters a = b = 148.49, c = 106.96 Å, γ = 120.0°. Assuming the presence of two molecules in the asymmetric unit, the calculated V M is 2.84 Å 3 Da −1 , which corresponds to a solvent content of 56.6%. A three-wavelength MAD data set was collected to 2.3 Å resolution from SeMet-substituted TM0222 crystals. Data sets were collected on the BL38B1 beamline at SPring-8, Japan

  9. Crystallization and preliminary X-ray crystallographic characterization of TrmFO, a folate-dependent tRNA methyltransferase from Thermotoga maritima

    International Nuclear Information System (INIS)

    Cicmil, Nenad

    2008-01-01

    T. maritima TrmFO was overexpressed, purified and crystallized. A diffraction data set was collected to a resolution of 2.6 Å. TrmFO, previously classified as GID, is a methyltransferase that catalyzes the formation of 5-methyluridine or ribothymidine (T) at position 54 in tRNA in some Gram-positive bacteria. To date, TrmFO is the only characterized tRNA methyltransferase that does not use S-adenosylmethionine as the methyl-group donor. Instead, the donor of the methyl group is N 5 ,N 10 -methylenetetrahydrofolate. The crystallization and preliminary X-ray crystallographic studies of TrmFO are reported here. The recombinant protein, cloned from Thermotoga maritima genomic DNA, was overproduced in Esherichia coli and crystallized in 25%(v/v) PEG 4000, 100 mM NaCl and sodium citrate buffer pH 5.0 at 291 K using the hanging-drop vapor-diffusion method. The plate-shaped crystals diffracted to 2.6 Å and belong to the orthorhombic space group P2 1 2 1 2 1 , with unit-cell parameters a = 79.94, b = 92.46, c = 127.20 Å

  10. Crystallization and preliminary X-ray crystallographic characterization of TrmFO, a folate-dependent tRNA methyltransferase from Thermotoga maritima

    Energy Technology Data Exchange (ETDEWEB)

    Cicmil, Nenad, E-mail: cicmil@uiuc.edu [Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801 (United States)

    2008-03-01

    T. maritima TrmFO was overexpressed, purified and crystallized. A diffraction data set was collected to a resolution of 2.6 Å. TrmFO, previously classified as GID, is a methyltransferase that catalyzes the formation of 5-methyluridine or ribothymidine (T) at position 54 in tRNA in some Gram-positive bacteria. To date, TrmFO is the only characterized tRNA methyltransferase that does not use S-adenosylmethionine as the methyl-group donor. Instead, the donor of the methyl group is N{sup 5},N{sup 10}-methylenetetrahydrofolate. The crystallization and preliminary X-ray crystallographic studies of TrmFO are reported here. The recombinant protein, cloned from Thermotoga maritima genomic DNA, was overproduced in Esherichia coli and crystallized in 25%(v/v) PEG 4000, 100 mM NaCl and sodium citrate buffer pH 5.0 at 291 K using the hanging-drop vapor-diffusion method. The plate-shaped crystals diffracted to 2.6 Å and belong to the orthorhombic space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 79.94, b = 92.46, c = 127.20 Å.

  11. Phenotypic Changes in Transgenic Tobacco Plants Overexpressing Vacuole-Targeted Thermotoga maritima BglB Related to Elevated Levels of Liberated Hormones

    Science.gov (United States)

    Nguyen, Quynh Anh; Lee, Dae-Seok; Jung, Jakyun; Bae, Hyeun-Jong

    2015-01-01

    The hyperthermostable β-glucosidase BglB of Thermotoga maritima was modified by adding a short C-terminal tetrapeptide (AFVY, which transports phaseolin to the vacuole, to its C-terminal sequence). The modified β-glucosidase BglB was transformed into tobacco (Nicotiana tabacum L.) plants. We observed a range of significant phenotypic changes in the transgenic plants compared to the wild-type (WT) plants. The transgenic plants had faster stem growth, earlier flowering, enhanced root systems development, an increased biomass biosynthesis rate, and higher salt stress tolerance in young plants compared to WT. In addition, programed cell death was enhanced in mature plants. Furthermore, the C-terminal AFVY tetrapeptide efficiently sorted T. maritima BglB into the vacuole, which was maintained in an active form and could perform its glycoside hydrolysis function on hormone conjugates, leading to elevated hormone [abscisic acid (ABA), indole 3-acetic acid (IAA), and cytokinin] levels that likely contributed to the phenotypic changes in the transgenic plants. The elevation of cytokinin led to upregulation of the transcription factor WUSCHELL, a homeodomain factor that regulates the development, division, and reproduction of stem cells in the shoot apical meristems. Elevation of IAA led to enhanced root development, and the elevation of ABA contributed to enhanced tolerance to salt stress and programed cell death. These results suggest that overexpressing vacuole-targeted T. maritima BglB may have several advantages for molecular farming technology to improve multiple targets, including enhanced production of the β-glucosidase BglB, increased biomass, and shortened developmental stages, that could play pivotal roles in bioenergy and biofuel production. PMID:26618153

  12. Improved Synthesis of 4-Cyanotryptophan and Other Tryptophan Analogues in Aqueous Solvent Using Variants of TrpB from Thermotoga maritima.

    Science.gov (United States)

    Boville, Christina E; Romney, David K; Almhjell, Patrick J; Sieben, Michaela; Arnold, Frances H

    2018-04-27

    The use of enzymes has become increasingly widespread in synthesis as chemists strive to reduce their reliance on organic solvents in favor of more environmentally benign aqueous media. With this in mind, we previously endeavored to engineer the tryptophan synthase β-subunit (TrpB) for production of noncanonical amino acids that had previously been synthesized through multistep routes involving water-sensitive reagents. This enzymatic platform proved effective for the synthesis of analogues of the amino acid tryptophan (Trp), which are frequently used in pharmaceutical synthesis as well as chemical biology. However, certain valuable compounds, such as the blue fluorescent amino acid 4-cyanotryptophan (4-CN-Trp), could only be made in low yield, even at elevated temperature (75 °C). Here, we describe the engineering of TrpB from Thermotoga maritima that improved synthesis of 4-CN-Trp from 24% to 78% yield. Remarkably, although the final enzyme maintains high thermostability ( T 50 = 93 °C), its temperature profile is shifted such that high reactivity is observed at ∼37 °C (76% yield), creating the possibility for in vivo 4-CN-Trp production. The improvements are not specific to 4-CN-Trp; a boost in activity at lower temperature is also demonstrated for other Trp analogues.

  13. Crystallization and preliminary crystallographic analysis of an esterese with a novel domein from the hyperthermophile Thermotoga maritima

    NARCIS (Netherlands)

    Sun, L.; Levisson, M.; Hendriks, S.N.A.; Akveld, T.; Kengen, S.W.M.; Dijkstra, B.W.; Oost, van der J.

    2007-01-01

    Esterase A4 (EA4) is a timer protein found in diapause eggs of the silkworm Bombyx mori. The gene for this metalloglycoprotein was cloned from B. mori eggs and expressed using a baculovirus expression system in silkworm pupae. Crystals of the purified protein have been grown that diffract to beyond

  14. Crystal structure of Thermotoga maritima TM0439: implications for the mechanism of bacterial GntR transcription regulators with Zn2+-binding FCD domains

    Energy Technology Data Exchange (ETDEWEB)

    Zheng, Meiying; Cooper, David; Grossoehmerb, Nickolas; Yu, Minmin; Hung, Li-Wei; Cieslik, Murcin; Derewendaro, Urszula; Lesley, Scott; Wilson, Ian; Giedrocb, David; Derewenda, Zygmunt

    2009-06-06

    The GntR superfamily of dimeric transcription factors, with more than 6200 members encoded in bacterial genomes, are characterized by N-terminal winged helix (WH) DNA-binding domains and diverse C-terminal, regulatory domains, which provide a basis for the classification of the constituent families. The largest of these families, FadR, contains nearly 3000 proteins with all a-helical regulatory domains classified into two related Pfam families: FadR{_}C and FCD. Only two crystal structures of the FadR family members, i.e. the E. coli FadR protein and the LldR from C. glutamicum, have been described to date in literature. Here we describe the crystal structure of TM0439, a GntR regulator with an FCD domain, found in the Thermotoga maritima genome. The FCD domain is similar to that of the LldR regulator, and contains a buried metal binding site. Using atomic absorption spectroscopy and Trp fluorescence, we show that the recombinant protein contains bound Ni{sup 2+} ions, but it is able to bind Zn{sup 2+} with K{sub D} < 70 nM . We conclude that Zn{sup 2+} is the likely physiological metal, where it may perform either or both structural and regulatory roles. Finally, we compare the TM0439 structure to two other FadR family structures recently deposited by Structural Genomics consortia. The results call for a revision in the classification of the FadR family of transcription factors.

  15. Over-expression of xylanolytic ∝-glucuronidase from Thermotoga ...

    African Journals Online (AJOL)

    The GH67∝-glucuronidase encoded by aguA of Thermotoga maritima is one of the most ... to date and thus has considerable potential in industrial application. ... Enzymatic hydrolysis of corncob xylan examined by HPLC showed that more ...

  16. Creation of metal-independent hyperthermophilic L-arabinose isomerase by homologous recombination.

    Science.gov (United States)

    Hong, Young-Ho; Lee, Dong-Woo; Pyun, Yu-Ryang; Lee, Sung Haeng

    2011-12-28

    Hyperthermophilic L-arabinose isomerases (AIs) are useful in the commercial production of D-tagatose as a low-calorie bulk sweetener. Their catalysis and thermostability are highly dependent on metals, which is a major drawback in food applications. To study the role of metal ions in the thermostability and catalysis of hyperthermophilic AI, four enzyme chimeras were generated by PCR-based hybridization to replace the variable N- and C-terminal regions of hyperthermophilic Thermotoga maritima AI (TMAI) and thermophilic Geobacillus stearothermophilus AI (GSAI) with those of the homologous mesophilic Bacillus halodurans AI (BHAI). Unlike Mn(2+)-dependent TMAI, the GSAI- and TMAI-based hybrids with the 72 C-terminal residues of BHAI were not metal-dependent for catalytic activity. By contrast, the catalytic activities of the TMAI- and GSAI-based hybrids containing the N-terminus (residues 1-89) of BHAI were significantly enhanced by metals, but their thermostabilities were poor even in the presence of Mn(2+), indicating that the effects of metals on catalysis and thermostability involve different structural regions. Moreover, in contrast to the C-terminal truncate (Δ20 residues) of GSAI, the N-terminal truncate (Δ7 residues) exhibited no activity due to loss of its native structure. The data thus strongly suggest that the metal dependence of the catalysis and thermostability of hyperthermophilic AIs evolved separately to optimize their activity and thermostability at elevated temperatures. This may provide effective target regions for engineering, thereby meeting industrial demands for the production of d-tagatose.

  17. Hydrogen production by hyperthermophilic and extremely thermophilic bacteria and archaea: mechanisms for reductant disposal.

    Science.gov (United States)

    Verhaart, Marcel R A; Bielen, Abraham A M; van der Oost, John; Stams, Alfons J M; Kengen, Servé W M

    2010-01-01

    Hydrogen produced from biomass by bacteria and archaea is an attractive renewable energy source. However, to make its application more feasible, microorganisms are needed with high hydrogen productivities. For several reasons, hyperthermophilic and extremely thermophilic bacteria and archaea are promising is this respect. In addition to the high polysaccharide-hydrolysing capacities of many of these organisms, an important advantage is their ability to use most of the reducing equivalents (e.g. NADH, reduced ferredoxin) formed during glycolysis for the production of hydrogen, enabling H2/hexose ratios of between 3.0 and 4.0. So, despite the fact that the hydrogen-yielding reactions, especially the one from NADH, are thermodynamically unfavourable, high hydrogen yields are obtained. In this review we focus on three different mechanisms that are employed by a few model organisms, viz. Caldicellulosiruptor saccharolyticus and Thermoanaerobacter tengcongensis, Thermotoga maritima, and Pyrococcus furiosus, to efficiently produce hydrogen. In addition, recent developments to improve hydrogen production by hyperthermophilic and extremely thermophilic bacteria and archaea are discussed.

  18. Cloning, expression and characterization of L-arabinose isomerase from Thermotoga neapolitana: bioconversion of D-galactose to D-tagatose using the enzyme.

    Science.gov (United States)

    Kim, Byoung-Chan; Lee, Yoon-Hee; Lee, Han-Seung; Lee, Dong-Woo; Choe, Eun-Ah; Pyun, Yu-Ryang

    2002-06-18

    Gene araA encoding an L-arabinose isomerase (AraA) from the hyperthermophile, Thermotoga neapolitana 5068 was cloned, sequenced, and expressed in Escherichia coli. The gene encoded a polypeptide of 496 residues with a calculated molecular mass of 56677 Da. The deduced amino acid sequence has 94.8% identical amino acids compared with the residues in a putative L-arabinose isomerase of Thermotoga maritima. The recombinant enzyme expressed in E. coli was purified to homogeneity by heat treatment, ion exchange chromatography and gel filtration. The thermophilic enzyme had a maximum activity of L-arabinose isomerization and D-galactose isomerization at 85 degrees C, and required divalent cations such as Co(2+) and Mn(2+) for its activity and thermostability. The apparent K(m) values of the enzyme for L-arabinose and D-galactose were 116 mM (v(max), 119 micromol min(-1) mg(-1)) and 250 mM (v(max), 14.3 micromol min(-1) mg(-1)), respectively, that were determined in the presence of both 1 mM Co(2+) and 1 mM Mn(2+). A 68% conversion of D-galactose to D-tagatose was obtained using the recombinant enzyme at the isomerization temperature of 80 degrees C.

  19. Domain-swapping of mesophilic xylanase with hyper-thermophilic glucanase

    Directory of Open Access Journals (Sweden)

    Liu Liangwei

    2012-06-01

    Full Text Available Abstract Background Domain fusion is limited at enzyme one terminus. The issue was explored by swapping a mesophilic Aspergillus niger GH11 xylanase (Xyn with a hyper-thermophilic Thermotoga maritima glucanase (Glu to construct two chimeras, Xyn-Glu and Glu-Xyn, with an intention to create thermostable xylanase containing glucanase activity. Results When expressed in E. coli BL21(DE3, the two chimeras exhibited bi-functional activities of xylanase and glucanase. The Xyn-Glu Xyn moiety had optimal reaction temperature (Topt at 50 °C and thermal in-activation half-life (t1/2 at 50 °C for 47.6 min, compared to 47 °C and 17.6 min for the Xyn. The Glu-Xyn Xyn moiety had equivalent Topt to and shorter t1/2 (5.2 min than the Xyn. Both chimera Glu moieties were more thermostable than the Glu, and the three enzyme Topt values were higher than 96 °C. The Glu-Xyn Glu moiety optimal pH was 5.8, compared to 3.8 for the Xyn-Glu Glu moiety and the Glu. Both chimera two moieties cooperated with each other in degrading substrates. Conclusions Domain-swapping created different effects on each moiety properties. Fusing the Glu domain at C-terminus increased the xylanase thermostability, but fusing the Glu domain at N-terminus decreased the xylanase thermostability. Fusing the Xyn domain at either terminus increased the glucanase thermostability, and fusing the Xyn domain at C-terminus shifted the glucanase pH property 2 units higher towards alkaline environments. Fusing a domain at C-terminus contributes more to enzyme catalytic activity; whereas, fusing a bigger domain at N-terminus disturbs enzyme substrate binding affinity.

  20. Genome sequence of the Thermotoga thermarum type strain (LA3(T)) from an African solfataric spring.

    Science.gov (United States)

    Göker, Markus; Spring, Stefan; Scheuner, Carmen; Anderson, Iain; Zeytun, Ahmet; Nolan, Matt; Lucas, Susan; Tice, Hope; Del Rio, Tijana Glavina; Cheng, Jan-Fang; Han, Cliff; Tapia, Roxanne; Goodwin, Lynne A; Pitluck, Sam; Liolios, Konstantinos; Mavromatis, Konstantinos; Pagani, Ioanna; Ivanova, Natalia; Mikhailova, Natalia; Pati, Amrita; Chen, Amy; Palaniappan, Krishna; Land, Miriam; Hauser, Loren; Chang, Yun-Juan; Jeffries, Cynthia D; Rohde, Manfred; Detter, John C; Woyke, Tanja; Bristow, James; Eisen, Jonathan A; Markowitz, Victor; Hugenholtz, Philip; Kyrpides, Nikos C; Klenk, Hans-Peter; Lapidus, Alla

    2014-06-15

    Thermotoga thermarum Windberger et al. 1989 is a member to the genomically well characterized genus Thermotoga in the phylum 'Thermotogae'. T. thermarum is of interest for its origin from a continental solfataric spring vs. predominantly marine oil reservoirs of other members of the genus. The genome of strain LA3T also provides fresh data for the phylogenomic positioning of the (hyper-)thermophilic bacteria. T. thermarum strain LA3(T) is the fourth sequenced genome of a type strain from the genus Thermotoga, and the sixth in the family Thermotogaceae to be formally described in a publication. Phylogenetic analyses do not reveal significant discrepancies between the current classification of the group, 16S rRNA gene data and whole-genome sequences. Nevertheless, T. thermarum significantly differs from other Thermotoga species regarding its iron-sulfur cluster synthesis, as it contains only a minimal set of the necessary proteins. Here we describe the features of this organism, together with the complete genome sequence and annotation. The 2,039,943 bp long chromosome with its 2,015 protein-coding and 51 RNA genes is a part of the Genomic Encyclopedia of Bacteria and Archaea project.

  1. TM0416, a Hyperthermophilic Promiscuous Nonphosphorylated Sugar Isomerase, Catalyzes Various C5 and C6 Epimerization Reactions.

    Science.gov (United States)

    Shin, Sun-Mi; Cao, Thinh-Phat; Choi, Jin Myung; Kim, Seong-Bo; Lee, Sang-Jae; Lee, Sung Haeng; Lee, Dong-Woo

    2017-05-15

    There is currently little information on nonphosphorylated sugar epimerases, which are of potential interest for producing rare sugars. We found a gene (the TM0416 gene) encoding a putative d-tagatose-3-epimerase-related protein from the hyperthermophilic bacterium Thermotoga maritima We overexpressed the TM0416 gene in Escherichia coli and purified the resulting recombinant protein for detailed characterization. Amino acid sequence alignment and a structural similarity search revealed that TM0416 is a putative nonphosphorylated sugar epimerase. The recombinant enzyme exhibited maximal C-3 epimerization of l-ribulose to l-xylulose at ∼80°C and pH 7 in the presence of 1 mM Mn 2+ In addition, this enzyme showed unusually high activity for the epimerization of d-tagatose to d-sorbose, with a conversion yield of 20% after 6 h at 80°C. Remarkably, the enzyme catalyzed the isomerization of d-erythrose or d-threose to d-erythrulose significantly, with conversion yields of 71% and 54.5%, respectively, after 6 h at 80°C at pH 7. To further investigate the substrate specificity of TM0416, we determined its crystal structures in complex with divalent metal ions and l-erythrulose at resolutions of 1.5 and 1.6 Å. Detailed inspection of the structural features and biochemical data clearly demonstrated that this metalloenzyme, with a freely accessible substrate-binding site and neighboring hydrophobic residues, exhibits different and promiscuous substrate preferences, compared with its mesophilic counterparts. Therefore, this study suggests that TM0416 can be functionally classified as a novel type of l-ribulose 3-epimerase (R3E) with d-erythrose isomerase activity. IMPORTANCE Rare sugars, which occur naturally in small amounts, have attracted considerable attention in the food and drug industries. However, there is little information on nonphosphorylated sugar epimerases, which might potentially be applied for the production of rare sugars. This study describes the

  2. Stability of Hyperthermophilic Proteins

    DEFF Research Database (Denmark)

    Stiefler-Jensen, Daniel

    stability by randomly generate mutants and lengthy screening processes to identify the best new mutants. However, with the increase in available genomic sequences of thermophilic or hyperthermophilic organisms a world of enzymes with intrinsic high stability are now available. As these organisms are adapted...... to life at high temperatures so are their enzymes, as a result the high stability is accompanied by low activity at moderate temperatures. Thus, much effort had been put into decoding the mechanisms behind the high stability of the thermophilic enzymes. The hope is to enable scientist to design enzymes...... in the high stability of hyperthermophilic enzymes. The thesis starts with an introduction to the field of protein and enzyme stability with special focus on the thermophilic and hyperthermophilic enzymes and proteins. After the introduction three original research manuscripts present the experimental data...

  3. Expression, purification, crystallization and preliminary X-ray diffraction analysis of Thermotoga neapolitana β-glucosidase B

    Energy Technology Data Exchange (ETDEWEB)

    Turner, Pernilla [Department of Biotechnology, Centre for Chemistry and Chemical Engineering, Lund University, Box 124, S-221 00 Lund (Sweden); Pramhed, Anna [Department of Molecular Biophysics, Centre for Chemistry and Chemical Engineering, Lund University, Box 124, S-221 00 Lund (Sweden); Kanders, Erik; Hedström, Martin; Karlsson, Eva Nordberg, E-mail: eva.nordberg-karlsson@biotek.lu.se [Department of Biotechnology, Centre for Chemistry and Chemical Engineering, Lund University, Box 124, S-221 00 Lund (Sweden); Logan, Derek T., E-mail: eva.nordberg-karlsson@biotek.lu.se [Department of Molecular Biophysics, Centre for Chemistry and Chemical Engineering, Lund University, Box 124, S-221 00 Lund (Sweden); Department of Biotechnology, Centre for Chemistry and Chemical Engineering, Lund University, Box 124, S-221 00 Lund (Sweden)

    2007-09-01

    Here, the expression, purification, crystallization and X-ray diffraction data of a family 3 β-glucosidase from the hyperthermophilic bacterium Thermotoga neapolitana are reported. β-Glucosidases belong to families 1, 3 and 9 of the glycoside hydrolases and act on cello-oligosaccharides. Family 1 and 3 enzymes are retaining and are reported to have transglycosylation activity, which can be used to produce oligosaccharides and glycoconjugates. Family 3 enzymes are less well characterized than their family 1 homologues and to date only two crystal structures have been solved. Here, the expression, purification, crystallization and X-ray diffraction data of a family 3 β-glucosidase from the hyperthermophilic bacterium Thermotoga neapolitana are reported. Crystals of selenomethionine-substituted protein have also been grown. The crystals belong to space group C222{sub 1}, with unit-cell parameters a = 74.9, b = 127.0, c = 175.2 Å. Native data have been collected to 2.4 Å resolution and the structure has been solved to 2.7 Å using the selenomethionine MAD method. Model building and refinement of the structure are under way.

  4. Biohydrogen Production from Glycerol using Thermotoga spp

    NARCIS (Netherlands)

    Maru, B.T.; Bielen, A.A.M.; Kengen, S.W.M.; Constantini, M.; Medina, F.

    2012-01-01

    Given the highly reduced state of carbon in glycerol and its availability as a substantial byproduct of biodiesel production, glycerol is of special interest for sustainable biofuel production. Glycerol was used as a substrate for biohydrogen production using the hyperthermophilic bacterium,

  5. Molecular biology of hyperthermophilic Archaea.

    Science.gov (United States)

    van der Oost, J; Ciaramella, M; Moracci, M; Pisani, F M; Rossi, M; de Vos, W M

    1998-01-01

    The sequences of a number of archaeal genomes have recently been completed, and many more are expected shortly. Consequently, the research of Archaea in general and hyperthermophiles in particular has entered a new phase, with many exciting discoveries to be expected. The wealth of sequence information has already led, and will continue to lead to the identification of many enzymes with unique properties, some of which have potential for industrial applications. Subsequent functional genomics will help reveal fundamental matters such as details concerning the genetic, biochemical and physiological adaptation of extremophiles, and hence give insight into their genomic evolution, polypeptide structure-function relations, and metabolic regulation. In order to optimally exploit many unique features that are now emerging, the development of genetic systems for hyperthermophilic Archaea is an absolute requirement. Such systems would allow the application of this class of Archaea as so-called "cell factories": (i) expression of certain archaeal enzymes for which no suitable conventional (mesophilic bacterial or eukaryal) systems are available, (ii) selection for thermostable variants of potentially interesting enzymes from mesophilic origin, and (iii) the development of in vivo production systems by metabolic engineering. An overview is given of recent insight in the molecular biology of hyperthermophilic Archaea, as well as of a number of promising developments that should result in the generation of suitable genetic systems in the near future.

  6. Regulation of transcription in hyperthermophilic archaea

    NARCIS (Netherlands)

    Brinkman, A.B.

    2002-01-01

    The aim of the research presented here was to insight in the mechanisms by which transcription in hyperthermophilic archaea is regulated. To accomplish this, we have aimed (I) to identify transcriptional regulatory proteins from hyperthermophilic archaea, (II) to characterize these

  7. Expression of Heterologous Cellulases in Thermotoga sp. Strain RQ2

    Directory of Open Access Journals (Sweden)

    Hui Xu

    2015-01-01

    Full Text Available The ability of Thermotoga spp. to degrade cellulose is limited due to a lack of exoglucanases. To address this deficiency, cellulase genes Csac_1076 (celA and Csac_1078 (celB from Caldicellulosiruptor saccharolyticus were cloned into T. sp. strain RQ2 for heterologous overexpression. Coding regions of Csac_1076 and Csac_1078 were fused to the signal peptide of TM1840 (amyA and TM0070 (xynB, resulting in three chimeric enzymes, namely, TM1840-Csac_1078, TM0070-Csac_1078, and TM0070-Csac_1076, which were carried by Thermotoga-E. coli shuttle vectors pHX02, pHX04, and pHX07, respectively. All three recombinant enzymes were successfully expressed in E. coli DH5α and T. sp. strain RQ2, rendering the hosts with increased endo- and/or exoglucanase activities. In E. coli, the recombinant enzymes were mainly bound to the bacterial cells, whereas in T. sp. strain RQ2, about half of the enzyme activities were observed in the culture supernatants. However, the cellulase activities were lost in T. sp. strain RQ2 after three consecutive transfers. Nevertheless, this is the first time heterologous genes bigger than 1 kb (up to 5.3 kb in this study have ever been expressed in Thermotoga, demonstrating the feasibility of using engineered Thermotoga spp. for efficient cellulose utilization.

  8. Enhancement of fermentative hydrogen production from green algal biomass of Thermotoga neapolitana by various pretreatment methods

    Energy Technology Data Exchange (ETDEWEB)

    Nguyen, Tam-Anh D.; Kim, Kyoung-Rok; Nguyen, Minh-Thu; Sim, Sang Jun [Department of Chemical Engineering, Sungkyunkwan University, Suwon 440-746 (Korea, Republic of); Kim, Mi Sun [Bioenergy Research Center, Korea Institute of Energy Research, Daejeon 305-343 (Korea, Republic of); Kim, Donhue [Department of Biochemical Engineering, Dongyang Mirae College, Seoul 152-714 (Korea, Republic of)

    2010-12-15

    Biomass of the green algae has been recently an attractive feedstock source for bio-fuel production because the algal carbohydrates can be derived from atmospheric CO{sub 2} and their harvesting methods are simple. We utilized the accumulated starch in the green alga Chlamydomonas reinhardtii as the sole substrate for fermentative hydrogen (H{sub 2}) production by the hyperthermophilic eubacterium Thermotoga neapolitana. Because of possessing amylase activity, the bacterium could directly ferment H{sub 2} from algal starch with H{sub 2} yield of 1.8-2.2 mol H{sub 2}/mol glucose and the total accumulated H{sub 2} level from 43 to 49% (v/v) of the gas headspace in the closed culture bottle depending on various algal cell-wall disruption methods concluding sonication or methanol exposure. Attempting to enhance the H{sub 2} production, two pretreatment methods using the heat-HCl treatment and enzymatic hydrolysis were applied on algal biomass before using it as substrate for H{sub 2} fermentation. Cultivation with starch pretreated by 1.5% HCl at 121 C for 20 min showed the total accumulative H{sub 2} yield of 58% (v/v). In other approach, enzymatic digestion of starch by thermostable {alpha}-amylase (Termamyl) applied in the SHF process significantly enhanced the H{sub 2} productivity of the bacterium to 64% (v/v) of total accumulated H{sub 2} level and a H{sub 2} yield of 2.5 mol H{sub 2}/mol glucose. Our results demonstrated that direct H{sub 2} fermentation from algal biomass is more desirably potential because one bacterial cultivation step was required that meets the cost-savings, environmental friendly and simplicity of H{sub 2} production. (author)

  9. Molecular characterization of hydrolytic enzymes from hyperthermophilic archaea

    NARCIS (Netherlands)

    Voorhorst, W.G.B.

    1998-01-01

    Hyperthermophiles are recently discovered microorganisms which are able to grow optimally above 85 °C. Most hyperthermophiles belong to the Archaea, the third domain of life. One of the main interests in hyperthermophiles to deepen the insight in the way their proteins

  10. (Hyper)thermophilic Enzymes: Production and Purification

    NARCIS (Netherlands)

    Falcicchio, P.; Levisson, M.; Kengen, S.W.M.; Koutsopoulos, S.

    2014-01-01

    The discovery of thermophilic and hyperthermophilic microorganisms, thriving at environmental temperatures near or above 100 °C, has revolutionized our ideas about the upper temperature limit at which life can exist. The characterization of (hyper)thermostable proteins has broadened our

  11. Phylogeny and molecular signatures for the phylum Thermotogae and its subgroups.

    Science.gov (United States)

    Gupta, Radhey S; Bhandari, Vaibhav

    2011-06-01

    Thermotogae species are currently identified mainly on the basis of their unique toga and distinct branching in the rRNA and other phylogenetic trees. No biochemical or molecular markers are known that clearly distinguish the species from this phylum from all other bacteria. The taxonomic/evolutionary relationships within this phylum, which consists of a single family, are also unclear. We report detailed phylogenetic analyses on Thermotogae species based on concatenated sequences for many ribosomal as well as other conserved proteins that identify a number of distinct clades within this phylum. Additionally, comprehensive analyses of protein sequences from Thermotogae genomes have identified >60 Conserved Signature Indels (CSI) that are specific for the Thermotogae phylum or its different subgroups. Eighteen CSIs in important proteins such as PolI, RecA, TrpRS and ribosomal proteins L4, L7/L12, S8, S9, etc. are uniquely present in various Thermotogae species and provide molecular markers for the phylum. Many CSIs were specific for a number of Thermotogae subgroups. Twelve of these CSIs were specific for a clade consisting of various Thermotoga species except Tt. lettingae, which was separated from other Thermotoga species by a long branch in phylogenetic trees; Fourteen CSIs were specific for a clade consisting of the Fervidobacterium and Thermosipho genera and eight additional CSIs were specific for the genus Thermosipho. In addition, the existence of a clade consisting of the deep branching species Petrotoga mobilis, Kosmotoga olearia and Thermotogales bacterium mesG1 was supported by seven CSIs. The deep branching of this clade was also supported by a number of CSIs that were present in various Thermotogae species, but absent in this clade and all other bacteria. Most of these clades were strongly supported by phylogenetic analyses based on two datasets of protein sequences and they identify potential higher taxonomic grouping (viz. families) within this phylum

  12. Hydrogen production from carrot pulp by the extreme thermophiles Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana

    NARCIS (Netherlands)

    Vrije, de G.J.; Budde, M.A.W.; Lips, S.J.J.; Bakker, R.R.; Mars, A.E.; Claassen, P.A.M.

    2010-01-01

    Hydrogen was produced from carrot pulp hydrolysate, untreated carrot pulp and (mixtures of) glucose and fructose by the extreme thermophiles Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana in pH-controlled bioreactors. Carrot pulp hydrolysate was obtained after enzymatic hydrolysis

  13. Phytochemical study, antioxidant and antibacterial activities of Stemodia maritima

    Directory of Open Access Journals (Sweden)

    Francisca R. L. da Silva

    2014-01-01

    Full Text Available Stemodinol, a new natural compound, together with known compounds including jaceidin, stemodin, stemodinoside B, isocrenatoside, verbascoside, crenatoside, and isoverbascoside, were isolated from Stemodia maritima Linn. The antioxidant (DPPH method and antimicrobial activities of stemodin, stemodinoside B, and crenatoside were investigated. Among the components tested, only crenatoside isolated from the roots showed a high antioxidant power. Stemodin and stemodinoside B exhibited antibacterial activities.

  14. Hyperthermophilic Archaeal Viruses as Novel Nanoplatforms

    DEFF Research Database (Denmark)

    Uldahl, Kristine Buch

    Viruses are the most abundant biological entities on earth, and with an estimated 1031 virus-like particles in the biosphere, viruses are virtually everywhere. Traditionally, the study of viruses has focused on their roles as infectious agents. However, over the last decades with the development...... presents an in depth investigation of the hyperthermophilic archaeal virus SMV. Decisive steps in the viral life-cycle are studied with focus on the early stages of infection. TEM observations suggest that SMV1 virions enter into host cells via a fusion entry mechanism, involving three distinct stages...

  15. Mesophilic and hyperthermophilic adenylate kinases differ in their tolerance to random fragmentation.

    Science.gov (United States)

    Segall-Shapiro, Thomas H; Nguyen, Peter Q; Dos Santos, Edgardo D; Subedi, Saurav; Judd, Justin; Suh, Junghae; Silberg, Jonathan J

    2011-02-11

    The extent to which thermostability influences the location of protein fragmentation sites that allow retention of function is not known. To evaluate this, we used a novel transposase-based approach to create libraries of vectors that express structurally-related fragments of Bacillus subtilis adenylate kinase (BsAK) and Thermotoga neapolitana adenylate kinase (TnAK) with identical modifications at their termini, and we selected for variants in each library that complement the growth of Escherichia coli with a temperature-sensitive adenylate kinase (AK). Mutants created using the hyperthermophilic TnAK were found to support growth with a higher frequency (44%) than those generated from the mesophilic BsAK (6%), and selected TnAK mutants complemented E. coli growth more strongly than homologous BsAK variants. Sequencing of functional clones from each library also identified a greater dispersion of fragmentation sites within TnAK. Nondisruptive fission sites were observed within the AMP binding and core domains of both AK homologs. However, only TnAK contained sites within the lid domain, which undergoes dynamic fluctuations that are critical for catalysis. These findings implicate the flexible lid domain as having an increased sensitivity to fission events at physiological temperatures. In addition, they provide evidence that comparisons of nondisruptive fission sites in homologous proteins could be useful for finding dynamic regions whose conformational fluctuations are important for function, and they show that the discovery of protein fragments that cooperatively function in mesophiles can be aided by the use of thermophilic enzymes as starting points for protein design. Copyright © 2010 Elsevier Ltd. All rights reserved.

  16. Hyperthermophilic endoglucanase for in planta lignocellulose conversion

    Directory of Open Access Journals (Sweden)

    Klose Holger

    2012-08-01

    Full Text Available Abstract Background The enzymatic conversion of lignocellulosic plant biomass into fermentable sugars is a crucial step in the sustainable and environmentally friendly production of biofuels. However, a major drawback of enzymes from mesophilic sources is their suboptimal activity under established pretreatment conditions, e.g. high temperatures, extreme pH values and high salt concentrations. Enzymes from extremophiles are better adapted to these conditions and could be produced by heterologous expression in microbes, or even directly in the plant biomass. Results Here we show that a cellulase gene (sso1354 isolated from the hyperthermophilic archaeon Sulfolobus solfataricus can be expressed in plants, and that the recombinant enzyme is biologically active and exhibits the same properties as the wild type form. Since the enzyme is inactive under normal plant growth conditions, this potentially allows its expression in plants without negative effects on growth and development, and subsequent heat-inducible activation. Furthermore we demonstrate that the recombinant enzyme acts in high concentrations of ionic liquids and can therefore degrade α-cellulose or even complex cell wall preparations under those pretreatment conditions. Conclusion The hyperthermophilic endoglucanase SSO1354 with its unique features is an excellent tool for advanced biomass conversion. Here we demonstrate its expression in planta and the possibility for post harvest activation. Moreover the enzyme is suitable for combined pretreatment and hydrolysis applications.

  17. Evolutionary insights from studies on viruses of hyperthermophilic archaea.

    Science.gov (United States)

    Prangishvili, David

    2003-05-01

    The morphological diversity of viruses which parasitize hyperthermophilic archaea thriving at temperatures > or = 80 degrees C appears to exceed that of viruses of prokaryotes living at lower temperatures. Based on assumptions of the existence of viruses in the prebiotic phase of evolution and hot origins of cellular life, we suggest that this remarkable diversity could have its source in ancestral diversity of viral morphotypes in hot environments. Attempts are made to trace evolutionary relationships of viruses of hyperthermophilic archaea with other viruses.

  18. (Hyper)thermophilic enzymes: production and purification.

    Science.gov (United States)

    Falcicchio, Pierpaolo; Levisson, Mark; Kengen, Servé W M; Koutsopoulos, Sotirios

    2014-01-01

    The discovery of thermophilic and hyperthermophilic microorganisms, thriving at environmental temperatures near or above 100 °C, has revolutionized our ideas about the upper temperature limit at which life can exist. The characterization of (hyper)thermostable proteins has broadened our understanding and presented new opportunities for solving one of the most challenging problems in biophysics: how is structural stability and biological function maintained at high temperatures where "normal" proteins undergo dramatic structural changes? In our laboratory we have purified and studied many thermostable and hyperthermostable proteins in an attempt to determine the molecular basis of heat stability. Here, we present methods to express such proteins and enzymes in E. coli and provide a general protocol for overproduction and purification. The ability to produce enzymes that retain their stability and activity at elevated temperatures creates exciting opportunities for a wide range of biocatalytic applications.

  19. Thermal-induced conformational changes in the product release area drive the enzymatic activity of xylanases 10B: Crystal structure, conformational stability and functional characterization of the xylanase 10B from Thermotoga petrophila RKU-1

    International Nuclear Information System (INIS)

    Santos, Camila Ramos; Meza, Andreia Navarro; Hoffmam, Zaira Bruna; Silva, Junio Cota; Alvarez, Thabata Maria; Ruller, Roberto; Giesel, Guilherme Menegon; Verli, Hugo; Squina, Fabio Marcio; Prade, Rolf Alexander; Murakami, Mario Tyago

    2010-01-01

    Research highlights: → The hyperthermostable xylanase 10B from Thermotoga petrophila RKU-1 produces exclusively xylobiose at the optimum temperature. → Circular dichroism spectroscopy suggests a coupling effect of temperature-induced structural changes with its enzymatic behavior. → Crystallographic and molecular dynamics studies indicate that conformational changes in the product release area modulate the enzyme action mode. -- Abstract: Endo-xylanases play a key role in the depolymerization of xylan and recently, they have attracted much attention owing to their potential applications on biofuels and paper industries. In this work, we have investigated the molecular basis for the action mode of xylanases 10B at high temperatures using biochemical, biophysical and crystallographic methods. The crystal structure of xylanase 10B from hyperthermophilic bacterium Thermotoga petrophila RKU-1 (TpXyl10B) has been solved in the native state and in complex with xylobiose. The complex crystal structure showed a classical binding mode shared among other xylanases, which encompasses the -1 and -2 subsites. Interestingly, TpXyl10B displayed a temperature-dependent action mode producing xylobiose and xylotriose at 20 o C, and exclusively xylobiose at 90 o C as assessed by capillary zone electrophoresis. Moreover, circular dichroism spectroscopy suggested a coupling effect of temperature-induced structural changes with this particular enzymatic behavior. Molecular dynamics simulations supported the CD analysis suggesting that an open conformational state adopted by the catalytic loop (Trp297-Lys326) provokes significant modifications in the product release area (+1,+2 and +3 subsites), which drives the enzymatic activity to the specific release of xylobiose at high temperatures.

  20. Thermal-induced conformational changes in the product release area drive the enzymatic activity of xylanases 10B: Crystal structure, conformational stability and functional characterization of the xylanase 10B from Thermotoga petrophila RKU-1

    Energy Technology Data Exchange (ETDEWEB)

    Santos, Camila Ramos; Meza, Andreia Navarro [Laboratorio Nacional de Biociencias (LNBio), Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP (Brazil); Hoffmam, Zaira Bruna; Silva, Junio Cota; Alvarez, Thabata Maria; Ruller, Roberto [Laboratorio Nacional de Ciencia e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP (Brazil); Giesel, Guilherme Menegon; Verli, Hugo [Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS (Brazil); Squina, Fabio Marcio [Laboratorio Nacional de Ciencia e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP (Brazil); Prade, Rolf Alexander [Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK (United States); Murakami, Mario Tyago, E-mail: mario.murakami@lnbio.org.br [Laboratorio Nacional de Biociencias (LNBio), Centro Nacional de Pesquisa em Energia e Materiais, Campinas, SP (Brazil)

    2010-12-10

    Research highlights: {yields} The hyperthermostable xylanase 10B from Thermotoga petrophila RKU-1 produces exclusively xylobiose at the optimum temperature. {yields} Circular dichroism spectroscopy suggests a coupling effect of temperature-induced structural changes with its enzymatic behavior. {yields} Crystallographic and molecular dynamics studies indicate that conformational changes in the product release area modulate the enzyme action mode. -- Abstract: Endo-xylanases play a key role in the depolymerization of xylan and recently, they have attracted much attention owing to their potential applications on biofuels and paper industries. In this work, we have investigated the molecular basis for the action mode of xylanases 10B at high temperatures using biochemical, biophysical and crystallographic methods. The crystal structure of xylanase 10B from hyperthermophilic bacterium Thermotoga petrophila RKU-1 (TpXyl10B) has been solved in the native state and in complex with xylobiose. The complex crystal structure showed a classical binding mode shared among other xylanases, which encompasses the -1 and -2 subsites. Interestingly, TpXyl10B displayed a temperature-dependent action mode producing xylobiose and xylotriose at 20 {sup o}C, and exclusively xylobiose at 90 {sup o}C as assessed by capillary zone electrophoresis. Moreover, circular dichroism spectroscopy suggested a coupling effect of temperature-induced structural changes with this particular enzymatic behavior. Molecular dynamics simulations supported the CD analysis suggesting that an open conformational state adopted by the catalytic loop (Trp297-Lys326) provokes significant modifications in the product release area (+1,+2 and +3 subsites), which drives the enzymatic activity to the specific release of xylobiose at high temperatures.

  1. Molecular characterisation of the thermostability and catalytic properties of enzymes from hyperthermophiles

    NARCIS (Netherlands)

    Lebbink, J.H.G.

    1999-01-01

    Hyperthermophilic organisms are able to survive and reproduce optimally between 80°C and 113°C. Most of them belong to the domain of the Archaea, although several hyperthermophilic Bacteria have been described. One of the major questions regarding hyperthermophiles concerns the molecular

  2. Diterpene and other constituents from Stemodia maritima (Scrophulariaceae)

    Energy Technology Data Exchange (ETDEWEB)

    Rodrigues, Francisco E.A.; Oliveira, Maria da Conceicao F. de; Vasconcelos, Jackson N.; Mafezoli, Jair; Arriaga, Angela M.C., E-mail: angelamcarriaga@yahoo.com.b [Universidade Federal do Ceara (DQOI/UFC), Fortaleza, CE (Brazil). Dept. de Quimica Organica e Inorganica. Curso de Pos-Graducao em Quimica; Lima, Jefferson Q. [Instituto Federal do Ceara, Juazeiro do Norte, CE (Brazil). Curso de Engenharia Ambiental; Santiago, Gilvandete M.P. [Universidade Federal do Ceara (DQOI/UFC), Fortaleza, CE (Brazil). Dept. de Farmacia; Braz-Filho, Raimundo [Universidade Estadual do Norte Fluminense Darcy Ribeiro (CCT/UENF), Campos dos Goytacazes, RJ (Brazil). Centro de Ciencias e Tecnologias

    2010-07-01

    A new diterpene, (5S{sup *},8S{sup *},9R{sup *},10S{sup *})-11{beta},12{beta}-epoxy-9{alpha}-hydroxy-19(4 -> 3)abeo-abieta-3,13-diene-19,18-olide, together with the known compounds stemodin, D-mannitol, betulinic acid, a mixture of 3{beta}-O-{beta}-D-glucopyranosyl-{beta}-sitosterol and 3{beta}-O-{beta}-D-glucopyranosylstigmasterol and 5,7,4'-trihydroxy-3,8,3'-trimethoxyflavone were isolated from the leaves and stems of Stemodia maritima. Structural elucidation of all compounds was based on interpretation of spectral data, mainly NMR (1D and 2D) and MS, including comparison with values described in the literature. (author)

  3. Diterpene and other constituents from Stemodia maritima (Scrophulariaceae)

    International Nuclear Information System (INIS)

    Rodrigues, Francisco E.A.; Oliveira, Maria da Conceicao F. de; Vasconcelos, Jackson N.; Mafezoli, Jair; Arriaga, Angela M.C.; Lima, Jefferson Q.; Santiago, Gilvandete M.P.; Braz-Filho, Raimundo

    2010-01-01

    A new diterpene, (5S * ,8S * ,9R * ,10S * )-11β,12β-epoxy-9α-hydroxy-19(4 -> 3)abeo-abieta-3,13-diene-19,18-olide, together with the known compounds stemodin, D-mannitol, betulinic acid, a mixture of 3β-O-β-D-glucopyranosyl-β-sitosterol and 3β-O-β-D-glucopyranosylstigmasterol and 5,7,4'-trihydroxy-3,8,3'-trimethoxyflavone were isolated from the leaves and stems of Stemodia maritima. Structural elucidation of all compounds was based on interpretation of spectral data, mainly NMR (1D and 2D) and MS, including comparison with values described in the literature. (author)

  4. Engineering of β-glycosidases from hyperthermophilic Archaea

    NARCIS (Netherlands)

    Kaper, T.

    2001-01-01

    Hyperthermophilic Archaea are microorganisms that grow optimally above 80°C. To be able to live at these temperature extremes their cell components display extreme resistance towards thermal degradation. This characteristic is an attractive feature

  5. A First Analysis of Metallome Biosignatures of Hyperthermophilic Archaea

    Directory of Open Access Journals (Sweden)

    Vyllinniskii Cameron

    2012-01-01

    Full Text Available To date, no experimental data has been reported for the metallome of hyperthermophilic microorganisms although their metal requirements for growth are known to be unique. Here, experiments were conducted to determine (i cellular trace metal concentrations of the hyperthermophilic Archaea Methanococcus jannaschii and Pyrococcus furiosus, and (ii a first estimate of the metallome for these hyperthermophilic species via ICP-MS. The metal contents of these cells were compared to parallel experiments using the mesophilic bacterium Escherichia coli grown under aerobic and anaerobic conditions. Fe and Zn were typically the most abundant metals in cells. Metal concentrations for E. coli grown aerobically decreased in the order Fe > Zn > Cu > Mo > Ni > W > Co. In contrast, M. jannaschii and P. furiosus show almost the reverse pattern with elevated Ni, Co, and W concentrations. Of the three organisms, a biosignature is potentially demonstrated for the methanogen M. jannaschii that may, in part, be related to the metallome requirements of methanogenesis. The bioavailability of trace metals more than likely has varied through time. If hyperthermophiles are very ancient, then the trace metal patterns observed here may begin to provide some insights regarding Earth's earliest cells and in turn, early Earth chemistry.

  6. Evolution of mal ABC transporter operons in the Thermococcales and Thermotogales

    Directory of Open Access Journals (Sweden)

    Gogarten J Peter

    2008-01-01

    Full Text Available Abstract Background The mal genes that encode maltose transporters have undergone extensive lateral transfer among ancestors of the archaea Thermococcus litoralis and Pyrococcus furiosus. Bacterial hyperthermophiles of the order Thermotogales live among these archaea and so may have shared in these transfers. The genome sequence of Thermotoga maritima bears evidence of extensive acquisition of archaeal genes, so its ancestors clearly had the capacity to do so. We examined deep phylogenetic relationships among the mal genes of these hyperthermophiles and their close relatives to look for evidence of shared ancestry. Results We demonstrate that the two maltose ATP binding cassette (ABC transporter operons now found in Tc. litoralis and P. furiosus (termed mal and mdx genes, respectively are not closely related to one another. The Tc. litoralis and P. furiosus mal genes are most closely related to bacterial mal genes while their respective mdx genes are archaeal. The genes of the two mal operons in Tt. maritima are not related to genes in either of these archaeal operons. They are highly similar to one another and belong to a phylogenetic lineage that includes mal genes from the enteric bacteria. A unique domain of the enteric MalF membrane spanning proteins found also in these Thermotogales MalF homologs supports their relatively close relationship with these enteric proteins. Analyses of genome sequence data from other Thermotogales species, Fervidobacterium nodosum, Thermosipho melanesiensis, Thermotoga petrophila, Thermotoga lettingae, and Thermotoga neapolitana, revealed a third apparent mal operon, absent from the published genome sequence of Tt. maritima strain MSB8. This third operon, mal3, is more closely related to the Thermococcales' bacteria-derived mal genes than are mal1 and mal2. F. nodosum, Ts. melanesiensis, and Tt. lettingae have only one of the mal1-mal2 paralogs. The mal2 operon from an unknown species of Thermotoga appears to

  7. Exceptionally diverse morphotypes and genomes of crenarchaeal hyperthermophilic viruses

    DEFF Research Database (Denmark)

    Prangishvili, D; Garrett, R A

    2004-01-01

    and Rudiviridae. They all have double-stranded DNA genomes and infect hyperthermophilic crenarchaea of the orders Sulfolobales and Thermoproteales. Representatives of the different viral families share a few homologous ORFs (open reading frames). However, about 90% of all ORFs in the seven sequenced genomes show...... no significant matches to sequences in public databases. This suggests that these hyperthermophilic viruses have exceptional biochemical solutions for biological functions. Specific features of genome organization, as well as strategies for DNA replication, suggest that phylogenetic relationships exist between...... crenarchaeal rudiviruses and the large eukaryal DNA viruses: poxviruses, the African swine fever virus and Chlorella viruses. Sequence patterns at the ends of the linear genome of the lipothrixvirus AFV1 are reminiscent of the telomeric ends of linear eukaryal chromosomes and suggest that a primitive telomeric...

  8. Identification of a mismatch-specific endonuclease in hyperthermophilic Archaea

    OpenAIRE

    Ishino, Sonoko; Nishi, Yuki; Oda, Soichiro; Uemori, Takashi; Sagara, Takehiro; Takatsu, Nariaki; Yamagami, Takeshi; Shirai, Tsuyoshi; Ishino, Yoshizumi

    2016-01-01

    The common mismatch repair system processed by MutS and MutL and their homologs was identified in Bacteria and Eukarya. However, no evidence of a functional MutS/L homolog has been reported for archaeal organisms, and it is not known whether the mismatch repair system is conserved in Archaea. Here, we describe an endonuclease that cleaves double-stranded DNA containing a mismatched base pair, from the hyperthermophilic archaeon Pyrococcus furiosus. The corresponding gene revealed that the act...

  9. A Novel Type of Polyhedral Viruses Infecting Hyperthermophilic Archaea.

    Science.gov (United States)

    Liu, Ying; Ishino, Sonoko; Ishino, Yoshizumi; Pehau-Arnaudet, Gérard; Krupovic, Mart; Prangishvili, David

    2017-07-01

    Encapsidation of genetic material into polyhedral particles is one of the most common structural solutions employed by viruses infecting hosts in all three domains of life. Here, we describe a new virus of hyperthermophilic archaea, Sulfolobus polyhedral virus 1 (SPV1), which condenses its circular double-stranded DNA genome in a manner not previously observed for other known viruses. The genome complexed with virion proteins is wound up sinusoidally into a spherical coil which is surrounded by an envelope and further encased by an outer polyhedral capsid apparently composed of the 20-kDa virion protein. Lipids selectively acquired from the pool of host lipids are integral constituents of the virion. None of the major virion proteins of SPV1 show similarity to structural proteins of known viruses. However, minor structural proteins, which are predicted to mediate host recognition, are shared with other hyperthermophilic archaeal viruses infecting members of the order Sulfolobales The SPV1 genome consists of 20,222 bp and contains 45 open reading frames, only one-fifth of which could be functionally annotated. IMPORTANCE Viruses infecting hyperthermophilic archaea display a remarkable morphological diversity, often presenting architectural solutions not employed by known viruses of bacteria and eukaryotes. Here we present the isolation and characterization of Sulfolobus polyhedral virus 1, which condenses its genome into a unique spherical coil. Due to the original genomic and architectural features of SPV1, the virus should be considered a representative of a new viral family, "Portogloboviridae." Copyright © 2017 American Society for Microbiology.

  10. Alpha-amylase from the Hyperthermophilic Archaeon Thermococcus thioreducens

    Science.gov (United States)

    Bernhardsdotter, E. C. M. J.; Pusey, M. L.; Ng, M. L.; Garriott, O. K.

    2003-01-01

    Extremophiles are microorganisms that thrive in, from an anthropocentric view, extreme environments such as hot springs. The ability of survival at extreme conditions has rendered enzymes from extremophiles to be of interest in industrial applications. One approach to producing these extremozymes entails the expression of the enzyme-encoding gene in a mesophilic host such as E.coli. This method has been employed in the effort to produce an alpha-amylase from a hyperthermophile (an organism that displays optimal growth above 80 C) isolated from a hydrothermal vent at the Rainbow vent site in the Atlantic Ocean. alpha-amylases catalyze the hydrolysis of starch to produce smaller sugars and constitute a class of industrial enzymes having approximately 25% of the enzyme market. One application for thermostable alpha-amylases is the starch liquefaction process in which starch is converted into fructose and glucose syrups. The a-amylase encoding gene from the hyperthermophile Thermococcus thioreducens was cloned and sequenced, revealing high similarity with other archaeal hyperthermophilic a-amylases. The gene encoding the mature protein was expressed in E.coli. Initial characterization of this enzyme has revealed an optimal amylolytic activity between 85-90 C and around pH 5.3-6.0.

  11. Dendrochronology of Atriplex portulacoides and Artemisia maritima in Wadden Sea salt marshes

    NARCIS (Netherlands)

    Decuyper, M.; Slim, P.A.; Loon-Steensma, van J.M.

    2014-01-01

    The study uses a rather unusual method, dendrochronology, to investigate the growth and survival of Atriplex portulacoides L. and Artemisia maritima L. on salt marshes at two field sites on the Dutch North Sea barrier islands of Terschelling and Ameland. By providing information on longevity of

  12. NaCl salinity affects lateral root development in Plantago maritima

    NARCIS (Netherlands)

    Rubinigg, M; Wenisch, J; Elzenga, JTM; Stulen, [No Value

    2004-01-01

    Root growth and morphology were assessed weekly in hydroponically-grown seedlings of the halophyte Plantago maritima L. during exposure to 0, 50, 100 and 200 mM NaCl for 21 d. Relative growth rate was reduced by 25% at 200 mM NaCl. The lower NaCl treatments did not affect relative growth rates.

  13. Reconstructed ancestral Myo-inositol-3-phosphate synthases indicate that ancestors of the Thermococcales and Thermotoga species were more thermophilic than their descendants.

    Science.gov (United States)

    Butzin, Nicholas C; Lapierre, Pascal; Green, Anna G; Swithers, Kristen S; Gogarten, J Peter; Noll, Kenneth M

    2013-01-01

    The bacterial genomes of Thermotoga species show evidence of significant interdomain horizontal gene transfer from the Archaea. Members of this genus acquired many genes from the Thermococcales, which grow at higher temperatures than Thermotoga species. In order to study the functional history of an interdomain horizontally acquired gene we used ancestral sequence reconstruction to examine the thermal characteristics of reconstructed ancestral proteins of the Thermotoga lineage and its archaeal donors. Several ancestral sequence reconstruction methods were used to determine the possible sequences of the ancestral Thermotoga and Archaea myo-inositol-3-phosphate synthase (MIPS). These sequences were predicted to be more thermostable than the extant proteins using an established sequence composition method. We verified these computational predictions by measuring the activities and thermostabilities of purified proteins from the Thermotoga and the Thermococcales species, and eight ancestral reconstructed proteins. We found that the ancestral proteins from both the archaeal donor and the Thermotoga most recent common ancestor recipient were more thermostable than their descendants. We show that there is a correlation between the thermostability of MIPS protein and the optimal growth temperature (OGT) of its host, which suggests that the OGT of the ancestors of these species of Archaea and the Thermotoga grew at higher OGTs than their descendants.

  14. Reconstructed ancestral Myo-inositol-3-phosphate synthases indicate that ancestors of the Thermococcales and Thermotoga species were more thermophilic than their descendants.

    Directory of Open Access Journals (Sweden)

    Nicholas C Butzin

    Full Text Available The bacterial genomes of Thermotoga species show evidence of significant interdomain horizontal gene transfer from the Archaea. Members of this genus acquired many genes from the Thermococcales, which grow at higher temperatures than Thermotoga species. In order to study the functional history of an interdomain horizontally acquired gene we used ancestral sequence reconstruction to examine the thermal characteristics of reconstructed ancestral proteins of the Thermotoga lineage and its archaeal donors. Several ancestral sequence reconstruction methods were used to determine the possible sequences of the ancestral Thermotoga and Archaea myo-inositol-3-phosphate synthase (MIPS. These sequences were predicted to be more thermostable than the extant proteins using an established sequence composition method. We verified these computational predictions by measuring the activities and thermostabilities of purified proteins from the Thermotoga and the Thermococcales species, and eight ancestral reconstructed proteins. We found that the ancestral proteins from both the archaeal donor and the Thermotoga most recent common ancestor recipient were more thermostable than their descendants. We show that there is a correlation between the thermostability of MIPS protein and the optimal growth temperature (OGT of its host, which suggests that the OGT of the ancestors of these species of Archaea and the Thermotoga grew at higher OGTs than their descendants.

  15. Model development and experimental validation of capnophilic lactic fermentation and hydrogen synthesis by Thermotoga neapolitana.

    Science.gov (United States)

    Pradhan, Nirakar; Dipasquale, Laura; d'Ippolito, Giuliana; Fontana, Angelo; Panico, Antonio; Pirozzi, Francesco; Lens, Piet N L; Esposito, Giovanni

    2016-08-01

    The aim of the present study was to develop a kinetic model for a recently proposed unique and novel metabolic process called capnophilic (CO2-requiring) lactic fermentation (CLF) pathway in Thermotoga neapolitana. The model was based on Monod kinetics and the mathematical expressions were developed to enable the simulation of biomass growth, substrate consumption and product formation. The calibrated kinetic parameters such as maximum specific uptake rate (k), semi-saturation constant (kS), biomass yield coefficient (Y) and endogenous decay rate (kd) were 1.30 h(-1), 1.42 g/L, 0.1195 and 0.0205 h(-1), respectively. A high correlation (>0.98) was obtained between the experimental data and model predictions for both model validation and cross validation processes. An increase of the lactate production in the range of 40-80% was obtained through CLF pathway compared to the classic dark fermentation model. The proposed kinetic model is the first mechanistically based model for the CLF pathway. This model provides useful information to improve the knowledge about how acetate and CO2 are recycled back by Thermotoga neapolitana to produce lactate without compromising the overall hydrogen yield. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Energetic and hydrogen limitations of thermophilic and hyperthermophilic methanogens

    Science.gov (United States)

    Stewart, L. C.; Holden, J. F.

    2013-12-01

    Deep-sea hydrothermal vents are a unique ecosystem, based ultimately not on photosynthesis but chemosynthetic primary production. This makes them an excellent analog environment for the early Earth, and for potential extraterrestrial habitable environments, such as those on Mars and Europa. The habitability of given vent systems for chemoautotrophic prokaryotes can be modeled energetically by estimating the available Gibbs energy for specific modes of chemoautotrophy, using geochemical data and mixing models for hydrothermal fluids and seawater (McCollom and Shock, 1997). However, modeling to date has largely not taken into account variation in organisms' energy demands in these environments. Controls on maintenance energies are widely assumed to be temperature-dependent, rising with increasing temperature optima (Tijhuis et al., 1993), and species-independent. The impacts of other environmental stressors and particular energy-gathering strategies on maintenance energies have not been investigated. We have undertaken culture-based studies of growth and maintenance energies in thermophilic and hyperthermophilic methanogenic (hydrogenotrophic) archaea from deep-sea hydrothermal vents to investigate potential controls on energy demands in hydrothermal vent microbes, and to quantify their growth and maintenance energies for future bioenergetic modeling. We have investigated trends in their growth energies over their full temperature range and a range of nitrogen concentrations, and in their maintenance energies at different hydrogen concentrations. Growth energies in these organisms appear to rise with temperature, but do not vary between hyperthermophilic and thermophilic methanogens. Nitrogen availability at tested levels (40μM - 9.4 mM) does not appear to affect growth energies in all but one tested organism. In continuous chemostat culture, specific methane production varied with hydrogen availability but was similar between a thermophilic and a hyperthermophilic

  17. Complete genome sequence of Hippea maritima type strain (MH2T)

    Energy Technology Data Exchange (ETDEWEB)

    Huntemann, Marcel [U.S. Department of Energy, Joint Genome Institute; Lu, Megan [Los Alamos National Laboratory (LANL); Nolan, Matt [U.S. Department of Energy, Joint Genome Institute; Lapidus, Alla L. [U.S. Department of Energy, Joint Genome Institute; Lucas, Susan [U.S. Department of Energy, Joint Genome Institute; Hammon, Nancy [U.S. Department of Energy, Joint Genome Institute; Deshpande, Shweta [U.S. Department of Energy, Joint Genome Institute; Cheng, Jan-Fang [U.S. Department of Energy, Joint Genome Institute; Tapia, Roxanne [Los Alamos National Laboratory (LANL); Han, Cliff [Los Alamos National Laboratory (LANL); Goodwin, Lynne A. [Los Alamos National Laboratory (LANL); Pitluck, Sam [U.S. Department of Energy, Joint Genome Institute; Liolios, Konstantinos [U.S. Department of Energy, Joint Genome Institute; Pagani, Ioanna [U.S. Department of Energy, Joint Genome Institute; Ivanova, N [U.S. Department of Energy, Joint Genome Institute; Ovchinnikova, Galina [U.S. Department of Energy, Joint Genome Institute; Pati, Amrita [U.S. Department of Energy, Joint Genome Institute; Chen, Amy [U.S. Department of Energy, Joint Genome Institute; Palaniappan, Krishna [U.S. Department of Energy, Joint Genome Institute; Land, Miriam L [ORNL; Hauser, Loren John [ORNL; Jeffries, Cynthia [Oak Ridge National Laboratory (ORNL); Detter, J. Chris [U.S. Department of Energy, Joint Genome Institute; Brambilla, Evelyne-Marie [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Rohde, Manfred [HZI - Helmholtz Centre for Infection Research, Braunschweig, Germany; Spring, Stefan [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Goker, Markus [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Woyke, Tanja [U.S. Department of Energy, Joint Genome Institute; Bristow, James [U.S. Department of Energy, Joint Genome Institute; Eisen, Jonathan [U.S. Department of Energy, Joint Genome Institute; Markowitz, Victor [U.S. Department of Energy, Joint Genome Institute; Hugenholtz, Philip [U.S. Department of Energy, Joint Genome Institute; Kyrpides, Nikos C [U.S. Department of Energy, Joint Genome Institute; Klenk, Hans-Peter [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Mavromatis, K [U.S. Department of Energy, Joint Genome Institute

    2011-01-01

    Hippea maritima (Miroshnichenko et al. 1999) is the type species of the genus Hippea, which belongs to the family Desulfurellaceae within the class Deltaproteobacteria. The anaerobic, moderately thermophilic marine sulfur-reducer was first isolated from shallow-water hot vents in Matipur Harbor, Papua New Guinea. H. maritima was of interest for genome se- quencing because of its isolated phylogenetic location, as a distant next neighbor of the ge- nus Desulfurella. Strain MH2T is the first type strain from the order Desulfurellales with a com- pletely sequenced genome. The 1,694,430 bp long linear genome with its 1,723 protein- coding and 57 RNA genes consists of one circular chromosome and is a part of the Genomic Encyclopedia of Bacteria and Archaea project.

  18. Rhizosphere O2 dynamics in young Zostera marina and Ruppia maritima

    DEFF Research Database (Denmark)

    Jovanovic, Zeljko; Pedersen, Mia Østergaard; Larsen, Morten

    2015-01-01

    Zostera marina and Ruppia maritima often share the same habitat, but R. maritima appears more resistant to environmental stress. We investigated the impact of light intensity and water column O2 concentrations on radial oxygen loss (ROL), in young specimens of Z. marina and R. maritima. Planar......, respectively. These values declined by 71 and 60% in darkness. However, both species were able to maintain ROL as long as ambient O2 levels remained >50% air saturation. The calculated ROL integrated over a 24 h cycle was 48.8 ± 10.6 nmol O2 plant−1 d−1 (n = 3) for R. maritima and 30% less for Z. marina...

  19. XX/XY system of sex determination in the geophilomorph centipede Strigamia maritima

    Czech Academy of Sciences Publication Activity Database

    Green, J. E.; Dalíková, Martina; Sahara, K.; Marec, František; Akam, M.

    2016-01-01

    Roč. 11, č. 2 (2016), č. článku e0150292. E-ISSN 1932-6203 R&D Projects: GA AV ČR IAA600960925; GA ČR(CZ) GA14-22765S Institutional support: RVO:60077344 Keywords : sex determination * Strigamia maritima * XX/XY system Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.806, year: 2016 http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0150292

  20. Contribution of Spartina maritima to the reduction of eutrophication in estuarine systems

    International Nuclear Information System (INIS)

    Sousa, Ana I.; Lillebo, Ana I.; Cacador, Isabel; Pardal, Miguel A.

    2008-01-01

    Salt marshes are among the most productive ecosystems in the world, performing important ecosystem functions, particularly nutrient recycling. In this study, a comparison is made between Mondego and Tagus estuaries in relation to the role of Spartina maritima in nitrogen retention capacity and cycling. Two mono-specific S. maritima stands per estuary were studied during 1 yr (biomass, nitrogen (N) pools, litter production, decomposition rates). Results showed that the oldest Tagus salt marsh population presented higher annual belowground biomass and N productions, and a slower decomposition rate for litter, contributing to the higher N accumulation in the sediment, whereas S. maritima younger marshes had higher aboveground biomass production. Detritus moved by tides represented a huge amount of aboveground production, probably significant when considering the N balance of these salt marshes. Results reinforce the functions of salt marshes as contributing to a reduction of eutrophication in transitional waters, namely through sedimentation processes. - The crucial capacity of salt marshes to retain nitrogen, thus reducing eutrophication, greatly depends on the salt marsh maturity, rather than the estuarine system

  1. Comparative assessment of LECA and Spartina maritima to remove emerging organic contaminants from wastewater.

    Science.gov (United States)

    Ferreira, Ana Rita; Guedes, Paula; Mateus, Eduardo P; Ribeiro, Alexandra B; Couto, Nazaré

    2017-03-01

    The present work aimed to evaluate the capacity of constructed wetlands (CWs) to remove three emerging organic contaminants with different physicochemical properties: caffeine (CAF), oxybenzone (MBPh), and triclosan (TCS). The simulated CWs were set up with a matrix of light expanded clay aggregates (LECA) and planted with Spartina maritima, a salt marsh plant. Controlled experiments were carried out in microcosms using deionized water and wastewater collected at a wastewater treatment plant (WWTP), with different contaminant mass ranges, for 3, 7, and 14 days. The effects of variables were tested isolatedly and together (LECA and/or S. maritima). The presence of LECA and/or S. maritima has shown higher removal (around 61-97%) of lipophilic compounds (MBPh and TCS) than the hydrophilic compound (CAF; around 19-85%). This was attributed to the fact that hydrophilic compounds are dissolved in the water column, whereas the lipophilic ones suffer sorption processes promoting their removal by plant roots and/or LECA. In the control (only wastewater), a decrease in the three contaminant levels was observed. Adsorption and bio/rhizoremediation are the strongest hypothesis to explain the decrease in contaminants in the tested conditions.

  2. Purification, crystallization and preliminary crytallographic analysis of phosphoglucose isomerase from the hyperthermophilic archaeon Pyrococcus furiosus

    NARCIS (Netherlands)

    Akerboom, A.P.; Turnbull, A.P.; Hargreaves, D.; Fischer, M.; Geus, de D.; Sedelnikova, S.E.; Berrisford, J.M.; Baker, P.J.; Verhees, C.H.; Oost, van der J.; Rice, D.W.

    2003-01-01

    The glycolytic enzyme phosphoglucose isomerase catalyses the reversible isomerization of glucose 6-phosphate to fructose 6-phosphate. The phosphoglucose isomerase from the hyperthermophilic archaeon Pyrococcus furiosus, which shows no sequence similarity to any known bacterial or eukaryotic

  3. Hydrogen production by hyperthermophilic and extremely thermophilic bacteria and archaea: mechanisms for reductant disposal

    NARCIS (Netherlands)

    Verhaart, M.R.A.; Bielen, A.A.M.; Oost, van der J.; Stams, A.J.M.; Kengen, S.W.M.

    2010-01-01

    Hydrogen produced from biomass by bacteria and archaea is an attractive renewable energy source. However, to make its application more feasible, microorganisms are needed with high hydrogen productivities. For several reasons, hyperthermophilic and extremely thermophilic bacteria and archaea are

  4. Scaffold diversification enhances effectiveness of a superlibrary of hyperthermophilic proteins.

    Science.gov (United States)

    Hussain, Mahmud; Gera, Nimish; Hill, Andrew B; Rao, Balaji M

    2013-01-18

    The use of binding proteins from non-immunoglobulin scaffolds has become increasingly common in biotechnology and medicine. Typically, binders are isolated from a combinatorial library generated by mutating a single scaffold protein. In contrast, here we generated a "superlibrary" or "library-of-libraries" of 4 × 10(8) protein variants by mutagenesis of seven different hyperthermophilic proteins; six of the seven proteins have not been used as scaffolds prior to this study. Binding proteins for five different model targets were successfully isolated from this library. Binders obtained were derived from five out of the seven scaffolds. Strikingly, binders from this modestly sized superlibrary have affinities comparable or higher than those obtained from a library with 1000-fold higher sequence diversity but derived from a single stable scaffold. Thus scaffold diversification, i.e., randomization of multiple different scaffolds, is a powerful alternate strategy for combinatorial library construction.

  5. The First Myriapod Genome Sequence Reveals Conservative Arthropod Gene Content and Genome Organisation in the Centipede Strigamia maritima

    Science.gov (United States)

    Chipman, Ariel D.; Ferrier, David E. K.; Brena, Carlo; Qu, Jiaxin; Hughes, Daniel S. T.; Schröder, Reinhard; Torres-Oliva, Montserrat; Znassi, Nadia; Jiang, Huaiyang; Almeida, Francisca C.; Alonso, Claudio R.; Apostolou, Zivkos; Aqrawi, Peshtewani; Arthur, Wallace; Barna, Jennifer C. J.; Blankenburg, Kerstin P.; Brites, Daniela; Capella-Gutiérrez, Salvador; Coyle, Marcus; Dearden, Peter K.; Du Pasquier, Louis; Duncan, Elizabeth J.; Ebert, Dieter; Eibner, Cornelius; Erikson, Galina; Evans, Peter D.; Extavour, Cassandra G.; Francisco, Liezl; Gabaldón, Toni; Gillis, William J.; Goodwin-Horn, Elizabeth A.; Green, Jack E.; Griffiths-Jones, Sam; Grimmelikhuijzen, Cornelis J. P.; Gubbala, Sai; Guigó, Roderic; Han, Yi; Hauser, Frank; Havlak, Paul; Hayden, Luke; Helbing, Sophie; Holder, Michael; Hui, Jerome H. L.; Hunn, Julia P.; Hunnekuhl, Vera S.; Jackson, LaRonda; Javaid, Mehwish; Jhangiani, Shalini N.; Jiggins, Francis M.; Jones, Tamsin E.; Kaiser, Tobias S.; Kalra, Divya; Kenny, Nathan J.; Korchina, Viktoriya; Kovar, Christie L.; Kraus, F. Bernhard; Lapraz, François; Lee, Sandra L.; Lv, Jie; Mandapat, Christigale; Manning, Gerard; Mariotti, Marco; Mata, Robert; Mathew, Tittu; Neumann, Tobias; Newsham, Irene; Ngo, Dinh N.; Ninova, Maria; Okwuonu, Geoffrey; Ongeri, Fiona; Palmer, William J.; Patil, Shobha; Patraquim, Pedro; Pham, Christopher; Pu, Ling-Ling; Putman, Nicholas H.; Rabouille, Catherine; Ramos, Olivia Mendivil; Rhodes, Adelaide C.; Robertson, Helen E.; Robertson, Hugh M.; Ronshaugen, Matthew; Rozas, Julio; Saada, Nehad; Sánchez-Gracia, Alejandro; Scherer, Steven E.; Schurko, Andrew M.; Siggens, Kenneth W.; Simmons, DeNard; Stief, Anna; Stolle, Eckart; Telford, Maximilian J.; Tessmar-Raible, Kristin; Thornton, Rebecca; van der Zee, Maurijn; von Haeseler, Arndt; Williams, James M.; Willis, Judith H.; Wu, Yuanqing; Zou, Xiaoyan; Lawson, Daniel; Muzny, Donna M.; Worley, Kim C.; Gibbs, Richard A.; Akam, Michael; Richards, Stephen

    2014-01-01

    Myriapods (e.g., centipedes and millipedes) display a simple homonomous body plan relative to other arthropods. All members of the class are terrestrial, but they attained terrestriality independently of insects. Myriapoda is the only arthropod class not represented by a sequenced genome. We present an analysis of the genome of the centipede Strigamia maritima. It retains a compact genome that has undergone less gene loss and shuffling than previously sequenced arthropods, and many orthologues of genes conserved from the bilaterian ancestor that have been lost in insects. Our analysis locates many genes in conserved macro-synteny contexts, and many small-scale examples of gene clustering. We describe several examples where S. maritima shows different solutions from insects to similar problems. The insect olfactory receptor gene family is absent from S. maritima, and olfaction in air is likely effected by expansion of other receptor gene families. For some genes S. maritima has evolved paralogues to generate coding sequence diversity, where insects use alternate splicing. This is most striking for the Dscam gene, which in Drosophila generates more than 100,000 alternate splice forms, but in S. maritima is encoded by over 100 paralogues. We see an intriguing linkage between the absence of any known photosensory proteins in a blind organism and the additional absence of canonical circadian clock genes. The phylogenetic position of myriapods allows us to identify where in arthropod phylogeny several particular molecular mechanisms and traits emerged. For example, we conclude that juvenile hormone signalling evolved with the emergence of the exoskeleton in the arthropods and that RR-1 containing cuticle proteins evolved in the lineage leading to Mandibulata. We also identify when various gene expansions and losses occurred. The genome of S. maritima offers us a unique glimpse into the ancestral arthropod genome, while also displaying many adaptations to its specific

  6. Hydrogen production from carrot pulp by the extreme thermophiles Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana

    Energy Technology Data Exchange (ETDEWEB)

    Vrije, Truus de; Budde, Miriam A.W.; Lips, Steef J.; Bakker, Robert R.; Mars, Astrid E.; Claassen, Pieternel A.M. [Wageningen UR, Food and Biobased Research, P.O. Box 17, 6700 AA Wageningen (Netherlands)

    2010-12-15

    Hydrogen was produced from carrot pulp hydrolysate, untreated carrot pulp and (mixtures of) glucose and fructose by the extreme thermophiles Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana in pH-controlled bioreactors. Carrot pulp hydrolysate was obtained after enzymatic hydrolysis of the polysaccharide fraction in carrot pulp. The main sugars in the hydrolysate were glucose, fructose, and sucrose. In fermentations with glucose hydrogen yields and productivities were similar for both strains. With fructose the hydrogen yield of C. saccharolyticus was reduced which might be related to uptake of glucose and fructose by different types of transport systems. With T. neapolitana the fructose consumption rate and consequently the hydrogen productivity were low. The hydrogen yields of both thermophiles were 2.7-2.8 mol H{sub 2}/mol hexose with 10 g/L sugars from carrot pulp hydrolysate. With 20 g/L sugars the yield of T. neapolitana was 2.4 mol H{sub 2}/mol hexose while the yield of C. saccharolyticus was reduced to 1.3 mol H{sub 2}/mol hexose due to high lactate production in the stationary growth phase. C. saccharolyticus was able to grow on carrot pulp and utilized soluble sugars and, after adaptation, pectin and some (hemi)cellulose. No growth was observed with T. neapolitana when using carrot pulp in agitated fermentations. Enzymatic hydrolysis of the polysaccharide fraction prior to fermentation increased the hydrogen yield with almost 10% to 2.3 g/kg of hydrolyzed carrot pulp. (author)

  7. Respiration of arsenate and selenate by hyperthermophilic archaea.

    Science.gov (United States)

    Huber, R; Sacher, M; Vollmann, A; Huber, H; Rose, D

    2000-10-01

    A novel, strictly anaerobic, hyperthermophilic, facultative organotrophic archaeon was isolated from a hot spring at Pisciarelli Solfatara, Naples, Italy. The rod-shaped cells grew chemolithoautotrophically with carbon dioxide as carbon source, hydrogen as electron donor and arsenate, thiosulfate or elemental sulfur as electron acceptor. H2S was formed from sulfur or thiosulfate, arsenite from arsenate. Organotrophically, the new isolate grew optimally in the presence of an inorganic electron acceptor like sulfur, selenate or arsenate. Cultures, grown on arsenate and thiosulfate or arsenate and L-cysteine, precipitated realgar (As2S2). During growth on selenate, elemental selenium was produced. The G+C content of the DNA was 58.3 mol%. Due to 16S rRNA gene sequence analysis combined with physiological and morphological criteria, the new isolate belongs to the Thermoproteales order. It represents a new species within the genus Pyrobaculum, the type species of which we name Pyrobaculum arsenaticum (type strain PZ6*, DSM 13514, ATCC 700994). Comparative studies with different Pyrobaculum-species showed, that Pyrobaculum aerophilum was also able to grow organotrophically under anaerobic culture conditions in the presence of arsenate, selenate and selenite. During growth on selenite, elemental selenium was formed as final product. In contrast to P. arsenaticum, P. aerophilum could use selenate or arsenate for lithoautotrophic growth with carbon dioxide and hydrogen.

  8. A new thermophilic nitrilase from an antarctic hyperthermophilic microorganism.

    Directory of Open Access Journals (Sweden)

    Geraldine V. Dennett

    2016-02-01

    Full Text Available Several environmental samples from Antarctica were collected and enriched to search for microorganisms with nitrilase activity. A new thermostable nitrilase from a novel hyperthermophilic archaea Pyrococcus sp. M24D13 was purified and characterized. The activity of this enzyme increased as the temperatures rise from 70 up to 85 °C. Its optimal activity occurred at 85 °C and pH 7.5. This new enzyme shows a remarkable resistance to thermal inactivation retaining more than 50% of its activity even after 8 h of incubation at 85 °C.In addition, this nitrilase is highly versatile demonstrating activity towards different substrates such as benzonitrile (60 mM, aromatic nitrile and butyronitrile (60 mM, aliphatic nitrile, with a specific activity of 3286.7 U mg-1 of protein and 4008.2 U mg-1 of protein respectively. Moreover the enzyme NitM24D13 also presents cyanidase activity.The apparent Michaelis-Menten constant (Km and Vmáx of this Nitrilase for benzonitrile were 0.3 mM and 333.3 µM min-1, respectively, and the specificity constant (kcat/Km for benzonitrile was 2.05×105 s-1 M-1.

  9. Alcohol dehydrogenases from thermophilic and hyperthermophilic archaea and bacteria.

    Science.gov (United States)

    Radianingtyas, Helia; Wright, Phillip C

    2003-12-01

    Many studies have been undertaken to characterise alcohol dehydrogenases (ADHs) from thermophiles and hyperthermophiles, mainly to better understand their activities and thermostability. To date, there are 20 thermophilic archaeal and 17 thermophilic bacterial strains known to have ADHs or similar enzymes, including the hypothetical proteins. Some of these thermophiles are found to have multiple ADHs, sometimes of different types. A rigid delineation of amino acid sequences amongst currently elucidated thermophilic ADHs and similar proteins is phylogenetically apparent. All are NAD(P)-dependent, with one exception that utilises the cofactor F(420) instead. Within the NAD(P)-dependent group, the thermophilic ADHs are orderly clustered as zinc-dependent ADHs, short-chain ADHs, and iron-containing/activated ADHs. Distance matrix calculations reveal that thermophilic ADHs within one type are homologous, with those derived from a single genus often showing high similarities. Elucidation of the enzyme activity and stability, coupled with structure analysis, provides excellent information to explain the relationship between them, and thermophilic ADHs diversity.

  10. The protective role of damsissa (Ambroosia Maritima) against gamma irradiation in albino rats

    International Nuclear Information System (INIS)

    Osman, O.A.; Mohamed, Y.S.

    2003-01-01

    The present work was directed to evaluate the effectiveness of treatment with damsissa (Ambrosia maritima) for thirty consecutive days pre- irradiation exposure in controlling the post-irradiation hazards in irradiated rats. Male albino rats (Spraue Dowley strain) weighing about 120+- 10 g were used and blood samples were collected from tails of animals thirty days after treatment with damsissa and seven days post irradiation. Blood samples were subjected to biochemical analysis such as liver functions, kidney function and lipid profile. Whole body gamma irradiation of rats at 6 Gy (single dose) caused significant decrease in the contents of total proteins accompanied by significant increase of urea level as recorded on the 7th days post irradiation. Data obtained in this study revealed that whole body gamma irradiation induced significant elevation in all tested blood lipid functions. There was significant increase of aspartate amino transferase (AST) and alanine amino transferase (ALT) whole alkaline phosphatase (ALP) showed statistical significant decrease as compared with the control group. Damisissa (Ambrosia maritima) treatment exerted noticeable amelioration in the the studied biochemical parameters of the irradiated albino rats. The mechanism of action of damsissa may be due to its anti-inflammatory properties against whole body gamma irradiation

  11. Effect of Gamma Rays and Salinity on Growth and Chemical Composition of Ambrosia maritima L. Plant

    International Nuclear Information System (INIS)

    Moemen, A.M.E.

    2012-01-01

    This work achieved to study the effects of, mixture of salt 2:2:1 (Na Cl-CaCl 2 and Mg SO 4 ), concentration of (0, 2000, 4000 and 6000 ppm). on growth characters, some chemical components and some active ingredients in shoots of Ambrosia maritima plants, at different stages of growth, during two seasons. Pots 30 cm in diameter were filled of sand-loamy soils in appropriate concentration, all pots were irrigated with tap water. The exposed damsisa seeds to gamma rays, doses (0, 20, 40, and 80 Gy) before sowing together with control non irradiated seeds were sown in saline soils (0, 2000, 4000 and 6000 ppm). Soil salinity treatments caused a decrease in plant height, number of leaves, content of damsin, and an increase in fresh weigh, dry weight, total sugars, total chlorophyll, amino acids and ambrosine content. Also, Gamma rays caused an increase in most of growth parameters and most of chemical composition. It was observed that 40 or 80 Gy was more effective. We investigated the combined effect of levels of salinity and doses of radiation used, this interference improve growth parameters and chemical composition in ambrosia maritima plants and caused ascertain the role of gamma irradiation in plants tolerance to soil salinity and alleviation their harmful effect on plants.

  12. Chemical composition and biological effects of Artemisia maritima and Artemisia nilagirica essential oils from wild plant of Western Himalaya

    Science.gov (United States)

    Artemisia species possess pharmacological properties that are used for medical purposes worldwide. In this paper, the essential oils from the aerial parts of A. nilagirica and A. maritima from the western Indian Himalaya region are described. The main compounds analyzed by simultaneous GC/MS and GC/...

  13. Identification of a mismatch-specific endonuclease in hyperthermophilic Archaea.

    Science.gov (United States)

    Ishino, Sonoko; Nishi, Yuki; Oda, Soichiro; Uemori, Takashi; Sagara, Takehiro; Takatsu, Nariaki; Yamagami, Takeshi; Shirai, Tsuyoshi; Ishino, Yoshizumi

    2016-04-20

    The common mismatch repair system processed by MutS and MutL and their homologs was identified in Bacteria and Eukarya. However, no evidence of a functional MutS/L homolog has been reported for archaeal organisms, and it is not known whether the mismatch repair system is conserved in Archaea. Here, we describe an endonuclease that cleaves double-stranded DNA containing a mismatched base pair, from the hyperthermophilic archaeon Pyrococcus furiosus The corresponding gene revealed that the activity originates from PF0012, and we named this enzyme Endonuclease MS (EndoMS) as the mismatch-specific Endonuclease. The sequence similarity suggested that EndoMS is the ortholog of NucS isolated from Pyrococcus abyssi, published previously. Biochemical characterizations of the EndoMS homolog from Thermococcus kodakarensis clearly showed that EndoMS specifically cleaves both strands of double-stranded DNA into 5'-protruding forms, with the mismatched base pair in the central position. EndoMS cleaves G/T, G/G, T/T, T/C and A/G mismatches, with a more preference for G/T, G/G and T/T, but has very little or no effect on C/C, A/C and A/A mismatches. The discovery of this endonuclease suggests the existence of a novel mismatch repair process, initiated by the double-strand break generated by the EndoMS endonuclease, in Archaea and some Bacteria. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

  14. Genome Sequence of a Hyperthermophilic Archaeon, Thermococcus nautili 30-1, That Produces Viral Vesicles.

    Science.gov (United States)

    Oberto, Jacques; Gaudin, Marie; Cossu, Matteo; Gorlas, Aurore; Slesarev, Alexeï; Marguet, Evelyne; Forterre, Patrick

    2014-03-27

    Thermococcus nautili 30-1 (formerly Thermococcus nautilus), an anaerobic hyperthermophilic marine archaeon, was isolated in 1999 from a deep-sea hydrothermal vent during the Amistad campaign. Here, we present the complete sequence of T. nautili, which is able to produce membrane vesicles containing plasmid DNA. This property makes T. nautili a model organism to study horizontal gene transfer.

  15. The activity of hyperthermophilic glycosynthases is significantly enhanced at acidic pH

    NARCIS (Netherlands)

    Perugino, G.; Trincone, A.; Giordano, A.; Oost, van der J.; Kaper, T.; Rossi, M.; Moracci, M.

    2003-01-01

    We have previously shown that the hyperthermophilic glycosynthase from Sulfolobus so fataricus (Ssbeta-glyE387G) can promote the synthesis of branched oligosaccharides from activated beta-glycosides, at pH 6.5, in the presence of 2 M sodium formate as an external nucleophile. In an effort to

  16. Improved oligosaccharide synthesis by protein engineering of b-glucosidase from hyperthermophilic Pyrococcus furiosus

    NARCIS (Netherlands)

    Hanson, T.; Kaper, T.; Oost, van der J.; Vos, de W.M.

    2001-01-01

    Enzymatic transglycosylation of lactose into oligosaccharides was studied using wild-type -glucosidase (CelB) and active site mutants thereof (M424K, F426Y, M424K/F426Y) and wild-type -mannosidase (BmnA) of the hyperthermophilic Pyrococcus furiosus. The effects of the mutations on kinetics, enzyme

  17. A novel rudivirus, ARV1, of the hyperthermophilic archaeal genus Acidianus

    DEFF Research Database (Denmark)

    Vestergaard, Gisle Alberg; Häring, Monika; Peng, Xu

    2005-01-01

    Virus ARV1, the first member of the family Rudiviridae infecting hyperthermophilic archaea of the genus Acidianus, was isolated from a hot spring in Pozzuoli, Italy. The rod-shaped virions, 610 +/- 50 nm long and 22 +/- 3 nm wide, are non-enveloped and carry a helical nucleoprotein core, with thr...

  18. Highlighting the mechanisms by which proline can confer tolerance to salt stress in cakile maritima

    International Nuclear Information System (INIS)

    Messedi, D.; Farhani, F.; Hamed, K.B.; Trabelsi, N.; Ksouri, R.; Chedly Abdelly, C.; Athar, H.U.R.

    2016-01-01

    Cakile maritima is an oleaginous halophyte growing in the sandy dunes along the Tunisian coast. In order to investigate the role of proline in inducing high salinity tolerance (200 and 400 mM NaCl) in this halophyte, we studied several aspects of the salt responses of C. maritma under exogenous proline supply (20 mM). Salinity levels above 100 mM, reduced growth, photosynthetic activity, and quantum yield of photosystem II (FPSII), while increasing the non photochemical quenching (NPQ). Significant inhibition of the linear electron transport rate (ETR) was also observed in plants grown at 400 mM NaCl. In addition, polyphenol content, total antioxidant and DPPH scavenging activities increased due to increasing salinity stress, and the concentration of malondialdehyde (MDA) also increased. The application of proline counteracted all these adverse effects of salt stress in plants grown at 200 mM NaCl, while it improved some of these physiological attributes at 400 mM NaCl. In addition, contribution of Na+ for the osmotic adjustment decreased in the leaves of salt treated plants supplied with proline exogenously. Exogenous application of proline induced the accumulation of potassium, proline and soluble carbohydrates in salt stressed plants, particularly at 400 mM. This explained the reason of growth enhancement induced by proline application. All together, our Results showed that the beneficial effect of exogenous proline on the response of C. maritima to salinity was due to its role in the protection of chloroplast structures, antioxidant defenses and osmotic adjustment. (author)

  19. Metabolism Dealing with Thermal Degradation of NAD+ in the Hyperthermophilic Archaeon Thermococcus kodakarensis.

    Science.gov (United States)

    Hachisuka, Shin-Ichi; Sato, Takaaki; Atomi, Haruyuki

    2017-10-01

    NAD + is an important cofactor for enzymatic oxidation reactions in all living organisms, including (hyper)thermophiles. However, NAD + is susceptible to thermal degradation at high temperatures. It can thus be expected that (hyper)thermophiles harbor mechanisms that maintain in vivo NAD + concentrations and possibly remove and/or reuse undesirable degradation products of NAD + Here we confirmed that at 85°C, thermal degradation of NAD + results mostly in the generation of nicotinamide and ADP-ribose, the latter known to display toxicity by spontaneously linking to proteins. The hyperthermophilic archaeon Thermococcus kodakarensis possesses a putative ADP-ribose pyrophosphatase (ADPR-PPase) encoded by the TK2284 gene. ADPR-PPase hydrolyzes ADP-ribose to ribose 5-phosphate (R5P) and AMP. The purified recombinant TK2284 protein exhibited activity toward ADP-ribose as well as ADP-glucose. Kinetic analyses revealed a much higher catalytic efficiency toward ADP-ribose, suggesting that ADP-ribose was the physiological substrate. To gain insight into the physiological function of TK2284, a TK2284 gene disruption strain was constructed and examined. Incubation of NAD + in the cell extract of the mutant strain at 85°C resulted in higher ADP-ribose accumulation and lower AMP production compared with those in experiments with the host strain cell extract. The mutant strain also exhibited lower cell yield and specific growth rates in a synthetic amino acid medium compared with those of the host strain. The results obtained here suggest that the ADPR-PPase in T. kodakarensis is responsible for the cleavage of ADP-ribose to R5P and AMP, providing a means to utilize the otherwise dead-end product of NAD + breakdown. IMPORTANCE Hyperthermophilic microorganisms living under high temperature conditions should have mechanisms that deal with the degradation of thermolabile molecules. NAD + is an important cofactor for enzymatic oxidation reactions and is susceptible to thermal

  20. Obligate sugar oxidation in Mesotoga spp., phylum Thermotogae, in the presence of either elemental sulfur or hydrogenotrophic sulfate-reducers as electron acceptor.

    Science.gov (United States)

    Fadhlaoui, Khaled; Ben Hania, Wagdi; Armougom, Fabrice; Bartoli, Manon; Fardeau, Marie-Laure; Erauso, Gaël; Brasseur, Gaël; Aubert, Corinne; Hamdi, Moktar; Brochier-Armanet, Céline; Dolla, Alain; Ollivier, Bernard

    2018-01-01

    Mesotoga prima strain PhosAc3 is a mesophilic representative of the phylum Thermotogae comprising only fermentative bacteria so far. We show that while unable to ferment glucose, this bacterium is able to couple its oxidation to reduction of elemental sulfur. We demonstrate furthermore that M. prima strain PhosAc3 as well as M. prima strain MesG1 and Mesotoga infera are able to grow in syntrophic association with sulfate-reducing bacteria (SRB) acting as hydrogen scavengers through interspecies hydrogen transfer. Hydrogen production was higher in M. prima strain PhosAc3 cells co-cultured with SRB than in cells cultured alone in the presence of elemental sulfur. We propose that the efficient sugar-oxidizing metabolism by M. prima strain PhosAc3 in syntrophic association with a hydrogenotrophic sulfate-reducing bacterium can be extrapolated to all members of the Mesotoga genus. Genome comparison of Thermotogae members suggests that the metabolic difference between Mesotoga and Thermotoga species (sugar oxidation versus fermentation) is mainly due to the absence of the bifurcating [FeFe]-hydrogenase in the former. Such an obligate oxidative process for using sugars, unusual within prokaryotes, is the first reported within the Thermotogae. It is hypothesized to be of primary ecological importance for growth of Mesotoga spp. in the environments that they inhabit. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

  1. Structural adaptation of the subunit interface of oligomeric thermophilic and hyperthermophilic enzymes.

    Science.gov (United States)

    Maugini, Elisa; Tronelli, Daniele; Bossa, Francesco; Pascarella, Stefano

    2009-04-01

    Enzymes from thermophilic and, particularly, from hyperthermophilic organisms are surprisingly stable. Understanding of the molecular origin of protein thermostability and thermoactivity attracted the interest of many scientist both for the perspective comprehension of the principles of protein structure and for the possible biotechnological applications through application of protein engineering. Comparative studies at sequence and structure levels were aimed at detecting significant differences of structural parameters related to protein stability between thermophilic and hyperhermophilic structures and their mesophilic homologs. Comparative studies were useful in the identification of a few recurrent themes which the evolution utilized in different combinations in different protein families. These studies were mostly carried out at the monomer level. However, maintenance of a proper quaternary structure is an essential prerequisite for a functional macromolecule. At the environmental temperatures experienced typically by hyper- and thermophiles, the subunit interactions mediated by the interface must be sufficiently stable. Our analysis was therefore aimed at the identification of the molecular strategies adopted by evolution to enhance interface thermostability of oligomeric enzymes. The variation of several structural properties related to protein stability were tested at the subunit interfaces of thermophilic and hyperthermophilic oligomers. The differences of the interface structural features observed between the hyperthermophilic and thermophilic enzymes were compared with the differences of the same properties calculated from pairwise comparisons of oligomeric mesophilic proteins contained in a reference dataset. The significance of the observed differences of structural properties was measured by a t-test. Ion pairs and hydrogen bonds do not vary significantly while hydrophobic contact area increases specially in hyperthermophilic interfaces. Interface

  2. Effets de dix traitements sur la germination des akènes d'Ambrosia maritima L

    Directory of Open Access Journals (Sweden)

    Schafer, JL.

    1989-01-01

    Full Text Available Effects of ten treatments on the germination of Ambrosia maritima L. seeds. Stratification of Ambrosia maritima seeds at + 10°C for 7 days appeared to be the best practical method to break their embryonic dormancy with 80 % germination occuring 18 days following the treatment, compared to 17 % for the control. This experiment confirms the susceptibility of embryonic dormancy to low temperatures. Treatments with waterat + 80°C or concentrated H2S04 for 15 mn and mechanical treatments eliminating the inhibition effect of seed integuments also gave higher results (43 to 53 % germination rate. However, germination in the control group was significantly higher than in the group of seeds subjected to dry heat. Although the first results obtained on the field from these trials were satisfactory, further research is needed to confirm them.

  3. Sulphate reduction and nitrogen fixation rates associated with roots, rhizomes and sediments from Zostera noltii and Spartina maritima meadows.

    Science.gov (United States)

    Nielsen, L B; Finster, K; Welsh, D T; Donelly, A; Herbert, R A; de Wit, R; Lomstein, B A

    2001-01-01

    Sulphate reduction rates (SRR) and nitrogen fixation rates (NFR) associated with isolated roots, rhizomes and sediment from the rhizosphere of the marine macrophytes Zostera noltii and Spartina maritima, and the presence and distribution of Bacteria on the roots and rhizomes, were investigated. Between 1% and 3% of the surface area of the roots and rhizomes of both macrophytes were colonized by Bacteria. Bacteria on the surfaces of S. maritima roots and rhizomes were evenly distributed, while the distribution of Bacteria on Z. noltii roots and rhizomes was patchy. Root- and rhizome-associated SRR and NFR were always higher than rates in the bulk sediment. In particular, nitrogen fixation associated with the roots and rhizomes was 41-650-fold higher than in the bulk sediment. Despite the fact that sulphate reduction was elevated on roots and rhizomes compared with bulk sediment, the contribution of plant-associated sulphate reduction to overall sulphate reduction was small (< or =11%). In contrast, nitrogen fixation associated with the roots and rhizomes accounted for 31% and 91% of the nitrogen fixed in the rhizosphere of Z. noltii and S. maritima respectively. In addition, plant-associated nitrogen fixation could supply 37-1,613% of the nitrogen needed by the sulphate-reducing community. Sucrose stimulated nitrogen fixation and sulphate reduction significantly in the root and rhizome compartments of both macrophytes, but not in the bulk sediment.

  4. Space agriculture for habitation on Mars with hyper-thermophilic aerobic composting bacteria

    Science.gov (United States)

    Kanazawa, S.; Ishikawa, Y.; Tomita-Yokotani, K.; Hashimoto, H.; Kitaya, Y.; Yamashita, M.; Nagatomo, M.; Oshima, T.; Wada, H.; Space Agriculture Task Force, J.

    Manned Mars exploration requires recycle of materials to support human life A conceptual design is developed for space agriculture which is driven by the biologically regenerative function Hyper-thermophilic aerobic composting bacterial ecology is the core of materials recycling system to process human metabolic waste and inedible biomass and convert them to fertilizer for plants cultivation A photosynthetic reaction of plants will be driven by solar energy Water will be recycled by cultivation of plants and passing it through plant bodies Sub-surface water and atmospheric carbon dioxide are the natural resource available on Mars and these resources will be converted to oxygen and foods We envision that the agricultural system will be scaled up by importing materials from Martian environment Excess oxygen will be obtained from growing trees for structural and other components Minor elements including N P K and other traces will be introduced as fertilizers or nutrients into the agricultural materials circulation Nitrogen will be collected from Martian atmosphere We will assess biological fixation of nitrogen using micro-organisms responsible in Earth biosphere Hyper-thermophilic aerobic bacterial ecology is effective to convert waste materials into useful forms to plants This microbial technology has been well established on ground for processing sewage and waste materials For instance the hyper-thermophilic bacterial system is applied to a composting machine in a size of a trash box in home kitchen Since such a home electronics

  5. Electrical current generation in microbial electrolysis cells by hyperthermophilic archaea Ferroglobus placidus and Geoglobus ahangari

    KAUST Repository

    Yilmazel, Yasemin D.

    2017-10-02

    Few microorganisms have been examined for current generation under thermophilic (40–65 °C) or hyperthermophilic temperatures (≥ 80 °C) in microbial electrochemical systems. Two iron-reducing archaea from the family Archaeoglobaceae, Ferroglobus placidus and Geoglobus ahangari, showed electro-active behavior leading to current generation at hyperthermophilic temperatures in single-chamber microbial electrolysis cells (MECs). A current density (j) of 0.68 ± 0.11 A/m2 was attained in F. placidus MECs at 85 °C, and 0.57 ± 0.10 A/m2 in G. ahangari MECs at 80 °C, with an applied voltage of 0.7 V. Cyclic voltammetry (CV) showed that both strains produced a sigmoidal catalytic wave, with a mid-point potential of − 0.39 V (vs. Ag/AgCl) for F. placidus and − 0.37 V for G. ahangari. The comparison of CVs using spent medium and turnover CVs, coupled with the detection of peaks at the same potentials in both turnover and non-turnover conditions, suggested that mediators were not used for electron transfer and that both archaea produced current through direct contact with the electrode. These two archaeal species, and other hyperthermophilic exoelectrogens, have the potential to broaden the applications of microbial electrochemical technologies for producing biofuels and other bioelectrochemical products under extreme environmental conditions.

  6. Understanding DNA Repair in Hyperthermophilic Archaea: Persistent Gaps and Other Reasons to Focus on the Fork

    Directory of Open Access Journals (Sweden)

    Dennis W. Grogan

    2015-01-01

    Full Text Available Although hyperthermophilic archaea arguably have a great need for efficient DNA repair, they lack members of several DNA repair protein families broadly conserved among bacteria and eukaryotes. Conversely, the putative DNA repair genes that do occur in these archaea often do not generate the expected phenotype when deleted. The prospect that hyperthermophilic archaea have some unique strategies for coping with DNA damage and replication errors has intellectual and technological appeal, but resolving this question will require alternative coping mechanisms to be proposed and tested experimentally. This review evaluates a combination of four enigmatic properties that distinguishes the hyperthermophilic archaea from all other organisms: DNA polymerase stalling at dU, apparent lack of conventional NER, lack of MutSL homologs, and apparent essentiality of homologous recombination proteins. Hypothetical damage-coping strategies that could explain this set of properties may provide new starting points for efforts to define how archaea differ from conventional models of DNA repair and replication fidelity.

  7. The Helicase Activity of Hyperthermophilic Archaeal MCM is Enhanced at High Temperatures by Lysine Methylation.

    Science.gov (United States)

    Xia, Yisui; Niu, Yanling; Cui, Jiamin; Fu, Yang; Chen, Xiaojiang S; Lou, Huiqiang; Cao, Qinhong

    2015-01-01

    Lysine methylation and methyltransferases are widespread in the third domain of life, archaea. Nevertheless, the effects of methylation on archaeal proteins wait to be defined. Here, we report that recombinant sisMCM, an archaeal homolog of Mcm2-7 eukaryotic replicative helicase, is methylated by aKMT4 in vitro. Mono-methylation of these lysine residues occurs coincidently in the endogenous sisMCM protein purified from the hyperthermophilic Sulfolobus islandicus cells as indicated by mass spectra. The helicase activity of mini-chromosome maintenance (MCM) is stimulated by methylation, particularly at temperatures over 70°C. The methylated MCM shows optimal DNA unwinding activity after heat-treatment between 76 and 82°C, which correlates well with the typical growth temperatures of hyperthermophilic Sulfolobus. After methylation, the half life of MCM helicase is dramatically extended at 80°C. The methylated sites are located on the accessible protein surface, which might modulate the intra- and inter- molecular interactions through changing the hydrophobicity and surface charge. Furthermore, the methylation-mimic mutants of MCM show heat resistance helicase activity comparable to the methylated MCM. These data provide the biochemical evidence that posttranslational modifications such as methylation may enhance kinetic stability of proteins under the elevated growth temperatures of hyperthermophilic archaea.

  8. Electrical current generation in microbial electrolysis cells by hyperthermophilic archaea Ferroglobus placidus and Geoglobus ahangari

    KAUST Repository

    Yilmazel, Yasemin D.; Zhu, Xiuping; Kim, Kyoung-Yeol; Holmes, Dawn E.; Logan, Bruce E.

    2017-01-01

    Few microorganisms have been examined for current generation under thermophilic (40–65 °C) or hyperthermophilic temperatures (≥ 80 °C) in microbial electrochemical systems. Two iron-reducing archaea from the family Archaeoglobaceae, Ferroglobus placidus and Geoglobus ahangari, showed electro-active behavior leading to current generation at hyperthermophilic temperatures in single-chamber microbial electrolysis cells (MECs). A current density (j) of 0.68 ± 0.11 A/m2 was attained in F. placidus MECs at 85 °C, and 0.57 ± 0.10 A/m2 in G. ahangari MECs at 80 °C, with an applied voltage of 0.7 V. Cyclic voltammetry (CV) showed that both strains produced a sigmoidal catalytic wave, with a mid-point potential of − 0.39 V (vs. Ag/AgCl) for F. placidus and − 0.37 V for G. ahangari. The comparison of CVs using spent medium and turnover CVs, coupled with the detection of peaks at the same potentials in both turnover and non-turnover conditions, suggested that mediators were not used for electron transfer and that both archaea produced current through direct contact with the electrode. These two archaeal species, and other hyperthermophilic exoelectrogens, have the potential to broaden the applications of microbial electrochemical technologies for producing biofuels and other bioelectrochemical products under extreme environmental conditions.

  9. XX/XY System of Sex Determination in the Geophilomorph Centipede Strigamia maritima.

    Directory of Open Access Journals (Sweden)

    Jack E Green

    Full Text Available We show that the geophilomorph centipede Strigamia maritima possesses an XX/XY system of sex chromosomes, with males being the heterogametic sex. This is, to our knowledge, the first report of sex chromosomes in any geophilomorph centipede. Using the recently assembled Strigamia genome sequence, we identified a set of scaffolds differentially represented in male and female DNA sequence. Using quantitative real-time PCR, we confirmed that three candidate X chromosome-derived scaffolds are present at approximately twice the copy number in females as in males. Furthermore, we confirmed that six candidate Y chromosome-derived scaffolds contain male-specific sequences. Finally, using this molecular information, we designed an X chromosome-specific DNA probe and performed fluorescent in situ hybridization against mitotic and meiotic chromosome spreads to identify the Strigamia XY sex-chromosome pair cytologically. We found that the X and Y chromosomes are recognizably different in size during the early pachytene stage of meiosis, and exhibit incomplete and delayed pairing.

  10. Kaempferol glycosides from Lobularia maritima and their potential role in plant interactions.

    Science.gov (United States)

    Fiorentino, Antonio; Ricci, Andreina; D'Abrosca, Brigida; Golino, Annunziata; Izzo, Angelina; Pascarella, Maria Teresa; Piccolella, Simona; Esposito, Assunta

    2009-02-01

    Six kaempferol glycosides, four of them characterized for the first time, were isolated from the leaf extract of Lobularia maritima. The structural elucidation was performed by a combined approach using Electrospray-Ionization Triple-Quadrupole Mass-Spectrometric (ESI/TQ/MS) techniques, and 1D- and 2D-NMR experiments (1H, 13C, DEPT, DQ-COSY, TOCSY, ROESY, NOESY, HSQC, HMBC, and HSQC-TOCSY). The isolated kaempferol derivatives have different disaccharide substituents at C(3) and four of them have a rhamnose unit at C(7). To evaluate their potential allelopathic role within the herbaceous plant community, the compounds, as well as the aglycone obtained from enzymatic hydrolysis, have been tested in vitro on three coexisting plant species, Dactylis hispanica, Petrorhagia velutina, and Phleum subulatum. The results obtained allow us to hypothesize that the type of the sugar modulates the biological response. The bioassay data, analyzed by a multivariate approach, and grouping the compounds on the basis of the number of sugar units and the nature of carbohydrates present in the disaccharide moiety, indicate a structure-activity relationship.

  11. Elementary analysis of Alternanthera Maritima and Blutaparon Portulacoides (Gomphreneae, Amaranthacear) by X-ray fluorescence

    International Nuclear Information System (INIS)

    Salvador, M.J.; Dias, D.A.; Zucchi, O.L.A.D.; Nascimento Filho, V.F.

    2002-01-01

    The phytochemical study and the evaluation of biological activities of plants species have been intensified in the last years. The modernization of analytical equipment allowed significant progress in the natural products chemistry. Most of the plants researches emphasize the isolation of secondary metabolites and the metal analysis is neglected. In this study, aqueous and ethanolic extracts and the intact plant of Alternanthera maritima (aerial parts and roots) and Blutaparon portulacoides (aerial parts and roots), species commonly found on the beaches of eastern coast of Brazil (Restinga de Marica, RJ), were selected for analysis by energy dispersive X-ray fluorescence (EDXRF). With the standard sample aid, the correlation between the elementary sensitivity and the atomic numbers of the elements was determined. The elements P, K, Ca, Ti, Fe, Ni, As, Zn, Br, Sr, Sn and Sb, supported in membranes, were detected in the two analyzed plants, but only the elements in the interval 26 (Fe) ≤ Z ≤ 30 (Zn) were quantified with elementary concentration between 8,7 and 895,0 μg.g -1 . (author)

  12. Production of D-tagatose at high temperatures using immobilized Escherichia coli cells expressing L-arabinose isomerase from Thermotoga neapolitana.

    Science.gov (United States)

    Hong, Young-Ho; Lee, Dong-Woo; Lee, Sang-Jae; Choe, Eun-Ah; Kim, Seong-Bo; Lee, Yoon-Hee; Cheigh, Chan-Ick; Pyun, Yu-Ryang

    2007-04-01

    Escherichia coli cells expressing L-arabinose isomerase from Thermotoga neapolitana (TNAI) were immobilized in calcium alginate beads. The resulting cell reactor (2.4 U, t (1/2) = 43 days at 70 degrees C) in a continuous recycling mode at 70 degrees C produced 49 and 38 g D-tagatose/l from 180 and 90 g D-galactose/l, respectively, within 12 h.

  13. EFFECT OF EXOGENOUS ABSCISIC ACID ON GROWTH AND BIOCHEMICAL CHANGES IN THE HALOPHYTE SUAEDA MARITIMA

    Directory of Open Access Journals (Sweden)

    Anbarasi G.

    2015-04-01

    Full Text Available Different types of phytohormones are being extensively used to alleviate the adverse effect of salinity stress on plant growth. Among those, Abscisic acid (ABA is a plant stress hormone and one of the most important signaling molecules in plants. Drought and salinity activate De-novo abscisic acid synthesis prevent further water loss by evaporation through stomata, mediated by changes in the guard cell turgor pressure. Under osmotic stress abscisic acid induce the accumulation of protein involved in the biosynthesis of osmolites which increasing the stress tolerance of plant. In addition, exogenous application of ABA enhances the tolerance of plants or plant cells to cold, heat, drought, anoxia and heavy metal stresses. This study was carried out to study the exogenous abscisic (ABA acid induced regulatory role on the growth, water content, protein content, chlorophyll content, osmolyte accumulation and protein profiling through SDS PAGE in a halophyte, Suaeda maritima. The osmolyte accumulation of proline and glycine betaine was found to be more in 50 µM ABA concentrations. The protein profiling through SDS PAGE revealed that ̴ 66KDa proteins was not expressed in the control plant and in 10μM ABA treated plants. Interestingly, the ABA treatment induced a new protein of 14.2KDa in 10μM concentration. The ABA treated plants with concentrations 50μM, 100μM and 150μM showed changes in the expression of protein in abundance than the control and 10μM ABA treated plants. The findings in this study indicate that among all the concentrations, 50μM ABA concentration treated plants exhibited higher growth rate.

  14. Armeria maritima from a calamine heap--initial studies on physiologic-metabolic adaptations to metal-enriched soil.

    Science.gov (United States)

    Olko, A; Abratowska, A; Zyłkowska, J; Wierzbicka, M; Tukiendorf, A

    2008-02-01

    Plants of Armeria maritima are found both on unpolluted sites and on soils strongly polluted with heavy metals. Seedlings of A. maritima from a zinc-lead calamine heap in ore-mining region (Bolesław population) and from unpolluted area (Manasterz population) were tested to determine the zinc, cadmium and lead tolerance. In hydroponic experiments Bolesław population was more tolerant to zinc, cadmium and lead. Localization of heavy metals in roots was determined using the histochemical method for detecting metal-complexes with dithizone. Their accumulation was found in root hairs, rhizoderma and at the surface of the central cylinder. Glutathione level in plants increased after metal treatment of both populations. However, its high level was not correlated with phytochelatin production. These metal-binding complexes were not detected in plants exposed to zinc, cadmium or lead. Changes of organic acids concentrations in Armeria treated with metals may suggest their role in metal translocation from roots to shoots. The content of organic acids, especially malate, decreased in the roots and increased in the leaves. These changes may be important in Pb-tolerance of Manasterz population and in Zn-, Cd-tolerance of calamine population from Bolesław.

  15. Synecology of Cutandia maritima (L. Barbey, a rare psammophytic species along the Montenegrin Coast (East Adriatic Coast

    Directory of Open Access Journals (Sweden)

    Stešević Danijela

    2017-12-01

    Full Text Available Cutandia maritima is a circum-Mediterranean species that inhabits sandy dunes along the coast line. It is fairly frequent on the western Adriatic coast but fairly rare and possibly even non-native in the east. In Croatia, it was discovered in 1990 in Crnika Bay on the island of Rab, which was considered until 2005 to be the only site on the eastern Adriatic coast from the Gulf of Trieste in the north to Corfu in the south. In 2009, the species was briefly reported for Velika plaža (Long Beach in Ulcinj (Montenegro but without details about the habitat type and synecology. The aim of this paper is thus to provide a deeper insight into the ecology and synecology of C. maritima in the eastern Adriatic part of the distribution area. On Velika plaža in Ulcinj, the species was found along the whole sea-inland gradient of sand dunes, in various types of vegetation: [1210] - annual vegetation of drift lines, [2110] - embryonic shifting dunes, [2120] - shifting dunes with Ammophila arenaria (white dunes, [2220] - dunes with Euphorbia terracina, [2130*] - fixed coastal dunes with herbaceous vegetation (grey dunes, and also [2190] - humid dune slacks.

  16. Biohydrogen production from untreated and hydrolyzed potato steam peels by the extreme thermophiles Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana

    Energy Technology Data Exchange (ETDEWEB)

    Mars, Astrid E.; Veuskens, Teun; Budde, Miriam A.W.; van Doeveren, Patrick F.N.M.; Lips, Steef J.; Bakker, Robert R.; de Vrije, Truus; Claassen, Pieternel A.M. [Wageningen UR, Food and Biobased Research, P.O. Box 17, 6700 AA Wageningen (Netherlands)

    2010-08-15

    Production of hydrogen by the extreme thermophiles Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana was studied in serum flasks and in pH-controlled bioreactors with glucose, and hydrolyzed and untreated potato steam peels (PSP) as carbon sources. Two types of PSP hydrolysates were used: one in which the starch in the PSP was liquefied with alpha-amylase, and one in which the liquefied starch was further hydrolyzed to glucose by amyloglucosidase. When the PSP hydrolysates or untreated PSP were added at circa 10-14 g/L of glucose units, both strains grew well and produced hydrogen with reasonable to high molar yields (2.4-3.8 moles H{sub 2}/mole glucose units), and no significant production of lactate. The hydrogen production rates and yields were similar with untreated PSP, hydrolyzed PSP, and pure glucose, showing that C. saccharolyticus and T. neapolitana are well equipped for the utilization of starch. When the concentrations of the substrates were increased, growth and hydrogen production of both strains were hampered. At substrate concentrations of circa 30-40 g/L of glucose units, the molar hydrogen yield of C. saccharolyticus was severely reduced due to the formation of high amounts of lactate, while T. neapolitana was unable to grow at all. The results showed that PSP and PSP hydrolysates are very suitable substrates for efficient fermentative hydrogen production at moderate substrate loadings. (author)

  17. Improving anaerobic sewage sludge digestion by implementation of a hyper-thermophilic prehydrolysis step

    DEFF Research Database (Denmark)

    Lu, Jingquan; Gavala, Hariklia N.; Skiadas, Ioannis V.

    2008-01-01

    The present study focuses on a two-step process for treatment and stabilisation of primary sludge. The process consists of a hyperthermophilic hydrolysis step operated at 70 degrees C and a hydraulic retention time (HRT) of 2 clays followed by a thermophilic (55 degrees C) anaerobic digestion step......) with and Without pre-treatment respectively) and up to 115% increase of the methane production rate. Finally it was shown that the extra energy requirements for the operation of a pre-treatment step would be covered by the energy Produced from the extra methane production and in addition there would...

  18. Space agriculture for habitation on Mars with hyper-thermophilic aerobic composting bacteria

    Science.gov (United States)

    Space Agriculture Task Force; Ishikawa, Y.; Tomita-Yokotani, K.; Hashimoto, H.; Kitaya, Y.; Yamashita, M.; Nagatomo, M.; Oshima, T.; Wada, H.

    Manned Mars exploration, especially for extended periods of time, will require recycle of materials to support human life. Here, a conceptual design is developed for a Martian agricultural system driven by biologically regenerative functions. One of the core biotechnologies function is the use of hyper-thermophilic aerobic composting bacterial ecology. These thermophilic bacteria can play an important role in increasing the effectiveness of the processing of human metabolic waste and inedible biomass and of converting them to fertilizer for the cultivation of plants. This microbial technology has been already well established for the purpose of processing sewage and waste materials for small local communities in Japan. One of the characteristics of the technology is that the metabolic heat release that occurs during bacterial fermentation raises the processing temperature sufficiently high at 80 100 °C to support hyper-thermophilic bacteria. Such a hyper-thermophilic system is found to have great capability of decomposing wastes including even their normally recalcitrant components, in a reasonably short period of time and of providing a better quality of fertilizer as an end-product. High quality compost has been shown to be a key element in creating a healthy regenerative food production system. In ground-based studies, the soil microbial ecology after the addition of high quality compost was shown to improve plant growth and promote a healthy symbiosis of arbuscular mycorrhizal fungi. Another advantage of such high processing temperature is the ability to sterilize the pathogenic organisms through the fermentation process and thus to secure the hygienic safety of the system. Plant cultivation is one of the other major systems. It should fully utilize solar energy received on the Martian surface for supplying energy for photosynthesis. Subsurface water and atmospheric carbon dioxide mined on Mars should be also used in the plant cultivation system. Oxygen and

  19. SEASONAL, SIZE-RELATED AND AGE-RELATED PATTERNS IN BODY-MASS AND COMPOSITION OF PURPLE SANDPIPERS CALIDRIS-MARITIMA IN BRITAIN

    NARCIS (Netherlands)

    SUMMERS, RW; UNDERHILL, LG; NICOLL, M; RAE, R; PIERSMA, T

    1992-01-01

    The masses Of 3229 Purple Sandpipers Calidris maritima from Britain were analysed for differences related to age, season and size. First-year birds were lighter by 2 g. There was only a slight increase in mass in mid-winter, in contrast to other waders wintering in Britain, suggesting that Purple

  20. Structural analysis of β-glucosidase mutants derived from a hyperthermophilic tetrameric structure

    International Nuclear Information System (INIS)

    Nakabayashi, Makoto; Kataoka, Misumi; Mishima, Yumiko; Maeno, Yuka; Ishikawa, Kazuhiko

    2014-01-01

    Substitutive mutations that convert a tetrameric β-glucosidase into a dimeric state lead to improvement of its crystal quality. β-Glucosidase from Pyrococcus furiosus (BGLPf) is a hyperthermophilic tetrameric enzyme which can degrade cellooligosaccharides to glucose under hyperthermophilic conditions and thus holds promise for the saccharification of lignocellulosic biomass at high temperature. Prior to the production of large amounts of this enzyme, detailed information regarding the oligomeric structure of the enzyme is required. Several crystals of BGLPf have been prepared over the past ten years, but its crystal structure had not been solved until recently. In 2011, the first crystal structure of BGLPf was solved and a model was constructed at somewhat low resolution (2.35 Å). In order to obtain more detailed structural data on BGLPf, the relationship between its tetrameric structure and the quality of the crystal was re-examined. A dimeric form of BGLPf was constructed and its crystal structure was solved at a resolution of 1.70 Å using protein-engineering methods. Furthermore, using the high-resolution crystal structural data for the dimeric form, a monomeric form of BGLPf was constructed which retained the intrinsic activity of the tetrameric form. The thermostability of BGLPf is affected by its oligomeric structure. Here, the biophysical and biochemical properties of engineered dimeric and monomeric BGLPfs are reported, which are promising prototype models to apply to the saccharification reaction. Furthermore, details regarding the oligomeric structures of BGLPf and the reasons why the mutations yielded improved crystal structures are discussed

  1. Proteomic Insights into Sulfur Metabolism in the Hydrogen-Producing Hyperthermophilic Archaeon Thermococcus onnurineus NA1

    Directory of Open Access Journals (Sweden)

    Yoon-Jung Moon

    2015-04-01

    Full Text Available The hyperthermophilic archaeon Thermococcus onnurineus NA1 has been shown to produce H2 when using CO, formate, or starch as a growth substrate. This strain can also utilize elemental sulfur as a terminal electron acceptor for heterotrophic growth. To gain insight into sulfur metabolism, the proteome of T. onnurineus NA1 cells grown under sulfur culture conditions was quantified and compared with those grown under H2-evolving substrate culture conditions. Using label-free nano-UPLC-MSE-based comparative proteomic analysis, approximately 38.4% of the total identified proteome (589 proteins was found to be significantly up-regulated (≥1.5-fold under sulfur culture conditions. Many of these proteins were functionally associated with carbon fixation, Fe–S cluster biogenesis, ATP synthesis, sulfur reduction, protein glycosylation, protein translocation, and formate oxidation. Based on the abundances of the identified proteins in this and other genomic studies, the pathways associated with reductive sulfur metabolism, H2-metabolism, and oxidative stress defense were proposed. The results also revealed markedly lower expression levels of enzymes involved in the sulfur assimilation pathway, as well as cysteine desulfurase, under sulfur culture condition. The present results provide the first global atlas of proteome changes triggered by sulfur, and may facilitate an understanding of how hyperthermophilic archaea adapt to sulfur-rich, extreme environments.

  2. Proteomic Insights into Sulfur Metabolism in the Hydrogen-Producing Hyperthermophilic Archaeon Thermococcus onnurineus NA1

    Science.gov (United States)

    Moon, Yoon-Jung; Kwon, Joseph; Yun, Sung-Ho; Lim, Hye Li; Kim, Jonghyun; Kim, Soo Jung; Kang, Sung Gyun; Lee, Jung-Hyun; Kim, Seung Il; Chung, Young-Ho

    2015-01-01

    The hyperthermophilic archaeon Thermococcus onnurineus NA1 has been shown to produce H2 when using CO, formate, or starch as a growth substrate. This strain can also utilize elemental sulfur as a terminal electron acceptor for heterotrophic growth. To gain insight into sulfur metabolism, the proteome of T. onnurineus NA1 cells grown under sulfur culture conditions was quantified and compared with those grown under H2-evolving substrate culture conditions. Using label-free nano-UPLC-MSE-based comparative proteomic analysis, approximately 38.4% of the total identified proteome (589 proteins) was found to be significantly up-regulated (≥1.5-fold) under sulfur culture conditions. Many of these proteins were functionally associated with carbon fixation, Fe–S cluster biogenesis, ATP synthesis, sulfur reduction, protein glycosylation, protein translocation, and formate oxidation. Based on the abundances of the identified proteins in this and other genomic studies, the pathways associated with reductive sulfur metabolism, H2-metabolism, and oxidative stress defense were proposed. The results also revealed markedly lower expression levels of enzymes involved in the sulfur assimilation pathway, as well as cysteine desulfurase, under sulfur culture condition. The present results provide the first global atlas of proteome changes triggered by sulfur, and may facilitate an understanding of how hyperthermophilic archaea adapt to sulfur-rich, extreme environments. PMID:25915030

  3. Insights into dynamics of mobile genetic elements in hyperthermophilic environments from five new Thermococcus plasmids.

    Directory of Open Access Journals (Sweden)

    Mart Krupovic

    Full Text Available Mobilome of hyperthermophilic archaea dwelling in deep-sea hydrothermal vents is poorly characterized. To gain insight into genetic diversity and dynamics of mobile genetic elements in these environments we have sequenced five new plasmids from different Thermococcus strains that have been isolated from geographically remote hydrothermal vents. The plasmids were ascribed to two subfamilies, pTN2-like and pEXT9a-like. Gene content and phylogenetic analyses illuminated a robust connection between pTN2-like plasmids and Pyrococcus abyssi virus 1 (PAV1, with roughly half of the viral genome being composed of genes that have homologues in plasmids. Unexpectedly, pEXT9a-like plasmids were found to be closely related to the previously sequenced plasmid pMETVU01 from Methanocaldococcus vulcanius M7. Our data suggests that the latter observation is most compatible with an unprecedented horizontal transfer of a pEXT9a-like plasmid from Thermococcales to Methanococcales. Gene content analysis revealed that thermococcal plasmids encode Hfq-like proteins and toxin-antitoxin (TA systems of two different families, VapBC and RelBE. Notably, although abundant in archaeal genomes, to our knowledge, TA and hfq-like genes have not been previously found in archaeal plasmids or viruses. Finally, the plasmids described here might prove to be useful in developing new genetic tools for hyperthermophiles.

  4. Crystal structure of a family 16 endoglucanase from the hyperthermophile Pyrococcus furiosus--structural basis of substrate recognition

    NARCIS (Netherlands)

    Ilari, A.; Fiorillo, A.; Angelaccio, S.; Florio, R.; Chiaraluce, R.; Oost, van der J.; Consalvi, V.

    2009-01-01

    Bacterial and archaeal endo-beta-1,3-glucanases that belong to glycoside hydrolase family 16 share a beta-jelly-roll fold, but differ significantly in sequence and in substrate specificity. The crystal structure of the laminarinase (EC 3.2.1.39) from the hyperthermophilic archaeon Pyrococcus

  5. Purification, crystallization and preliminary crystallographic analysis of GTP-binding protein from the hyperthermophilic archaeon Sulfolobus solfataricus

    NARCIS (Netherlands)

    Wu Hao,; Sun, L.; Brouns, S.J.J.; Fu, S.; Akerboom, A.P.; Li, X.; Oost, van der J.

    2007-01-01

    A predicted GTP-binding protein from the hyperthermophilic archaeon Sulfolobus solfataricus, termed SsGBP, has been cloned and overexpressed in Escherichia coli. The purified protein was crystallized using the hanging-drop vapour-diffusion technique in the presence of 0.05 M cadmium sulfate and 0.8

  6. Obligate sugar oxidation in Mesotoga spp., phylum Thermotogae, in the presence of either elemental sulfur or hydrogenotrophic sulfate-reducers as electron acceptor

    OpenAIRE

    Fadhlaoui, K.; Ben Hania, W.; Armougom, Fabrice; Bartoli, M.; Fardeau, Marie-Laure; Erauso, G.; Brasseur, G.; Aubert, C.; Hamdi, M.; Brochier-Armanet, C.; Dolla, A.; Ollivier, Bernard

    2018-01-01

    Mesotoga prima strain PhosAc3 is a mesophilic representative of the phylum Thermotogae comprising only fermentative bacteria so far. We show that while unable to ferment glucose, this bacterium is able to couple its oxidation to reduction of elemental sulfur. We demonstrate furthermore that M. prima strain PhosAc3 as well as M. prima strain MesG1 and Mesotoga infera are able to grow in syntrophic association with sulfate-reducing bacteria (SRB) acting as hydrogen scavengers through interspeci...

  7. Efficient hydrogen production from the lignocellulosic energy crop Miscanthus by the extreme thermophilic bacteria Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana

    Directory of Open Access Journals (Sweden)

    de Vrije Truus

    2009-06-01

    Full Text Available Abstract Background The production of hydrogen from biomass by fermentation is one of the routes that can contribute to a future sustainable hydrogen economy. Lignocellulosic biomass is an attractive feedstock because of its abundance, low production costs and high polysaccharide content. Results Batch cultures of Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana produced hydrogen, carbon dioxide and acetic acid as the main products from soluble saccharides in Miscanthus hydrolysate. The presence of fermentation inhibitors, such as furfural and 5-hydroxylmethyl furfural, in this lignocellulosic hydrolysate was avoided by the mild alkaline-pretreatment conditions at a low temperature of 75°C. Both microorganisms simultaneously and completely utilized all pentoses, hexoses and oligomeric saccharides up to a total concentration of 17 g l-1 in pH-controlled batch cultures. T. neapolitana showed a preference for glucose over xylose, which are the main sugars in the hydrolysate. Hydrogen yields of 2.9 to 3.4 mol H2 per mol of hexose, corresponding to 74 to 85% of the theoretical yield, were obtained in these batch fermentations. The yields were higher with cultures of C. saccharolyticus compared to T. neapolitana. In contrast, the rate of substrate consumption and hydrogen production was higher with T. neapolitana. At substrate concentrations exceeding 30 g l-1, sugar consumption was incomplete, and lower hydrogen yields of 2.0 to 2.4 mol per mol of consumed hexose were obtained. Conclusion Efficient hydrogen production in combination with simultaneous and complete utilization of all saccharides has been obtained during the growth of thermophilic bacteria on hydrolysate of the lignocellulosic feedstock Miscanthus. The use of thermophilic bacteria will therefore significantly contribute to the energy efficiency of a bioprocess for hydrogen production from biomass.

  8. Hyperthermophilic Archaeon Thermococcus kodakarensis Utilizes a Four-Step Pathway for NAD+ Salvage through Nicotinamide Deamination.

    Science.gov (United States)

    Hachisuka, Shin-Ichi; Sato, Takaaki; Atomi, Haruyuki

    2018-06-01

    Many organisms possess pathways that regenerate NAD + from its degradation products, and two pathways are known to salvage NAD + from nicotinamide (Nm). One is a four-step pathway that proceeds through deamination of Nm to nicotinic acid (Na) by Nm deamidase and phosphoribosylation to nicotinic acid mononucleotide (NaMN), followed by adenylylation and amidation. Another is a two-step pathway that does not involve deamination and directly proceeds with the phosphoribosylation of Nm to nicotinamide mononucleotide (NMN), followed by adenylylation. Judging from genome sequence data, the hyperthermophilic archaeon Thermococcus kodakarensis is supposed to utilize the four-step pathway, but the fact that the adenylyltransferase encoded by TK0067 recognizes both NMN and NaMN also raises the possibility of a two-step salvage mechanism. Here, we examined the substrate specificity of the recombinant TK1676 protein, annotated as nicotinic acid phosphoribosyltransferase. The TK1676 protein displayed significant activity toward Na and phosphoribosyl pyrophosphate (PRPP) and only trace activity with Nm and PRPP. We further performed genetic analyses on TK0218 (quinolinic acid phosphoribosyltransferase) and TK1650 (Nm deamidase), involved in de novo biosynthesis and four-step salvage of NAD + , respectively. The ΔTK0218 mutant cells displayed growth defects in a minimal synthetic medium, but growth was fully restored with the addition of Na or Nm. The ΔTK0218 ΔTK1650 mutant cells did not display growth in the minimal medium, and growth was restored with the addition of Na but not Nm. The enzymatic and genetic analyses strongly suggest that NAD + salvage in T. kodakarensis requires deamination of Nm and proceeds through the four-step pathway. IMPORTANCE Hyperthermophiles must constantly deal with increased degradation rates of their biomolecules due to their high growth temperatures. Here, we identified the pathway that regenerates NAD + from nicotinamide (Nm) in the

  9. Ten years of demographic and genetic monitoring of Stachys maritima in Catalonia (2001-2010. Implications for a recovery plan

    Directory of Open Access Journals (Sweden)

    Massó, S.

    2010-12-01

    Full Text Available Stachys maritima is a species typical of the coastal dunes, with a wide distribution within the Mediterranean Basin. In spite of this, the species shows a clear regression. In Catalonia, it has been observed an important reduction of its populations since early 20th century, where it has disappeared from several localities in which it was relatively common (Tarragonès, Barcelonès. Herein we present the results of the demographic monitoring of the species during the last 10 years (2001-2010 in the known localities in Catalonia. Besides corroborating the disappearance (northern Sant Martí d’Empúries, the re-discovering (Llobregat Delta beach and the detection of new populations (inner dunes of the Montgrí, a large year-to-year fluctuation of the monitored populations is stated; the possible reasons are discussed. In addition, the present work also includes the results of the allozyme diversity analysis of the new detected populations as well as the rediscoveries of the period 2004-2008, which were not included in a former study of genetic diversity carried out in 2002-2003. It is necessary to emphasize that the contribution of the new populations to the genetic diversity of Stachys maritima is very small, which can be attributed to their limited population size and /or to founder effects. Despite that the species is included in the Annex 2 (“En Perill d’Extinció” within the Catàleg de Flora Amenaçada de Catalunya (Catalogue of Endangered Flora of Catalonia, and some “soft” conservation measures have been applied at local level (signposting of the beach accesses, environmental education, etc. coupled with other more significant measures (e.g. translocation of individuals discovered in an artificial sandbank, it would be necessary the coordinated action and the scientific support of any initiative of conservation that could be carried out. The general frame to initiate actions of conservation should be the recovery plan of

  10. Biotypes of scentless chamomile Matricaria maritima (L.) ssp. inodora (L.) Dostal and common poppy Papaver rhoeas (L.) resistant to tribenuron methyl, in Poland

    OpenAIRE

    Adamczewski Kazimierz; Kierzek Roman; Matysiak Kinga

    2014-01-01

    Scentless chamomile Matricaria maritima (L.) ssp. inodora (L.) Dostal and common poppy Papaver rhoeas (L.) are species which very often infest winter cereal and winter rape crops. Inhibitors of acetolactate synthase (ALS) are commonly used for control of these weeds. The herbicides are characterised by a single site of action in the plant, which has an influence on selection of the weed population and may result in a rapid development of resistance. In 2012, five seed samples of scen...

  11. Spatial genetic structure in Beta vulgaris subsp. maritima and Beta macrocarpa reveals the effect of contrasting mating system, influence of marine currents, and footprints of postglacial recolonization routes.

    Science.gov (United States)

    Leys, Marie; Petit, Eric J; El-Bahloul, Yasmina; Liso, Camille; Fournet, Sylvain; Arnaud, Jean-François

    2014-05-01

    Understanding the factors that contribute to population genetic divergence across a species' range is a long-standing goal in evolutionary biology and ecological genetics. We examined the relative importance of historical and ecological features in shaping the present-day spatial patterns of genetic structure in two related plant species, Beta vulgaris subsp. maritima and Beta macrocarpa. Using nuclear and mitochondrial markers, we surveyed 93 populations from Brittany (France) to Morocco - the southern limit of their species' range distribution. Whereas B. macrocarpa showed a genotypic structure and a high level of genetic differentiation indicative of selfing, the population genetic structure of B. vulgaris subsp. maritima was consistent with an outcrossing mating system. We further showed (1) a strong geographic clustering in coastal B. vulgaris subsp. maritima populations that highlighted the influence of marine currents in shaping different lineages and (2) a peculiar genetic structure of inland B. vulgaris subsp. maritima populations that could indicate the admixture of distinct evolutionary lineages and recent expansions associated with anthropogenic disturbances. Spatial patterns of nuclear diversity and differentiation also supported a stepwise recolonization of Europe from Atlantic-Mediterranean refugia after the last glacial period, with leading-edge expansions. However, cytoplasmic diversity was not impacted by postglacial recolonization: stochastic long-distance seed dispersal mediated by major oceanic currents may mitigate the common patterns of reduced cytoplasmic diversity observed for edge populations. Overall, the patterns we documented here challenge the general view of reduced genetic diversity at the edge of a species' range distribution and provide clues for understanding how life-history and major geographic features interact to shape the distribution of genetic diversity.

  12. Inhibitory Effects of Urginea maritima (L. Baker, Zhumeria majdae Rech. F. and Wendelbo and Physalis divaricata D. Don Ethanolic Extracts on Mushroom Tyrosinase

    Directory of Open Access Journals (Sweden)

    Foroogh Namjoyan, Alireza Jahangiri, Mohammad Ebrahim Azemi, Hamideh Mousavi

    2016-06-01

    Full Text Available Background: Tyrosinase is a key enzyme in melanin synthesis from tyrosine. To prevent or treat pigmentation disorders, tyrosinase inhibitors have been used increasingly for medicinal and cosmetic products. The aim of this study is to evaluate inhibitory effects of Urginea maritima (L. Baker, Zhumeria majdae Rech.f. & Wendelbo and Physalis divaricata D.Don on mushroom tyrosinase. Methods: The inhibitory activities of the hydroalcoholic extracts of plants against oxidation of L-DOPA (as a substrate by mushroom tyrosinase were investigated. The amount of formed DOPAchrome was determined at 475 nm as optical density. Results: The extracts showed anti-tyrosinase activity weaker than positive control (Kojic acid. The inhibitory activity of tested plants: U.maritima, Z.majdae and P.divaricata against mushroom tyrosinase were 38.61, 29.70 and 25.74 % at 1.67 mg/mL, respectively. Conclusion: The most tyrosinase inhibitory activity was seen for U.maritima. However more investigations on human tyrosinase, toxicological and clinical studies are needed to confirm its activity.

  13. Evolution and thermodynamics of the slow unfolding of hyperstable monomeric proteins

    Directory of Open Access Journals (Sweden)

    Koga Yuichi

    2010-07-01

    Full Text Available Abstract Background The unfolding speed of some hyperthermophilic proteins is dramatically lower than that of their mesostable homologs. Ribonuclease HII from the hyperthermophilic archaeon Thermococcus kodakaraensis (Tk-RNase HII is stabilized by its remarkably slow unfolding rate, whereas RNase HI from the thermophilic bacterium Thermus thermophilus (Tt-RNase HI unfolds rapidly, comparable with to that of RNase HI from Escherichia coli (Ec-RNase HI. Results To clarify whether the difference in the unfolding rate is due to differences in the types of RNase H or differences in proteins from archaea and bacteria, we examined the equilibrium stability and unfolding reaction of RNases HII from the hyperthermophilic bacteria Thermotoga maritima (Tm-RNase HII and Aquifex aeolicus (Aa-RNase HII and RNase HI from the hyperthermophilic archaeon Sulfolobus tokodaii (Sto-RNase HI. These proteins from hyperthermophiles are more stable than Ec-RNase HI over all the temperature ranges examined. The observed unfolding speeds of all hyperstable proteins at the different denaturant concentrations studied are much lower than those of Ec-RNase HI, which is in accordance with the familiar slow unfolding of hyperstable proteins. However, the unfolding rate constants of these RNases H in water are dispersed, and the unfolding rate constant of thermophilic archaeal proteins is lower than that of thermophilic bacterial proteins. Conclusions These results suggest that the nature of slow unfolding of thermophilic proteins is determined by the evolutionary history of the organisms involved. The unfolding rate constants in water are related to the amount of buried hydrophobic residues in the tertiary structure.

  14. "Hot cores" in proteins: Comparative analysis of the apolar contact area in structures from hyper/thermophilic and mesophilic organisms

    Directory of Open Access Journals (Sweden)

    Bossa Francesco

    2008-02-01

    Full Text Available Abstract Background A wide variety of stabilizing factors have been invoked so far to elucidate the structural basis of protein thermostability. These include, amongst the others, a higher number of ion-pairs interactions and hydrogen bonds, together with a better packing of hydrophobic residues. It has been frequently observed that packing of hydrophobic side chains is improved in hyperthermophilic proteins, when compared to their mesophilic counterparts. In this work, protein crystal structures from hyper/thermophilic organisms and their mesophilic homologs have been compared, in order to quantify the difference of apolar contact area and to assess the role played by the hydrophobic contacts in the stabilization of the protein core, at high temperatures. Results The construction of two datasets was carried out so as to satisfy several restrictive criteria, such as minimum redundancy, resolution and R-value thresholds and lack of any structural defect in the collected structures. This approach allowed to quantify with relatively high precision the apolar contact area between interacting residues, reducing the uncertainty due to the position of atoms in the crystal structures, the redundancy of data and the size of the dataset. To identify the common core regions of these proteins, the study was focused on segments that conserve a similar main chain conformation in the structures analyzed, excluding the intervening regions whose structure differs markedly. The results indicated that hyperthermophilic proteins underwent a significant increase of the hydrophobic contact area contributed by those residues composing the alpha-helices of the structurally conserved regions. Conclusion This study indicates the decreased flexibility of alpha-helices in proteins core as a major factor contributing to the enhanced termostability of a number of hyperthermophilic proteins. This effect, in turn, may be due to an increased number of buried methyl groups in

  15. Elementary analysis of Alternanthera Maritima and Blutaparon Portulacoides (Gomphreneae, Amaranthacear) by X-ray fluorescence; Analise elementar de Alternanthera Maritima e Blutaparon Portulacoides (Gomphreneae, Amaranthacear) por fluorescencia de raios-X

    Energy Technology Data Exchange (ETDEWEB)

    Salvador, M J; Dias, D A; Zucchi, O L.A.D. [Sao Paulo Univ., Ribeirao Preto, SP (Brazil). Faculdade de Ciencias Farmaceuticas. Dept. de Fisica e Quimica; br, mjsalva@fcfrp usp; Nascimento Filho, V F [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil). Lab. de Instrumentacao Nuclear

    2002-07-01

    The phytochemical study and the evaluation of biological activities of plants species have been intensified in the last years. The modernization of analytical equipment allowed significant progress in the natural products chemistry. Most of the plants researches emphasize the isolation of secondary metabolites and the metal analysis is neglected. In this study, aqueous and ethanolic extracts and the intact plant of Alternanthera maritima (aerial parts and roots) and Blutaparon portulacoides (aerial parts and roots), species commonly found on the beaches of eastern coast of Brazil (Restinga de Marica, RJ), were selected for analysis by energy dispersive X-ray fluorescence (EDXRF). With the standard sample aid, the correlation between the elementary sensitivity and the atomic numbers of the elements was determined. The elements P, K, Ca, Ti, Fe, Ni, As, Zn, Br, Sr, Sn and Sb, supported in membranes, were detected in the two analyzed plants, but only the elements in the interval 26 (Fe) {<=} Z {<=} 30 (Zn) were quantified with elementary concentration between 8,7 and 895,0 {mu}g.g{sup -1}. (author)

  16. Improved methane production from sugarcane vinasse with filter cake in thermophilic UASB reactors, with predominance of Methanothermobacter and Methanosarcina archaea and Thermotogae bacteria.

    Science.gov (United States)

    Barros, Valciney Gomes de; Duda, Rose Maria; Vantini, Juliana da Silva; Omori, Wellington Pine; Ferro, Maria Inês Tiraboschi; Oliveira, Roberto Alves de

    2017-11-01

    Biogas production from sugarcane vinasse has enormous economic, energy, and environmental management potential. However, methane production stability and biodigested vinasse quality remain key issues, requiring better nutrient and alkalinity availability, operational strategies, and knowledge of reactor microbiota. This study demonstrates increased methane production from vinasse through the use of sugarcane filter cake and improved effluent recirculation, with elevated organic loading rates (OLR) and good reactor stability. We used UASB reactors in a two-stage configuration, with OLRs up to 45gCODL -1 d -1 , and obtained methane production as high as 3LL -1 d -1 . Quantitative PCR indicated balanced amounts of bacteria and archaea in the sludge (10 9 -10 10 copiesg -1 VS), and of the predominant archaea orders, Methanobacteriales and Methanosarcinales (10 6 -10 8 copiesg -1 VS). 16S rDNA sequencing also indicated the thermophilic Thermotogae as the most abundant class of bacteria in the sludge. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. The seasonal dormancy pattern and germination of Matricaria maritima subsp. inodora (L. Dostal seeds in hydrotime model terms

    Directory of Open Access Journals (Sweden)

    Anna Bochenek

    2011-01-01

    Full Text Available Changes in hydrotime model parameters were determined in Matricaria maritima L. subsp. inodora seeds during burial in a field in order to describe the seasonal dormancy pattern. Seeds were exhumed at regular intervals over a year and incubated at different water potentials at 19°C. Germination time courses were analyzed to determine hydrotime population parameters. Values of ѱb(50, ѲH and σѱb varied each month. Mean base water potential values in seeds exhumed each month were related to precipitation over 20 days before their exhumation. Soil temperature could be a trend-controlling factor of this relationship. The seeds were in deep dormancy after remaining 80-90 days in soil below or above limit temperature 15°C. The application of the hydrotime model to describe and predict seasonal dormancy patterns of weed seed is promising, especially for species with a considerable diversification of life strategies and ecophysiological flexibility of diaspores. It could also suggest mechanisms of seasonal dormancy changes of seeds in natural conditions and provide a basis for their examination. One of advantages of the dormancy pattern description of weed seeds remaining in a soil bank by means of threshold models is its simplicity.

  18. Big wigs and small wigs: Time, sex, size and shelter affect cohabitation in the maritime earwig (Anisolabis maritima.

    Directory of Open Access Journals (Sweden)

    Nicole L Hack

    Full Text Available Animal aggregations can occur for a variety of abiotic factors, such as resource limitation, or biotic factors, including group foraging and protection from predators. In our study, we examined whether time, sex, body size or shelter availability affected aggregation behavior of the maritime earwig, Anisolabis maritima (Order Dermaptera, an insect found globally at high densities under driftwood. Specifically, we monitored the distribution of two individuals in arenas with either two shelters (no habitat limitation or one shelter (habitat limitation to determine their propensity for cohabitation at times of peak activity and times of quiescence. Females, whose high levels of aggression are often associated with maternal care, were particularly averse to cohabitation, whereas males were generally more tolerant of other earwigs. Females initially preferred not to cohabitate when placed with a male, but were more tolerant of cohabitation later, regardless of the number of shelters. Same-sex pairs, on the other hand, were less likely to cohabitate with only one shelter present, but males were again more tolerant of conspecifics than females regardless of habitat limitation. When competition for one shelter did not lead to cohabitation, the lone occupant was more likely to be the larger individual in same-sex trials and females in mixed-sex trials. Understanding the tolerance for close proximity under these varying conditions may provide insight into aggregative behavior and spatial distribution patterns in the maritime earwig.

  19. CMS-G from Beta vulgaris ssp. maritima is maintained in natural populations despite containing an atypical cytochrome c oxidase.

    Science.gov (United States)

    Meyer, Etienne H; Lehmann, Caroline; Boivin, Stéphane; Brings, Lea; De Cauwer, Isabelle; Bock, Ralph; Kühn, Kristina; Touzet, Pascal

    2018-02-23

    While mitochondrial mutants of the respiratory machinery are rare and often lethal, cytoplasmic male sterility (CMS), a mitochondrially inherited trait that results in pollen abortion, is frequently encountered in wild populations. It generates a breeding system called gynodioecy. In Beta vulgaris ssp. maritima , a gynodioecious species, we found CMS-G to be widespread across the distribution range of the species. Despite the sequencing of the mitochondrial genome of CMS-G, the mitochondrial sterilizing factor causing CMS-G is still unknown. By characterizing biochemically CMS-G, we found that the expression of several mitochondrial proteins is altered in CMS-G plants. In particular, Cox1, a core subunit of the cytochrome c oxidase (complex IV), is larger but can still assemble into complex IV. However, the CMS-G-specific complex IV was only detected as a stabilized dimer. We did not observe any alteration of the affinity of complex IV for cytochrome c ; however, in CMS-G, complex IV capacity is reduced. Our results show that CMS-G is maintained in many natural populations despite being associated with an atypical complex IV. We suggest that the modified complex IV could incur the associated cost predicted by theoretical models to maintain gynodioecy in wild populations. © 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

  20. Biohydrogen production from pig slurry in a CSTR reactor system with mixed cultures under hyper-thermophilic temperature (70 oC)

    International Nuclear Information System (INIS)

    Kotsopoulos, Thomas A.; Fotidis, Ioannis A.; Tsolakis, Nikolaos; Martzopoulos, Gerassimos G.

    2009-01-01

    A continuous stirred tank reactor (CSTR) (750 cm 3 working volume) was operated with pig slurry under hyper-thermophilic (70 o C) temperature for hydrogen production. The hydraulic retention time (HRT) was 24 h and the organic loading rate was 24.9 g d -1 of volatile solid (VS). The inoculum used in the hyper-thermophilic reactor was sludge obtained from a mesophilic methanogenic reactor. The continuous feeding with active biomass (inoculum) from the mesophilic methanogenic reactor was necessary in order to achieve hydrogen production. The hyper-thermophilic reactor started to produce hydrogen after a short adapted period of 4 days. During the steady state period the mean hydrogen yield was 3.65 cm 3 g -1 of volatile solid added. The high operation temperature of the reactor enhanced the hydrolytic activity in pig slurry and increased the volatile fatty acids (VFA) production. The short HRT (24 h) and the hyper-thermophilic temperature applied in the reactor were enough to prevent methanogenesis. No pre-treatment methods or other control methods for preventing methanogenesis were necessary. Hyper-thermophilic hydrogen production was demonstrated for the first time in a CSTR system, fed with pig slurry, using mixed culture. The results indicate that this system is a promising one for biohydrogen production from pig slurry.

  1. Isolation of a hyperthermophilic archaeum predicted by in situ RNA analysis.

    Science.gov (United States)

    Huber, R; Burggraf, S; Mayer, T; Barns, S M; Rossnagel, P; Stetter, K O

    1995-07-06

    A variety of hyperthermophilic bacteria and archaea have been isolated from high-temperature environments by plating and serial dilutions. However, these techniques allow only the small percentage of organisms able to form colonies, or those that are predominant within environmental samples, to be obtained in pure culture. Recently, in situ 16S ribosomal RNA analyses of samples from the Obsidian hot pool at Yellowstone National Park, Wyoming, revealed a variety of archaeal sequences, which were all different from those of previously isolated species. This suggests substantial diversity of archaea with so far unknown morphological, physiological and biochemical features, which may play an important part within high-temperature ecosystems. Here we describe a procedure to obtain pure cultures of unknown organisms harbouring specific 16S rRNA sequences identified previously within the environment. It combines visual recognition of single cells by phylogenetic staining and cloning by 'optical tweezers'. Our result validates polymerase chain reaction data on the existence of large archael communities.

  2. A virus of hyperthermophilic archaea with a unique architecture among DNA viruses.

    Science.gov (United States)

    Rensen, Elena Ilka; Mochizuki, Tomohiro; Quemin, Emmanuelle; Schouten, Stefan; Krupovic, Mart; Prangishvili, David

    2016-03-01

    Viruses package their genetic material in diverse ways. Most known strategies include encapsulation of nucleic acids into spherical or filamentous virions with icosahedral or helical symmetry, respectively. Filamentous viruses with dsDNA genomes are currently associated exclusively with Archaea. Here, we describe a filamentous hyperthermophilic archaeal virus, Pyrobaculum filamentous virus 1 (PFV1), with a type of virion organization not previously observed in DNA viruses. The PFV1 virion, 400 ± 20 × 32 ± 3 nm, contains an envelope and an inner core consisting of two structural units: a rod-shaped helical nucleocapsid formed of two 14-kDa major virion proteins and a nucleocapsid-encompassing protein sheath composed of a single major virion protein of 18 kDa. The virion organization of PFV1 is superficially similar to that of negative-sense RNA viruses of the family Filoviridae, including Ebola virus and Marburg virus. The linear dsDNA of PFV1 carries 17,714 bp, including 60-bp-long terminal inverted repeats, and contains 39 predicted ORFs, most of which do not show similarities to sequences in public databases. PFV1 is a lytic virus that completely disrupts the host cell membrane at the end of the infection cycle.

  3. Random mutagenesis of the hyperthermophilic archaeon Pyrococcus furiosus using in vitro mariner transposition and natural transformation.

    Science.gov (United States)

    Guschinskaya, Natalia; Brunel, Romain; Tourte, Maxime; Lipscomb, Gina L; Adams, Michael W W; Oger, Philippe; Charpentier, Xavier

    2016-11-08

    Transposition mutagenesis is a powerful tool to identify the function of genes, reveal essential genes and generally to unravel the genetic basis of living organisms. However, transposon-mediated mutagenesis has only been successfully applied to a limited number of archaeal species and has never been reported in Thermococcales. Here, we report random insertion mutagenesis in the hyperthermophilic archaeon Pyrococcus furiosus. The strategy takes advantage of the natural transformability of derivatives of the P. furiosus COM1 strain and of in vitro Mariner-based transposition. A transposon bearing a genetic marker is randomly transposed in vitro in genomic DNA that is then used for natural transformation of P. furiosus. A small-scale transposition reaction routinely generates several hundred and up to two thousands transformants. Southern analysis and sequencing showed that the obtained mutants contain a single and random genomic insertion. Polyploidy has been reported in Thermococcales and P. furiosus is suspected of being polyploid. Yet, about half of the mutants obtained on the first selection are homozygous for the transposon insertion. Two rounds of isolation on selective medium were sufficient to obtain gene conversion in initially heterozygous mutants. This transposition mutagenesis strategy will greatly facilitate functional exploration of the Thermococcales genomes.

  4. Cloning and Characterization of an Alpha-amylase Gene from the Hyperthermophilic Archaeon Thermococcus Thioreducens

    Science.gov (United States)

    Bernhardsdotter, Eva C. M. J.; Pusey, Marc L.; Ng, Joseph D.; Garriott, Owen K.

    2004-01-01

    The gene encoding an extracellular a-amylase, TTA, from the hyperthermophilic archaeon Thermococcus thioreducens was cloned and expressed in Escherichia coli. Primary structural analysis revealed high similarity with other a-amylases from the Thermococcus and Pyrococcus genera, as well as the four highly conserved regions typical for a-amylases. The 1374 bp gene encodes a protein of 457 amino acids, of which 435 constitute the mature protein preceded by a 22 amino acid signal peptide. The molecular weight of the purified recombinant enzyme was estimated to be 43 kDa by denaturing gel electrophoresis. Maximal enzymatic activity of recombinant TTA was observed at 90 C and pH 5.5 in the absence of exogenous Ca(2+), and the enzyme was considerably stable even after incubation at 90 C for 2 hours. The thermostability at 90 and 102 C was enhanced in the presence of 5 mM Ca(2+). The extraordinarily high specific activity (about 7.4 x 10(exp 3) U/mg protein at 90 C, pH 5.5 with soluble starch as substrate) together with its low pH optimum makes this enzyme an interesting candidate for starch processing applications.

  5. AFV1, a novel virus infecting hyperthermophilic archaea of the genus acidianus

    International Nuclear Information System (INIS)

    Bettstetter, Marcus; Peng Xu; Garrett, Roger A.; Prangishvili, David

    2003-01-01

    We describe a novel virus, AFV1, of the hyperthermophilic archaeal genus Acidianus. Filamentous virions are covered with a lipid envelope and contain at least five different proteins with molecular masses in the range of 23-130 kDa and a 20.8-kb-long linear double-stranded DNA. The virus has been assigned to the family Lipothrixviridae on the basis of morphotypic characteristics. Host range is confined to several strains of Acidianus and the virus persists in its hosts in a stable carrier state. The latent period of virus infection is about 4 h. Viral DNA was sequenced and sequence similarities were found to the lipothrixvirus SIFV, the rudiviruses SIRV1 and SIRV2, as well as to conjugative plasmids and chromosomes of the genus Sulfolobus. Exceptionally for the linear genomes of archaeal viruses, many short direct repeats, with the sequence TTGTT or close variants thereof, are closely clustered over 300 bp at each end of the genome. They are reminiscent of the telomeric ends of linear eukaryal chromosomes

  6. Survival of thermophilic and hyperthermophilic microorganisms after exposure to UV-C, ionizing radiation and desiccation.

    Science.gov (United States)

    Beblo, Kristina; Douki, Thierry; Schmalz, Gottfried; Rachel, Reinhard; Wirth, Reinhard; Huber, Harald; Reitz, Günther; Rettberg, Petra

    2011-11-01

    In this study, we investigated the ability of several (hyper-) thermophilic Archaea and phylogenetically deep-branching thermophilic Bacteria to survive high fluences of monochromatic UV-C (254 nm) and high doses of ionizing radiation, respectively. Nine out of fourteen tested microorganisms showed a surprisingly high tolerance against ionizing radiation, and two species (Aquifex pyrophilus and Ignicoccus hospitalis) were even able to survive 20 kGy. Therefore, these species had a comparable survivability after exposure to ionizing radiation such as Deinococcus radiodurans. In contrast, there was nearly no difference in survival of the tested strains after exposure to UV-C under anoxic conditions. If the cells had been dried in advance of UV-C irradiation, they were more sensitive to UV-C radiation compared with cells irradiated in liquid suspension; this effect could be reversed by the addition of protective material like sulfidic ores before irradiation. By exposure to UV-C, photoproducts were formed in the DNA of irradiated Archaea and Bacteria. The distribution of the main photoproducts was species specific, but the amount of the photoproducts was only partly dependent on the applied fluence. Overall, our results show that tolerance to radiation seems to be a common phenomenon among thermophilic and hyperthermophilic microorganisms.

  7. The complete genome sequence of hyperthermophile Dictyoglomus turgidum DSM 6724™ reveals a specialized carbohydrate fermentor

    Directory of Open Access Journals (Sweden)

    Phillip Brumm

    2016-12-01

    Full Text Available Here we report the complete genome sequence of the chemoorganotrophic, extremely thermophilic bacterium, Dictyoglomus turgidum, which is a Gram negative, strictly anaerobic bacterium. D. turgidum and D. thermophilum together form the Dictyoglomi phylum. The two Dictyoglomus genomes are highly syntenic, and both are distantly related to Caldicellulosiruptor spp. D. turgidum is able to grow on a wide variety of polysaccharide substrates due to significant genomic commitment to glycosyl hydrolases, sixteen of which were cloned and expressed in our study. The GH5, GH10 and GH42 enzymes characterized in this study suggest that D. turgidum can utilize most plant-based polysaccharides except crystalline cellulose. The DNA polymerase I enzyme was also expressed and characterized. The pure enzyme showed improved amplification of long PCR targets compared to Taq polymerase. The genome contains a full complement of DNA modifying enzymes, and an unusually high copy number (4 of a new, ancestral family of polB type nucleotidyltransferases designated as MNT (minimal nucleotidyltransferases. Considering its optimal growth at 72ºC, D. turgidum has an anomalously low G+C content of 39.9% that may account for the presence of reverse gyrase, usually associated with hyperthermophiles.

  8. Activation and thermostabilization effects of cyclic 2, 3-diphosphoglycerate on enzymes from the hyperthermophilic Methanopyrus kandleri.

    Science.gov (United States)

    Shima, S; Hérault, D A; Berkessel, A; Thauer, R K

    1998-11-01

    Enzymes involved in methane formation from carbon dioxide and dihydrogen in Methanopyrus kandleri require high concentrations (> 1 M) of lyotropic salts such as K2HPO4/KH2PO4 or (NH4)2SO4 for activity and for thermostability. The requirement correlates with high intracellular concentrations of cyclic 2,3-diphosphoglycerate (cDPG; approximately 1 M) in this hyperthermophilic organism. We report here on the effects of potassium cDPG on the activity and thermostability of the two methanogenic enzymes cyclohydrolase and formyltransferase and show that at cDPG concentrations prevailing in the cells the investigated enzymes are highly active and completely thermostable. At molar concentrations also the potassium salts of phosphate and of 2,3-bisphosphoglycerate, the biosynthetic precursor of cDPG, were found to confer activity and thermostability to the enzymes. Thermodynamic arguments are discussed as to why cDPG, rather than these salts, is present in high concentrations in the cells of Mp. kandleri.

  9. Membrane homeoviscous adaptation in the piezo-hyperthermophilic archaeon Thermococcus barophilus

    Directory of Open Access Journals (Sweden)

    Anaïs eCario

    2015-10-01

    Full Text Available The archaeon Thermococcus barophilus, one of the most extreme members of hyperthermophilic piezophiles known thus far, is able to grow at temperatures up to 103°C and pressures up to 80MPa. We analyzed the membrane lipids of T. barophilus by HPLC-MS as a function of pressure and temperature. In contrast to previous reports, we show that under optimal growth conditions (40 MPa, 85°C the membrane spanning tetraether lipid GDGT-0 (sometimes called caldarchaeol is a major membrane lipid of T. barophilus together with archaeol. Increasing pressure and decreasing temperature lead to an increase of the proportion of archaeol and, reversely, a higher proportion of GDGT-0 is observed under low pressure and high temperature conditions. Noticeably, pressure and temperature fluctuations also impact the level of unsaturation of non-polar lipids with an irregular polyisoprenoid carbon skeleton (polyunsaturated lycopane derivatives, suggesting a structural role for these neutral lipids in the membrane of T. barophilus. Whether these apolar lipids insert in the membrane or not remains to be addressed. However, our results raise questions about the structure of the membrane in this archaeon and other archaeon harboring a mixture of di- and tetraether lipids.

  10. Pyrobaculum calidifontis sp. nov., a novel hyperthermophilic archaeon that grows in atmospheric air

    Directory of Open Access Journals (Sweden)

    Taku Amo

    2002-01-01

    Full Text Available A novel, facultatively aerobic, heterotrophic hyperthermophilic archaeon was isolated from a terrestrial hot spring in the Philippines. Cells of the new isolate, strain VA1, were rod-shaped with a length of 1.5 to 10 μm and a width of 0.5 to 1.0 μm. Isolate VA1 grew optimally at 90 to 95 °C and pH 7.0 under atmospheric air. Oxygen served as a final electron acceptor under aerobic growth conditions, and vigorous shaking of the medium significantly enhanced growth. Elemental sulfur inhibited cell growth under aerobic growth conditions, whereas thiosulfate stimulated cell growth. Under anaerobic growth conditions, nitrate served as a final electron acceptor, but nitrite or sulfur-containing compounds such as elemental sulfur, thiosulfate, sulfate and sulfite could not act as final electron acceptors. The G+C content of the genomic DNA was 51 mol%. Phylogenetic analysis based on 16S rRNA sequences indicated that strain VA1 exhibited close relationships to species of the genus Pyrobaculum. A DNA–DNA hybridization study revealed a low level of similarity (≤ 18% between strain VA1 and previously described members of the genus Pyrobaculum. Physiological characteristics also indicated that strain VA1 was distinct from these Pyrobaculum species. Our results indicate that isolate VA1 represents a novel species, named Pyrobaculum calidifontis.

  11. ORF Alignment: NC_000853 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ... Proteomics Target Tm0979_1_87; Northeast Structural ... Genomics Target Vt98. pdb|1X9A|A Chain... ... Thermotoga Maritima. Ontario Center For Structural ... Proteomics Target Tm0979_1_87; Northeast Stru

  12. Differences between the rhizosphere microbiome of Beta vulgaris ssp. maritima - ancestor of all beet crops - and modern sugar beets

    Directory of Open Access Journals (Sweden)

    Christin eZachow

    2014-08-01

    Full Text Available The structure and function of the plant microbiome is driven by plant species and prevailing environmental conditions. Effectuated by breeding efforts, modern crops diverge genetically and phenotypically from their wild relatives but little is known about consequences for the associated microbiota. Therefore, we studied bacterial rhizosphere communities associated with the wild beet B. vulgaris ssp. maritima grown in their natural habitat soil from coastal drift lines (CS and modern sugar beets (Beta vulgaris ssp. vulgaris cultivated in CS and potting soil (PS under greenhouse conditions. Analysis of 16S rRNA gene fingerprints and pyrosequencing-based amplicon libraries revealed plant genotype- and soil-specific microbiomes. Wild beet plants harbor distinct operational taxonomic units (OTUs and a more diverse bacterial community than the domesticated sugar beet plants. Although the rhizospheres of both plant genotypes were dominated by Proteobacteria and Planctomycetes, 47.4% of dominant OTUs were additionally detected in the wild beet rhizosphere. Analysis of the cultivable fraction confirmed these plant genotype-specific differences at functional level. The proportion of isolates displayed in vitro activity against phytopathogens was lower for wild beet (≤45.8% than for sugar beet (≤57.5%. Conversely, active isolates from the wild beet exhibited stronger ability to cope with abiotic stresses. From all samples, active isolates of Stenotrophomonas rhizophila were frequently identified. In addition, soil type-specific impacts on the composition of bacterial communities were found: Acidobacteria, Chloroflexi, and Planctomycetes were only detected in plants cultivated in CS; whereas Bacteroidetes and Proteobacteria dominated in PS. Overall, in comparison to modern sugar beets, wild beets were associated with taxonomically and functionally distinct microbiomes.

  13. Structural and genomic properties of the hyperthermophilic archaeal virus ATV with an extracellular stage of the reproductive cycle

    DEFF Research Database (Denmark)

    Prangishvili, David; Vestergaard, Gisle Alberg; Häring, Monika

    2006-01-01

    A novel virus, ATV, of the hyperthermophilic archaeal genus Acidianus has the unique property of undergoing a major morphological development outside of, and independently of, the host cell. Virions are extruded from host cells as lemon-shaped tail-less particles, after which they develop long...... tails at each pointed end, at temperatures close to that of the natural habitat, 85 degrees C. The extracellularly developed tails constitute tubes, which terminate in an anchor-like structure that is not observed in the tail-less particles. A thin filament is located within the tube, which exhibits...... can be interrupted by different stress factors....

  14. A Novel Process Configuration for Anaerobic Digestion of Source-Sorted Household Waste Using Hyper-Thermophilic Post-Treatment

    DEFF Research Database (Denmark)

    Hartmann, H.; Ahring, Birgitte Kiær

    2005-01-01

    A novel reactor configuration was investigated for anaerobic digestion (AD) of the organic fraction of municipal solid waste (OFMSW). An anaerobic hyper-thermophilic (68°C) reactor R68 was implemented as a post–treatment step for the effluent of a thermophilic reactor R1 (55°C) in order to enhance...... hydrolysis of recalcitrant organic matter, improve sanitation and ease the stripping of ammonia from the reactor. The efficiency of the combined system was studied in terms of methane yield, volatile solids (VS) reduction and volatile fatty acid (VFA) production at different hydraulic retention times (HRT...

  15. Molecular Studies of Filamentous and Biofilm-Forming Hyperthermophilic Communities in Yellowstone National Park

    Science.gov (United States)

    Summons, R. E.; Meyer-Dombard, D. R.; Bradley, A. S.; Dibbell, A. K.; Fredricks, H. F.; Hinrichs, K.; Jahnke, L. L.; Shock, E.; Amend, J. P.

    2005-12-01

    The Aquificales, the most deeply-branching order of Bacteria in the phylogenetic tree of life, comprises eight recognized thermophilic genera, including Aquifex, Hydrogenobacter, and Thermocrinis. The common metabolism for these Bacteria, when grown in culture, is the oxidation of hydrogen with molecular oxygen (Knallgas reaction). Aquificales have been identified by molecular techniques (16S rRNA gene surveys, fluorescent in situ hybridization) in Yellowstone National Park (YNP), sea vent chimneys and fluids, and many other terrestrial and marine locations. In situ, Aquificales can reside as biofilms on vent sinters but they also commonly form filamentous communities, otherwise known as pink streamers, which attach to solid substrates. Initial 16S rRNA gene surveys conducted on streamer communities from Octopus Spring YNP indicated that these were low diversity ecosystems dominated by a few phylotypes including Thermocrinis sp., Thermotoga sp. and one other bacterial clade (Reysenbach et al 1994). Archaea were notable for their absence. In one of the first geobiological studies of pink streamers and vent biofilms in Yellowstone National Park, Jahnke and coworkers (2001) used classical lipidological techniques to compare Aquificales cultures with environmental samples to show that YNP pink filaments were more phylogenetically diverse and physiologically more complex than the early genomic studies indicated. The presence of archaeol, the range and structures of other lipids and a wide dispersion in the carbon isotopic signatures of biomass and individual lipids (-15 to -27%) showed that Archaea were present in pink filament communities and that there was, at least, one additional bacterial group besides the dominant Aquificales component. New molecular studies that comprise analyses of 16S rRNA genes and total lipid extracts by liquid chromatography and mass spectrometry and chemical degradation with gas chromatography and mass spectrometry now show that Crenarchaea

  16. A proposal to rename the hyperthermophile Pyrococcus woesei as Pyrococcus furiosus subsp. woesei

    Directory of Open Access Journals (Sweden)

    Wirojne Kanoksilapatham

    2004-01-01

    Full Text Available Pyrococcus species are hyperthermophilic members of the order Thermococcales, with optimal growth temperatures approaching 100 °C. All species grow heterotrophically and produce H2 or, in the presence of elemental sulfur (S°, H2S. Pyrococcus woesei and P. furiosus were isolated from marine sediments at the same Vulcano Island beach site and share many morphological and physiological characteristics. We report here that the rDNA operons of these strains have identical sequences, including their intergenic spacer regions and part of the 23S rRNA. Both species grow rapidly and produce H2 in the presence of 0.1% maltose and 10–100 µM sodium tungstate in S°-free medium. However,P. woesei shows more extensive autolysis than P. furiosus in the stationary phase. Pyrococcusfuriosus and P. woesei share three closely related families of insertion sequences (ISs. A Southern blot performed with IS probes showed extensive colinearity between the genomes of P. woesei and P. furiosus. Cloning and sequencing of ISs that were in different contexts in P. woesei and P. furiosus revealed that the napA gene in P. woesei is disrupted by a type III IS element, whereas in P. furiosus, this gene is intact. A type I IS element, closely linked to the napA gene, was observed in the same context in both P. furiosus and P. woesei genomes. Our results suggest that the IS elements are implicated in genomic rearrangements and reshuffling in these closely related strains. We propose to rename P. woesei a subspecies of P. furiosus based on their identical rDNA operon sequences, many common IS elements that are shared genomic markers, and the observation that all P. woesei nucleotide sequences deposited in GenBank to date are > 99% identical to P. furiosus sequences.

  17. A proposal to rename the hyperthermophile Pyrococcus woesei as Pyrococcus furiosus subsp. woesei.

    Science.gov (United States)

    Kanoksilapatham, Wirojne; González, Juan M; Maeder, Dennis L; DiRuggiero, Jocelyne; Robb, Frank T

    2004-10-01

    Pyrococcus species are hyperthermophilic members of the order Thermococcales, with optimal growth temperatures approaching 100 degrees C. All species grow heterotrophically and produce H2 or, in the presence of elemental sulfur (S(o)), H2S. Pyrococcus woesei and P. furiosus were isolated from marine sediments at the same Vulcano Island beach site and share many morphological and physiological characteristics. We report here that the rDNA operons of these strains have identical sequences, including their intergenic spacer regions and part of the 23S rRNA. Both species grow rapidly and produce H2 in the presence of 0.1% maltose and 10-100 microM sodium tungstate in S(o)-free medium. However, P. woesei shows more extensive autolysis than P. furiosus in the stationary phase. Pyrococcus furiosus and P. woesei share three closely related families of insertion sequences (ISs). A Southern blot performed with IS probes showed extensive colinearity between the genomes of P. woesei and P. furiosus. Cloning and sequencing of ISs that were in different contexts in P. woesei and P. furiosus revealed that the napA gene in P. woesei is disrupted by a type III IS element, whereas in P. furiosus, this gene is intact. A type I IS element, closely linked to the napA gene, was observed in the same context in both P. furiosus and P. woesei genomes. Our results suggest that the IS elements are implicated in genomic rearrangements and reshuffling in these closely related strains. We propose to rename P. woesei a subspecies of P. furiosus based on their identical rDNA operon sequences, many common IS elements that are shared genomic markers, and the observation that all P. woesei nucleotide sequences deposited in GenBank to date are > 99% identical to P. furiosus sequences.

  18. Aerobic lineage of the oxidative stress response protein rubrerythrin emerged in an ancient microaerobic, (hyperthermophilic environment

    Directory of Open Access Journals (Sweden)

    Juan Pablo Cardenas

    2016-11-01

    Full Text Available Rubrerythrins (RBRs are non-heme di-iron proteins belonging to the ferritin-like superfamily (FLSF. They are involved in oxidative stress defense as peroxide scavengers in a wide range of organisms. The vast majority of RBRs, including classical forms of this protein, contain a C-terminal rubredoxin-like domain involved in electron transport that is used during catalysis in anaerobic conditions. Rubredoxin is an ancient and large protein family of short length (<100 residues that contains a Fe-S center involved in electron transfer. However, functional forms of the enzyme lacking the rubredoxin-like domain have been reported (e.g., sulerythrin and ferriperoxin. In this study, phylogenomic evidence is presented that suggests that a complete lineage of rubrerythrins, lacking the rubredoxin-like domain, arose in an ancient microaerobic and (hyperthermophilic environments in the ancestors of the Archaea Thermoproteales and Sulfolobales. This lineage (termed the aerobic-type lineage subsequently evolved to become adapted to environments with progressively lower temperatures and higher oxygen concentrations via the acquisition of two co-localized genes, termed DUF3501 and RFO, encoding a conserved protein of unknown function and a predicted Fe-S oxidoreductase respectively. Proposed Horizontal Gene Transfer (HGT events from these archaeal ancestors to Bacteria expanded the opportunities for further evolution of this RBR including adaption to lower temperatures. The second lineage (termed the cyanobacterial lineage is proposed to have evolved in cyanobacterial ancestors, maybe in direct response to the production of oxygen via oxygenic photosynthesis during the Great Oxygen Event (GOE. It is hypothesized that both lineages of RBR emerged in a largely anaerobic world with whiffs of oxygen and that their subsequent independent evolutionary trajectories allowed microorganisms to transition from this anaerobic world to an aerobic one.

  19. Heterologous Production of an Energy-Conserving Carbon Monoxide Dehydrogenase Complex in the Hyperthermophile Pyrococcus furiosus

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    Gerrit Jan Schut

    2016-01-01

    Full Text Available Carbon monoxide (CO is an important intermediate in anaerobic carbon fixation pathways in acetogenesis and methanogenesis. In addition, some anaerobes can utilize CO as an energy source. In the hyperthermophilic archaeon Thermococcus onnurineus, which grows optimally at 80°C, CO oxidation and energy conservation is accomplished by a respiratory complex encoded by a 16-gene cluster containing a carbon monoxide dehydrogenase, a membrane-bound [NiFe]-hydrogenase and a Na+/H+ antiporter module. This complex oxidizes CO, evolves CO2 and H2, and generates a Na+ motive force that is used to conserve energy by a Na+-dependent ATP synthase. Herein we used a bacterial artificial chromosome to insert the 13.2 kb gene cluster encoding the CO-oxidizing respiratory complex of T. onnurineus into the genome of the heterotrophic archaeon, Pyrococcus furiosus, which grows optimally at 100°C. P. furiosus is normally unable to utilize CO, however, the recombinant strain readily oxidized CO and generated H2 at 80°C. Moreover, CO also served as an energy source and allowed the P. furiosus strain to grow with a limiting concentration of sugar or with peptides as the carbon source. Moreover, CO oxidation by P. furiosus was also coupled to the re-utilization, presumably for biosynthesis, of acetate generated by fermentation. The functional transfer of CO utilization between Thermococcus and Pyrococcus species demonstrated herein is representative of the horizontal gene transfer of an environmentally-relevant metabolic capability. The transfer of CO utilizing, hydrogen-producing genetic modules also has applications for biohydrogen production and a CO-based industrial platform for various thermophilic organisms.

  20. Continuous Hydrogen Production from Agricultural Wastewaters at Thermophilic and Hyperthermophilic Temperatures.

    Science.gov (United States)

    Ramos, Lucas Rodrigues; Silva, Edson Luiz

    2017-06-01

    The objective of this study was to investigate the effects of hydraulic retention time (HRT) (8 to 0.5 h) and temperature (55 to 75 °C) in two anaerobic fluidized bed reactors (AFBR) using cheese whey (AFBR-CW = 10,000 mg sugars L -1 ) and vinasse (AFBR-V = 10,000 mg COD L -1 ) as substrates. Decreasing the HRT to 0.5 h increased the hydrogen production rates in both reactors, with maximum values of 5.36 ± 0.81 L H 2 h -1 L -1 in AFBR-CW and 0.71 ± 0.16 L H 2 h -1 L -1 in AFBR-V. The optimal conditions for hydrogen production were the HRT of 4 h and temperature of 65 °C in AFBR-CW, observing maximum hydrogen yield (HY) of 5.51 ± 0.37 mmol H 2 g COD -1 . Still, the maximum HY in AFBR-V was 1.64 ± 0.22 mmol H 2 g COD -1 at 4 h and 55 °C. However, increasing the temperature to 75 °C reduced the hydrogen production in both reactors. Methanol and butyric, acetic, and lactic acids were the main metabolites at temperatures of 55 and 65 °C, favoring the butyric and acetic metabolic pathways of hydrogen production. The increased productions of lactate, propionate, and methanol at 75 °C indicate that the hydrogen-producing bacteria in the thermophilic inoculum were inhibited under hyperthermophilic conditions.

  1. Overexpression, purification and crystallization of tyrosyl-tRNA synthetase from the hyperthermophilic archaeon Aeropyrum pernix K1

    International Nuclear Information System (INIS)

    Iwaki, Jun; Suzuki, Ryuichiro; Fujimoto, Zui; Momma, Mitsuru; Kuno, Atsushi; Hasegawa, Tsunemi

    2005-01-01

    Tyrosyl-tRNA synthetase from the hyperthermophilic archaeon A. pernix K1 was cloned, purified and crystallized. The crystals belonged to the tetragonal space group P4 3 2 1 2, with unit-cell parameters a = b = 66.1, c = 196.2 Å, and diffracted to beyond 2.15 Å resolution at 100 K. Hyperthermophilic archaeal tyrosyl-tRNA synthetase from Aeropyrum pernix K1 was cloned and overexpressed in Escherichia coli. The expressed protein was purified by Cibacron Blue affinity chromatography following heat treatment at 363 K. Crystals suitable for X-ray diffraction studies were obtained under optimized crystallization conditions in the presence of 1.5 M ammonium sulfate using the hanging-drop vapour-diffusion method. The crystals belonged to the tetragonal space group P4 3 2 1 2, with unit-cell parameters a = b = 66.1, c = 196.2 Å, and diffracted to beyond 2.15 Å resolution at 100 K

  2. Bio-hydrolysis and bio-hydrogen production from food waste by thermophilic and hyperthermophilic anaerobic process.

    Science.gov (United States)

    Algapani, Dalal E; Qiao, Wei; Su, Min; di Pumpo, Francesca; Wandera, Simon M; Adani, Fabrizio; Dong, Renjie

    2016-09-01

    High-temperature pretreatment plays a key role in the anaerobic digestion of food waste (FW). However, the suitable temperature is not yet determined. In this work, a long-term experiment was conducted to compare hydrolysis, acidogenesis, acetogenesis, and hydrogen production at 55°C and 70°C, using real FW in CSTR reactors. The results obtained indicated that acidification was the rate-limiting step at both temperatures with similar process kinetics characterizations. However, the thermophilic pretreatment was more advantageous than the hyperthermophilic with suspended solids solubilization of 47.7% and 29.5% and total VFA vs. soluble COD ratio of 15.2% and 4.9%, for thermophilic and hyperthermophilic treatment, respectively, with a hydrolytic reaction time (HRT) of 10days and an OLR of 14kgCOD/m(3)d. Moreover, stable hydrogen yield (70.7ml-H2/gVSin) and content in off gas (58.6%) was achieved at HRT 5days, pH 5.5, and temperature of 55°C, as opposed to 70°C. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. An Integrative Genomic Island Affects the Adaptations of Piezophilic Hyperthermophilic Archaeon Pyrococcus yayanosii to High Temperature and High Hydrostatic Pressure

    Directory of Open Access Journals (Sweden)

    Zhen Li

    2016-11-01

    Full Text Available Deep-sea hydrothermal vent environments are characterized by high hydrostatic pressure and sharp temperature and chemical gradients. Horizontal gene transfer is thought to play an important role in the microbial adaptation to such an extreme environment. In this study, a 21.4-kb DNA fragment was identified as a genomic island, designated PYG1, in the genomic sequence of the piezophilic hyperthermophile Pyrococcus yayanosii. According to the sequence alignment and functional annotation, the genes in PYG1 could tentatively be divided into five modules, with functions related to mobility, DNA repair, metabolic processes and the toxin-antitoxin system. Integrase can mediate the site-specific integration and excision of PYG1 in the chromosome of P. yayanosii A1. Gene replacement of PYG1 with a SimR cassette was successful. The growth of the mutant strain ∆PYG1 was compared with its parent strain P. yayanosii A2 under various stress conditions, including different pH, salinity, temperature and hydrostatic pressure. The ∆PYG1 mutant strain showed reduced growth when grown at 100 °C, while the biomass of ∆PYG1 increased significantly when cultured at 80 MPa. Differential expression of the genes in module Ⅲ of PYG1 was observed under different temperature and pressure conditions. This study demonstrates the first example of an archaeal integrative genomic island that could affect the adaptation of the hyperthermophilic piezophile P. yayanosii to high temperature and high hydrostatic pressure.

  4. Microbiological evidence for Fe(III) reduction on early Earth

    Science.gov (United States)

    Vargas, Madeline; Kashefi, Kazem; Blunt-Harris, Elizabeth L.; Lovley, Derek R.

    1998-09-01

    It is generally considered that sulphur reduction was one of the earliest forms of microbial respiration, because the known microorganisms that are most closely related to the last common ancestor of modern life are primarily anaerobic, sulphur-reducing hyperthermophiles. However, geochemical evidence indicates that Fe(III) is more likely than sulphur to have been the first external electron acceptor of global significance in microbial metabolism. Here we show that Archaea and Bacteria that are most closely related to the last common ancestor can reduce Fe(III) to Fe(II) and conserve energy to support growth from this respiration. Surprisingly, even Thermotoga maritima, previously considered to have only a fermentative metabolism, could grow as a respiratory organism when Fe(III) was provided as an electron acceptor. These results provide microbiological evidence that Fe(III) reduction could have been an important process on early Earth and suggest that microorganisms might contribute to Fe(III) reduction in modern hot biospheres. Furthermore, our discovery that hyperthermophiles that had previously been thought to require sulphur for cultivation can instead be grown without the production of toxic and corrosive sulphide, should aid biochemical investigations of these poorly understood organisms.

  5. The 60 kDa heat shock proteins in the hyperthermophilic archaeon Sulfolobus shibatae.

    Science.gov (United States)

    Kagawa, H K; Osipiuk, J; Maltsev, N; Overbeek, R; Quaite-Randall, E; Joachimiak, A; Trent, J D

    1995-11-10

    One of the most abundant proteins in the hyperthermophilic archaeon Sulfolobus shibatae is the 59 kDa heat shock protein (TF55) that is believed to form a homo-oligomeric double ring complex structurally similar to the bacterial chaperonins. We discovered a second protein subunit in the S. shibatae ring complex (referred to as alpha) that is stoichiometric with TF55 (renamed beta). The gene and flanking regions of alpha were cloned and sequenced and its inferred amino acid sequence has 54.4% identity and 74.4% similarity to beta. Transcription start sites for both alpha and beta were mapped and three potential transcription regulatory regions were identified. Northern analyses of cultures shifted from normal growth temperatures (70 to 75 degrees C) to heat shock temperatures (85 to 90 degrees C) indicated that the levels of alpha and beta mRNAs increased during heat shock, but at all temperatures their relative proportions remained constant. Monitoring protein synthesis by autoradiography of total proteins from cultures pulse labeled with L(-)[35S]methionine at normal and heat shock temperatures indicated significant increases in alpha and beta synthesis during heat shock. Under extreme heat shock conditions (> or = 90 degrees C) alpha and beta appeared to be the only two proteins synthesized. The purified alpha and beta subunits combined to form high molecular mass complexes with similar mobilities on native polyacrylamide gels to the complexes isolated directly from cells. Equal proportions of the two subunits gave the greatest yield of the complex, which we refer to as a "rosettasome". It is argued that the rosettasome consists of two homo-oligomeric rings; one of alpha and the other of beta. Polyclonal antibodies against alpha and beta from S. shibatae cross-reacted with proteins of similar molecular mass in 10 out of the 17 archaeal species tested, suggesting that the two rosettasome proteins are highly conserved among the archaea. The archaeal sequences were

  6. Thermococcus prieurii sp. nov., a hyperthermophilic archaeon isolated from a deep-sea hydrothermal vent.

    Science.gov (United States)

    Gorlas, Aurore; Alain, Karine; Bienvenu, Nadège; Geslin, Claire

    2013-08-01

    A novel hyperthermophilic, anaerobic archaeon, strain Bio-pl-0405IT2(T), was isolated from a hydrothermal chimney sample collected from the East Pacific Rise at 2700 m depth in the 'Sarah Spring' area (7° 25' 24" S 107° 47' 66" W). Cells were irregular, motile cocci (0.8-1.5 µm in diameter) and divided by constriction. Growth was observed at temperatures between 60 °C and 95 °C with an optimum at 80 °C. The pH range for growth was between pH 4.0 and pH 8.0 with an optimum around pH 7.0. Strain Bio-pl-0405IT2(T) grew at salt concentrations of 1-5 % (w/v) NaCl with an optimum at 2 %. The novel isolate grew by fermentation or sulphur respiration on a variety of organic compounds. It was a chemoorganoheterotrophic archaeon growing preferentially with yeast extract, peptone and tryptone as carbon and energy sources and sulphur and organic compounds as electron acceptors; it also grew on maltose and starch. Sulphur or l-cystine were required for growth and were reduced to hydrogen sulfide. The strain was resistant to rifampicin, chloramphenicol, vancomycin and kanamycin (all at 100 µg ml(-1)) but was sensitive to tetracycline. The G+C content of its genomic DNA was 53.6 mol%. Phylogenetic analysis of the almost complete 16S rRNA gene sequence (1450 bp) of strain Bio-pl-0405IT2(T) showed that the novel isolate belonged to the genus Thermococcus. DNA-DNA hybridization values with the two closest relatives Thermococcus hydrothermalis AL662(T) and Thermococcus celer JCM 8558(T) were below the threshold value of 70 %. On the basis of the physiological and genotypic distinctness, we propose a novel species, Thermococcus prieurii sp. nov. The type strain is Bio-pl-0405IT2(T) ( = CSUR P577(T)= JCM 16307(T)).

  7. Expression, purification, crystallization and preliminary X-ray analysis of a nucleoside kinase from the hyperthermophile Methanocaldococcus jannaschii

    International Nuclear Information System (INIS)

    Arnfors, Linda; Hansen, Thomas; Meining, Winfried; Schönheit, Peter; Ladenstein, Rudolf

    2005-01-01

    Nucleoside kinase from the hyperthermophilic archaeon M. jannaschii is a member of the PFK-B family which belongs to the ribokinase superfamily. Here, its expression, purification, crystallization and preliminary X-ray analysis are described. Methanocaldococcus jannaschii nucleoside kinase (MjNK) is an ATP-dependent non-allosteric phosphotransferase that shows high catalytic activity for guanosine, inosine and cytidine. MjNK is a member of the phosphofructokinase B family, but participates in the biosynthesis of nucleoside monophosphates rather than in glycolysis. MjNK was crystallized as the apoenzyme as well as in complex with an ATP analogue and Mg 2+ . The latter crystal form was also soaked with fructose-6-phosphate. Synchrotron-radiation data were collected to 1.70 Å for the apoenzyme crystals and 1.93 Å for the complex crystals. All crystals exhibit orthorhombic symmetry; however, the apoenzyme crystals contain one monomer per asymmetric unit whereas the complex crystals contain a dimer

  8. A synthetic arabinose-inducible promoter confers high levels of recombinant protein expression in hyperthermophilic archaeon Sulfolobus islandicus

    DEFF Research Database (Denmark)

    Peng, Nan; Deng, Ling; Mei, Yuxia

    2012-01-01

    Despite major progresses in genetic studies of hyperthermophilic archaea, recombinant protein production in these organisms always suffers from low yields and a robust expression system is still in great demand. Here we report a versatile vector that confers high levels of protein expression...... to remove the peptide tags from expressed recombinant proteins. While pEXA employed an araS promoter for protein expression, pSeSD utilized P(araS-SD), an araS derivative promoter carrying an engineered ribosome-binding site (RBS; a Shine-Dalgarno [SD] sequence). We found that P(araS-SD) directed high...... levels of target gene expression. More strikingly, N-terminal amino acid sequencing of recombinant proteins unraveled that the protein synthesized from pEXA-N-lacS lacked the designed 6×His tag and that translation initiation did not start at the ATG codon of the fusion gene. Instead, it started...

  9. Metagenomics of Kamchatkan hot spring filaments reveal two new major (hyper)thermophilic lineages related to Thaumarchaeota.

    Science.gov (United States)

    Eme, Laura; Reigstad, Laila J; Spang, Anja; Lanzén, Anders; Weinmaier, Thomas; Rattei, Thomas; Schleper, Christa; Brochier-Armanet, Céline

    2013-06-01

    Based on phylogenetic analyses and gene distribution patterns of a few complete genomes, a new distinct phylum within the Archaea, the Thaumarchaeota, has recently been proposed. Here we present analyses of six archaeal fosmid sequences derived from a microbial hot spring community in Kamchatka. The phylogenetic analysis of informational components (ribosomal RNAs and proteins) reveals two major (hyper-)thermophilic clades ("Hot Thaumarchaeota-related Clade" 1 and 2, HTC1 and HTC2) related to Thaumarchaeota, representing either deep branches of this phylum or a new archaeal phylum and provides information regarding the ancient evolution of Archaea and their evolutionary links with Eukaryotes. Copyright © 2013 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  10. A ocorrência do mutualismo facultativo entre Dyckia maritima Backer (Bromeliaceae e o cupim Cortaritermes silvestrii (Holmgren, Nasutitermitinae, em afloramentos rochosos no Parque Estadual de Itapuã, Viamão, RS The occurrence of facultative mutualism between Dyckia maritima Backer (Bromeliaceae and the termite Cortaritermes silvestrii (Holmgren, Nasutitermitinae, on rock outcrops in Itapuã State Park, Viamão, RS

    Directory of Open Access Journals (Sweden)

    Celso Copstein Waldemar

    2003-03-01

    Full Text Available A presença de colônias de C. silvestrii é comum nos lajeados existentes em Itapuã. Na estação Morro da Grota1, 92,0 % dos termiteiros situados na rocha exposta e em ilhas de vegetação estão associados a D. maritima. Esta convivência ocorre em 31,2 % das ilhas na qual esta bromélia se faz presente. Nas ilhas, a comparação entre os substratos aonde D. maritima vegeta, o solo litólico húmico existente sob o manto do musgo Campylopus spp. e o substrato constituído pelo cupinzeiro indica que este último possui os teores mais elevados dos nutrientes P, K, Ca, Mg, Zn e Mn, maior CTC e maiores teores de partículas finas, principalmente o silte. O estabelecimento de D. maritima sobre os termiteiros de grande porte aumenta o seu valor de cobertura em ilhas de vegetação quando comparado com ilhas sem termiteiros ou com termiteiros de pequeno porte em áreas entre 2,7 a 8,0 m². Este fato é atribuído à melhoria físico-química do substrato e ao aumento de superfície e volume aptos a serem colonizados pela bromélia e proporciona maior competitividade em relação a outras espécies vegetais. As características apresentadas pela interação entre este cupim e D. maritima, pela primeira vez descrita na literatura, permitem indicar esta relação ecológica como mutualismo facultativo. Inferimos que o conjunto de observações apresentado constitui um modelo temporal de crescimento deste mutualismo, cujas fases inicial e tardia estão descritas neste trabalho.The presence of colonies of C. silvestrii is common, both on the rock surface and at islands of vegetation. At Morro da Grota1 station, 92,0 % of the termite nests on rocky outcrops and at island of vegetation are associated with this bromeliad. These nests are associated with D. maritima, in 31,2 % of the islands where this bromeliad occurs. At these island communities, the comparison between the substrata where D. maritima occurs, the litolic Waldemar & Irgang: Mutualismo

  11. Deletion of the topoisomerase III gene in the hyperthermophilic archaeon Sulfolobus islandicus results in slow growth and defects in cell cycle control

    DEFF Research Database (Denmark)

    Li, Xiyang; Guo, Li; Deng, Ling

    2011-01-01

    Topoisomerase III (topo III), a type IA topoisomerase, is widespread in hyperthermophilic archaea. In order to interrogate the in vivo role of archaeal topo III, we constructed and characterized a topo III gene deletion mutant of Sulfolobus islandicus. The mutant was viable but grew more slowly...... results suggest that the enzyme may serve roles in chromosomal segregation and control of the level of supercoiling in the cell....

  12. Caractérisation de nouvelles molécules et variabilité chimique de trois plantes du continuum Corse-Sardaigne: Chamaemelum mixtum, Anthemis maritima et Eryngium maritimum.

    OpenAIRE

    Darriet , Florent

    2011-01-01

    Corsica is a Mediterranean mountain island which displays many aromatic and medicinal plants liable to produce essential oils potentially used in different domains. For their valorization, the knowledge of the oil chemical compositions is an essential challenge. This thesis deals with the chemical compositions of the essential oils and the volatile fractions of three coastal plants from Corsica and Sardinia: Chamaemelum mixtum, Anthemis maritima and Erygium maritimum. There are two aims: (i) ...

  13. Isolation and characterization of the first xylanolytic hyperthermophilic euryarchaeon Thermococcus sp. strain 2319x1 and its unusual multidomain glycosidase

    Directory of Open Access Journals (Sweden)

    Sergey N Gavrilov

    2016-05-01

    Full Text Available Enzymes from (hyperthermophiles Thermozymes offer a great potential for biotechnological applications. Thermophilic adaptation does not only provide stability towards high temperature but is also often accompanied by a higher resistance to other harsh physicochemical conditions, which are also frequently employed in industrial processes, such as the presence of e.g. denaturing agents as well as low or high pH of the medium. In order to find new thermostable, xylan degrading hydrolases with potential for biotechnological application we used an in situ enrichment strategy incubating Hungate tubes with xylan as the energy substrate in a hot vent located in the tidal zone of Kunashir Island (Kuril archipelago. Using this approach a hyperthermophilic euryarchaeon, designated Thermococcus sp. strain 2319x1, growing on xylan as sole energy and carbon source was isolated. The organism grows optimally at 85°C and pH 7.0 on a variety of natural polysaccharides including xylan, carboxymethyl cellulose (CMC, amorphous cellulose (AMC, xyloglucan, and chitin. The protein fraction extracted from the cells surface with Twin 80 exhibited endoxylanase, endoglucanase and xyloglucanase activities. The genome of Thermococcus sp. strain 2319x1 was sequenced and assembled into one circular chromosome. Within the newly sequenced genome, a gene, encoding a novel type of glycosidase (143 kDa with a unique five-domain structure, was identified. It consists of three glycoside hydrolase (GH domains and two carbohydrate-binding modules (CBM with the domain order GH5-12-12-CBM2-2 (N- to C-terminal direction. The full length protein, as well as truncated versions, were heterologously expressed in Escherichia coli and their activity was analyzed. The full length multidomain glycosidase (MDG was able to hydrolyze various polysaccharides, with the highest activity for barley β-glucan (β-1,3/1,4-glucoside, followed by that for carboxymethyl cellulose (β-1,4-glucoside

  14. Thermodesulfobacterium geofontis sp. nov., a hyperthermophilic, sulfate-reducing bacterium isolated from Obsidian Pool, Yellowstone National Park.

    Science.gov (United States)

    Hamilton-Brehm, Scott D; Gibson, Robert A; Green, Stefan J; Hopmans, Ellen C; Schouten, Stefan; van der Meer, Marcel T J; Shields, John P; Damsté, Jaap S S; Elkins, James G

    2013-03-01

    A novel sulfate-reducing bacterium designated OPF15(T) was isolated from Obsidian Pool, Yellowstone National Park, Wyoming. The phylogeny of 16S rRNA and functional genes (dsrAB) placed the organism within the family Thermodesulfobacteriaceae. The organism displayed hyperthermophilic temperature requirements for growth with a range of 70-90 °C and an optimum of 83 °C. Optimal pH was around 6.5-7.0 and the organism required the presence of H2 or formate as an electron donor and CO2 as a carbon source. Electron acceptors supporting growth included sulfate, thiosulfate, and elemental sulfur. Lactate, acetate, pyruvate, benzoate, oleic acid, and ethanol did not serve as electron donors. Membrane lipid analysis revealed diacyl glycerols and acyl/ether glycerols which ranged from C14:0 to C20:0. Alkyl chains present in acyl/ether and diether glycerol lipids ranged from C16:0 to C18:0. Straight, iso- and anteiso-configurations were found for all lipid types. The presence of OPF15(T) was also shown to increase cellulose consumption during co-cultivation with Caldicellulosiruptor obsidiansis, a fermentative, cellulolytic extreme thermophile isolated from the same environment. On the basis of phylogenetic, phenotypic, and structural analyses, Thermodesulfobacterium geofontis sp. nov. is proposed as a new species with OPF15(T) representing the type strain.

  15. Influence of C-terminal tail deletion on structure and stability of hyperthermophile Sulfolobus tokodaii RNase HI.

    Science.gov (United States)

    Chen, Lin; Zhang, Ji-Long; Zheng, Qing-Chuan; Chu, Wen-Ting; Xue, Qiao; Zhang, Hong-Xing; Sun, Chia-Chung

    2013-06-01

    The C-terminus tail (G144-T149) of the hyperthermophile Sulfolobus tokodaii (Sto-RNase HI) plays an important role in this protein's hyperstabilization and may therefore be a good protein stability tag. Detailed understanding of the structural and dynamic effects of C-terminus tail deletion is required for gaining insights into the thermal stability mechanism of Sto-RNase HI. Focused on Sulfolobus tokodaii RNase HI (Sto-RNase HI) and its derivative lacking the C-terminal tail (ΔC6 Sto-RNase HI) (PDB codes: 2EHG and 3ALY), we applied molecular dynamics (MD) simulations at four different temperatures (300, 375, 475, and 500 K) to examine the effect of the C-terminal tail on the hyperstabilization of Sto-RNase HI and to investigate the unfolding process of Sto-RNase HI and ΔC6 Sto-RNase HI. The simulations suggest that the C-terminal tail has significant impact in hyperstabilization of Sto-RNase HI and the unfolding of these two proteins evolves along dissimilar pathways. Essential dynamics analysis indicates that the essential subspaces of the two proteins at different temperatures are non-overlapping within the trajectories and they exhibit different directions of motion. Our work can give important information to understand the three-state folding mechanism of Sto-RNase HI and to offer alternative strategies to improve the protein stability.

  16. Improving the Thermostability and Optimal Temperature of a Lipase from the Hyperthermophilic Archaeon Pyrococcus furiosus by Covalent Immobilization

    Directory of Open Access Journals (Sweden)

    Roberta V. Branco

    2015-01-01

    Full Text Available A recombinant thermostable lipase (Pf2001Δ60 from the hyperthermophilic Archaeon Pyrococcus furiosus (PFUL was immobilized by hydrophobic interaction on octyl-agarose (octyl PFUL and by covalent bond on aldehyde activated-agarose in the presence of DTT at pH = 7.0 (one-point covalent attachment (glyoxyl-DTT PFUL and on glyoxyl-agarose at pH 10.2 (multipoint covalent attachment (glyoxyl PFUL. The enzyme’s properties, such as optimal temperature and pH, thermostability, and selectivity, were improved by covalent immobilization. The highest enzyme stability at 70°C for 48 h incubation was achieved for glyoxyl PFUL (around 82% of residual activity, whereas glyoxyl-DTT PFUL maintained around 69% activity, followed by octyl PFUL (27% remaining activity. Immobilization on glyoxyl-agarose improved the optimal temperature to 90°C, while the optimal temperature of octyl PFUL was 70°C. Also, very significant changes in activity with different substrates were found. In general, the covalent bond derivatives were more active than octyl PFUL. The E value also depended substantially on the derivative and the conditions used. It was observed that the reaction of glyoxyl-DTT PFUL using methyl mandelate as a substrate at pH 7 presented the best results for enantioselectivity E=22 and enantiomeric excess (ee (% = 91.

  17. Survival of thermophilic and hyper-thermophilic microorganisms after exposure to UV-C, ionizing radiation and desiccation

    International Nuclear Information System (INIS)

    Beblo, K.; Wirth, R.; Huber, H.; Douki, T.; Schmalz, G.; Rachel, R.

    2011-01-01

    In this study, we investigated the ability of several (hyper-) thermophilic Archaea and phylo-genetically deep-branching thermophilic Bacteria to survive high fluences of monochromatic UV-C (254 nm) and high doses of ionizing radiation, respectively. Nine out of fourteen tested microorganisms showed a surprisingly high tolerance against ionizing radiation, and two species (Aquifex pyrophilus and Ignicoccus hospitalis) were even able to survive 20 kGy. Therefore, these species had a comparable survivability after exposure to ionizing radiation such as Deinococcus radiodurans. In contrast, there was nearly no difference in survival of the tested strains after exposure to UV-C under anoxic conditions. If the cells had been dried in advance of UV-C irradiation, they were more sensitive to UV-C radiation compared with cells irradiated in liquid suspension; this effect could be reversed by the addition of protective material like sulfidic ores before irradiation. By exposure to UV-C, photoproducts were formed in the DNA of irradiated Archaea and Bacteria. The distribution of the main photoproducts was species specific, but the amount of the photoproducts was only partly dependent on the applied fluence. Overall, our results show that tolerance to radiation seems to be a common phenomenon among thermophilic and hyper-thermophilic microorganisms. (authors)

  18. Biochemical, transcriptional and translational evidences of the phenol-meta-degradation pathway by the hyperthermophilic Sulfolobus solfataricus 98/2.

    Directory of Open Access Journals (Sweden)

    Alexia Comte

    Full Text Available Phenol is a widespread pollutant and a model molecule to study the biodegradation of monoaromatic compounds. After a first oxidation step leading to catechol in mesophilic and thermophilic microorganisms, two main routes have been identified depending on the cleavage of the aromatic ring: ortho involving a catechol 1,2 dioxygenase (C12D and meta involving a catechol 2,3 dioxygenase (C23D. Our work aimed at elucidating the phenol-degradation pathway in the hyperthermophilic archaea Sulfolobus solfataricus 98/2. For this purpose, the strain was cultivated in a fermentor under different substrate and oxygenation conditions. Indeed, reducing dissolved-oxygen concentration allowed slowing down phenol catabolism (specific growth and phenol-consumption rates dropped 55% and 39%, respectively and thus, evidencing intermediate accumulations in the broth. HPLC/Diode Array Detector and LC-MS analyses on culture samples at low dissolved-oxygen concentration (DOC  =  0.06 mg x L(-1 suggested, apart for catechol, the presence of 2-hydroxymuconic acid, 4-oxalocrotonate and 4-hydroxy-2-oxovalerate, three intermediates of the meta route. RT-PCR analysis on oxygenase-coding genes of S. solfataricus 98/2 showed that the gene coding for the C23D was expressed only on phenol. In 2D-DIGE/MALDI-TOF analysis, the C23D was found and identified only on phenol. This set of results allowed us concluding that S. solfataricus 98/2 degrade phenol through the meta route.

  19. Stability of the 'L12 stalk' in ribosomes from mesophilic and (hyper)thermophilic Archaea and Bacteria.

    Science.gov (United States)

    Shcherbakov, D; Dontsova, M; Tribus, M; Garber, M; Piendl, W

    2006-01-01

    The ribosomal stalk complex, consisting of one molecule of L10 and four or six molecules of L12, is attached to 23S rRNA via protein L10. This complex forms the so-called 'L12 stalk' on the 50S ribosomal subunit. Ribosomal protein L11 binds to the same region of 23S rRNA and is located at the base of the 'L12 stalk'. The 'L12 stalk' plays a key role in the interaction of the ribosome with translation factors. In this study stalk complexes from mesophilic and (hyper)thermophilic species of the archaeal genus Methanococcus and from the Archaeon Sulfolobus solfataricus, as well as from the Bacteria Escherichia coli, Geobacillus stearothermophilus and Thermus thermophilus, were overproduced in E.coli and purified under non-denaturing conditions. Using filter-binding assays the affinities of the archaeal and bacterial complexes to their specific 23S rRNA target site were analyzed at different pH, ionic strength and temperature. Affinities of both archaeal and bacterial complexes for 23S rRNA vary by more than two orders of magnitude, correlating very well with the growth temperatures of the organisms. A cooperative effect of binding to 23S rRNA of protein L11 and the L10/L12(4) complex from mesophilic and thermophilic Archaea was shown to be temperature-dependent.

  20. Overexpression, purification, crystallization and preliminary crystallographic studies of a hyperthermophilic adenylosuccinate synthetase from Pyrococcus horikoshii OT3

    International Nuclear Information System (INIS)

    Wang, Xiaoying; Akasaka, Ryogo; Takemoto, Chie; Morita, Satoshi; Yamaguchi, Machiko; Terada, Takaho; Shirozu, Mikako; Yokoyama, Shigeyuki; Chen, Shilin; Si, Shuyi; Xie, Yong

    2011-01-01

    A hyperthermophilic adenylosuccinate synthetase from P. horikoshii OT3, which is 90–120 amino acids shorter than those from the vast majority of organisms, was expressed, purified and crystallized and X-ray diffraction data were collected to 2.5 Å resolution. Adenylosuccinate synthetase (AdSS) is a ubiquitous enzyme that catalyzes the first committed step in the conversion of inosine monophosphate (IMP) to adenosine monophosphate (AMP) in the purine-biosynthetic pathway. Although AdSS from the vast majority of organisms is 430–457 amino acids in length, AdSS sequences isolated from thermophilic archaea are 90–120 amino acids shorter. In this study, crystallographic studies of a short AdSS sequence from Pyrococcus horikoshii OT3 (PhAdSS) were performed in order to reveal the unusual structure of AdSS from thermophilic archaea. Crystals of PhAdSS were obtained by the microbatch-under-oil method and X-ray diffraction data were collected to 2.50 Å resolution. The crystal belonged to the trigonal space group P3 2 12, with unit-cell parameters a = b = 57.2, c = 107.9 Å. There was one molecule per asymmetric unit, giving a Matthews coefficient of 2.17 Å 3 Da −1 and an approximate solvent content of 43%. In contrast, the results of native polyacrylamide gel electrophoresis and analytical ultracentrifugation showed that the recombinant PhAdSS formed a dimer in solution

  1. Elucidating the transcription cycle of the UV-inducible hyperthermophilic archaeal virus SSV1 by DNA microarrays

    International Nuclear Information System (INIS)

    Froels, Sabrina; Gordon, Paul M.K.; Panlilio, Mayi Arcellana; Schleper, Christa; Sensen, Christoph W.

    2007-01-01

    The spindle-shaped Sulfolobus virus SSV1 was the first of a series of unusual and uniquely shaped viruses isolated from hyperthermophilic Archaea. Using whole-genome microarrays we show here that the circular 15.5 kb DNA genome of SSV1 exhibits a chronological regulation of its transcription upon UV irradiation, reminiscent to the life cycles of bacteriophages and eukaryotic viruses. The transcriptional cycle starts with a small UV-specific transcript and continues with early transcripts on both its flanks. The late transcripts appear after the onset of viral replication and are extended to their full lengths towards the end of the approximately 8.5 h cycle. While we detected only small differences in genome-wide analysis of the host Sulfolobus solfataricus comparing infected versus uninfected strains, we found a marked difference with respect to the strength and speed of the general UV response of the host. Models for the regulation of the virus cycle, and putative functions of genes in SSV1 are presented

  2. Unusual Starch Degradation Pathway via Cyclodextrins in the Hyperthermophilic Sulfate-Reducing Archaeon Archaeoglobus fulgidus Strain 7324▿

    Science.gov (United States)

    Labes, Antje; Schönheit, Peter

    2007-01-01

    The hyperthermophilic archaeon Archaeoglobus fulgidus strain 7324 has been shown to grow on starch and sulfate and thus represents the first sulfate reducer able to degrade polymeric sugars. The enzymes involved in starch degradation to glucose 6-phosphate were studied. In extracts of starch-grown cells the activities of the classical starch degradation enzymes, α-amylase and amylopullulanase, could not be detected. Instead, evidence is presented here that A. fulgidus utilizes an unusual pathway of starch degradation involving cyclodextrins as intermediates. The pathway comprises the combined action of an extracellular cyclodextrin glucanotransferase (CGTase) converting starch to cyclodextrins and the intracellular conversion of cyclodextrins to glucose 6-phosphate via cyclodextrinase (CDase), maltodextrin phosphorylase (Mal-P), and phosphoglucomutase (PGM). These enzymes, which are all induced after growth on starch, were characterized. CGTase catalyzed the conversion of starch to mainly β-cyclodextrin. The gene encoding CGTase was cloned and sequenced and showed highest similarity to a glucanotransferase from Thermococcus litoralis. After transport of the cyclodextrins into the cell by a transport system to be defined, these molecules are linearized via a CDase, catalyzing exclusively the ring opening of the cyclodextrins to the respective maltooligodextrins. These are degraded by a Mal-P to glucose 1-phosphate. Finally, PGM catalyzes the conversion of glucose 1-phosphate to glucose 6-phosphate, which is further degraded to pyruvate via the modified Embden-Meyerhof pathway. PMID:17921308

  3. Palaeococcus helgesonii sp. nov., a facultatively anaerobic, hyperthermophilic archaeon from a geothermal well on Vulcano Island, Italy.

    Science.gov (United States)

    Amend, Jan P; Meyer-Dombard, D'Arcy R; Sheth, Seema N; Zolotova, Natalya; Amend, Andrea C

    2003-06-01

    A novel, hyperthermophilic archaeon was isolated from a shallow geothermal well that taps marine waters on the Island of Vulcano in the southern Tyrrhenian Sea, Italy. The cells were irregular cocci, 0.6-1.5 microm in diameter, with multiple polar flagella. Growth was observed at temperatures from 45 to 85 degrees C (optimum at approximately 80 degrees C), pH 5-8 (optimum at 6.5), and 0.5-6.0% NaCl (optimum at approximately 2.8%). The minimum doubling time was 50 min. The isolate was obligately chemoheterotrophic, utilizing complex organic compounds including yeast or beef extract, peptone, tryptone, or casein for best growth. The presence of elemental sulfur enhanced growth. The isolate grew anaerobically as well as microaerobically. The G+C content of the genomic DNA was 42.5 mol%. The 16S rRNA sequence indicated that the new isolate was a member of the Thermococcales within the euryarchaeota, representing the second species in the genus Palaeococcus. Its physiology and phylogeny differed in several key characteristics from those of Palaeococcus ferrophilus, justifying the establishment of a new species; the name Palaeococcus helgesonii sp. nov. is proposed, type strain PI1 (DSM 15127).

  4. Aglycone specificity of Thermotoga neapolitana β-glucosidase 1A modified by mutagenesis, leading to increased catalytic efficiency in quercetin-3-glucoside hydrolysis

    Directory of Open Access Journals (Sweden)

    Lindahl Sofia

    2011-02-01

    Full Text Available Abstract Background The thermostable β-glucosidase (TnBgl1A from Thermotoga neapolitana is a promising biocatalyst for hydrolysis of glucosylated flavonoids and can be coupled to extraction methods using pressurized hot water. Hydrolysis has however been shown to be dependent on the position of the glucosylation on the flavonoid, and e.g. quercetin-3-glucoside (Q3 was hydrolysed slowly. A set of mutants of TnBgl1A were thus created to analyse the influence on the kinetic parameters using the model substrate para-nitrophenyl-β-D-glucopyranoside (pNPGlc, and screened for hydrolysis of Q3. Results Structural analysis pinpointed an area in the active site pocket with non-conserved residues between specificity groups in glycoside hydrolase family 1 (GH1. Three residues in this area located on β-strand 5 (F219, N221, and G222 close to sugar binding sub-site +2 were selected for mutagenesis and amplified in a protocol that introduced a few spontaneous mutations. Eight mutants (four triple: F219L/P165L/M278I, N221S/P165L/M278I, G222Q/P165L/M278I, G222Q/V203M/K214R, two double: F219L/K214R, N221S/P342L and two single: G222M and N221S were produced in E. coli, and purified to apparent homogeneity. Thermostability, measured as Tm by differential scanning calorimetry (101.9°C for wt, was kept in the mutated variants and significant decrease (ΔT of 5 - 10°C was only observed for the triple mutants. The exchanged residue(s in the respective mutant resulted in variations in KM and turnover. The KM-value was only changed in variants mutated at position 221 (N221S and was in all cases monitored as a 2-3 × increase for pNPGlc, while the KM decreased a corresponding extent for Q3. Turnover was only significantly changed using pNPGlc, and was decreased 2-3 × in variants mutated at position 222, while the single, double and triple mutated variants carrying a mutation at position 221 (N221S increased turnover up to 3.5 × compared to the wild type. Modelling

  5. Identification of a novel amino acid racemase from a hyperthermophilic archaeon Pyrococcus horikoshii OT-3 induced by D-amino acids.

    Science.gov (United States)

    Kawakami, Ryushi; Ohmori, Taketo; Sakuraba, Haruhiko; Ohshima, Toshihisa

    2015-08-01

    To date, there have been few reports analyzing the amino acid requirement for growth of hyperthermophilic archaea. We here found that the hyperthermophilic archaeon Pyrococcus horikoshii OT-3 requires Thr, Leu, Val, Phe, Tyr, Trp, His and Arg in the medium for growth, and shows slow growth in medium lacking Met or Ile. This largely corresponds to the presence, or absence, of genes related to amino acid biosynthesis in its genome, though there are exceptions. The amino acid requirements were dramatically lost by addition of D-isomers of Met, Leu, Val, allo-Ile, Phe, Tyr, Trp and Arg. Tracer analysis using (14)C-labeled D-Trp showed that D-Trp in the medium was used as a protein component in the cells, suggesting the presence of D-amino acid metabolic enzymes. Pyridoxal 5'-phosphate (PLP)-dependent racemase activity toward Met, Leu and Phe was detected in crude extract of P. horikoshii and was enhanced in cells grown in the medium supplemented with D-amino acids, especially D-allo-Ile. The gene encoding the racemase was narrowed down to one open reading frame on the basis of enzyme purification from P. horikoshii cells, and the recombinant enzyme exhibited PLP-dependent racemase activity toward several amino acids, including Met, Leu and Phe, but not Pro, Asp or Glu. This is the first report showing the presence in a hyperthermophilic archaeon of a PLP-dependent amino acid racemase with broad substrate specificity that is likely responsible for utilization of D-amino acids for growth.

  6. Structural and genomic properties of the hyperthermophilic archaeal virus ATV with an extracellular stage of the reproductive cycle.

    Science.gov (United States)

    Prangishvili, David; Vestergaard, Gisle; Häring, Monika; Aramayo, Ricardo; Basta, Tamara; Rachel, Reinhard; Garrett, Roger A

    2006-06-23

    A novel virus, ATV, of the hyperthermophilic archaeal genus Acidianus has the unique property of undergoing a major morphological development outside of, and independently of, the host cell. Virions are extruded from host cells as lemon-shaped tail-less particles, after which they develop long tails at each pointed end, at temperatures close to that of the natural habitat, 85 degrees C. The extracellularly developed tails constitute tubes, which terminate in an anchor-like structure that is not observed in the tail-less particles. A thin filament is located within the tube, which exhibits a periodic structure. Tail development produces a one half reduction in the volume of the virion, concurrent with a slight expansion of the virion surface. The circular, double-stranded DNA genome contains 62,730 bp and is exceptional for a crenarchaeal virus in that it carries four putative transposable elements as well as genes, which previously have been associated only with archaeal self-transmissable plasmids. In total, it encodes 72 predicted proteins, including 11 structural proteins with molecular masses in the range of 12 to 90 kDa. Several of the larger proteins are rich in coiled coil and/or low complexity sequence domains, which are unusual for archaea. One protein, in particular P800, resembles an intermediate filament protein in its structural properties. It is modified in the two-tailed, but not in the tail-less, virion particles and it may contribute to viral tail development. Exceptionally for a crenarchaeal virus, infection with ATV results either in viral replication and subsequent cell lysis or in conversion of the infected cell to a lysogen. The lysogenic cycle involves integration of the viral genome into the host chromosome, probably facilitated by the virus-encoded integrase and this process can be interrupted by different stress factors.

  7. Crystal structure of hyperthermophilic esterase EstE1 and the relationship between its dimerization and thermostability properties

    Directory of Open Access Journals (Sweden)

    Koh Eunhee

    2007-07-01

    Full Text Available Abstract Background EstE1 is a hyperthermophilic esterase belonging to the hormone-sensitive lipase family and was originally isolated by functional screening of a metagenomic library constructed from a thermal environmental sample. Dimers and oligomers may have been evolutionally selected in thermophiles because intersubunit interactions can confer thermostability on the proteins. The molecular mechanisms of thermostabilization of this extremely thermostable esterase are not well understood due to the lack of structural information. Results Here we report for the first time the 2.1-Å resolution crystal structure of EstE1. The three-dimensional structure of EstE1 exhibits a classic α/β hydrolase fold with a central parallel-stranded beta sheet surrounded by alpha helices on both sides. The residues Ser154, Asp251, and His281 form the catalytic triad motif commonly found in other α/β hydrolases. EstE1 exists as a dimer that is formed by hydrophobic interactions and salt bridges. Circular dichroism spectroscopy and heat inactivation kinetic analysis of EstE1 mutants, which were generated by structure-based site-directed mutagenesis of amino acid residues participating in EstE1 dimerization, revealed that hydrophobic interactions through Val274 and Phe276 on the β8 strand of each monomer play a major role in the dimerization of EstE1. In contrast, the intermolecular salt bridges contribute less significantly to the dimerization and thermostability of EstE1. Conclusion Our results suggest that intermolecular hydrophobic interactions are essential for the hyperthermostability of EstE1. The molecular mechanism that allows EstE1 to endure high temperature will provide guideline for rational design of a thermostable esterase/lipase using the lipolytic enzymes showing structural similarity to EstE1.

  8. Thermococcus sulfurophilus sp. nov., a New Hyperthermophilic, Sulfur-Reducing Archaeon Isolated from Deep-Sea Hydrothermal Vent

    Science.gov (United States)

    Pikuta, Elena V.; Hoover, Richard B.; Whitman, William B.; Marsic, Damien; Garriott, Owen; Six, N. Frank (Technical Monitor)

    2002-01-01

    A new hyperthermophilic, anaerobic, sulfur-reducing, organo-heterotrophic archaeon, strain OGL-20P, was isolated from "black smoker" chimney material at the Rainbow hydrothermal vent site in the Atlantic Ocean (36.2 N; 33.9 W). The cells of strain OGL-20P have irregular coccoid shape and are motile with a single flagellum. Growth occurs within pH range of 5.5-8.2 (optimal at pH 7.0-7.2), salinity range of 1-5% NaCl (optimal concentration 3% NaCl wt/vol), and temperature range of +55 C to +94 C (optimal growth at +83 C to +85 C). Strain OGL-20P is resistant to freezing (at -20 C). New isolate is strictly anaerobic with sulfur-type of respiration. A limited number of compounds are utilized as electron donors, including peptone, becto-tryptone, casamino-acids, and yeast extract but does not grow with separate amino acids. Sulfur and Iron can be used as electron acceptors; but not sulfate, sulfite, thiosulfate or nitrate. Strain OGL-20P is resistant to chloramphenicol, kanamycin, and gentamycin. Growth of str. OGL20P is inhibited by tetracyclin but not by Na2MoO4. The G+C content of DNA is 57.2 mol%. The 16S ribosomal RNA sequence analysis allows one to classify strain OGL-20P as a representative of a now species of Thermococcus genus. The name Thermococcus sulfurophilus op. nov., was suggested for the new isolate, type strain OGL-20P (sup T) (= ATCC BAA_394 (sup T) = DSM...(supT)).

  9. Crystal structure of THEP1 from the hyperthermophile Aquifex aeolicus: a variation of the RecA fold

    Directory of Open Access Journals (Sweden)

    Wittinghofer Alfred

    2005-03-01

    Full Text Available Abstract Background aaTHEP1, the gene product of aq_1292 from Aquifex aeolicus, shows sequence homology to proteins from most thermophiles, hyperthermophiles, and higher organisms such as man, mouse, and fly. In contrast, there are almost no homologous proteins in mesophilic unicellular microorganisms. aaTHEP1 is a thermophilic enzyme exhibiting both ATPase and GTPase activity in vitro. Although annotated as a nucleotide kinase, such an activity could not be confirmed for aaTHEP1 experimentally and the in vivo function of aaTHEP1 is still unknown. Results Here we report the crystal structure of selenomethionine substituted nucleotide-free aaTHEP1 at 1.4 Å resolution using a multiple anomalous dispersion phasing protocol. The protein is composed of a single domain that belongs to the family of 3-layer (α/β/α-structures consisting of nine central strands flanked by six helices. The closest structural homologue as determined by DALI is the RecA family. In contrast to the latter proteins, aaTHEP1 possesses an extension of the β-sheet consisting of four additional β-strands. Conclusion We conclude that the structure of aaTHEP1 represents a variation of the RecA fold. Although the catalytic function of aaTHEP1 remains unclear, structural details indicate that it does not belong to the group of GTPases, kinases or adenosyltransferases. A mainly positive electrostatic surface indicates that aaTHEP1 might be a DNA/RNA modifying enzyme. The resolved structure of aaTHEP1 can serve as paradigm for the complete THEP1 family.

  10. Thermococcus Thioreducens sp. Nov., a Novel Hyperthermophilic, Obligately Sulfur-reducing Archaeon from a Deep-sea Hydrothermal Vent

    Science.gov (United States)

    Pikuta, Elena V.; Marsic, Damien; Itoh, Takashi; Bej, Asim K.; Tang, Jane; Whitman, William B.; Ng, Joseph D.; Garriott, Owen K.; Hoover, Richard B.

    2007-01-01

    A hyperthermophilic, sulfur-reducing, organo-heterotrophic archaeon, strain OGL-20P was isolated from black smoker chimney material from the Rainbow hydrothermal vent site on the Mid-Atlantic Ridge (36.2 N, 33.9 W). The cells of strain OGL-20P(sup T) have an irregular coccoid shape and are motile with a single flagellum. Growth was observed within the pH range 5.0-8.5 (optimum pH 7.0), NaCl concentration range 1-5 % (w/v) (optimum 3%), and temperature range 55-94 C (optimum 83-85 C). The novel isolate is strictly anaerobic and obligately dependent upon elemental sulfur as an electron acceptor, but it does not reduce sulfate, sulfite, thiosulfate, iron (III) or nitrate. Proteolysis products (peptone, bacto-tryptone, casamino-acids, and yeast extract) are utilized as substrates during sulfur-reduction. Strain OGL-20P(sup T) is resistant to ampicillin, chloramphenicol, kanamycin, and gentamycin, but sensitive to tetracycline and rifampicin. The G+C content of DNA is 52.9 mol%. The 16S rRNA gene sequence analysis revealed that strain OGL-20P(sup T) is closely related to Thermococcus coalescens and related species, but no significant homology by DNA-DNA hybridization was observed between those species and the new isolate. On the basis of physiological and molecular properties of the new isolate, we conclude that strain OGL-20P(sup T) represents a new separate species within the genus Thermococcus, and propose the name Thermococcus thioreducens sp. nov. The type strain is OGL-20P(sup T) (= ATCC BAA-394(sup T) = JCM 12859(sup T) = DSM 14981(sup T)).

  11. Contrasting effects of ecosystem engineering by the cordgrass Spartina maritima and the sandprawn Callianassa kraussi in a marine-dominated lagoon

    Science.gov (United States)

    Pillay, D.; Branch, G. M.; Dawson, J.; Henry, D.

    2011-01-01

    Ecosystem engineering by plants and animals significantly influences community structure and the physico-chemical characteristics of marine habitats. In this paper we document the contrasting effects of ecosystem engineering by the cordgrass Spartina maritima and the burrowing sandprawn Callianassa kraussi on physico-chemical characteristics, microflora, macrofaunal community structure and morphological attributes in the high shore intertidal sandflats of Langebaan Lagoon, a marine-dominated system on the west coast of South Africa. Comparisons were made at six sites in the lagoon within Spartina and Callianassa beds, and in a "bare zone" of sandflat between these two habitats that lacks both sandprawns and cordgrass. Sediments in Spartina habitats were consolidated by the root-shoot systems of the cordgrass, leading to low sediment penetrability, while sediments in beds of C. kraussi were more penetrable, primarily due to the destabilising effects of sandprawn bioturbation. Sediments in the "bare zone" had intermediate to low values of penetrability. Sediment organic content was lowest in bare zones and greatest in Spartina beds, while sediment chl- a levels were greatest on bare sand, but were progressively reduced in the Spartina and Callianassa beds. These differences among habitats induced by ecosystem engineering in turn affected the macrofauna. Community structure was different between all three habitats sampled, with species richness being surprisingly greater in Callianassa beds than either the bare zone or Spartina beds. In general, the binding of surface sediments by the root systems of Spartina favoured rigid-bodied, surface-dwelling and tube-building species, while the destabilising effect of bioturbation by C. kraussi favoured burrowing species. The contrasting effects of these ecosystem engineers suggest that they play important roles in increasing habitat heterogeneity. Importantly, the role of bioturbation by C. kraussi in enhancing macrofaunal

  12. Chemical Composition and Biological Studies of the Essential Oil from Aerial Parts of Beta vulgaris subsp. maritima (L.) Arcang. Growing in Tunisia.

    Science.gov (United States)

    Zardi-Bergaoui, Afifa; Ben Nejma, Aymen; Harzallah-Skhiri, Fethia; Flamini, Guido; Ascrizzi, Roberta; Ben Jannet, Hichem

    2017-10-01

    The chemical composition, antioxidant, cytotoxic, anticholinesterase and anti-tyrosinase activities of the hydrodistilled essential oil of the aerial parts of Beta vulgaris subsp. maritime (L.) Arcang. from Tunisia have been evaluated. The chemical composition of the oil (yield 0.037% [w/w]), determined by GC-FID and GC/MS is reported for the first time. Twenty five components, accounting for 98.1% of the total oil have been identified. The oil was characterized by a high proportion of oxygenated sesquiterpenes (39.2%), followed by sesquiterpene hydrocarbons (30.3%) and one apocarotenoids (26.3%). The main compounds were γ-irone (26.3%), α-cadinol (12.1%), T-cadinol (10.6%), bicyclogermacrene (10.4%) and δ-cadinene (6.0%). The isolated oil was tested for its antioxidant activity using the DPPH · , ABTS +· , catalase, and paraoxonase assays and also for its cytotoxic, anticholinesterase, and anti-tyrosinase activities. The essential oil exhibited high antioxidant activity (IC 50  = 0.055 ± 0.006 mg/ml) and important result oncatalase (524.447 ± 2.58 Units/mg protein). Furthermore, it exerted a significant cytotoxic effect against A549 cell line, with IC 50  = 42.44 ± 1.40 μg/ml. The results indicate that the essential oil of B. vulgaris subsp. maritima (L.) Arcang. aerial parts may be used in future as an alternative to synthetic antioxidant agents, with potential application in the food and pharmaceutical industries. © 2017 Wiley-VHCA AG, Zurich, Switzerland.

  13. Comparative study of antibacterial and antifugal activity of callus culture and adult plants extracts from Alternanthera maritima (Amaranthaceae Estudo comparativo da atividade antibacteriana e antifúngica de extratos obtidos da cultura de calos e da planta adulta de Alternanthera maritima (Amaranthaceae

    Directory of Open Access Journals (Sweden)

    Marcos J. Salvador

    2004-06-01

    Full Text Available The aim of this study was to evaluate the antibacterial and antifungal activity of callus culture (two different hormonal combination culture medium and adult plants (two collect extracts from Alternanthera maritima (Amaranthaceae investigating the maintenance of antimicrobial activity in vivo and in vitro. The antibacterial and antifungal activity was determined by the agar-well diffusion method against thirty strains of microorganisms including Gram-positive and Gram-negative bacteria, yeasts and dermatophytes. All the organic crude extracts studied were bioactive. Extracts of aerial parts and roots of adult plants collected during the same period of years of 1995 and 1998 (Restinga de Maricá (RJ, collect 1 and 2 inhibited the growth of several microorganisms (bacteria, yeasts and dermatophytes with inhibition halo between 6 and 20 mm. Plant cell callus culture extracts obtained from two culture conditions were also bioactive. Thus, the positive results suggest that the A. maritima extracts should be further studied to determine the bioactive chemical compounds as well as to understand the possible mechanisms of action and evaluate their toxicity looking toward a pharmaceutical employment.Neste estudo procedeu-se a avaliação da atividade antibacteriana e antifúngica dos extratos brutos de Alternanthera maritima (Amaranthaceae planta in natura de duas coletas distintas e obtidos por cultura de células buscando-se averiguar a manutenção da atividade antimicrobiana dos extratos obtidos da planta in vivo e in vitro. A ação antibacteriana e antifúngica foi determinada pelo método de difusão em ágar (técnica do poço utilizando-se trinta cepas de microrganismos indicadores (bactérias Gram-positivas e Gram-negativas, leveduras e dermatófitos. Todos os extratos obtidos com solventes orgânicos avaliados apresentaram-se bioativos com halos de inibição de 6 a 20 mm. Os extratos da planta in natura das duas coletas (Restinga de Marica

  14. Microfluidic glycosyl hydrolase screening for biomass-to-biofuel conversion.

    Science.gov (United States)

    Bharadwaj, Rajiv; Chen, Zhiwei; Datta, Supratim; Holmes, Bradley M; Sapra, Rajat; Simmons, Blake A; Adams, Paul D; Singh, Anup K

    2010-11-15

    The hydrolysis of biomass to fermentable sugars using glycosyl hydrolases such as cellulases and hemicellulases is a limiting and costly step in the conversion of biomass to biofuels. Enhancement in hydrolysis efficiency is necessary and requires improvement in both enzymes and processing strategies. Advances in both areas in turn strongly depend on the progress in developing high-throughput assays to rapidly and quantitatively screen a large number of enzymes and processing conditions. For example, the characterization of various cellodextrins and xylooligomers produced during the time course of saccharification is important in the design of suitable reactors, enzyme cocktail compositions, and biomass pretreatment schemes. We have developed a microfluidic-chip-based assay for rapid and precise characterization of glycans and xylans resulting from biomass hydrolysis. The technique enables multiplexed separation of soluble cellodextrins and xylose oligomers in around 1 min (10-fold faster than HPLC). The microfluidic device was used to elucidate the mode of action of Tm_Cel5A, a novel cellulase from hyperthermophile Thermotoga maritima . The results demonstrate that the cellulase is active at 80 °C and effectively hydrolyzes cellodextrins and ionic-liquid-pretreated switchgrass and Avicel to glucose, cellobiose, and cellotriose. The proposed microscale approach is ideal for quantitative large-scale screening of enzyme libraries for biomass hydrolysis, for development of energy feedstocks, and for polysaccharide sequencing.

  15. A Robust and Versatile Method of Combinatorial Chemical Synthesis of Gene Libraries via Hierarchical Assembly of Partially Randomized Modules

    Science.gov (United States)

    Popova, Blagovesta; Schubert, Steffen; Bulla, Ingo; Buchwald, Daniela; Kramer, Wilfried

    2015-01-01

    A major challenge in gene library generation is to guarantee a large functional size and diversity that significantly increases the chances of selecting different functional protein variants. The use of trinucleotides mixtures for controlled randomization results in superior library diversity and offers the ability to specify the type and distribution of the amino acids at each position. Here we describe the generation of a high diversity gene library using tHisF of the hyperthermophile Thermotoga maritima as a scaffold. Combining various rational criteria with contingency, we targeted 26 selected codons of the thisF gene sequence for randomization at a controlled level. We have developed a novel method of creating full-length gene libraries by combinatorial assembly of smaller sub-libraries. Full-length libraries of high diversity can easily be assembled on demand from smaller and much less diverse sub-libraries, which circumvent the notoriously troublesome long-term archivation and repeated proliferation of high diversity ensembles of phages or plasmids. We developed a generally applicable software tool for sequence analysis of mutated gene sequences that provides efficient assistance for analysis of library diversity. Finally, practical utility of the library was demonstrated in principle by assessment of the conformational stability of library members and isolating protein variants with HisF activity from it. Our approach integrates a number of features of nucleic acids synthetic chemistry, biochemistry and molecular genetics to a coherent, flexible and robust method of combinatorial gene synthesis. PMID:26355961

  16. Growth of Thermophilic and Hyperthermophilic Fe(III)-Reducing Microorganisms on a Ferruginous Smectite as the Sole Electron Acceptor▿

    Science.gov (United States)

    Kashefi, Kazem; Shelobolina, Evgenya S.; Elliott, W. Crawford; Lovley, Derek R.

    2008-01-01

    Recent studies have suggested that the structural Fe(III) within phyllosilicate minerals, including smectite and illite, is an important electron acceptor for Fe(III)-reducing microorganisms in sedimentary environments at moderate temperatures. The reduction of structural Fe(III) by thermophiles, however, has not previously been described. A wide range of thermophilic and hyperthermophilic Archaea and Bacteria from marine and freshwater environments that are known to reduce poorly crystalline Fe(III) oxides were tested for their ability to reduce structural (octahedrally coordinated) Fe(III) in smectite (SWa-1) as the sole electron acceptor. Two out of the 10 organisms tested, Geoglobus ahangari and Geothermobacterium ferrireducens, were not able to conserve energy to support growth by reduction of Fe(III) in SWa-1 despite the fact that both organisms were originally isolated with solid-phase Fe(III) as the electron acceptor. The other organisms tested were able to grow on SWa-1 and reduced 6.3 to 15.1% of the Fe(III). This is 20 to 50% less than the reported amounts of Fe(III) reduced in the same smectite (SWa-1) by mesophilic Fe(III) reducers. Two organisms, Geothermobacter ehrlichii and archaeal strain 140, produced copious amounts of an exopolysaccharide material, which may have played an active role in the dissolution of the structural iron in SWa-1 smectite. The reduction of structural Fe(III) in SWa-1 by archaeal strain 140 was studied in detail. Microbial Fe(III) reduction was accompanied by an increase in interlayer and octahedral charges and some incorporation of potassium and magnesium into the smectite structure. However, these changes in the major element chemistry of SWa-1 smectite did not result in the formation of an illite-like structure, as reported for a mesophilic Fe(III) reducer. These results suggest that thermophilic Fe(III)-reducing organisms differ in their ability to reduce and solubilize structural Fe(III) in SWa-1 smectite and that SWa-1

  17. Physiological and molecular studies of the resistance to ionizing radiations of hyper-thermophilic archaea isolated from deep ocean hydrothermal sources

    International Nuclear Information System (INIS)

    Jolivet, E.

    2002-10-01

    In this study, we have first tested in vivo the effect of gamma irradiation on Pyrococcus abyssi, a hyper-thermophilic archaeon, isolated from a deep-sea hydrothermal vent. We have shown that this strain was as radioresistant as P. furiosus but less than Deinococcus radiodurans. The rates of double stranded breaks provoked into DNA following irradiation were monitored by the pulsed-field gel electrophoresis technique (P.F.G.E.) with P. abyssi, P. furiosus, D. radiodurans and Escherichia coli. Results clearly showed that all these rates were similar suggesting that no specific DNA protection system exits in Pyrococcus species. The growth of P. abyssi was efficiently recovered within two hours following the exposure to 2.5 kGy of gamma irradiation. As revealed by P.F.G.E., genomic DNA of P. abyssi totally fragmented after irradiation was efficiently restored within two hours presumably by inter chromosomal homologous recombination. The DNA replication in P. abyssi cells following irradiation at 2.5 kGy was blocked for 90 minutes that corresponds to the decay for repairing damaged DNA. Moreover, following irradiation P. abyssi actively expulse damaged DNA material before DNA replication resumes, preventing the amplification of genetic mutations. We have also showed that at least a subset cf P. abyssi DNA repair and replication proteins, such as RadA, RPA-41 and RFC-S. were constitutively expressed in chromatin bound forms in stationary phase cells. Our results were in agreement with the view that P. abyssi contains a very efficient DNA repair system, which is continuously ready to counteract the DNA damaged caused by the high temperature and/or ionizing radiation. For the first time, three novel hyper-thermophilic archaea species from deep-sea hydrothermal vents more radioresistant than P. abyssi were isolated and characterized, after 'y-irradiation exposures of some enrichment cultures. Thermococcus marinus, Thermococcus radiophilus and Thermococcus gammafolerans

  18. Co-expression of the Thermotoga neapolitana aglB gene with an upstream 3'-coding fragment of the malG gene improves enzymatic characteristics of recombinant AglB cyclomaltodextrinase.

    Science.gov (United States)

    Lunina, Natalia A; Agafonova, Elena V; Chekanovskaya, Lyudmila A; Dvortsov, Igor A; Berezina, Oksana V; Shedova, Ekaterina N; Kostrov, Sergey V; Velikodvorskaya, Galina A

    2007-07-01

    A cluster of Thermotoga neapolitana genes participating in starch degradation includes the malG gene of sugar transport protein and the aglB gene of cyclomaltodextrinase. The start and stop codons of these genes share a common overlapping sequence, aTGAtg. Here, we compared properties of expression products of three different constructs with aglB from T. neapolitana. The first expression vector contained the aglB gene linked to an upstream 90-bp 3'-terminal region of the malG gene with the stop codon overlapping with the start codon of aglB. The second construct included the isolated coding sequence of aglB with two tandem potential start codons. The expression product of this construct in Escherichia coli had two tandem Met residues at its N terminus and was characterized by low thermostability and high tendency to aggregate. In contrast, co-expression of aglB and the 3'-terminal region of malG (the first construct) resulted in AglB with only one N-terminal Met residue and a much higher specific activity of cyclomaltodextrinase. Moreover, the enzyme expressed by such a construct was more thermostable and less prone to aggregation. The third construct was the same as the second one except that it contained only one ATG start codon. The product of its expression had kinetic and other properties similar to those of the enzyme with only one N-terminal Met residue.

  19. Computational genomics of hyperthermophiles

    NARCIS (Netherlands)

    Werken, van de H.J.G.

    2008-01-01

    With the ever increasing number of completely sequenced prokaryotic genomes and the subsequent use of functional genomics tools, e.g. DNA microarray and proteomics, computational data analysis and the integration of microbial and molecular data is inevitable. This thesis describes the computational

  20. Flow synthesis of phenylserine using threonine aldolase (TA) immobilized on Eupergit support

    NARCIS (Netherlands)

    Tibhe, J.; Fu, Hui; Noel, T.; Wang, Q.; Meuldijk, J.; Hessel, V.

    2013-01-01

    Threonine aldolase (TA) from Thermotoga maritima was immobilized on Eupergit support by both a direct and an indirect method. The incubation time for the direct immobilization method was optimized for the highest amount of enzyme on the support. By introducing the immobilized TA in a packed-bed

  1. Bifunctional xylanases and their potential use in biotechnology

    Digital Repository Service at National Institute of Oceanography (India)

    Khandeparker, R.; Numan, M.Th.

    . J Chromatography 919:389–394 33. Hong SY, Lee JS, Cho KM, Math RK, Kim YH, Hong SJ, Cho YU, Kim H, Yun HD (2006) Assembling a novel bifunctional cel- lulase–xylanase from Thermotoga maritima by end-to-end fusion. Biotechnol Lett 28:1857–1862 34...

  2. Dicty_cDB: Contig-U11847-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 00512_699( AE000512 |pid:none) Thermotoga maritima MSB8, comple... 42 0.065 JC2551( JC2551 ) tropomyosin alpha chain - axolotl...AltName: Full=Myosin heavy chai... 42 0.11 JC6199( JC6199 ) alpha-tropomyosin S-1 - axolotl &U33450_1(U33450

  3. ORF Alignment: NC_000853 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_000853 gi|15643179 >1v7zA 3 256 7 270 9e-43 ... ref|NP_228223.1| creatinine amidoh...ydrolase, putative [Thermotoga maritima MSB8] ... gb|AAD35498.1| creatinine amidohydrolase, putative

  4. Morphology and genome organization of the virus PSV of the hyperthermophilic archaeal genera Pyrobaculum and Thermoproteus: a novel virus family, the Globuloviridae.

    Science.gov (United States)

    Häring, Monika; Peng, Xu; Brügger, Kim; Rachel, Reinhard; Stetter, Karl O; Garrett, Roger A; Prangishvili, David

    2004-06-01

    A novel virus, termed Pyrobaculum spherical virus (PSV), is described that infects anaerobic hyperthermophilic archaea of the genera Pyrobaculum and Thermoproteus. Spherical enveloped virions, about 100 nm in diameter, contain a major multimeric 33-kDa protein and host-derived lipids. A viral envelope encases a superhelical nucleoprotein core containing linear double-stranded DNA. The PSV infection cycle does not cause lysis of host cells. The viral genome was sequenced and contains 28337 bp. The genome is unique for known archaeal viruses in that none of the genes, including that encoding the major structural protein, show any significant sequence matches to genes in public sequence databases. Exceptionally for an archaeal double-stranded DNA virus, almost all the recognizable genes are located on one DNA strand. The ends of the genome consist of 190-bp inverted repeats that contain multiple copies of short direct repeats. The two DNA strands are probably covalently linked at their termini. On the basis of the unusual morphological and genomic properties of this DNA virus, we propose to assign PSV to a new viral family, the Globuloviridae.

  5. Distribution and phylogenies of enzymes of the Embden-Meyerhof-Parnas pathway from archaea and hyperthermophilic bacteria support a gluconeogenic origin of metabolism

    Directory of Open Access Journals (Sweden)

    Ron S. Ronimus

    2003-01-01

    Full Text Available Enzymes of the gluconeogenic/glycolytic pathway (the Embden-Meyerhof-Parnas (EMP pathway, the reductive tricarboxylic acid cycle, the reductive pentose phosphate cycle and the Entner-Doudoroff pathway are widely distributed and are often considered to be central to the origins of metabolism. In particular, several enzymes of the lower portion of the EMP pathway (the so-called trunk pathway, including triosephosphate isomerase (TPI; EC 5.3.1.1, glyceraldehyde-3-phosphate dehydrogenase (GAPDH; EC 1.2.1.12/13, phosphoglycerate kinase (PGK; EC 2.7.2.3 and enolase (EC 4.2.1.11, are extremely well conserved and universally distributed among the three domains of life. In this paper, the distribution of enzymes of gluconeogenesis/glycolysis in hyperthermophiles—microorganisms that many believe represent the least evolved organisms on the planet—is reviewed. In addition, the phylogenies of the trunk pathway enzymes (TPIs, GAPDHs, PGKs and enolases are examined. The enzymes catalyzing each of the six-carbon transformations in the upper portion of the EMP pathway, with the possible exception of aldolase, are all derived from multiple gene sequence families. In contrast, single sequence families can account for the archaeal and hyperthermophilic bacterial enzyme activities of the lower portion of the EMP pathway. The universal distribution of the trunk pathway enzymes, in combination with their phylogenies, supports the notion that the EMP pathway evolved in the direction of gluconeogenesis, i.e., from the bottom up.

  6. Improvement and characterization of a hyperthermophilic glucose isomerase from Thermoanaerobacter ethanolicus and its application in production of high fructose corn syrup.

    Science.gov (United States)

    Liu, Zhi-Qiang; Zheng, Wei; Huang, Jian-Feng; Jin, Li-Qun; Jia, Dong-Xu; Zhou, Hai-Yan; Xu, Jian-Miao; Liao, Cheng-Jun; Cheng, Xin-Ping; Mao, Bao-Xing; Zheng, Yu-Guo

    2015-08-01

    High fructose corn syrup (HFCS) is an alternative of liquid sweetener to sucrose that is isomerized by commercial glucose isomerase (GI). One-step production of 55 % HFCS by thermostable GI has been drawn more and more attentions. In this study, a new hyperthermophilic GI from Thermoanaerobacter ethanolicus CCSD1 (TEGI) was identified by genome mining, and then a 1317 bp fragment encoding the TEGI was synthesized and expressed in Escherichia coli BL21(DE3). To improve the activity of TEGI, two amino acid residues, Trp139 and Val186, around the active site and substrate-binding pocket based on the structural analysis and molecular docking were selected for site-directed mutagenesis. The specific activity of mutant TEGI-W139F/V186T was 2.3-fold and the value of k cat/K m was 1.86-fold as compared to the wild type TEGI, respectively. Thermostability of mutant TEGI-W139F/V186T at 90 °C for 24 h showed 1.21-fold extension than that of wild type TEGI. During the isomerization of glucose to fructose, the yield of fructose could maintain above 55.4 % by mutant TEGI-W139F/V186T as compared to 53.8 % by wild type TEGI at 90 °C. This study paved foundation for the production of 55 % HFCS using the thermostable TEGI.

  7. Fiscal 1999 achievement report on research and development project on intellectual infrastructure creation and utilization technologies. Development of efficient protein expression system (Development of efficient protein expression system utilizing protein folding mechanism of hyperthermophilic bacteria); 1999 nendo kokoritsu tanpakushitsu hatsugen system no kaihatsu seika hokokusho. Chokonetsukin no tanpakushitsu oritatami kiko wo riyoshita kokoritsu tanpakushitsu hatsugen system no kaihatsu

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2000-03-01

    Efforts were exerted to achieve efficient expression of proteins of hyperthermophilic bacteria, hyperthermophilic archaeabacteria in particular, using a heterogene expression system in which Escherichia coli was the host. In an effort to search for genes related to protein folding and to elucidate the mechanism of folding, chaperonin and prefoldin subunit genes, out of various factors participating in protein folding in hyperthermophilic archaeabacteria, were cloned, and expressed in Escherichia coli. As a system for analyzing protein folding reaction, an experimental system was established on a substrate comprising isopropyl malate dehydrogenase, citrate synthase, glucose dehydrogenase, and a green fluorescent protein. Studies were further conducted to elucidate the mechanism of expression of enzyme genes in Escherichia coli for the establishment of a mass production method for useful enzymes. Also carried out was the research and development of an element technology evaluation system involving protein expression. (NEDO)

  8. Sulfur-inhibited Thermosphaera aggregans sp. nov., a new genus of hyperthermophilic archaea isolated after its prediction from environmentally derived 16S rRNA sequences.

    Science.gov (United States)

    Huber, R; Dyba, D; Huber, H; Burggraf, S; Rachel, R

    1998-01-01

    Recently, a new procedure was developed which allowed for the first time the isolation of a hyperthermophilic archaeum tracked by 165 rRNA analysis from a terrestrial hot solfataric spring ('Obsidian Pool', Yellowstone National Park, WY, USA). This novel isolate is characterized here. Cells are round cocci with a diameter of 0.2-0.8 micron, occurring singly, in pairs, short chains and in grape-like aggregates. The aggregates exhibit a weak bluish-green fluorescence under UV radiation at 420 nm. The new isolate is an anaerobic obligate heterotroph, using preferentially yeast extract for growth. The metabolic products include CO2, H2, acetate and isovalerate. Growth is observed between 65 and 90 degrees C (optimum: 85 degrees C), from pH 5.0 to 7.0 (optimum: 6.5) and up to 0.7% NaCl. The apparent activation energy for growth is about 149 kJ mol-1. Elemental sulfur or hydrogen inhibits growth. The core lipids consist mainly of acyclic and cyclic glycerol diphytanyl tetraethers. The cell envelope contains a cytoplasmic membrane covered by an amorphous layer of unknown composition; there is no evidence for a regularly arrayed surface-layer protein. The G + C content is 46 mol%. On the basis of 165 rRNA sequence comparisons in combination with morphological, physiological and biochemical properties, the isolate represents a new genus within the Desulfurococcaceae, which has been named Thermosphaera. The type species is Thermosphaera aggregans, the type strain is isolate M11TLT (= DSM 11486T).

  9. Deletion of acetyl-CoA synthetases I and II increases production of 3-hydroxypropionate by the metabolically-engineered hyperthermophile Pyrococcus furiosus.

    Science.gov (United States)

    Thorgersen, Michael P; Lipscomb, Gina L; Schut, Gerrit J; Kelly, Robert M; Adams, Michael W W

    2014-03-01

    The heterotrophic, hyperthermophilic archaeon Pyrococcus furiosus is a new addition to the growing list of genetically-tractable microorganisms suitable for metabolic engineering to produce liquid fuels and industrial chemicals. P. furiosus was recently engineered to generate 3-hydroxypropionate (3-HP) from CO₂ and acetyl-CoA by the heterologous-expression of three enzymes from the CO₂ fixation cycle of the thermoacidophilic archaeon Metallosphaera sedula using a thermally-triggered induction system. The acetyl-CoA for this pathway is generated from glucose catabolism that in wild-type P. furiosus is converted to acetate with concurrent ATP production by the heterotetrameric (α₂β₂) acetyl-CoA synthetase (ACS). Hence ACS in the engineered 3-HP production strain (MW56) competes with the heterologous pathway for acetyl-CoA. Herein we show that strains of MW56 lacking the α-subunit of either of the two ACSs previously characterized from P. furiosus (ACSI and ACSII) exhibit a three-fold increase in specific 3-HP production. The ΔACSIα strain displayed only a minor defect in growth on either maltose or peptides, while no growth defect on these substrates was observed with the ΔACSIIα strain. Deletion of individual and multiple ACS subunits was also shown to decrease CoA release activity for several different CoA ester substrates in addition to acetyl-CoA, information that will be extremely useful for future metabolic engineering endeavors in P. furiosus. Copyright © 2014 International Metabolic Engineering Society. All rights reserved.

  10. Crystal structure of the NADP+ and tartrate-bound complex of L-serine 3-dehydrogenase from the hyperthermophilic archaeon Pyrobaculum calidifontis.

    Science.gov (United States)

    Yoneda, Kazunari; Sakuraba, Haruhiko; Araki, Tomohiro; Ohshima, Toshihisa

    2018-05-01

    A gene encoding L-serine dehydrogenase (L-SerDH) that exhibits extremely low sequence identity to the Agrobacterium tumefaciens L-SerDH was identified in the hyperthermophilic archaeon Pyrobaculum calidifontis. The predicted amino acid sequence showed 36% identity with that of Pseudomonas aeruginosa L-SerDH, suggesting that P. calidifontis L-SerDH is a novel type of L-SerDH, like Ps. aeruginosa L-SerDH. The overexpressed enzyme appears to be the most thermostable L-SerDH described to date, and no loss of activity was observed by incubation for 30 min at temperatures up to 100 °C. The enzyme showed substantial reactivity towards D-serine, in addition to L-serine. Two different crystal structures of P. calidifontis L-SerDH were determined using the Se-MAD and MR method: the structure in complex with NADP + /sulfate ion at 1.18 Å and the structure in complex with NADP + /L-tartrate (substrate analog) at 1.57 Å. The fold of the catalytic domain showed similarity with that of Ps. aeruginosa L-SerDH. However, the active site structure significantly differed between the two enzymes. Based on the structure of the tartrate, L- and D-serine and 3-hydroxypropionate molecules were modeled into the active site and the substrate binding modes were estimated. A structural comparison suggests that the wide cavity at the substrate binding site is likely responsible for the high reactivity of the enzyme toward both L- and D-serine enantiomers. This is the first description of the structure of the novel type of L-SerDH with bound NADP + and substrate analog, and it provides new insight into the substrate binding mechanism of L-SerDH. The results obtained here may be very informative for the creation of L- or D-serine-specific SerDH by protein engineering.

  11. Hydrogen production from paper sludge hydrolysate

    NARCIS (Netherlands)

    Kádár, Z.; Vrije, de G.J.; Budde, M.A.W.; Szengyel, Z.; Reczey, K.; Claassen, P.A.M.

    2003-01-01

    The main objective of this study was to develop a system for the production of 'renewable' hydrogen. Paper sludge is a solid industrial waste yielding mainly cellulose, which can be used, after hydrolysis, as a feedstock in anaerobic fermentation by (hyper)thermophilic organisms, such as Thermotoga

  12. Fiscal 1999 R and D project report on intellectual base creation and use technology. Development of the efficient expression system of proteins. Part 1 (Development of the system for hyperthermophilic proteins); 1999 nendo kokoritsu tanpakushitsu hatsugen system no kaihatsu gyomu seika hokokusho. 1. Chokonetsukin yurai tanpakushitsu wo kokoritsu ni hatsugensuru system no kaihatsu

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2000-03-01

    R and D were made on the efficient expression system of hyperthermophilic proteins. Hyperthermophilic strains living in the limited tropical zone of the earth can produce heat- resistant enzyme group with activity even at more than 90 degrees C. To utilize the effective information obtained from analysis of these genomes for industries, the base arrangement of all genomes of P.horikoshii OT3 has been opened. For the efficient expression of hyperthermophilic proteins in Escherichia coli, enzyme PhFEN was improved. For Bacillus strains, new host strains were screened. Expression of several genes from hyperthermophile, P.horikoshii OT3 was tried to be expressed in T.thermophilus using expression vector pTEV131. 8 genes were selected to be expressed using T.thermophilus as a host for independent insertion of every gene. 7 genes except the gene encoding DNA polymerase I were introduced into T.thermophilus as expression plasmid, and 5 genes were also expressed active oxygen. This R and D can largely contribute to development of genome informatics technology based on DNA analysis data. (NEDO)

  13. Structural studies of substrate and product complexes of 5-aminolaevulinic acid dehydratase from humans, Escherichia coli and the hyperthermophile Pyrobaculum calidifontis.

    Science.gov (United States)

    Mills-Davies, N; Butler, D; Norton, E; Thompson, D; Sarwar, M; Guo, J; Gill, R; Azim, N; Coker, A; Wood, S P; Erskine, P T; Coates, L; Cooper, J B; Rashid, N; Akhtar, M; Shoolingin-Jordan, P M

    2017-01-01

    A number of X-ray analyses of an enzyme involved in a key early stage of tetrapyrrole biosynthesis are reported. Two structures of human 5-aminolaevulinate dehydratase (ALAD), native and recombinant, have been determined at 2.8 Å resolution, showing that the enzyme adopts an octameric quaternary structure in accord with previously published analyses of the enzyme from a range of other species. However, this is in contrast to the finding that a disease-related F12L mutant of the human enzyme uniquely forms hexamers [Breinig et al. (2003), Nature Struct. Biol. 10, 757-763]. Monomers of all ALADs adopt the TIM-barrel fold; the subunit conformation that assembles into the octamer includes the N-terminal tail of one monomer curled around the (α/β) 8 barrel of a neighbouring monomer. Both crystal forms of the human enzyme possess two monomers per asymmetric unit, termed A and B. In the native enzyme there are a number of distinct structural differences between the A and B monomers, with the latter exhibiting greater disorder in a number of loop regions and in the active site. In contrast, the second monomer of the recombinant enzyme appears to be better defined and the active site of both monomers clearly possesses a zinc ion which is bound by three conserved cysteine residues. In native human ALAD, the A monomer also has a ligand resembling the substrate ALA which is covalently bound by a Schiff base to one of the active-site lysines (Lys252) and is held in place by an ordered active-site loop. In contrast, these features of the active-site structure are disordered or absent in the B subunit of the native human enzyme. The octameric structure of the zinc-dependent ALAD from the hyperthermophile Pyrobaculum calidifontis is also reported at a somewhat lower resolution of 3.5 Å. Finally, the details are presented of a high-resolution structure of the Escherichia coli ALAD enzyme co-crystallized with a noncovalently bound moiety of the product, porphobilinogen (PBG

  14. Modos de vida maritima en Europa

    DEFF Research Database (Denmark)

    Højrup, Thomas; Schriewer, Klaus

    2013-01-01

    Analyse og syntese af de biologiske livsformers betydning som mulighedsbetingelse for de anvendte teknologier og fangstmåders betydning som mulighedsbetingelse for de to produktionsmåders sameksistens i euroæisk fiskeri i 500 år og deres betydning som mulighedsbetingelser for de sameksisterende k...

  15. AcEST: DK952773 [AcEST

    Lifescience Database Archive (English)

    Full Text Available 0Y1|P1254_THEMA Phosphorylated carbohydrates phosphatase T... 58 4e-08 sp|P77247|YNIC_ECOLI Phosphatase yniC...hydrates phosphatase TM_1254 OS=Thermotoga maritima GN=TM_1254 PE=1 SV=1 Length = 2...PELYLLAAKNLGVSPAECLAFEDSVNGSIAAKRAGMKCVI 183 Query: 515 TKKRV 529 +V Sbjct: 184 VPNKV 188 >sp|Q9X0Y1|P1254_THEMA Phosphorylated carbo

  16. AcEST: DK952111 [AcEST

    Lifescience Database Archive (English)

    Full Text Available P1254_THEMA Phosphorylated carbohydrates phosphatase T... 33 0.91 sp|P97347|RPTN_MOUSE Repetin OS=Mus muscul...----RPGVEAYLNAAKDLGLKIGL 106 Query: 582 CSTSNELAVS 611 S+S+ VS Sbjct: 107 ASSSDYKWVS 116 >sp|Q9X0Y1|P1254_THEMA Phosphorylated carboh...ydrates phosphatase TM_1254 OS=Thermotoga maritima GN=TM_1254 PE=1 SV=1 Length = 21

  17. Development of soluble and immobilized biocatalysts based on a recombinant thermostable ß-fructosidase enabling complete sucrose inversion at pasteurization temperatures

    OpenAIRE

    Menéndez, Carmen; Martínez, Duniesky; Trujillo, Luis E; Ramírez, Ricardo; Sobrino, Alina; Cutiño-Ávila, Bessy V; Basabe, Liliana; del Monte-Martínez, Alberto; Pérez, Enrique R; Hernández, Lázaro

    2014-01-01

    Biocatalysts for the industrial production of invert sugar are preferred to stably operate at high sucrose concentrations and pasteurization temperatures. Thermotoga maritima ß-fructosidase (BfrA) is more thermostable and less susceptible to substrate inhibition than the current commercial invertase from Saccharomyces cerevisiae. In this research, the non-saccharolytic host Pichia pastoris was engineered for BfrA production. Fed-batch fermentation of the recombinant yeast for 72 h using cane ...

  18. Saccharolobus caldissimus gen. nov., sp. nov., a facultatively anaerobic iron-reducing hyperthermophilic archaeon isolated from an acidic terrestrial hot spring, and reclassification of Sulfolobus solfataricus as Saccharolobus solfataricus comb. nov. and Sulfolobus shibatae as Saccharolobus shibatae comb. nov.

    Science.gov (United States)

    Sakai, Hiroyuki D; Kurosawa, Norio

    2018-04-01

    A novel hyperthermophilic archaeon of strain HS-3 T , belonging to the family Sulfolobaceae, was isolated from an acidic terrestrial hot spring in Hakone Ohwaku-dani, Japan. Based on 16S rRNA gene sequence analysis, the closest phylogenetic relatives of strain HS-3 T were, first, Sulfolobus solfataricus (96.4 %) and, second, Sulfolobus shibatae (96.2 %), indicating that the strain belongs to the genus Sulfolobus. However, the sequence similarity to the type species of the genus Sulfolobus (Sulfolobus acidocaldarius) was remarkably low (91.8 %). In order to determine whether strain HS-3 T belongs to the genus Sulfolobus, its morphological, biochemical and physiological characteristics were examined in parallel with those of S. solfataricus and S. shibatae. Although there were some differences in chemolithotrophic growth between strain HS-3 T , S. solfataricus and S. shibatae, their temperature, pH and facultatively anaerobic characteristics of growth, and their utilization of various sugars were almost identical. In contrast, the utilization of various sugars by S. acidocaldarius was quite different from that of HS-3 T , S. solfataricus and S. shibatae. Phylogenetic evidence based on the 16S and the 23S rRNA gene sequences also clearly distinguished the monophyletic clade composed of strain HS-3 T , S. solfataricus, and S. shibatae from S. acidocaldarius. Based on these results, we propose a new genus and species, Saccharolobus caldissimus gen. nov., sp. nov., for strain HS-3 T , as well as two reclassifications, Saccharolobus solfataricus comb. nov. and Saccharolobus shibatae comb. nov. The type strain of Saccharolobus caldissimus is HS-3 T (=JCM 32116 T and InaCC Ar80 T ). The type species of the genus is Saccharolobus solfataricus.

  19. Crystal structure of the regulatory subunit of archaeal initiation factor 2B (aIF2B) from hyperthermophilic archaeon Pyrococcus horikoshii OT3: a proposed structure of the regulatory subcomplex of eukaryotic IF2B

    International Nuclear Information System (INIS)

    Kakuta, Yoshimitsu; Tahara, Maino; Maetani, Shigehiro; Yao, Min; Tanaka, Isao; Kimura, Makoto

    2004-01-01

    Eukaryotic translation initiation factor 2B (eIF2B) is the guanine-nucleotide exchange factor for eukaryotic initiation factor 2 (eIF2). eIF2B is a heteropentameric protein composed of α-ε subunits. The α, β, and δ subunits form a regulatory subcomplex, while the γ and ε form a catalytic subcomplex. Archaea possess homologues of α, β, and δ subunits of eIF2B. Here, we report the three-dimensional structure of an archaeal regulatory subunit (aIF2Bα) from the hyperthermophilic archaeon Pyrococcus horikoshii OT3 determined by X-ray crystallography at 2.2 A resolution. aIF2Bα consists of two subdomains, an N-domain (residues 1-95) and a C-domain (residues 96-276), connected by a long α-helix (α5: 78-106). The N-domain contains a five helix bundle structure, while the C-domain folds into the α/β structure, thus showing similarity to D-ribose-5-phosphate isomerase structure. The presence of two molecules in the crystallographic asymmetric unit and the gel filtration analysis suggest a dimeric structure of aIF2Bα in solution, interacting with each other by C-domains. Furthermore, the crystallographic 3-fold symmetry generates a homohexameric structure of aIF2Bα; the interaction is primarily mediated by the long α-helix at the N-domains. This structure suggests an architecture of the three subunits, α, β, and δ, in the regulatory subcomplex within eIF2B

  20. Fiscal 2000 achievement report on project for research and development of technologies for intelligent infrastructure creation and utilization. 'Development of high-efficiency protein expression system - 1 Development of system capable of high-efficiency expression of hyperthermophile-derived protein'; 2000 nendo chiteki kiban sose riyo gijutsu kenkyu kaihatsu gyomu seika hokokusho. Kokoritsu tanpakushitsu hatsugen system no kaihatsu -1 (Cho konetsukin yurai tanpakushitsu wo kokoritsu ni hatsugen suru system no kaihatsu)

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2001-03-01

    Research and development was conducted aiming at the establishment of a system to enable the high-efficiency expression of the gene products of P. horikoshii OT3 and A. pernix K1. In an effort to develop a high-efficiency protein expression system with Escherichia coli acting as the host, studies were made about the expression of hyperthermophile protein by arginine rare codon elimination, and Ph FEN (flap endonuclease) was successfully overexpressed. In the development of Bacillus strains, screening was conducted for novel hosts, and a library was constructed for a screening task suitable for hyperthermophile-derived protein production. A system was also constructed capable of the high-throughput expression of various kinds of genes using Bacillus brevis. In the study of the expression of hyperthermophile-derived genes using T. thermophilus, promoter replacement resulted in an approximately 2-fold increase in representation at the maximum. Moreover, studies were made about the length at which foreign genes were efficiently incorporated into the T. thermophilus genome. (NEDO)

  1. Pretreatment of Miscanthus for hydrogen production by Thermotoga elfii

    NARCIS (Netherlands)

    Vrije, de T.; Haas, de G.G.; Tan, G.B.; Keijsers, E.R.P.; Claassen, P.A.M.

    2002-01-01

    Pretreatment methods for the production of fermentable substrates from Miscanthus, a lignocellulosic biomass, were investigated. Results demonstrated an inverse relationship between lignin content and the efficiency of enzymatic hydrolysis of polysaccharides. High delignification values were

  2. Over-expression of xylanolytic α-glucuronidase from Thermotoga ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-07-18

    Jul 18, 2008 ... ... at http://www.academicjournals.org/AJB. ISSN 1684–5315 © 2008 Academic Journals .... version 6.0 of the sequence analysis software package (Lynnon. Biosoft, USA). .... the xylan supernatant by nylon fabric. The resulting ...

  3. Cell architecture and flagella of hyperthermophilic Archaea

    OpenAIRE

    Bellack, Annett

    2011-01-01

    Earlier studies indicated that flagella might play a crucial role in motility, adhesion, and cell-cell contacts of Archaea. Thus, the ultrastructural and functional characterization of flagella and their anchoring in the cell are crucial for understanding the archaeal cell organization in general. To address this topic, Pyrococcus furiosus was chosen as a suitable model organism. However, in the course of this study, morphological changes of this strain, cultured continuously for several y...

  4. Structure and mechanism of a bacterial t6A biosynthesis system

    OpenAIRE

    Luthra, Amit; Swinehart, William; Bayooz, Susan; Phan, Phuc; Stec, Boguslaw; Iwata-Reuyl, Dirk; Swairjo, Manal A

    2018-01-01

    Abstract The universal N(6)-threonylcarbamoyladenosine (t6A) modification at position 37 of ANN-decoding tRNAs is central to translational fidelity. In bacteria, t6A biosynthesis is catalyzed by the proteins TsaB, TsaC/TsaC2, TsaD and TsaE. Despite intense research, the molecular mechanisms underlying t6A biosynthesis are poorly understood. Here, we report biochemical and biophysical studies of the t6A biosynthesis system from Thermotoga maritima. Small angle X-ray scattering analysis reveals...

  5. GTPase activity, structure, and mechanical properties of filaments assembled from bacterial cytoskeleton protein MreB.

    Science.gov (United States)

    Esue, Osigwe; Wirtz, Denis; Tseng, Yiider

    2006-02-01

    MreB, a major component of the recently discovered bacterial cytoskeleton, displays a structure homologous to its eukaryotic counterpart actin. Here, we study the assembly and mechanical properties of Thermotoga maritima MreB in the presence of different nucleotides in vitro. We found that GTP, not ADP or GDP, can mediate MreB assembly into filamentous structures as effectively as ATP. Upon MreB assembly, both GTP and ATP release the gamma phosphate at similar rates. Therefore, MreB is an equally effective ATPase and GTPase. Electron microscopy and quantitative rheology suggest that the morphologies and micromechanical properties of filamentous ATP-MreB and GTP-MreB are similar. In contrast, mammalian actin assembly is favored in the presence of ATP over GTP. These results indicate that, despite high structural homology of their monomers, T. maritima MreB and actin filaments display different assembly, morphology, micromechanics, and nucleotide-binding specificity. Furthermore, the biophysical properties of T. maritima MreB filaments, including high rigidity and propensity to form bundles, suggest a mechanism by which MreB helical structure may be involved in imposing a cylindrical architecture on rod-shaped bacterial cells.

  6. In silico method for modelling metabolism and gene product expression at genome scale

    Energy Technology Data Exchange (ETDEWEB)

    Lerman, Joshua A.; Hyduke, Daniel R.; Latif, Haythem; Portnoy, Vasiliy A.; Lewis, Nathan E.; Orth, Jeffrey D.; Rutledge, Alexandra C.; Smith, Richard D.; Adkins, Joshua N.; Zengler, Karsten; Palsson, Bernard O.

    2012-07-03

    Transcription and translation use raw materials and energy generated metabolically to create the macromolecular machinery responsible for all cellular functions, including metabolism. A biochemically accurate model of molecular biology and metabolism will facilitate comprehensive and quantitative computations of an organism's molecular constitution as a function of genetic and environmental parameters. Here we formulate a model of metabolism and macromolecular expression. Prototyping it using the simple microorganism Thermotoga maritima, we show our model accurately simulates variations in cellular composition and gene expression. Moreover, through in silico comparative transcriptomics, the model allows the discovery of new regulons and improving the genome and transcription unit annotations. Our method presents a framework for investigating molecular biology and cellular physiology in silico and may allow quantitative interpretation of multi-omics data sets in the context of an integrated biochemical description of an organism.

  7. γ-irradiation resistance and UV-sensitivity of extremely thermophilic archebacteria and eubacteria

    International Nuclear Information System (INIS)

    Kopylov, V.M.; Bonch-Osmolovskaya, E.A.; Svetlichnyi, V.A.; Miroshnichenko, M.L.; Skobkin, V.S.

    1993-01-01

    Cells of extremely thermophilic sulfur-dependent archebacteria Desulfurococcus amylolyticus Z533 and Thermococcus stelleri K15 are resistant to γ-irradiation. These archebacteria survive γ-irradiation at a dose of up to 5 kGy but are no longer viable after 8-9 kGy. Comparison of the survival profiles showed that archebacteria are 12 to 25 times more resistant to γ-irradiation at moderate doses (LD 50 and LD 90 ) than E. coli K12 but are 2 to 2.5 times more sensitive than D. radiodurans. γ-irradiation at a dose of 1 to 2.5 kGy killed extremely thermophilic anaerobic eubacteria Thermotoga maritima 2706 and Thermodesulfobacterium P. All extreme thermophiles studied were more sensitive to UV-irradiation than E. coli

  8. Converting a Natural Protein Compartment into a Nanofactory for the Size-Constrained Synthesis of Antimicrobial Silver Nanoparticles.

    Science.gov (United States)

    Giessen, Tobias W; Silver, Pamela A

    2016-12-16

    Engineered biological systems are used extensively for the production of high value and commodity organics. On the other hand, most inorganic nanomaterials are still synthesized via chemical routes. By engineering cellular compartments, functional nanoarchitectures can be produced under environmentally sustainable conditions. Encapsulins are a new class of microbial nanocompartments with promising applications in nanobiotechnology. Here, we engineer the Thermotoga maritima encapsulin EncTm to yield a designed compartment for the size-constrained synthesis of silver nanoparticles (Ag NPs). These Ag NPs exhibit uniform shape and size distributions as well as long-term stability. Ambient aqueous conditions can be used for Ag NP synthesis, while no reducing agents or solvents need to be added. The antimicrobial activity of the synthesized protein-coated or shell-free Ag NPs is superior to that of silver nitrate and citrate-capped Ag NPs. This study establishes encapsulins as an engineerable platform for the synthesis of biogenic functional nanomaterials.

  9. The Deinococcus-Thermus phylum and the effect of rRNA composition on phylogenetic tree construction

    Science.gov (United States)

    Weisburg, W. G.; Giovannoni, S. J.; Woese, C. R.

    1989-01-01

    Through comparative analysis of 16S ribosomal RNA sequences, it can be shown that two seemingly dissimilar types of eubacteria Deinococcus and the ubiquitous hot spring organism Thermus are distantly but specifically related to one another. This confirms an earlier report based upon 16S rRNA oligonucleotide cataloging studies (Hensel et al., 1986). Their two lineages form a distinctive grouping within the eubacteria that deserved the taxonomic status of a phylum. The (partial) sequence of T. aquaticus rRNA appears relatively close to those of other thermophilic eubacteria. e.g. Thermotoga maritima and Thermomicrobium roseum. However, this closeness does not reflect a true evolutionary closeness; rather it is due to a "thermophilic convergence", the result of unusually high G+C composition in the rRNAs of thermophilic bacteria. Unless such compositional biases are taken into account, the branching order and root of phylogenetic trees can be incorrectly inferred.

  10. Dicty_cDB: Contig-U05179-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 07 AE000512_160( AE000512 |pid:none) Thermotoga maritima MSB8, comple... 59 3e-07 AM494956_53( AM494956 |pid...5462316567 Global-Ocean-Sampling_GS-31-01-01-1... 36 4.7 3 ( CU675067 ) Synthetic construct Homo sapiens gat...4 0.15 2 ( EK365242 ) 1095469394395 Global-Ocean-Sampling_GS-31-01-01-1... 50 0.19 1 ( DQ832718 ) Phytophtho... F... 38 0.55 2 ( EK172602 ) 1095458082313 Global-Ocean-Sampling_GS-31-01-01-1...... 34 0.55 2 ( EK226550 ) 1095460160991 Global-Ocean-Sampling_GS-31-01-01-1... 34 0.55 2 ( ES217923 ) MpFVN_ag2_E11 Myzus persicae, li

  11. Boosting dark fermentation with co-cultures of extreme thermophiles for biohythane production from garden waste.

    Science.gov (United States)

    Abreu, Angela A; Tavares, Fábio; Alves, Maria Madalena; Pereira, Maria Alcina

    2016-11-01

    Proof of principle of biohythane and potential energy production from garden waste (GW) is demonstrated in this study in a two-step process coupling dark fermentation and anaerobic digestion. The synergistic effect of using co-cultures of extreme thermophiles to intensify biohydrogen dark fermentation is demonstrated using xylose, cellobiose and GW. Co-culture of Caldicellulosiruptor saccharolyticus and Thermotoga maritima showed higher hydrogen production yields from xylose (2.7±0.1molmol(-1) total sugar) and cellobiose (4.8±0.3molmol(-1) total sugar) compared to individual cultures. Co-culture of extreme thermophiles C. saccharolyticus and Caldicellulosiruptor bescii increased synergistically the hydrogen production yield from GW (98.3±6.9Lkg(-1) (VS)) compared to individual cultures and co-culture of T. maritima and C. saccharolyticus. The biochemical methane potential of the fermentation end-products was 322±10Lkg(-1) (CODt). Biohythane, a biogas enriched with 15% hydrogen could be obtained from GW, yielding a potential energy generation of 22.2MJkg(-1) (VS). Copyright © 2016 Elsevier Ltd. All rights reserved.

  12. The assembly of MreB, a prokaryotic homolog of actin.

    Science.gov (United States)

    Esue, Osigwe; Cordero, Maria; Wirtz, Denis; Tseng, Yiider

    2005-01-28

    MreB, a major component of the bacterial cytoskeleton, exhibits high structural homology to its eukaryotic counterpart actin. Live cell microscopy studies suggest that MreB molecules organize into large filamentous spirals that support the cell membrane and play a key shape-determining function. However, the basic properties of MreB filament assembly remain unknown. Here, we studied the assembly of Thermotoga maritima MreB triggered by ATP in vitro and compared it to the well-studied assembly of actin. These studies show that MreB filament ultrastructure and polymerization depend crucially on temperature as well as the ions present on solution. At the optimal growth temperature of T. maritima, MreB assembly proceeded much faster than that of actin, without nucleation (or nucleation is highly favorable and fast) and with little or no contribution from filament end-to-end annealing. MreB exhibited rates of ATP hydrolysis and phosphate release similar to that of F-actin, however, with a critical concentration of approximately 3 nm, which is approximately 100-fold lower than that of actin. Furthermore, MreB assembled into filamentous bundles that have the ability to spontaneously form ring-like structures without auxiliary proteins. These findings suggest that despite high structural homology, MreB and actin display significantly different assembly properties.

  13. Bacterial actin MreB assembles in complex with cell shape protein RodZ.

    Science.gov (United States)

    van den Ent, Fusinita; Johnson, Christopher M; Persons, Logan; de Boer, Piet; Löwe, Jan

    2010-03-17

    Bacterial actin homologue MreB is required for cell shape maintenance in most non-spherical bacteria, where it assembles into helical structures just underneath the cytoplasmic membrane. Proper assembly of the actin cytoskeleton requires RodZ, a conserved, bitopic membrane protein that colocalises to MreB and is essential for cell shape determination. Here, we present the first crystal structure of bacterial actin engaged with a natural partner and provide a clear functional significance of the interaction. We show that the cytoplasmic helix-turn-helix motif of Thermotoga maritima RodZ directly interacts with monomeric as well as filamentous MreB and present the crystal structure of the complex. In vitro and in vivo analyses of mutant T. maritima and Escherichia coli RodZ validate the structure and reveal the importance of the MreB-RodZ interaction in the ability of cells to propagate as rods. Furthermore, the results elucidate how the bacterial actin cytoskeleton might be anchored to the membrane to help constrain peptidoglycan synthesis in the periplasm.

  14. Primordial-like enzymes from bacteria with reduced genomes.

    Science.gov (United States)

    Ferla, Matteo P; Brewster, Jodi L; Hall, Kelsi R; Evans, Gary B; Patrick, Wayne M

    2017-08-01

    The first cells probably possessed rudimentary metabolic networks, built using a handful of multifunctional enzymes. The promiscuous activities of modern enzymes are often assumed to be relics of this primordial era; however, by definition these activities are no longer physiological. There are many fewer examples of enzymes using a single active site to catalyze multiple physiologically-relevant reactions. Previously, we characterized the promiscuous alanine racemase (ALR) activity of Escherichia coli cystathionine β-lyase (CBL). Now we have discovered that several bacteria with reduced genomes lack alr, but contain metC (encoding CBL). We characterized the CBL enzymes from three of these: Pelagibacter ubique, the Wolbachia endosymbiont of Drosophila melanogaster (wMel) and Thermotoga maritima. Each is a multifunctional CBL/ALR. However, we also show that CBL activity is no longer required in these bacteria. Instead, the wMel and T. maritima enzymes are physiologically bi-functional alanine/glutamate racemases. They are not highly active, but they are clearly sufficient. Given the abundance of the microorganisms using them, we suggest that much of the planet's biochemistry is carried out by enzymes that are quite different from the highly-active exemplars usually found in textbooks. Instead, primordial-like enzymes may be an essential part of the adaptive strategy associated with streamlining. © 2017 The Authors. Molecular Microbiology Published by John Wiley & Sons Ltd.

  15. Enige opmerkingen over de aanpassing van de zeeraket (Cakile maritima Scop.) aan het strand

    NARCIS (Netherlands)

    Veldkamp, J.F.

    1971-01-01

    It has been assumed that the sea-rockets, Cakile Mill., are adapted to life on the beach. It is here suggested that the habit of the plants, their annual cycle, the structure of the fruit and the consequent distribution by the sea cannot be regarded as typical, special adaptations to the sandy

  16. Effect of NaCl salinity on nitrate uptake in Plantago maritima L.

    NARCIS (Netherlands)

    Rubinigg, Michael; Posthumus, F.S; Elzenga, J.T.M.; Stulen, I.

    2005-01-01

    Exposure of plants to NaCl salinity reduces the rate of nitrate net uptake by the roots. Previous studies showed that this effect was due to a reduced nitrate influx, which could only partially be explained by a lower demand of nitrate for growth. In the present work we tested the hypothesis that

  17. Restoration Potential of Ruppia Maritima and Potamogeton Perfoliatus by Seed in the Mid-Chesapeake Bay

    National Research Council Canada - National Science Library

    Ailstock, Steve

    2004-01-01

    ... in the mesohaline reaches of the mid-Chesapeake Bay. Once reproductive potential by seed is defined for healthy populations of these species, their life cycles can be evaluated to identify nondestructive methods of harvesting seeds for restoration projects...

  18. How do Purple Sandpipers Calidris maritima survive the winter north of the Arctic circle?

    NARCIS (Netherlands)

    Summers, R.W.; Piersma, T.; Strann, K.B.; Wiersma, P.

    1998-01-01

    Winter north of the Arctic circle in northern Norway is colder, windier and there is less solar radiation than in eastern Scotland, at a latitude 13 degrees further south. We predicted from equations derived from heated taxidermic mounts that the maintenance metabolism (Basal Metabolic Rate plus

  19. Thermostability promotes the cooperative function of split adenylate kinases.

    Science.gov (United States)

    Nguyen, Peter Q; Liu, Shirley; Thompson, Jeremy C; Silberg, Jonathan J

    2008-05-01

    Proteins can often be cleaved to create inactive polypeptides that associate into functional complexes through non-covalent interactions, but little is known about what influences the cooperative function of the ensuing protein fragments. Here, we examine whether protein thermostability affects protein fragment complementation by characterizing the function of split adenylate kinases from the mesophile Bacillus subtilis (AKBs) and the hyperthermophile Thermotoga neapolitana (AKTn). Complementation studies revealed that the split AKTn supported the growth of Escherichia coli with a temperature-sensitive AK, but not the fragmented AKBs. However, weak complementation occurred when the AKBs fragments were fused to polypeptides that strongly associate, and this was enhanced by a Q16L mutation that thermostabilizes the full-length protein. To examine how the split AK homologs differ in structure and function, their catalytic activity, zinc content, and circular dichroism spectra were characterized. The reconstituted AKTn had higher levels of zinc, greater secondary structure, and >10(3)-fold more activity than the AKBs pair, albeit 17-fold less active than full-length AKTn. These findings provide evidence that the design of protein fragments that cooperatively function can be improved by choosing proteins with the greatest thermostability for bisection, and they suggest that this arises because hyperthermophilic protein fragments exhibit greater residual structure compared to their mesophilic counterparts.

  20. Functional characterization and crystal structure of thermostable amylase from Thermotoga petrophila, reveals high thermostability and an unusual form of dimerization

    DEFF Research Database (Denmark)

    Hameed, Uzma; Price, Ian; Ikram-Ul-Haq

    2017-01-01

    and able to hydrolyze starch into dextrin between 90 and 100°C, with optimum activity at 98°C and pH8.5. The activity increased in the presence of Rb(1+), K(1+) and Ca(2+) ions, whereas other ions inhibited activity. The crystal structure of Tp-AmyS at 1.7Å resolution showed common features of the GH-13...... of salivary amylase from a previous crystal structure, and thus could be a functional feature of some amylases....

  1. Efficient hydrogen production from the lignocellulosic energy crop Miscanthus by the extreme thermophilic bacteria Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana

    NARCIS (Netherlands)

    Vrije, de G.J.; Bakker, R.R.; Budde, M.A.W.; Lai, M.H.; Mars, A.E.; Claassen, P.A.M.

    2009-01-01

    The production of hydrogen from biomass by fermentation is one of the routes that can contribute to a future sustainable hydrogen economy. Lignocellulosic biomass is an attractive feedstock because of its abundance, low production costs and high polysaccharide content. Batch cultures of

  2. Thermotoga lettingae sp. nov. : a novel thermophilic, methanol-degrading bacterium isolated from a thermophilic anaerobic reactor

    NARCIS (Netherlands)

    Balk, M.; Weijma, J.; Stams, A.J.M.

    2002-01-01

    A novel, anaerobic, non-spore-forming, mobile, Gram-negative, thermophilic bacterium, strain TMO(T), was isolated from a thermophilic sulfate-reducing bioreactor operated at 65 degrees C with methanol as the sole substrate. The G C content of the DNA of strain TMO(T) was 39.2 molÐThe optimum pH,

  3. A selection that reports on protein-protein interactions within a thermophilic bacterium.

    Science.gov (United States)

    Nguyen, Peter Q; Silberg, Jonathan J

    2010-07-01

    Many proteins can be split into fragments that exhibit enhanced function upon fusion to interacting proteins. While this strategy has been widely used to create protein-fragment complementation assays (PCAs) for discovering protein-protein interactions within mesophilic organisms, similar assays have not yet been developed for studying natural and engineered protein complexes at the temperatures where thermophilic microbes grow. We describe the development of a selection for protein-protein interactions within Thermus thermophilus that is based upon growth complementation by fragments of Thermotoga neapolitana adenylate kinase (AK(Tn)). Complementation studies with an engineered thermophile (PQN1) that is not viable above 75 degrees C because its adk gene has been replaced by a Geobacillus stearothermophilus ortholog revealed that growth could be restored at 78 degrees C by a vector that coexpresses polypeptides corresponding to residues 1-79 and 80-220 of AK(Tn). In contrast, PQN1 growth was not complemented by AK(Tn) fragments harboring a C156A mutation within the zinc-binding tetracysteine motif unless these fragments were fused to Thermotoga maritima chemotaxis proteins that heterodimerize (CheA and CheY) or homodimerize (CheX). This enhanced complementation is interpreted as arising from chemotaxis protein-protein interactions, since AK(Tn)-C156A fragments having only one polypeptide fused to a chemotaxis protein did not complement PQN1 to the same extent. This selection increases the maximum temperature where a PCA can be used to engineer thermostable protein complexes and to map protein-protein interactions.

  4. Stability of endoglucanases from mesophilic fungus and thermophilic bacterium in acidified polyols.

    Science.gov (United States)

    Chong, Barrie Fong; Harrison, Mark D; O'Hara, Ian M

    2014-01-01

    Recent developments in chemical pretreatments of lignocellulosic biomass using polyols as co-solvents (e.g., glycerol and ethylene glycol) at temperatures less than 100°C may allow the effective use of thermostable and non-thermostable cellulases in situ during the saccharification process. The potential of biomass saccharifying enzymes, endoglucanases (EG) from a thermophilic bacterium (Thermotoga maritima) and a mesophilic fungus (Trichoderma longibrachiatum), to retain their activity in aqueous buffer, acidified glycerol, and acidified ethylene glycol used as co-solvents at pretreatment temperatures at or below 100°C were examined. The results show that despite its origin, T. longibrachiatum EG (Tl-EG) retained 75% of its activity after exposure to 100°C for 5 min in aqueous buffer while T. maritima EG (Tm-EG) retained only 5% activity. However, at 90°C both enzymes retained over 87% of their activity. In acidified (0.1% (w/w) H2SO4) glycerol, Tl-EG retained similar activity (80%) to that obtained in glycerol alone, while Tm-EG retained only 35%. With acidified ethylene glycol under these conditions, both Tl-EG and Tm-EG retained 36% of their activity. The results therefore show that Tl-EG is more stable in both acidified glycerol and ethylene glycol than Tm-EG. A preliminary kinetic study showed that pure glycerol improved the thermal stability of Tl-EG but destabilized Tm-EG, relative to the buffer solution. The half-lives of both Tl-EG and Tm-EG are 4.5 min in acidified glycerol, indicating that the effectiveness of these enzymes under typical pretreatment times of greater than 15 min will be considerably diminished. Attempts have been made to explain the differences in the results obtained between the two enzymes. Copyright © 2014 Elsevier Inc. All rights reserved.

  5. Non-complexed four cascade enzyme mixture: simple purification and synergetic co-stabilization.

    Directory of Open Access Journals (Sweden)

    Suwan Myung

    Full Text Available Cell-free biosystems comprised of synthetic enzymatic pathways would be a promising biomanufacturing platform due to several advantages, such as high product yield, fast reaction rate, easy control and access, and so on. However, it was essential to produce (purified enzymes at low costs and stabilize them for a long time so to decrease biocatalyst costs. We studied the stability of the four recombinant enzyme mixtures, all of which originated from thermophilic microorganisms: triosephosphate isomerase (TIM from Thermus thermophiles, fructose bisphosphate aldolase (ALD from Thermotoga maritima, fructose bisphosphatase (FBP from T. maritima, and phosphoglucose isomerase (PGI from Clostridium thermocellum. It was found that TIM and ALD were very stable at evaluated temperature so that they were purified by heat precipitation followed by gradient ammonia sulfate precipitation. In contrast, PGI was not stable enough for heat treatment. In addition, the stability of a low concentration PGI was enhanced by more than 25 times in the presence of 20 mg/L bovine serum albumin or the other three enzymes. At a practical enzyme loading of 1000 U/L for each enzyme, the half-life time of free PGI was prolong to 433 h in the presence of the other three enzymes, resulting in a great increase in the total turn-over number of PGI to 6.2×10(9 mole of product per mole of enzyme. This study clearly suggested that the presence of other proteins had a strong synergetic effect on the stabilization of the thermolabile enzyme PGI due to in vitro macromolecular crowding effect. Also, this result could be used to explain why not all enzymes isolated from thermophilic microorganisms are stable in vitro because of a lack of the macromolecular crowding environment.

  6. Complete Genome Sequence of the Hyperthermophilic Sulfate-Reducing Bacterium Thermodesulfobacterium geofontis OPF15T.

    Science.gov (United States)

    Elkins, James G; Hamilton-Brehm, Scott D; Lucas, Susan; Han, James; Lapidus, Alla; Cheng, Jan-Fang; Goodwin, Lynne A; Pitluck, Sam; Peters, Lin; Mikhailova, Natalia; Davenport, Karen W; Detter, John C; Han, Cliff S; Tapia, Roxanne; Land, Miriam L; Hauser, Loren; Kyrpides, Nikos C; Ivanova, Natalia N; Pagani, Ioanna; Bruce, David; Woyke, Tanja; Cottingham, Robert W

    2013-04-11

    Thermodesulfobacterium geofontis OPF15(T) (ATCC BAA-2454, JCM 18567) was isolated from Obsidian Pool, Yellowstone National Park, and grows optimally at 83°C. The 1.6-Mb genome sequence was finished at the Joint Genome Institute and has been deposited for future genomic studies pertaining to microbial processes and nutrient cycles in high-temperature environments.

  7. A GH57 4-alpha-glucanotransferase of hyperthermophilic origin with potential for alkyl glycoside production

    NARCIS (Netherlands)

    Paul, Catherine J.; Leemhuis, Hans; Dobruchowska, Justyna M.; Grey, Carl; Onnby, Linda; van Leeuwen, Sander S.; Dijkhuizen, Lubbert; Karlsson, Eva Nordberg

    4-alpha-Glucanotransferase (GTase) enzymes (EC 2.4.1.25) modulate the size of alpha-glucans by cleaving and reforming alpha-1,4 glycosidic bonds in alpha-glucans, an essential process in starch and glycogen metabolism in plants and microorganisms. The glycoside hydrolase family 57 enzyme (GTase57)

  8. How hyperthermophiles adapt to change their lives : DNA exchange in extreme conditions

    NARCIS (Netherlands)

    van Wolferen, Marleen; Ajon, Malgorzata; Driessen, Arnold J. M.; Albers, Sonja-Verena; Ajon, Małgorzata; Huang, L.

    Transfer of DNA has been shown to be involved in genome evolution. In particular with respect to the adaptation of bacterial species to high temperatures, DNA transfer between the domains of bacteria and archaea seems to have played a major role. In addition, DNA exchange between similar species

  9. Crystallization of [Fe4S3]-ferredoxin from the hyperthermophile archaeon pyrococcus furiosus

    DEFF Research Database (Denmark)

    Nielsen, Michael Ericsson Skovbo; Harris, Pernille; Christensen, Hans Erik Mølager

    2003-01-01

    Recombinant Pyrococcus furiosus ferredoxin with a [Fe3S4]-cluster was crystallized through steps of optimization and X-ray diffraction data were collected from several crystal forms. Flat plate-like crystals were grown by hanging-drop vapour diffusion. The precipitant used was 30% PEG 400; the p...

  10. DHAP-dependent aldolases from (hyper)thermophiles: biochemistry and applications

    NARCIS (Netherlands)

    Falcicchio, P.; Wolterink-van Loo, S.; Franssen, M.C.R.; Oost, van der J.

    2014-01-01

    Generating new carbon-carbon (C-C) bonds in an enantioselective way is one of the big challenges in organic synthesis. Aldolases are a natural tool for stereoselective C-C bond formation in a green and sustainable way. This review will focus on thermophilic aldolases in general and on

  11. Maillard reactions and increased enzyme inactivation during oligosaccharide synthesis by a hyperthermophilic glycosidase

    NARCIS (Netherlands)

    Bruins, M.E.; Hellemond, van E.W.; Janssen, A.E.M.; Boom, R.M.

    2003-01-01

    The thermostable Pyrococcus furiosus beta-glycosidase was used for oligosaccharide production from lactose in a kinetically controlled reaction. Our experiments showed that higher temperatures are beneficial for the absolute as well as relative oligosaccharide yield. However, at reaction

  12. Production of Recombinant and Tagged Proteins in the Hyperthermophilic Archaeon Sulfolobus solfataricus

    NARCIS (Netherlands)

    Albers, S.-V.; Jonuscheit, M.; Dinkelaker, S.; Urich, T.; Kletzin, A.; Tampé, R.; Driessen, A.J.M.; Schleper, C.

    Many systems are available for the production of recombinant proteins in bacterial and eukaryotic model organisms, which allow us to study proteins in their native hosts and to identify protein-protein interaction partners. In contrast, only a few transformation systems have been developed for

  13. Extensive Lysine Methylation in Hyperthermophilic Crenarchaea: Potential Implications for Protein Stability and Recombinant Enzymes

    Directory of Open Access Journals (Sweden)

    Catherine H. Botting

    2010-01-01

    Full Text Available In eukarya and bacteria, lysine methylation is relatively rare and is catalysed by sequence-specific lysine methyltransferases that typically have only a single-protein target. Using RNA polymerase purified from the thermophilic crenarchaeum Sulfolobus solfataricus, we identified 21 methyllysines distributed across 9 subunits of the enzyme. The modified lysines were predominantly in α-helices and showed no conserved sequence context. A limited survey of the Thermoproteus tenax proteome revealed widespread modification with 52 methyllysines in 30 different proteins. These observations suggest the presence of an unusual lysine methyltransferase with relaxed specificity in the crenarchaea. Since lysine methylation is known to enhance protein thermostability, this may be an adaptation to a thermophilic lifestyle. The implications of this modification for studies and applications of recombinant crenarchaeal enzymes are discussed.

  14. 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.

  15. Hyperthermophilic Composting Accelerates the Removal of Antibiotic Resistance Genes and Mobile Genetic Elements in Sewage Sludge

    NARCIS (Netherlands)

    Liao, Hanpeng; Lu, Xiaomei; Rensing, Christopher; Friman, Ville Petri; Geisen, Stefan; Chen, Zhi; Yu, Zhen; Wei, Zhong; Zhou, Shungui; Zhu, Yongguan

    2018-01-01

    Composting is an efficient way to convert organic waste into fertilizers. However, waste materials often contain large amounts of antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs) that can reduce the efficacy of antibiotic treatments when transmitted to humans. Because

  16. Improvement of the catalytic efficiency of a hyperthermophilic xylanase from Bispora sp. MEY-1.

    Directory of Open Access Journals (Sweden)

    Xiaoyu Wang

    Full Text Available Extremophilic xylanases have attracted great scientific and industrial interest. In this study, a GH10 xylanase-encoding gene, Xyl10E, was cloned from Bispora sp. MEY-1 and expressed in Pichia pastoris GS115. Deduced Xyl10E shares the highest identities of 62% and 57% with characterized family GH10 xylanases from Talaromyces leycettanus and Penicillium canescens (structure 4F8X, respectively. Xyl10E was most active at 93 to 95°C and pH 4.0, retained more than 75% or 48% of the initial activity when heated at 80°C or 90°C for 30 min, respectively, and hardly lost activity at pH 1.0 to 7.0, but was completely inhibited by SDS. Two residues, A160 and A161, located on loop 4, were identified to play roles in catalysis. Mutants A160D/E demonstrated higher affinity to substrate with lower Km values, while mutants A161D/E mainly displayed elevated Vmax values. All of these mutants had significantly improved catalytic efficiency. According to the molecular dynamics simulation, the mutation of A160E was able to affect the important substrate binding site Y204 and then improve the substrate affinity, and the mutation of A161D was capable of forming a hydrogen bond with the substrate to promote the substrate binding or accelerate the product release. This study introduces a highly thermophilic fungal xylanase and reveals the importance of loop 4 for catalytic efficiency.

  17. A virus of hyperthermophilic archaea with a unique architecture among DNA viruses

    NARCIS (Netherlands)

    Rensen, Elena Ilka; Mochizuki, Tomohiro; Quemin, Emmanuelle; Schouten, S.; Krupovic, Mart; Prangishvili, David

    2016-01-01

    Viruses package their genetic material in diverse ways. Most known strategies include encapsulation of nucleic acids into spherical or filamentous virions with icosahedral or helical symmetry, respectively. Filamentous viruses with dsDNA genomes are currently associated exclusively with Archaea.

  18. Crystal Structure of Hyperthermophilic Endo-β-1,4-glucanase

    Science.gov (United States)

    Zheng, Baisong; Yang, Wen; Zhao, Xinyu; Wang, Yuguo; Lou, Zhiyong; Rao, Zihe; Feng, Yan

    2012-01-01

    Endo-β-1,4-glucanase from thermophilic Fervidobacterium nodosum Rt17-B1 (FnCel5A), a new member of glycosyl hydrolase family 5, is highly thermostable and exhibits the highest activity on carboxymethylcellulose among the reported homologues. To understand the structural basis for the thermostability and catalytic mechanism, we report here the crystal structures of FnCel5A and the complex with glucose at atomic resolution. FnCel5A exhibited a (β/α)8-barrel structure typical of clan GH-A of the glycoside hydrolase families with a large and deep catalytic pocket located in the C-terminal end of the β-strands that may permit substrate access. A comparison of the structure of FnCel5A with related structures from thermopile Clostridium thermocellum, mesophile Clostridium cellulolyticum, and psychrophile Pseudoalteromonas haloplanktis showed significant differences in intramolecular interactions (salt bridges and hydrogen bonds) that may account for the difference in their thermostabilities. The substrate complex structure in combination with a mutagenesis analysis of the catalytic residues implicates a distinctive catalytic module Glu167-His226-Glu283, which suggests that the histidine may function as an intermediate for the electron transfer network between the typical Glu-Glu catalytic module. Further investigation suggested that the aromatic residues Trp61, Trp204, Phe231, and Trp240 as well as polar residues Asn51, His127, Tyr228, and His235 in the active site not only participated in substrate binding but also provided a unique microenvironment suitable for catalysis. These results provide substantial insight into the unique characteristics of FnCel5A for catalysis and adaptation to extreme temperature. PMID:22128157

  19. Hyperthermophilic enzymes - stability, activity and implementation strategies for high temperature applications

    NARCIS (Netherlands)

    Unsworth, L.D.; Oost, van der J.; Koutsopoulos, S.

    2007-01-01

    Current theories agree that there appears to be no unique feature responsible for the remarkable heat stability properties of hyperthermostable proteins. A concerted action of structural, dynamic and other physicochemical attributes are utilized to ensure the delicate balance between stability and

  20. Metagenomic analyses of novel viruses and plasmids from a cultured environmental sample of hyperthermophilic neutrophiles

    DEFF Research Database (Denmark)

    Garrett, Roger Antony; Prangishvili, David; Shah, Shiraz Ali

    2010-01-01

    Two novel viral genomes and four plasmids were assembled from an environmental sample collected from a hot spring at Yellowstone National Park, USA, and maintained anaerobically in a bioreactor at 85°C and pH 6. The double-stranded DNA viral genomes are linear (22.7 kb) and circular (17.7 kb...... respectively. Strategies are considered for assembling genomes of smaller genetic elements from complex environmental samples, and for establishing possible host identities on the basis of sequence similarity to host CRISPR immune systems....

  1. The ABC of ABC-transport in the hyperthermophilic archaeon Pyrococcus furiosus

    NARCIS (Netherlands)

    Koning, S

    2003-01-01

    Living organisms of our earth can be divided into two groups, the prokaryotes and the eukaryotes. Eukaryotic cells have a nucleus, a special compartment in the cell, where the genetic material, the DNA is located. The DNA in the prokaryotic cell is floating freely in the cell. The eukaryotes, that

  2. A novel carbohydrate-binding surface layer protein from the hyperthermophilic archaeon Pyrococcus horikoshii.

    Science.gov (United States)

    Goda, Shuichiro; Koga, Tomoyuki; Yamashita, Kenichiro; Kuriura, Ryo; Ueda, Toshifumi

    2018-04-08

    In Archaea and Bacteria, surface layer (S-layer) proteins form the cell envelope and are involved in cell protection. In the present study, a putative S-layer protein was purified from the crude extract of Pyrococcus horikoshii using affinity chromatography. The S-layer gene was cloned and expressed in Escherichia coli. Isothermal titration calorimetry analyses showed that the S-layer protein bound N-acetylglucosamine and induced agglutination of the gram-positive bacterium Micrococcus lysodeikticus. The protein comprised a 21-mer structure, with a molecular mass of 1,340 kDa, as determined using small-angle X-ray scattering. This protein showed high thermal stability, with a midpoint of thermal denaturation of 79 °C in dynamic light scattering experiments. This is the first description of the carbohydrate-binding archaeal S-layer protein and its characteristics.

  3. Characterization of sodium dodecyl sulfate-resistant proteolytic activity in the hyperthermophilic archaebacterium Pyrococcus furiosus

    Energy Technology Data Exchange (ETDEWEB)

    Blumentals, I.I.; Robinson, A.S.; Kelly, R.M. (Johns Hopkins Univ., Baltimore, MD (USA))

    1990-07-01

    Cell extracts from Pyrococcus furiosus were found to contain five proteases, two of which (S66 and S102) are resistant to sodium dodecyl sulfate (SDS) denaturation. Cell extracts incubated at 98{degree}C in the presence of 1% SDS for 24 h exhibited substantial cellular proteolysis such that only four proteins could be visualized by amido black-Coomassie brilliant blue staining of SDS-polyacrylamide gels. The SDS-treated extract retained 19% of the initial proteolytic activity as represented by two proteases, S66 (66 kilodaltons (kDa)) and S102 (102 kDa). Immunoblot analysis with guinea pig sera containing antibodies against protease S66 indicated that S66 is related neither to S102 nor to the other proteases. The results of this analysis also suggest that S66 might be the hydrolysis product of a 200-kDa precursor which does not have proteolytic activity. The 24-h SDS-treated extract showed unusually thermostable proteolytic activity; the measured half-life at 98{degree}C was found to be 33 h. Proteases S66 and S102 were also resistant to denaturation by 8 M urea, 80 mM dithiothreitol, and 5% {beta}-mercaptoethanol. Purified protease S66 was inhibited by phenylmethylsulfonyl fluoride and diisopropyl fluorophosphate but not by EDTA, ethylene glycol-bis({beta}-aminoethyl ether)-N,N,N{prime},N{prime}-tetraacetic acid, or iodoacetic acid. These results indicate that S66 is a serine protease. Amino acid ester hydrolysis studies showed that protease S66 was hydrolytically active towards N-benzoyl-L-arginine ethyl ester.

  4. Tax incidence on services rendered on the high seas; Incidencia de ISS sobre servicos prestados em aguas maritimas

    Energy Technology Data Exchange (ETDEWEB)

    Paco, Daniel Hora do; Giamattey, Ricardo Henrique Dionisio; Miranda, Thales Ribamar [PETROBRAS S.A., Rio de Janeiro, RJ (Brazil)

    2008-07-01

    This paper analyze the legal aspects of the incidence of ISSQN tax (Tax on Services of any Nature), on the services provide on the high seas. Also comment the controversy surrounding the active tax competency (municipality who may be due to the tax) for the charging of the incident ISSQN on the services provided on the high seas, if overcome the arguments in favor of non-levy of the tax.

  5. Ventilation and air conditioning systems in maritime productions units; Panorama dos sistemas de VAC em unidades maritimas de producao

    Energy Technology Data Exchange (ETDEWEB)

    Guedes, Fernando Pedrosa; Sztajnbok, Ernani Luis [PETROBRAS, Rio de Janeiro, RJ (Brazil); Padua, Carlos Eduardo Dantas de; Passos, Alfredo Silveira [DUOVAC Engenharia Ltda. (Brazil)

    2004-07-01

    In an Offshore Stationary Production Unit (SPU), the adequate project of the Ventilation and Air Conditioning (VAC) System is not only a thermal comfort requirement but part of the essential safety services of the installation and complement for area classification requirements associated with electrical equipment. The VAC installations are sometimes the object of complaints by onboard team. Problems such as unsatisfactory system performance, high noise levels in the accommodation quarters, offices and other areas and the discomfort caused by unbalanced ventilation and air conditioning systems, are some of the most frequent complaints. Air Conditioning systems are classified as Direct and Indirect Expansion. Decentralized systems with Indirect Expansion has been adopted in PETROBRAS projects. This conception is not used in VAC Systems for platforms installed in North Sea, where the use of Centralized Systems with Direct Expansion are more common. The objective of this work is to compare the VAC conception projects, analyzing their advantages and disadvantages . The evaluation of VAC System in PETROBRAS project, and their steps in SPU development, is also scope of this paper. (author)

  6. The first myriapod genome sequence reveals conservative arthropod gene content and genome organisation in the centipede Strigamia maritima

    NARCIS (Netherlands)

    Chipman, Ariel D; Ferrier, David E K; Brena, Carlo; Qu, Jiaxin; Hughes, Daniel S T; Schröder, Reinhard; Torres-Oliva, Montserrat; Znassi, Nadia; Jiang, Huaiyang; Almeida, Francisca C; Alonso, Claudio R; Apostolou, Zivkos; Aqrawi, Peshtewani; Arthur, Wallace; Barna, Jennifer C J; Blankenburg, Kerstin P; Brites, Daniela; Capella-Gutiérrez, Salvador; Coyle, Marcus; Dearden, Peter K; Du Pasquier, Louis; Duncan, Elizabeth J; Ebert, Dieter; Eibner, Cornelius; Erikson, Galina; Evans, Peter D; Extavour, Cassandra G; Francisco, Liezl; Gabaldón, Toni; Gillis, William J; Goodwin-Horn, Elizabeth A; Green, Jack E; Griffiths-Jones, Sam; Grimmelikhuijzen, Cornelis J P; Gubbala, Sai; Guigó, Roderic; Han, Yi; Hauser, Frank; Havlak, Paul; Hayden, Luke; Helbing, Sophie; Holder, Michael; Hui, Jerome H L; Hunn, Julia P; Hunnekuhl, Vera S; Jackson, LaRonda; Javaid, Mehwish; Jhangiani, Shalini N; Jiggins, Francis M; Jones, Tamsin E; Kaiser, Tobias S; Kalra, Divya; Kenny, Nathan J; Korchina, Viktoriya; Kovar, Christie L; Kraus, F Bernhard; Lapraz, François; Lee, Sandra L; Lv, Jie; Mandapat, Christigale; Manning, Gerard; Mariotti, Marco; Mata, Robert; Mathew, Tittu; Neumann, Tobias; Newsham, Irene; Ngo, Dinh N; Ninova, Maria; Okwuonu, Geoffrey; Ongeri, Fiona; Palmer, William J; Patil, Shobha; Patraquim, Pedro; Pham, Christopher; Pu, Ling-Ling; Putman, Nicholas H; Rabouille, Catherine; Ramos, Olivia Mendivil; Rhodes, Adelaide C; Robertson, Helen E; Robertson, Hugh M; Ronshaugen, Matthew; Rozas, Julio; Saada, Nehad; Sánchez-Gracia, Alejandro; Scherer, Steven E; Schurko, Andrew M; Siggens, Kenneth W; Simmons, DeNard; Stief, Anna; Stolle, Eckart; Telford, Maximilian J; Tessmar-Raible, Kristin; Thornton, Rebecca; van der Zee, Maurijn; von Haeseler, Arndt; Williams, James M; Willis, Judith H; Wu, Yuanqing; Zou, Xiaoyan; Lawson, Daniel; Muzny, Donna M; Worley, Kim C; Gibbs, Richard A; Akam, Michael; Richards, Stephen

    2014-01-01

    Myriapods (e.g., centipedes and millipedes) display a simple homonomous body plan relative to other arthropods. All members of the class are terrestrial, but they attained terrestriality independently of insects. Myriapoda is the only arthropod class not represented by a sequenced genome. We present

  7. Local and global influences on population declines of coastal waders: Purple Sandpiper Calidris maritima numbers in the Moray Firth, Scotland

    Science.gov (United States)

    Summers, Ron W.; Foster, Simon; Swann, Bob; Etheridge, Brian

    2012-05-01

    Declines in numbers by several wader species in Britain have been linked to climate change, but the mechanism for the declines has rarely been explored. Britain lies at the northern end of the East Atlantic Flyway, and supports 1.3 million out of the Flyway's 8.5 million coastal waders (Charadrii) in winter and the Purple Sandpiper is one of the species whose numbers have declined. Here, we examine the dynamics of the decline as observed in the Moray Firth, northeast Scotland, investigating whether the decline was due to poorer apparent survival (return rate) or poorer recruitment of young birds. The maximum number in the Moray Firth declined from 860 in 1987/88 to 236 in 2006/07, with some increase during winters 2007/08 and 2008/09. At the three main high-tide roosts (Balintore, Lossiemouth and Buckie) the maximum combined number declined from 574 to 90. Changes in survival and recruitment (percentage of first-year birds) were examined at these roosts from captured samples, which were ringed and recaptured. There were no significant changes between winters in survival rates, nor were there differences between the survival rates of age groups (first-year and adult) or bill size groups, which represented birds of different sex and breeding origin. Annual survival estimates for the three roosts ranged from 72 to 77%. The percentage of first-year birds varied among roosts and years; the lowest values were during the late 1980s/early 1990s and early 2000s. A free-running population model incorporating varying percentages of first-year birds and constant mortality for each roost provided a plausible explanation for the decline. Although modelled numbers followed the observed pattern, a discrepancy in one year was carried forward in subsequent years, so that the fit with the observed numbers was parallel rather than similar. However, it seems that the decline in numbers was largely due to poorer recruitment. We discuss whether breeding success had declined, whether the population had responded to changes in the local sewage treatment systems, which could affect invertebrate food for Purple Sandpipers, or whether fewer birds chose to winter in Scotland. The Moray Firth population is derived from Norway and possibly Canada, and there is evidence that the Norwegian population was disproportionately affected. The reason for poor recruitment requires further study, and other wader species require examination to test if poor recruitment is a common feature of decline in numbers.

  8. Crystallization and preliminary X-ray analysis of membrane-bound pyrophosphatases.

    Science.gov (United States)

    Kellosalo, Juho; Kajander, Tommi; Honkanen, Riina; Goldman, Adrian

    2013-02-01

    Membrane-bound pyrophosphatases (M-PPases) are enzymes that enhance the survival of plants, protozoans and prokaryotes in energy constraining stress conditions. These proteins use pyrophosphate, a waste product of cellular metabolism, as an energy source for sodium or proton pumping. To study the structure and function of these enzymes we have crystallized two membrane-bound pyrophosphatases recombinantly produced in Saccharomyces cerevisae: the sodium pumping enzyme of Thermotoga maritima (TmPPase) and the proton pumping enzyme of Pyrobaculum aerophilum (PaPPase). Extensive crystal optimization has allowed us to grow crystals of TmPPase that diffract to a resolution of 2.6 Å. The decisive step in this optimization was in-column detergent exchange during the two-step purification procedure. Dodecyl maltoside was used for high temperature solubilization of TmPPase and then exchanged to a series of different detergents. After extensive screening, the new detergent, octyl glucose neopentyl glycol, was found to be the optimal for TmPPase but not PaPPase.

  9. Catalytic properties of thermophilic lactate dehydrogenase and halophilic malate dehydrogenase at high temperature and low water activity.

    Science.gov (United States)

    Hecht, K; Wrba, A; Jaenicke, R

    1989-07-15

    Thermophilic lactate dehydrogenases from Thermotoga maritima and Bacillus stearothermophilus are stable up to temperature limits close to the optimum growth temperature of their parent organisms. Their catalytic properties are anomalous in that Km shows a drastic increase with increasing temperature. At low temperatures, the effect levels off. Extreme halophilic malate dehydrogenase from Halobacterium marismortui exhibits a similar anomaly. Increasing salt concentration (NaCl) leads to an optimum curve for Km, oxaloacctate while Km, NADH remains constant. Previous claims that the activity of halophilic malate dehydrogenase shows a maximum at 1.25 M NaCl are caused by limiting substrate concentration; at substrate saturation, specific activity of halophilic malate dehydrogenase reaches a constant value at ionic strengths I greater than or equal to 1 M. Non-halophilic (mitochondrial) malate dehydrogenase shows Km characteristics similar to those observed for the halophilic enzyme. The drastic decrease in specific activity of the mitochondrial enzyme at elevated salt concentrations is caused by the salt-induced increase in rigidity of the enzyme, rather than gross structural changes.

  10. Cloning, overexpression, purification, crystallization and preliminary X-ray diffraction analysis of an inositol monophosphatase family protein (SAS2203) from Staphylococcus aureus MSSA476

    International Nuclear Information System (INIS)

    Bhattacharyya, Sudipta; Dutta, Debajyoti; Ghosh, Ananta Kumar; Das, Amit Kumar

    2011-01-01

    The cloning, overexpression, purification, crystallization and preliminary X-ray diffraction analysis of an inositol monophosphatase family protein (SAS2203) from S. aureus MSSA476 is reported. The gene product of the sas2203 ORF of Staphylococcus aureus MSSA476 encodes a 30 kDa molecular-weight protein with a high sequence resemblance (29% identity) to tetrameric inositol monophosphatase from Thermotoga maritima. The protein was cloned, expressed, purified to homogeneity and crystallized. Crystals appeared in several conditions and good diffraction-quality crystals were obtained from 0.2 M Li 2 SO 4 , 20% PEG 3350, 0.1 M HEPES pH 7.0 using the sitting-drop vapour-diffusion method. A complete diffraction data set was collected to 2.6 Å resolution using a Rigaku MicroMax-007 HF Cu Kα X-ray generator and a Rigaku R-AXIS IV ++ detector. The diffraction data were consistent with the orthorhombic space group P2 1 2 1 2 1 , with unit-cell parameters a = 49.98, b = 68.35, c = 143.79 Å, α = β = γ = 90°, and the crystal contained two molecules in the asymmetric unit

  11. Flow synthesis of phenylserine using threonine aldolase immobilized on Eupergit support

    Directory of Open Access Journals (Sweden)

    Jagdish D. Tibhe

    2013-10-01

    Full Text Available Threonine aldolase (TA from Thermotoga maritima was immobilized on an Eupergit support by both a direct and an indirect method. The incubation time for the direct immobilization method was optimized for the highest amount of enzyme on the support. By introducing the immobilized TA in a packed-bed microreactor, a flow synthesis of phenylserine was developed, and the effects of temperature and residence time were studied in particular. Calculations of the Damköhler number revealed that no mass transfer limitations are given in the micro-interstices of the packed bed. The yield does not exceed 40% and can be rationalized by the natural equilibrium as well as product inhibition which was experimentally proven. The flow synthesis with the immobilized enzyme was compared with the corresponding transformation conducted with the free enzyme. The product yield was further improved by operating under slug flow conditions which is related to the very short residence time distribution. In all cases 20% diastereomeric excess (de and 99% enantiomeric excess (ee were observed. A continuous run of the reactant solution was carried out for 10 hours in order to check enzyme stability at higher temperature. Stable operation was achieved at 20 minute residence time. Finally, the productivity of the reactor was calculated, extrapolated to parallel run units, and compared with data collected previously.

  12. Identification of a Minimal Peptide Tag for in Vivo and in Vitro Loading of Encapsulin.

    Science.gov (United States)

    Cassidy-Amstutz, Caleb; Oltrogge, Luke; Going, Catherine C; Lee, Antony; Teng, Poh; Quintanilla, David; East-Seletsky, Alexandra; Williams, Evan R; Savage, David F

    2016-06-21

    The encapsulation of enzymes and other proteins within a proteinaceous shell has been observed in many bacteria and archaea, but the function and utility of many such compartments are enigmatic. Efforts to study these functions have been complicated by the size and complexity of traditional protein compartments. One potential system for investigating the effect of compartmentalization is encapsulin, a large and newly discovered class of protein shells that are typically composed of two proteins: a protomer that assembles into the icosahedral shell and a cargo protein packaged inside. Encapsulins are some of the simplest known protein shell systems and readily self-assemble in vivo. Systematic characterization of the effects of compartmentalization requires the ability to load a wide range of cargo proteins. Here, we demonstrate that foreign cargo can be loaded into the encapsulin from Thermotoga maritima both in vivo and in vitro by fusion of the cargo protein with a short C-terminal peptide present in the native cargo. To facilitate biochemical characterization, we also develop a simple and rapid purification protocol and demonstrate the thermal and pH stability of the shell. Efforts to study the biophysical effects of protein encapsulation have been problematic in complex compartments, but the simplicity of assembling and loading encapsulin makes it an ideal system for future experiments exploring the effects of encapsulation on proteins.

  13. Flow synthesis of phenylserine using threonine aldolase immobilized on Eupergit support

    Science.gov (United States)

    Tibhe, Jagdish D; Fu, Hui; Noël, Timothy; Wang, Qi; Meuldijk, Jan

    2013-01-01

    Summary Threonine aldolase (TA) from Thermotoga maritima was immobilized on an Eupergit support by both a direct and an indirect method. The incubation time for the direct immobilization method was optimized for the highest amount of enzyme on the support. By introducing the immobilized TA in a packed-bed microreactor, a flow synthesis of phenylserine was developed, and the effects of temperature and residence time were studied in particular. Calculations of the Damköhler number revealed that no mass transfer limitations are given in the micro-interstices of the packed bed. The yield does not exceed 40% and can be rationalized by the natural equilibrium as well as product inhibition which was experimentally proven. The flow synthesis with the immobilized enzyme was compared with the corresponding transformation conducted with the free enzyme. The product yield was further improved by operating under slug flow conditions which is related to the very short residence time distribution. In all cases 20% diastereomeric excess (de) and 99% enantiomeric excess (ee) were observed. A continuous run of the reactant solution was carried out for 10 hours in order to check enzyme stability at higher temperature. Stable operation was achieved at 20 minute residence time. Finally, the productivity of the reactor was calculated, extrapolated to parallel run units, and compared with data collected previously. PMID:24204429

  14. Reconstruction of the Chemotaxis Receptor-Kinase Assembly

    International Nuclear Information System (INIS)

    Park, S.; Borbat, P.; Gonzalez-Bonet, G.; Bhatnagar, J.; Pollard, A.; Freed, J.; Bilwes, A.; Crane, B.

    2006-01-01

    In bacterial chemotaxis, an assembly of transmembrane receptors, the CheA histidine kinase and the adaptor protein CheW processes environmental stimuli to regulate motility. The structure of a Thermotoga maritima receptor cytoplasmic domain defines CheA interaction regions and metal ion-coordinating charge centers that undergo chemical modification to tune receptor response. Dimeric CheA-CheW, defined by crystallography and pulsed ESR, positions two CheWs to form a cleft that is lined with residues important for receptor interactions and sized to clamp one receptor dimer. CheW residues involved in kinase activation map to interfaces that orient the CheW clamps. CheA regulatory domains associate in crystals through conserved hydrophobic surfaces. Such CheA self-contacts align the CheW receptor clamps for binding receptor tips. Linking layers of ternary complexes with close-packed receptors generates a lattice with reasonable component ratios, cooperative interactions among receptors and accessible sites for modification enzymes

  15. The actin-like MreB proteins in Bacillus subtilis: a new turn.

    Science.gov (United States)

    Chastanet, Arnaud; Carballido-Lopez, Rut

    2012-06-01

    A decade ago, two breakthrough descriptions were reported: 1) the first helix-like protein localization pattern of MreB and its paralog Mbl in Bacillus subtilis and 2) the crystal structure of Thermotoga maritima MreB1, which was remarkably similar to that of actin. These discoveries strongly stimulated the field of bacterial development, leading to the identification of many new cytoskeletal proteins (1) and the publication of many studies describing the helical patterns of protein, DNA and even lipid domains. However, today, new breakthroughs are shaking up what had become a dogma. Instead of helical structures, MreBs appear to form discrete patches that move circumferentially around the cell, questioning the idea of MreB cables forming an actin-like cytoskeleton. Furthermore, increasing evidence of biochemical properties that are unlike the properties of actin suggest that the molecular behavior of MreB proteins may be different. The aim of this review is to summarize the current knowledge of the so-called "actin-like" MreB cytoskeleton through a discussion of the model Gram-positive bacterium B. subtilis and the most recent findings in this rapidly evolving research field.

  16. MreB and MurG as scaffolds for the cytoplasmic steps of peptidoglycan biosynthesis.

    Science.gov (United States)

    Favini-Stabile, Sandy; Contreras-Martel, Carlos; Thielens, Nicole; Dessen, Andréa

    2013-12-01

    Peptidoglycan is a major determinant of cell shape in bacteria, and its biosynthesis involves the concerted action of cytoplasmic, membrane-associated and periplasmic enzymes. Within the cytoplasm, Mur enzymes catalyse the first steps leading to peptidoglycan precursor biosynthesis, and have been suggested as being part of a multicomponent complex that could also involve the transglycosylase MurG and the cytoskeletal protein MreB. In order to initialize the characterization of a potential Mur interaction network, we purified MurD, MurE, MurF, MurG and MreB from Thermotoga maritima and characterized their interactions using membrane blotting and surface plasmon resonance. MurD, MurE and MurF all recognize MurG and MreB, but not each other, while the two latter proteins interact. In addition, we solved the crystal structures of MurD, MurE and MurF, which indicate that their C-termini display high conformational flexibilities. The differences in Mur conformations could be important parameters for the stability of an intracytoplasmic murein biosynthesis complex. © 2013 Society for Applied Microbiology and John Wiley & Sons Ltd.

  17. An Aromatic Cap Seals the Substrate Binding Site in an ECF-Type S Subunit for Riboflavin

    Energy Technology Data Exchange (ETDEWEB)

    Karpowich, Nathan K.; Song, Jinmei; Wang, Da-Neng

    2016-06-13

    ECF transporters are a family of active membrane transporters for essential micronutrients, such as vitamins and trace metals. Found exclusively in archaea and bacteria, these transporters are composed of four subunits: an integral membrane substrate-binding subunit (EcfS), a transmembrane coupling subunit (EcfT), and two ATP-binding cassette ATPases (EcfA and EcfA'). We have characterized the structural basis of substrate binding by the EcfS subunit for riboflavin from Thermotoga maritima, TmRibU. TmRibU binds riboflavin with high affinity, and the protein–substrate complex is exceptionally stable in solution. The crystal structure of riboflavin-bound TmRibU reveals an electronegative binding pocket at the extracellular surface in which the substrate is completely buried. Analysis of the intermolecular contacts indicates that nearly every available substrate hydrogen bond is satisfied. A conserved aromatic residue at the extracellular end of TM5, Tyr130, caps the binding site to generate a substrate-bound, occluded state, and non-conservative mutation of Tyr130 reduces the stability of this conformation. Using a novel fluorescence binding assay, we find that an aromatic residue at this position is essential for high-affinity substrate binding. Comparison with other S subunit structures suggests that TM5 and Loop5-6 contain a dynamic, conserved motif that plays a key role in gating substrate entry and release by S subunits of ECF transporters.

  18. A comparative molecular dynamics study of thermophilic and mesophilic β-fructosidase enzymes.

    Science.gov (United States)

    Mazola, Yuliet; Guirola, Osmany; Palomares, Sucel; Chinea, Glay; Menéndez, Carmen; Hernández, Lázaro; Musacchio, Alexis

    2015-09-01

    Arabidopsis thaliana cell wall invertase 1 (AtcwINV1) and Thermotoga maritima β-fructosidase (BfrA) are among the best structurally studied members of the glycoside hydrolase family 32. Both enzymes hydrolyze sucrose as the main substrate but differ strongly in their thermal stability. Mesophilic AtcwINV1 and thermophilic BfrA have divergent sequence similarities in the N-terminal five bladed β-propeller catalytic domain (31 %) and the C-terminal β-sandwich domain (15 %) of unknown function. The two enzymes were subjected to 200 ns molecular dynamics simulations at 300 K (27 °C) and 353 K (80 °C). Regular secondary structure regions, but not loops, in AtcwINV1 and BfrA showed no significant fluctuation differences at both temperatures. BfrA was more rigid than AtcwINV1 at 300 K. The simulation at 353 K did not alter the structural stability of BfrA, but did increase the overall flexibility of AtcwINV1 exhibiting the most fluctuating regions in the β-propeller domain. The simulated heat treatment also increased the gyration radius and hydrophobic solvent accessible surface area of the plant enzyme, consistent with the initial steps of an unfolding process. The preservation of the conformational rigidity of BfrA at 353 K is linked to the shorter size of the protein loops. Shortening of BfrA loops appears to be a key mechanism for thermostability.

  19. Thermophilic enzymes and their applications in biocatalysis: a robust aldo-keto reductase.

    Science.gov (United States)

    Willies, Simon; Isupov, Misha; Littlechild, Jennifer

    2010-09-01

    Extremophiles are providing a good source of novel robust enzymes for use in biocatalysis for the synthesis of new drugs. This is particularly true for the enzymes from thermophilic organisms which are more robust than their mesophilic counterparts to the conditions required for industrial bio-processes. This paper describes a new aldo-keto reductase enzyme from a thermophilic eubacteria, Thermotoga maritima which can be used for the production of primary alcohols. The enzyme has been cloned and over-expressed in Escherichia coli and has been purified and subjected to full biochemical characterization. The aldo-keto reductase can be used for production of primary alcohols using substrates including benzaldehyde, 1,2,3,6-tetrahydrobenzaldehyde and para-anisaldehyde. It is stable up to 80 degrees C, retaining over 60% activity for 5 hours at this temperature. The enzyme at pH 6.5 showed a preference for the forward, carbonyl reduction. The enzyme showed moderate stability with organic solvents, and retained 70% activity in 20% (v/v) isopropanol or DMSO. These properties are favourable for its potential industrial applications.

  20. High sensitive RNA detection by one-step RT-PCR using the genetically engineered variant of DNA polymerase with reverse transcriptase activity from hyperthermophilies.

    Science.gov (United States)

    Okano, Hiroyuki; Baba, Misato; Kawato, Katsuhiro; Hidese, Ryota; Yanagihara, Itaru; Kojima, Kenji; Takita, Teisuke; Fujiwara, Shinsuke; Yasukawa, Kiyoshi

    2018-03-01

    One-step RT-PCR has not been widely used even though some thermostable DNA polymerases with reverse transcriptase (RT) activity were developed from bacterial and archaeal polymerases, which is owing to low cDNA synthesis activity from RNA. In the present study, we developed highly-sensitive one-step RT-PCR using the single variant of family A DNA polymerase with RT activity, K4pol L329A (L329A), from the hyperthermophilic bacterium Thermotoga petrophila K4 or the 16-tuple variant of family B DNA polymerase with RT activity, RTX, from the hyperthermophilic archaeon Thermococcus kodakarensis. Optimization of reaction condition revealed that the activities for cDNA synthesis and PCR of K4pol L329A and RTX were highly affected by the concentrations of MgCl 2 and Mn(OCOCH 3 ) 2 as well as those of K4pol L329A or RTX. Under the optimized condition, 300 copies/μl of target RNA in 10 μl reaction volumes were successfully detected by the one-step RT-PCR with K4pol L329A or RTX, which was almost equally sensitive enough compared with the current RT-PCR condition using retroviral RT and thermostable DNA polymerase. Considering that K4pol L329A and RTX are stable even at 90-100°C, our results suggest that the one-step RT-PCR with K4pol L329A or RTX is more advantageous than the current one. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  1. Introducing capnophilic lactic fermentation in a combined dark-photo fermentation process: a route to unparalleled H2 yields.

    Science.gov (United States)

    Dipasquale, L; Adessi, A; d'Ippolito, G; Rossi, F; Fontana, A; De Philippis, R

    2015-01-01

    Two-stage process based on photofermentation of dark fermentation effluents is widely recognized as the most effective method for biological production of hydrogen from organic substrates. Recently, it was described an alternative mechanism, named capnophilic lactic fermentation, for sugar fermentation by the hyperthermophilic bacterium Thermotoga neapolitana in CO2-rich atmosphere. Here, we report the first application of this novel process to two-stage biological production of hydrogen. The microbial system based on T. neapolitana DSM 4359(T) and Rhodopseudomonas palustris 42OL gave 9.4 mol of hydrogen per mole of glucose consumed during the anaerobic process, which is the best production yield so far reported for conventional two-stage batch cultivations. The improvement of hydrogen yield correlates with the increase in lactic production during capnophilic lactic fermentation and takes also advantage of the introduction of original conditions for culturing both microorganisms in minimal media based on diluted sea water. The use of CO2 during the first step of the combined process establishes a novel strategy for biohydrogen technology. Moreover, this study opens the way to cost reduction and use of salt-rich waste as feedstock.

  2. Crystallization and preliminary X-ray characterization of a ferritin from the hyperthermophilic archaeon and anaerobe Pyrococcus furiosus

    International Nuclear Information System (INIS)

    Matias, Pedro M.; Tatur, Jana; Carrondo, Maria Arménia; Hagen, Wilfred R.

    2005-01-01

    Ferritin from P. furiosus crystallizes in space group C222 1 , with unit-cell parameters a = 258.1, b = 340.1, c = 266.5 Å and 36 monomers in the asymmetric unit, corresponding to one and a half 24-mers. Crystals of the title protein have been produced and preliminary structural analysis has been carried out. The crystals belong to the orthorhombic space group C222 1 , with unit-cell parameters a = 258.1, b = 340.1, c = 266.5 Å. The protein forms a 24-mer of 20 kDa subunits, which assemble with 432 non-crystallographic symmetry. A total of 36 monomers are found in the asymmetric unit, corresponding to one and a half 24-mers

  3. Crystallization and quaternary structure analysis of an Lrp-like regulatory protein from the hyperthermophile Pyrococcus furiosus

    NARCIS (Netherlands)

    Sedelnikova, S.E.; Smits, S.H.J.; Leonard, P.M.; Brinkman, A.B.; Oost, van der J.; Rafferty, J.B.

    2001-01-01

    The LrpA transcriptional regulator from Pyrococcus furiosus, a member of the leucine-responsive regulatory protein (Lrp) family, has been crystallized by the hanging-drop method of vapour diffusion using ammonium sulfate as the precipitant. The crystals belong to the tetragonal system and are in

  4. Molecular evolution of the hyperthermophilic archaea of the Pyrococcus genus: analysis of adaptation to different environmental conditions

    Directory of Open Access Journals (Sweden)

    Afonnikov Dmitry A

    2009-12-01

    Full Text Available Abstract Background Prokaryotic microorganisms are able to survive and proliferate in severe environmental conditions. The increasing number of complete sequences of prokaryotic genomes has provided the basis for studying the molecular mechanisms of their adaptation at the genomic level. We apply here a computer-based approach to compare the genomes and proteomes from P. furiosus, P. horikoshii, and P. abyssi to identify features of their molecular evolution related to adaptation strategy to diverse environmental conditions. Results Phylogenetic analysis of rRNA genes from 26 Pyrococcus strains suggested that the divergence of P. furiosus, P. horikoshii and P. abyssi might have occurred from ancestral deep-sea organisms. It was demonstrated that the function of genes that have been subject to positive Darwinian selection is closely related to abiotic and biotic conditions to which archaea managed to become adapted. Divergence of the P. furiosus archaea might have been due to loss of some genes involved in cell motility or signal transduction, and/or to evolution under positive selection of the genes for translation machinery. In the course of P. horikoshii divergence, positive selection was found to operate mainly on the transcription machinery; divergence of P. abyssi was related with positive selection for the genes mainly involved in inorganic ion transport. Analysis of radical amino acid replacement rate in evolving P. furiosus, P. horikoshii and P. abyssi showed that the fixation rate was higher for radical substitutions relative to the volume of amino acid side-chain. Conclusions The current results give due credit to the important role of hydrostatic pressure as a cause of variability in the P. furiosus, P. horikoshii and P. abyssi genomes evolving in different habitats. Nevertheless, adaptation to pressure does not appear to be the sole factor ensuring adaptation to environment. For example, at the stage of the divergence of P. horikoshii and P. abyssi, an essential evolutionary role may be assigned to changes in the trophic chain, namely, acquisition of a consumer status at a high (P. horikoshii or low level (P. abyssi.

  5. Structure of the large terminase from a hyperthermophilic virus reveals a unique mechanism for oligomerization and ATP hydrolysis.

    Science.gov (United States)

    Xu, Rui-Gang; Jenkins, Huw T; Antson, Alfred A; Greive, Sandra J

    2017-12-15

    The crystal structure of the large terminase from the Geobacillus stearothermophilus bacteriophage D6E shows a unique relative orientation of the N-terminal adenosine triphosphatase (ATPase) and C-terminal nuclease domains. This monomeric 'initiation' state with the two domains 'locked' together is stabilized via a conserved C-terminal arm, which may interact with the portal protein during motor assembly, as predicted for several bacteriophages. Further work supports the formation of an active oligomeric state: (i) AUC data demonstrate the presence of oligomers; (ii) mutational analysis reveals a trans-arginine finger, R158, indispensable for ATP hydrolysis; (iii) the location of this arginine is conserved with the HerA/FtsK ATPase superfamily; (iv) a molecular docking model of the pentamer is compatible with the location of the identified arginine finger. However, this pentameric model is structurally incompatible with the monomeric 'initiation' state and is supported by the observed increase in kcat of ATP hydrolysis, from 7.8 ± 0.1 min-1 to 457.7 ± 9.2 min-1 upon removal of the C-terminal nuclease domain. Taken together, these structural, biophysical and biochemical data suggest a model where transition from the 'initiation' state into a catalytically competent pentameric state, is accompanied by substantial domain rearrangements, triggered by the removal of the C-terminal arm from the ATPase active site. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

  6. Hydrogen production and enzyme activities in the hyperthermophile Thermococcus paralvinellae grown on maltose, tryptone and agricultural waste

    Directory of Open Access Journals (Sweden)

    Sarah A. Hensley

    2016-02-01

    Full Text Available Thermococcus may be an important alternative source of H2 in the hot subseafloor in otherwise low H2 environments such as some hydrothermal vents and oil reservoirs. It may also be useful in industry for rapid agricultural waste treatment and concomitant H2 production. Thermococcus paralvinellae grown at 82°C without sulfur produced up to 5 mmol of H2 L-1 at rates of 5-36 fmol H2 cell-1 h-1 on 0.5% (wt vol-1 maltose, 0.5% (wt vol-1 tryptone, and 0.5% maltose + 0.05% tryptone media. Two potentially inhibiting conditions, the presence of 10 mM acetate and low pH (pH 5 in maltose-only medium, did not significantly affect growth or H2 production. Growth rates, H2 production rates, and cell yields based on H2 production were the same as those for Pyrococcus furiosus grown at 95°C on the same media for comparison. Acetate, butyrate, succinate, isovalerate and formate were also detected as end products. After 100 h, T. paralvinellae produced up to 5 mmol of H2 L-1 of medium when grown on up to 70% (vol vol-1 waste milk from cows undergoing treatment for mastitis with the bacterial antibiotic Ceftiofur and from untreated cows. The amount of H2 produced by T. paralvinellae increased with increasing waste concentrations, but decreased in P. furiosus cultures supplemented with waste milk above 1% concentration. All mesophilic bacteria from the waste milk that grew on Luria Bertani, Sheep’s Blood (selective for Staphylococcus, the typical cause of mastitis, and MacConkey (selective for Gram-negative enteric bacteria agar plates were killed by heat during incubation at 82°C. Ceftiofur, which is heat labile, was below the detection limit following incubation at 82°C. T. paralvinellae also produced up to 6 mmol of H2 L-1 of medium when grown on 0.1-10% (wt vol-1 spent brewery grain while P. furiosus produced < 1 mmol of H2 L-1. Twelve of 13 enzyme activities in T. paralvinellae showed significant (p<0.05 differences across six different growth conditions; however, methyl viologen-dependent membrane hydrogenase activity remained constant across all media types. The results demonstrate the potential of at least some Thermococcus species to produce H2 if protein and α-glucosides are present as substrates.

  7. Thioredoxin-linked redox control of metabolism in Methanocaldococcus jannaschii, an evolutionarily deeply-rooted hyperthermophilic methanogenic archaeon

    Science.gov (United States)

    Thioredoxin (Trx), a small redox protein, controls multiple processes in eukaryotes and bacteria by changing the thiol redox status of selected proteins. We have investigated this aspect in methanarchaea. These ancient methanogens produce methane almost exclusively from H2 plus CO2 carried approxima...

  8. Comparison of NMR and crystal structures highlights conformational isomerism in protein active sites

    International Nuclear Information System (INIS)

    Serrano, Pedro; Pedrini, Bill; Geralt, Michael; Jaudzems, Kristaps; Mohanty, Biswaranjan; Horst, Reto; Herrmann, Torsten; Elsliger, Marc-André; Wilson, Ian A.; Wüthrich, Kurt

    2010-01-01

    Tools for systematic comparisons of NMR and crystal structures developed by the JCSG were applied to two proteins with known functions: the T. maritima anti-σ factor antagonist TM1081 and the mouse γ-glutamylamine cyclotransferase A2LD1 (gi:13879369). In an attempt to exploit the complementarity of crystal and NMR data, the combined use of the two structure-determination techniques was explored for the initial steps in the challenge of searching proteins of unknown functions for putative active sites. The JCSG has recently developed a protocol for systematic comparisons of high-quality crystal and NMR structures of proteins. In this paper, the extent to which this approach can provide function-related information on the two functionally annotated proteins TM1081, a Thermotoga maritima anti-σ factor antagonist, and A2LD1 (gi:13879369), a mouse γ-glutamylamine cyclotransferase, is explored. The NMR structures of the two proteins have been determined in solution at 313 and 298 K, respectively, using the current JCSG protocol based on the software package UNIO for extensive automation. The corresponding crystal structures were solved by the JCSG at 100 K and 1.6 Å resolution and at 100 K and 1.9 Å resolution, respectively. The NMR and crystal structures of the two proteins share the same overall molecular architectures. However, the precision of the structure determination along the amino-acid sequence varies over a significantly wider range in the NMR structures than in the crystal structures. Thereby, in each of the two NMR structures about 65% of the residues have displacements below the average and in both proteins the less well ordered residues include large parts of the active sites, in addition to some highly solvent-exposed surface areas. Whereas the latter show increased disorder in the crystal and in solution, the active-site regions display increased displacements only in the NMR structures, where they undergo local conformational exchange on the

  9. Maritime prerequisites for development of infrastructure for liquefied natural gas (LNG / LBG); Maritima foerutsaettningar foer utbyggnad av infrastruktur foer flytande gas (LNG/LBG)

    Energy Technology Data Exchange (ETDEWEB)

    Gahnstroem, Johan; Molitor, Edvard; Raggl, Karl-Johan; Sandkvist, Jim [SSPA Sweden AB, Goeteborg (Sweden)

    2011-06-15

    This study has provided an initial picture of where the most interesting ports and areas available for future expansion of a maritime infrastructure for LNG. On the basis of supplying vessels with LNG as fuel, from a long term perspective, we recommend locating LNG terminals in or near major ports and around the big ship routes. Given the current age distribution of ships operating waters of the Baltic Sea, almost 20% of the vessels are 30-40 years old and likely to be replaced by 2015 - 2020. Thus, there is a potential for newly built ships will be equipped with LNG operation. Selected criteria s; Size of the LNG terminal and hence the need for the size of the fairway and the area of land. Proximity to traffic routes with much ship traffic. Proximity to the major port. Proximity to consumers on the land side. On the basis of selected criteria and analyzed for possible location of the terminal it can be noted that a number of Swedish ports are found suitable. For example, ports of Sundsvall, Gothenburg and Helsingborg has been identified as suitable, but with different starting point and different types and sizes of terminals possible.

  10. The first myriapod genome sequence reveals conservative arthropod gene content and genome organisation in the centipede Strigamia maritima

    DEFF Research Database (Denmark)

    Chipman, Ariel D.; Ferrier, David E.K.; Brena, Carlo

    2014-01-01

    phylogeny several particular molecular mechanisms and traits emerged. For example, we conclude that juvenile hormone signalling evolved with the emergence of the exoskeleton in the arthropods and that RR-1 containing cuticle proteins evolved in the lineage leading to Mandibulata. We also identify when...

  11. Interactions between indole-3-acetic acid (IAA) with a lectin from Canavalia maritima seeds reveal a new function for lectins in plant physiology.

    Science.gov (United States)

    Delatorre, Plinio; Silva-Filho, José Caetano; Rocha, Bruno Anderson Matias; Santi-Gadelha, Tatiane; da Nóbrega, Raphael Batista; Gadelha, Carlos Alberto Almeida; do Nascimento, Kyria Santiago; Nagano, Celso Shiniti; Sampaio, Alexandre Holanda; Cavada, Benildo Sousa

    2013-09-01

    Indole-3-acetic acid (IAA) bound is considered a storage molecule and is inactive. However, some studies have proposed an additional possible regulatory mechanism based on the ability of lectins to form complexes with IAA. We report the first crystal structure of ConM in complex with IAA at 2.15 Å resolution. Based on a tetrameric model of the complex, we hypothesize how the lectin controls the availability of IAA during the early seedling stages, indicating a possible new physiological role for these proteins. A free indole group is also bound to the protein. The ConM interaction with different forms of IAA is a strategy to render the phytohormone unavailable to the cell. Thus, this new physiological role proposed for legume lectins might be a novel mechanism by which IAA levels are decreased in addition to the destruction and formation of new complexes in the later stages of seed germination. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  12. Haplotype Detection from Next-Generation Sequencing in High-Ploidy-Level Species: 45S rDNA Gene Copies in the Hexaploid Spartina maritima

    Czech Academy of Sciences Publication Activity Database

    Boutte, J.; Aliaga, B.; Lima, O.; de Carvalho, J.F.; Ainouche, A.; Macas, Jiří; Rousseau-Gueutin, M.; Coriton, O.; Ainouche, M.; Salmon, A.

    2016-01-01

    Roč. 6, č. 1 (2016), s. 29-40 ISSN 2160-1836 Institutional support: RVO:60077344 Keywords : poaceae * duplication * paralogy * bioinformatics * polyploidy Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.861, year: 2016

  13. Monobuoy monitoring system - project, installation and preliminary results; Sistema de monitoracao de monoboias maritimas - desenvolvimento, instalacao e analise preliminar de resultados

    Energy Technology Data Exchange (ETDEWEB)

    Lima, Daniel L; Machado Filho, Remo Z [PETROBRAS, Rio de Janeiro, RJ (Brazil). Centro de Pesquisas; Medeiros, Junior, Adory J [PETROBRAS Transporte S.A. (TRANSPETRO), Rio de Janeiro, RJ (Brazil)

    2005-07-01

    A monobuoy monitoring system was developed and installed on 2004 in two oil terminals of TRANSPETRO. The main purpose of this system is to assure that monobuoys will be assisted during offloading operations, which will reduce the risk of accidents and detect immediately damages on mooring lines and equipment. The system comprises a Control Module on terminal site and a Remote Monitoring Unit, installed on each monobuoy. The link between modules is provided by an UHF radio-modem. The parameters acquired are pitch, roll, heave, geographic position, internal variables, and, if available, pressure and temperature of lines, traction on mooring system, etc. Positioning information is processed by the PETRONAV Positioning System. The whole process is registered and monitored on terminal with graphical interface. This article describes the development and installation of the system and the preliminary analysis of data produced since the system was installed. (author)

  14. Crystal complexes of a predicted S-adenosylmethionine-dependent methyltransferase reveal a typical AdoMet binding domain and a substrate recognition domain

    Energy Technology Data Exchange (ETDEWEB)

    Miller, D.J.; Ouellette, N.; Evodokimova, E.; Savchenko, A.; Edwards, A.; Anderson, W.F. (Toronto); (NWU)

    2010-03-08

    S-adenosyl-L-methionine-dependent methyltransferases (MTs) are abundant, and highly conserved across phylogeny. These enzymes use the cofactor AdoMet to methylate a wide variety of molecular targets, thereby modulating important cellular and metabolic activities. Thermotoga maritima protein 0872 (TM0872) belongs to a large sequence family of predicted MTs, ranging phylogenetically from relatively simple bacteria to humans. The genes for many of the bacterial homologs are located within operons involved in cell wall synthesis and cell division. Despite preliminary biochemical studies in E. coli and B. subtilis, the substrate specificity of this group of more than 150 proteins is unknown. As part of the Midwest Center for Structural Genomics initiative (www.mcsg.anl.gov), we have determined the structure of TM0872 in complexes with AdoMet and with S-adenosyl-L-homocysteine (AdoHcy). As predicted, TM0872 has a typical MT domain, and binds endogenous AdoMet, or co-crystallized AdoHcy, in a manner consistent with other known MT structures. In addition, TM0872 has a second domain that is novel among MTs in both its location in the sequence and its structure. The second domain likely acts in substrate recognition and binding, and there is a potential substrate-binding cleft spanning the two domains. This long and narrow cleft is lined with positively charged residues which are located opposite the S{sup +}-CH{sub 3} bond, suggesting that a negatively charged molecule might be targeted for catalysis. However, AdoMet and AdoHcy are both buried, and access to the methyl group would presumably require structural rearrangement. These TM0872 crystal structures offer the first structural glimpses at this phylogenetically conserved sequence family.

  15. Structural Diversity Within the Mononuclear and Binuclear Active Sites of N-Acetyl-D-Glucosamine-6-Phosphate Deacetylase

    Energy Technology Data Exchange (ETDEWEB)

    Hall,R.; Brown, S.; Fedorov, A.; Fedorov, E.; Xu, C.; Babbitt, P.; Almo, S.; Raushel, F.

    2007-01-01

    NagA catalyzes the hydrolysis of N-acetyl-D-glucosamine-6-phosphate to D-glucosamine-6-phosphate and acetate. X-ray crystal structures of NagA from Escherichia coli were determined to establish the number and ligation scheme for the binding of zinc to the active site and to elucidate the molecular interactions between the protein and substrate. The three-dimensional structures of the apo-NagA, Zn-NagA, and the D273N mutant enzyme in the presence of a tight-binding N-methylhydroxyphosphinyl-D-glucosamine-6-phosphate inhibitor were determined. The structure of the Zn-NagA confirms that this enzyme binds a single divalent cation at the beta-position in the active site via ligation to Glu-131, His-195, and His-216. A water molecule completes the ligation shell, which is also in position to be hydrogen bonded to Asp-273. In the structure of NagA bound to the tight binding inhibitor that mimics the tetrahedral intermediate, the methyl phosphonate moiety has displaced the hydrolytic water molecule and is directly coordinated to the zinc within the active site. The side chain of Asp-273 is positioned to activate the hydrolytic water molecule via general base catalysis and to deliver this proton to the amino group upon cleavage of the amide bond of the substrate. His-143 is positioned to help polarize the carbonyl group of the substrate in conjunction with Lewis acid catalysis by the bound zinc. The inhibitor is bound in the {alpha}-configuration at the anomeric carbon through a hydrogen bonding interaction of the hydroxyl group at C-1 with the side chain of His-251. The phosphate group of the inhibitor attached to the hydroxyl at C-6 is ion paired with Arg-227 from the adjacent subunit. NagA from Thermotoga maritima was shown to require a single divalent cation for full catalytic activity.

  16. Responses of Microbial Community Composition to Temperature Gradient and Carbon Steel Corrosion in Production Water of Petroleum Reservoir

    Directory of Open Access Journals (Sweden)

    Xiao-Xiao Li

    2017-12-01

    Full Text Available Oil reservoir production systems are usually associated with a temperature gradient and oil production facilities frequently suffer from pipeline corrosion failures. Both bacteria and archaea potentially contribute to biocorrosion of the oil production equipment. Here the response of microbial populations from the petroleum reservoir to temperature gradient and corrosion of carbon steel coupons were investigated under laboratory condition. Carbon steel coupons were exposed to production water from a depth of 1809 m of Jiangsu petroleum reservoir (China and incubated for periods of 160 and 300 days. The incubation temperatures were set at 37, 55, and 65°C to monitoring mesophilic, thermophilic and hyperthermophilic microorganisms associated with anaerobic carbon steel corrosion. The results showed that corrosion rate at 55°C (0.162 ± 0.013 mm year-1 and 37°C (0.138 ± 0.008 mm year-1 were higher than that at 65°C (0.105 ± 0.007 mm year-1, and a dense biofilm was observed on the surface of coupons under all biotic incubations. The microbial community analysis suggests a high frequency of bacterial taxa associated with families Porphyromonadaceae, Enterobacteriaceae, and Spirochaetaceae at all three temperatures. While the majority of known sulfate-reducing bacteria, in particular Desulfotignum, Desulfobulbus and Desulfovibrio spp., were predominantly observed at 37°C; Desulfotomaculum spp., Thermotoga spp. and Thermanaeromonas spp. as well as archaeal members closely related to Thermococcus and Archaeoglobus spp. were substantially enriched at 65°C. Hydrogenotrophic methanogens of the family Methanobacteriaceae were dominant at both 37 and 55°C; acetoclastic Methanosaeta spp. and methyltrophic Methanolobus spp. were enriched at 37°C. These observations show that temperature changes significantly alter the microbial community structure in production fluids and also affected the biocorrosion of carbon steel under anaerobic conditions.

  17. Molecular and biochemical analyses of the GH44 module of CbMan5B/Cel44A, a bifunctional enzyme from the hyperthermophilic bacterium Caldicellulosiruptor bescii.

    Science.gov (United States)

    Ye, Libin; Su, Xiaoyun; Schmitz, George E; Moon, Young Hwan; Zhang, Jing; Mackie, Roderick I; Cann, Isaac K O

    2012-10-01

    A large polypeptide encoded in the genome of the thermophilic bacterium Caldicellulosiruptor bescii was determined to consist of two glycoside hydrolase (GH) modules separated by two carbohydrate-binding modules (CBMs). Based on the detection of mannanase and endoglucanase activities in the N-terminal GH5 and the C-terminal GH44 module, respectively, the protein was designated CbMan5B/Cel44A. A GH5 module with >99% identity from the same organism was characterized previously (X. Su, R. I. Mackie, and I. K. Cann, Appl. Environ. Microbiol. 78:2230-2240, 2012); therefore, attention was focused on CbMan5A/Cel44A-TM2 (or TM2), which harbors the GH44 module and the two CBMs. On cellulosic substrates, TM2 had an optimal temperature and pH of 85°C and 5.0, respectively. Although the amino acid sequence of the GH44 module of TM2 was similar to those of other GH44 modules that hydrolyzed cello-oligosaccharides, cellulose, lichenan, and xyloglucan, it was unique that TM2 also displayed modest activity on mannose-configured substrates and xylan. The TM2 protein also degraded Avicel with higher specific activity than activities reported for its homologs. The GH44 catalytic module is composed of a TIM-like domain and a β-sandwich domain, which consists of one β-sheet at the N terminus and nine β-sheets at the C terminus. Deletion of one or more β-sheets from the β-sandwich domain resulted in insoluble proteins, suggesting that the β-sandwich domain is essential for proper folding of the polypeptide. Combining TM2 with three other endoglucanases from C. bescii led to modest synergistic activities during degradation of cellulose, and based on our results, we propose a model for cellulose hydrolysis and utilization by C. bescii.

  18. Structural analysis by reductive cleavage with LiAlH4 of an allyl ether choline-phospholipid, archaetidylcholine, from the hyperthermophilic methanoarchaeon Methanopyrus kandleri

    Directory of Open Access Journals (Sweden)

    Masateru Nishihara

    2002-01-01

    Full Text Available A choline-containing phospholipid (PL-4 in Methanopyrus kandleri cells was identified as archaetidylcholine, which has been described by Sprott et al. (1997. The PL-4 consisted of a variety of molecular species differing in hydrocarbon composition. Most of the PL-4 was acid-labile because of its allyl ether bond. The identity of PL-4 was confirmed by thin-layer chromatography (TLC followed by positive staining with Dragendorff-reagent and fast-atom bombardment–mass spectrometry. A new method of LiAlH4 hydrogenolysis was developed to cleave allyl ether bonds and recover the corresponding hydrocarbons. We confirmed the validity of the LiAlH4 method in a study of the model compound synthetic unsaturated archaetidic acid (2,3-di-O-geranylgeranyl-sn-glycerol-1-phosphate. Saturated ether bonds were not cleaved by the LiAlH4 method. The hydrocarbons formed following LiAlH4 hydrogenolysis of PL-4 were identified by gas–liquid chromatography and mass spectrometry. Four kinds of hydrocarbons with one to four double bonds were detected: 47% of the hydrocarbons had four double bonds; 11% had three double bonds; 14% had two double bonds; 7% had one double bond; and 6% were saturated species. The molecular species composition of PL-4 was also estimated based on acid lability: 77% of the molecular species had two acid-labile hydrocarbons; 11% had one acid-labile and one acid-stable hydrocarbon; and 11% had two acid-stable hydrocarbons. To our knowledge, this is the first report of a specific chemical degradation method for the structural analysis of allyl ether phospholipid in archaea.

  19. Aromatic residues located close to the active center are essential for the catalytic reaction of flap endonuclease-1 from hyperthermophilic archaeon Pyrococcus horikoshii.

    Science.gov (United States)

    Matsui, Eriko; Abe, Junko; Yokoyama, Hideshi; Matsui, Ikuo

    2004-04-16

    Flap endonuclease-1 (FEN-1) possessing 5'-flap endonuclease and 5'-->3' exonuclease activity plays important roles in DNA replication and repair. In this study, the kinetic parameters of mutants at highly conserved aromatic residues, Tyr33, Phe35, Phe79, and Phe278-Phe279, in the vicinity of the catalytic centers of FEN-1 were examined. The substitution of these aromatic residues with alanine led to a large reduction in kcat values, although these mutants retained Km values similar to that of the wild-type enzyme. Notably, the kcat of Y33A and F79A decreased 333-fold and 71-fold, respectively, compared with that of the wild-type enzyme. The aromatic residues Tyr33 and Phe79, and the aromatic cluster Phe278-Phe279 mainly contributed to the recognition of the substrates without the 3' projection of the upstream strand (the nick, 5'-recess-end, single-flap, and pseudo-Y substrates) for the both exo- and endo-activities, but played minor roles in recognizing the substrates with the 3' projection (the double flap substrate and the nick substrate with the 3' projection). The replacement of Tyr33, Phe79, and Phe278-Phe279, with non-charged aromatic residues, but not with aliphatic hydrophobic residues, recovered the kcat values almost fully for the substrates without the 3' projection of the upstream strand, suggesting that the aromatic groups of Tyr33, Phe79, and Phe278-Phe279 might be involved in the catalytic reaction, probably via multiple stacking interactions with nucleotide bases. The stacking interactions of Tyr33 and Phe79 might play important roles in fixing the template strand and the downstream strand, respectively, in close proximity to the active center to achieve the productive transient state leading to the hydrolysis.

  20. Photobiosynthesis of stable and functional silver/silver chloride nanoparticles with hydrolytic activity using hyperthermophilic β-glucosidases with industrial potential.

    Science.gov (United States)

    Araújo, Juscemácia N; Tofanello, Aryane; da Silva, Viviam M; Sato, Juliana A P; Squina, Fabio M; Nantes, Iseli L; Garcia, Wanius

    2017-09-01

    The β-glucosidases are important enzymes employed in a large number of processes and industrial applications, including biofuel production from biomass. Therefore, in this study, we reported for the first time the photobiosynthesis of stable and functional silver/silver chloride nanoparticles (Ag/AgCl-NPs) using two hyperthermostable bacterial β-glucosidases with industrial potential. The syntheses were straightforward and rapid processes carried out by mixing β-glucosidase and silver nitrate (in buffer 10mM Tris-HCl, pH 8) under irradiation with light (over a wavelength range of 450-600nm), therefore, compatible with the green chemistry procedure. Synthesized Ag/AgCl-NPs were characterized using a series of physical techniques. Absorption spectroscopy showed a strong absorption band centered at 460nm due to surface plasmon resonance of the Ag-NPs. X-ray diffraction analysis revealed that the Ag/AgCl-NPs were purely crystalline in nature. Under electron microscopy, Ag/AgCl-NPs of variable diameter ranging from 10 to 100nm can be visualized. Furthermore, electron microscopy, zeta potential and Fourier transform infrared spectroscopy results confirmed the presence of β-glucosidases coating and stabilizing the Ag/AgCl-NPs. Finally, the results showed that the enzymatic activities were maintained in the β-glucosidases assisted Ag/AgCl-NPs. The information described here should provide a useful basis for future studies of β-glucosidases assisted Ag/AgCl-NPs, including biotechnological applications. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Improving the Catalytic Activity of Hyperthermophilic Pyrococcus horikoshii Prolidase for Detoxification of Organophosphorus Nerve Agents over a Broad Range of Temperatures

    Science.gov (United States)

    2011-01-01

    affinity for metal, and increased thermostability compared to P. furiosus prolidase, Pf prol (PF1343). To obtain a better enzyme for OP nerve agent...decontamination and to investigate the structural factors that may influence protein thermostability and thermoactivity, randomly mutated Ph1prol enzymes ...Introduction Pyrococcus horikoshii and Pyrococcus furiosus are both hyper- thermophilic archaea, growing optimally at 98 –100◦C that were isolated from a

  2. Comparison of NMR and crystal structures for the proteins TM1112 and TM1367

    International Nuclear Information System (INIS)

    Mohanty, Biswaranjan; Serrano, Pedro; Pedrini, Bill; Jaudzems, Kristaps; Geralt, Michael; Horst, Reto; Herrmann, Torsten; Elsliger, Marc-André; Wilson, Ian A.; Wüthrich, Kurt

    2010-01-01

    NMR structures of the proteins TM1112 and TM1367 solved by the JCSG in solution at 298 K could be superimposed with the corresponding crystal structures at 100 K with r.m.s.d. values of <1.0 Å for the backbone heavy atoms. For both proteins the structural differences between multiple molecules in the asymmetric unit of the crystals correlated with structural variations within the bundles of conformers used to represent the NMR solution structures. A recently introduced JCSG NMR structure-determination protocol, which makes use of the software package UNIO for extensive automation, was further evaluated by comparison of the TM1112 structure obtained using these automated methods with another NMR structure that was independently solved in another PSI center, where a largely interactive approach was applied. The NMR structures of the TM1112 and TM1367 proteins from Thermotoga maritima in solution at 298 K were determined following a new protocol which uses the software package UNIO for extensive automation. The results obtained with this novel procedure were evaluated by comparison with the crystal structures solved by the JCSG at 100 K to 1.83 and 1.90 Å resolution, respectively. In addition, the TM1112 solution structure was compared with an NMR structure solved by the NESG using a conventional largely interactive methodology. For both proteins, the newly determined NMR structure could be superimposed with the crystal structure with r.m.s.d. values of <1.0 Å for the backbone heavy atoms, which provided a starting platform to investigate local structure variations, which may arise from either the methods used or from the different chemical environments in solution and in the crystal. Thereby, these comparative studies were further explored with the use of reference NMR and crystal structures, which were computed using the NMR software with input of upper-limit distance constraints derived from the molecular models that represent the results of structure

  3. Bayesian spatial modeling applied to environmental monitoring due to the use of different drilling fluids in the maritime exploration activity; Modelagem espacial Bayesiana aplicada ao monitoramento ambiental decorrente do uso de diferentes fluidos de perfuracao na atividade exploratoria maritima

    Energy Technology Data Exchange (ETDEWEB)

    Pulgati, Fernando H. [Universidade Federal do Rio de Janeiro (UFRJ), RJ (Brazil). Coordenacao dos Programas de Pos-graduacao de Engenharia (COPPE). Lab. de Modelagem de Bacias (LAB2M); Zouain, Ricardo N.A. [Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, RS (Brazil). Inst. de Geociencias. Centro de Estudos de Geologia Costeira e Oceanica; Fachel, Jandyra M.G. [Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, RS (Brazil). Inst. de Matematica; Landau, Luiz [Universidade Federal do Rio de Janeiro (UFRJ), RJ (Brazil). Coordenacao dos Programas de Pos-graduacao de Engenharia (COPPE). Lab. de Metodos Computacionais em Engenharia (LAMCE)

    2004-07-01

    Controlling and monitoring environmental researches have accompanied the development of offshore exploration drill activities aimed at finding oil and gas reserves, as there has been an increase in the environmental demands and restrictions. Three stages of the drilling process were isolated and the effects of different fluids were measured using Bayesian spatial models. The probable impact of the use of non-aqueous fluid (NAF) was measured through changes observed in sea sediments in three different occasions: previous to the activity, one (1) month after the end of the activity, and one (1) year after the end of the activity. BACI (Before-After Control Impact) design, which allows the control of temporal and spatial variation components, was chosen. (author)

  4. Diagnosis of solid waste of oil and natural gas exploration and production activities in Brazil offshore sedimentary basins; Diagnostico dos residuos solidos das atividades de exploracao e producao de petroleo e gas natural em bacias sedimentares maritimas no Brasil

    Energy Technology Data Exchange (ETDEWEB)

    Koehler, Pedro Henrique Wisniewski; Mendonca; Gilberto Moraes de

    2012-07-01

    The objective of this study is to analyze the generation and disposal of solid waste from the exploration and production activities of oil and natural gas in Brazilian waters. We used data from the implementation reports of pollution control project of the activities licensed by IBAMA. During 2009 the activities related to exploration and production of offshore oil and gas produced a total of 44,437 tons of solid waste, with the main waste generated corresponding to: oily waste (16,002 t); Metal uncontaminated (11,085 t); contaminated waste (5630 t), non recycling waste (4935 t); Wood uncontaminated (1,861 t), chemicals (1,146 t). Considering the total waste generated by activities during the period analyzed, it was observed that 54.3% are made up of waste Class I (hazardous waste), 27.9% of Class II wastes (waste non-hazardous non-inert); and 17.8% of waste Class IIB (non-hazardous and inert waste). The results obtained in this work enabled the scenario of waste generation by the E and P offshore activities. As a result, the survey serves as a starting point for monitoring the progress in implementing the projects sought Pollution Control of licensed projects, as well as support the monitoring of reflexes arising from the intensification of activities in certain regions. (author)

  5. Technology platforms and tools invigorate the economy. situation maritime Spanish Technology Platform; Las plataformas tecnologicas como herramientas dinamizadoras de la economia. Situacion de la plataforma tecnologica maritima espanola (PTME)

    Energy Technology Data Exchange (ETDEWEB)

    Ruiz de Leon Loriga, C.

    2012-07-01

    The Ministry of Industry, Energy and Tourisme and the Ministry of Education and Science, through Fundacion INNOVAMAR, promote and help in 2005 the creation of the Spanish Maritime Technology Platform, (SMTP). Is a forum where all stake holders from the maritime sector define and share a common vision and a Strategic Research Agenda, driving the necessary innovation efforts forward. The Challenges 2020: a competitive maritime industry, safe, sustainable and efficient operations, adaptation of the demand, transversal areas. (Author)

  6. Low level of gene flow from cultivated beets (¤Beta vulgaris¤ L. ssp. ¤vulgaris¤) into Danish populations of sea beet (¤Beta vulgaris¤ L. ssp. ¤maritima¤ (L.) Arcangeli)

    DEFF Research Database (Denmark)

    Andersen, N.S.; Siegismund, H.R.; Meyer, V.

    2005-01-01

    Gene flow from sugar beets to sea beets occurs in the seed propagation areas in southern Europe. Some seed propagation also takes place in Denmark, but here the crop-wild gene flow has not been investigated. Hence, we studied gene flow to sea beet populations from sugar beet lines used in Danish ...

  7. Oil seepage detection technique as a tool to hydrocarbon prospecting in offshore Campos Basin-Brazil; Deteccao de exsudacoes de oleo como uma ferramenta de prospeccao de hidrocarbonetos na regiao maritima da Bacia de Campos - Brasil

    Energy Technology Data Exchange (ETDEWEB)

    Castilho, Jose G.; Brito, Ademilson F. [Universidade Federal do Rio de Janeiro (UFRJ), RJ (Brazil). Coordenacao dos Programas de Pos-graduacao de Engenharia (COPPE). Lab. de Modelagem de Bacias (LAB2M); Landau, Luiz [Universidade Federal do Rio de Janeiro (UFRJ), RJ (Brazil). Coordenacao dos Programas de Pos-graduacao de Engenharia (COPPE). Lab. de Metodos Computacionais em Engenharia (LAMCE)

    2004-07-01

    With a proven capacity to identify oil slicks in offshore regions, RADARSAT-1 imagery can be useful for oil exploration purposes. The paper discusses the seepage detection method at Campos Basin, offshore Rio de Janeiro State, which is responsible for 80% of the Brazilian production of oil and gas. It is known that the horizontal migration of petroleum can occurs over tens or even hundreds of kilometers, where the source rock placed in more deep locations can be linked with shallow reservoirs or traps and even reach the ocean. It means that seepage can provide information for risking petroleum charge at basin scales, and cannot have a direct relation with the geographical position of the interpreted seeps and possible filled prospects. A good understanding of the geology, and hence the petroleum systems of a basin is the key to use seepage in exploration. The work is divided into three main steps. First step were select oil seepages interpreted at Campos Basin where is found several giant petroleum fields. Second, the geology of the study area and its structural and stratigraphic features were analyzed, in order to identify possible migration pathways related to faults generated by halokinesis. Another important aspect is the presence of 'windows' or ducts in the evaporates beds allowing the contact between the section that contains source rocks and the turbidities reservoirs, that contain the majority of the oil discovers. All these features were interpreted based on a regional dip seismic line (203 - 76), and a geologic cross section with E-W orientation, showing the structure of the Marlim Field. Finally, all the information was integrated in a Geographical Information System (GIS), and then analyzed in an interdisciplinary environment, with the intention to link possible routes of oil migration to post-evaporites reservoirs or to interpreted seeps. (author)

  8. Microbial Geochemistry in Shallow-Sea Hydrothermal Systems

    Science.gov (United States)

    Amend, J. P.; Pichler, T.

    2006-12-01

    Shallow-sea hydrothermal systems are far more ubiquitous than generally recognized. Approximately 50-60 systems are currently known, occurring world-wide in areas of high heat flow, such as, volcanic island arcs, near-surface mid-ocean ridges, and intraplate oceanic volcanoes. In contrast to deep-sea systems, shallow- sea vent fluids generally include a meteoric component, they experience phase separation near the sediment- water interface, and they discharge into the photic zone (thermophilic bacteria and archaea. Perhaps because deep-sea smokers and continental hot springs are visually more stunning, shallow-sea systems are often overlooked study sites. We will discuss their particular features that afford unique opportunities in microbial geochemistry. Two of the better studied examples are at Vulcano Island (Italy) and Ambitle Island (Papua New Guinea). The vents and sediment seeps at Vulcano are the "type locality" for numerous cultured hyperthermophiles, including the bacteria Aquifex and Thermotoga, the crenarchaeon Pyrodictium, and the Euryarchaeota Archaeoglobus and Pyrococcus. Isotope-labeled incubation experiments of heated sediments and an array of culturing studies have shown that simple organic compounds are predominantly fermented or anaerobically respired with sulfate. 16S rRNA gene surveys, together with fluorescent in situ hybridization studies, demonstrated the dominance of key thermophilic bacteria and archaea (e.g., Aquificales, Thermotogales, Thermococcales, Archaeoglobales) in the sediments and the presence of a broad spectrum of mostly uncultured crenarchaeota in several vent waters, sediment samples, and geothermal wells. Thermodynamic modeling quantified potential energy yields from aerobic and anaerobic respiration reactions and fermentation reactions. In contrast to their deep-sea counterparts, shallow-sea hydrothermal systems are often characterized by high arsenic concentrations of more than 500-times seawater levels. The arsenic

  9. Comparative microbial analysis before and after foaming incidents in biogas reactors

    DEFF Research Database (Denmark)

    Kougias, Panagiotis; De Francisci, Davide; Treu, Laura

    2014-01-01

    biosurfactants (Lactobacillus, Bacillus, Pseudomonas, Thermotoga), others contain mycolic acid in their cell wall (Thermoactinomyces, Pseudonocardia) or decrease the surface tension of the media (Micrococcus, Streptococcus). Frankia, Dialister and Paenibacillus are known to be correlated to this phenomenon...

  10. Stabilization of enzymes against thermal stress and freeze-drying by mannosylglycerate

    NARCIS (Netherlands)

    Ramos, A.; Raven, N.; Sharp, R.J.; Bartolucci, S.; Rossi, M.; Cannio, R.; Lebbink, J.; Oost, van der J.; Vos, de W.M.; Santos, H.

    1997-01-01

    2-O-(beta)-Mannosylglycerate, a solute that accumulates in some (hyper)thermophilic organisms, was purified from Pyrococcus furiosus cells, and its effect on enzyme stabilization in vitro was assessed. Enzymes from hyperthermophilic, thermophilic, and mesophilic sources were examined. The

  11. Meri ottab, meri andab / Juhan Kreem

    Index Scriptorium Estoniae

    Kreem, Juhan, 1971-

    2014-01-01

    Arvustus: Shipwreck heritage : digitizing and opening access to maritime history sources = Laevavrakid : digitaliseerimine ja avatud ligipääs mereajalooallikatele = Skeppsvrak : digitalisering och förmedling av det maritima kulturarvet. Tallinn, 2013

  12. Some extinct plant taxa on the territory of Novi Sad and their vulnerability status in Vojvodina and Serbia

    Directory of Open Access Journals (Sweden)

    Đakić Žarko S.

    2012-01-01

    Full Text Available Natural habitats on the territory of Novi Sad are almost fully destroyed today, as well as their characteristic plant taxa. The reason for disappearance of natural habitats is the development of suburban communities, which is an irreversible process. Plant taxa, specific for wet, salty, and sandy ecosystems grew on those habitats twenty years ago and earlier. This paper presents the overview of 9 taxa (Suaeda maritima subsp. maritima, Androsace elongata subsp. elongata, Cirsium boujartii subsp. boujartii, Aster sedifolius subsp. canus, Blackstonia perfoliata subsp. serotina, Plantago maritima subsp. maritima, Salvia nutans, Allium angulosum, and Typha schuttleworthii. These taxa presented integral parts of autochthonous flora of Novi Sad. Since some of these taxa were found in the field 21 years ago and some even 93 years ago, they are extinct from the flora of Novi Sad.

  13. Ability of salt marsh plants for TBT remediation in sediments

    OpenAIRE

    Carvalho, P. N.; Basto, M. C.; Moreira da Silva, M.; Machado, A.; Bordalo, A.; Vasconcelos, M. T.

    2010-01-01

    The capability of Halimione portulacoides, Spartina maritima, and Sarcocornia fruticosa (halophytes very commonly found in salt marshes from Mediterranean areas) for enhancing remediation of tributyltin (TBT) from estuarine sediments was investigated, using different experimental conditions.

  14. Spatial and temporal habitat partitioning by zooplankton in the Bornholm Basin (central Baltic Sea)

    DEFF Research Database (Denmark)

    Schultz, J.; Peck, M.A.; Barz, K.

    2012-01-01

    taxa (Bosmina coregoni maritima, Acartia spp., Pseudocalanus spp., Temora longicornis, Synchaeta spp.) contributed >10% to the zooplankton community composition. The appearance of cladocerans was mainly correlated with the phenology of thermocline development in the spring. The cladoceran B. coregoni...

  15. Agronomy of strip intercropping broccoli with alyssum for biological control of aphids

    Science.gov (United States)

    Organic broccoli growers in California typically control aphids by intercropping broccoli with strips of alyssum (Lobularia maritima (L.) Desv.) which attracts hoverflies (Diptera: Syrphidae) that are important predators of aphids. A three year study with transplanted organic broccoli in Salinas, ...

  16. Produzione di bioidrogeno in dark fermentation da scarti dell'industria agroalimentale mediante l'impiego di batteri ipertermofili

    OpenAIRE

    Alberini, Andrea

    2013-01-01

    La presente tesi di dottorato ha come argomento la produzione d’idrogeno per via fermentativa sfruttando il metabolismo anaerobico di particolari batteri estremofili del genere Thermotoga. In questo lavoro, svolto in seno al progetto Bio-Hydro, sfruttando reattori batch da 116 mL, è stato selezionato il ceppo migliore di Thermotoga fra i quatto ceppi testati: T. neapolitana. Una volta individuato il candidato batterico migliore è stato individuato il valore ottimale di pH (8.5 a t.amb) per la...

  17. The catalytic potency of ß-glucosidase from Pyroccus furiosus in the direct glucosylation reaction

    NARCIS (Netherlands)

    Roode, de B.M.; Meer, van der T.D.; Kaper, T.; Franssen, M.C.R.; Padt, van der A.; Oost, van der J.; Boom, R.M.

    2001-01-01

    Enzymes from extremophiles operate at conditions that are different from their `normal' counterparts, and are therefore a useful extension of the enzyme toolbox. In this paper, the direct glucosylation reaction mediated by a hyperthermophilic -glucosidase from Pyrocuccus furiosus was investigated.

  18. The Peculiar Glycolytic Pathway in Hyperthermophylic Archaea : Understanding Its Whims by Experimentation In Silico

    NARCIS (Netherlands)

    Zhang, Y.; Kouril, T.; Snoep, J.L.; Siebers, B.; Barberis, M.; Westerhoff, H.V.

    2017-01-01

    Mathematical models are key to systems biology where they typically describe the topology and dynamics of biological networks, listing biochemical entities and their relationships with one another. Some (hyper)thermophilic Archaea contain an enzyme, called non-phosphorylating

  19. Functional analysis of thermostable proteins involved in carbohydrate metabolism

    NARCIS (Netherlands)

    Akerboom, A.P.

    2007-01-01

    Thermostable proteins can resist temperature stress whilst keeping their integrity and functionality. In many cases, thermostable proteins originate from hyperthermophilic microorganisms that thrive in extreme environments. These systems are generally located close to geothermal (volcanic) activity,

  20. Novel class III phosphoribosyl diphosphate synthase: structure and properties of the tetrameric, phosphate-activated, non-allosterically inhibited enzyme from Methanocaldococcus jannaschii

    DEFF Research Database (Denmark)

    Kadziola, Anders; Jepsen, Clemens H; Johansson, Eva

    2005-01-01

    The prs gene encoding phosphoribosyl diphosphate (PRPP) synthase of the hyperthermophilic autotrophic methanogenic archaeon Methanocaldococcus jannaschii has been cloned and expressed in Escherichia coli. Subsequently, M.jannaschii PRPP synthase has been purified, characterised, crystallised, and...

  1. Cloning, Sequencing, and Expression of the Gene Encoding Cyclic 2,3-Diphosphoglycerate Synthetase, the Key Enzyme of Cyclic 2,3-Diphosphoglycerate Metabolism in Methanothermus fervidus

    OpenAIRE

    Matussek, Karl; Moritz, Patrick; Brunner, Nina; Eckerskorn, Christoph; Hensel, Reinhard

    1998-01-01

    Cyclic 2,3-diphosphoglycerate synthetase (cDPGS) catalyzes the synthesis of cyclic 2,3-diphosphoglycerate (cDPG) by formation of an intramolecular phosphoanhydride bond in 2,3-diphosphoglycerate. cDPG is known to be accumulated to high intracellular concentrations (>300 mM) as a putative thermoadapter in some hyperthermophilic methanogens. For the first time, we have purified active cDPGS from a methanogen, the hyperthermophilic archaeon Methanothermus fervidus, sequenced the coding gene, and...

  2. AcEST: BP918158 [AcEST

    Lifescience Database Archive (English)

    Full Text Available inase OS=Thermotoga petrophila... 40 0.12 tr|B7RFK1|B7RFK1_9THEM Guanylate kinase OS=Marinitoga piezophila.....7RFK1|B7RFK1_9THEM Guanylate kinase OS=Marinitoga piezophila KA3 GN=gmk PE=4 SV=1 Length = 207 Score = 39.7

  3. Distinctive properties of high hydrogen producing extreme thermophiles, Caldicellulosiruptor saccharolyticus and Thermotaga elfii

    NARCIS (Netherlands)

    Niel, van E.W.J.; Budde, M.A.W.; Haas, de G.G.; Wal, van der F.J.; Claassen, P.A.M.; Stams, A.J.M.

    2002-01-01

    Growth and hydrogen production by two extreme thermophiles during sugar fermentation was investigated. In cultures of Caldicellulosiruptor saccharolyticus grown on sucrose and Thermotoga elfii grown on glucose stoichiometries of 3.3 mol of hydrogen and 2 mol of acetate per mol C6-sugar unit were

  4. [The floristic diversity of the psammophyte vegetation in the region of Tlemcen (north-west Algeria)].

    Science.gov (United States)

    Stambouli-Meziane, Hassiba; Bouazza, M; Thinon, Michel

    2009-08-01

    This study is devoted to the analysis of the psammophyte of the coastal and semi-continental dunes in Tlemcen. Interesting results have been obtained, in particular, on the biological and ecological aspects of the psammophyte. The interpretation from Factoriel analysis of correspondences enabled us to identify the different phytosociological classes (Cakiletea maritimae, Ammophiletea, Quercetea ilicis, Therobrachypodietea and Stellarietea mediae). Some of these classes (Cakiletea maritimae and Ammophiletea) inhabit, exceedingly well, the embryonic dunes. Some species (Therobrachypodietea) colonize the quickset dunes. Lastly, some others (Quercetea ilicis) settle in the more mature and stable dunes. By using the phytosociological and phytodynamical data, we have been able to understand the vegetation and its diversity.

  5. Coping with low nutrient availability and inundation: root growth responses of three halophytic grass species from different elevations along a flooding gradient

    NARCIS (Netherlands)

    Bouma, T.J.; Koutstaal, B.P.; Van Dongen, M.; Nielsen, K.F.

    2001-01-01

    We describe the responses of three halophytic grass species that dominate the low (Spartina anglica), middle (Puccinellia maritima) and high (Elymus pycnanthus) parts of a salt marsh, to soil conditions that are believed to favour contrasting root-growth strategies. Our hypotheses were: (1)

  6. SPINIFICI-SCAEVOLETEA SERICEAE, A NEW VEGETATION CLASS FOR PSAMMOPHYTIC DUNE VEGETATION IN THAILAND

    Directory of Open Access Journals (Sweden)

    S. PIGNATII

    1996-04-01

    Full Text Available This is a short account on the coastal dune vegetation of the Gulf of Siam in Thailand. Vegetation is mainly composed by succulent creeping plants with herbaceous habit as to Canavalia maritima (Papilionaceae and Iponwea pes-caprae (Convolvulaceae and the robust stoloniferous grass Spinijex littoreus, the last having an important function for the fonnation of coastal dunes.

  7. SPINIFICI-SCAEVOLETEA SERICEAE, A NEW VEGETATION CLASS FOR PSAMMOPHYTIC DUNE VEGETATION IN THAILAND

    Directory of Open Access Journals (Sweden)

    S. PIGNATII

    1996-01-01

    Full Text Available This is a short account on the coastal dune vegetation of the Gulf of Siam in Thailand. Vegetation is mainly composed by succulent creeping plants with herbaceous habit as to Canavalia maritima (Papilionaceae and Iponwea pes-caprae (Convolvulaceae and the robust stoloniferous grass Spinijex littoreus, the last having an important function for the fonnation of coastal dunes.

  8. New floristic records in the Balkans: 2

    DEFF Research Database (Denmark)

    2006-01-01

    caespitosa subsp. alpina (34), Plantago maritima subsp. serpentina (38), Thymus callieri subsp. callieri (31); Montenegro - Asperula hercegovina (73); Serbia - Allium paniculatum subsp. villosum (98), Viola obliqua (57); Turkey-in-Europe - Chamaecytisus jankae (37). subsp. (98), (57); Turkey-in-Europe - (37...

  9. U.S. Navy Aeromedical Reference and Waiver Guide

    Science.gov (United States)

    2010-05-05

    Passion flower) – Piper methysticum (Kava-Kava) – Psilocybe semilanceata (magic mushrooms) – Rauwolfia serpentina (Indian snakeroot) – Rauwolfia... serpentina (Indian Snakeroot) – Scilla maritima (White Squill) – Scopolia carniolica (Scopolia)* U.S. Navy Aeromedical Reference and Waiver Guide...be sedatives: – Valeriana officinalis (Valerian) – Rauwolfia serpentina (Indian snakeroot) – Atropa belladonna (Deadly Nightshade)* – Chelidonium

  10. Salt stress in Plantago - The role of membranes, channels and pumps

    NARCIS (Netherlands)

    Prins, HBA

    1995-01-01

    In the present article the cellular mechanism of Na+ transport across the plasma membrane and tonoplast of root cells of Plantago media (salt sensitive) and Plantago maritima (salt tolerant) is discussed based on findings obtained mainly by patch clamp technique. It is conluded that the combination

  11. The Security of the South Atlantic: Is It a Case for ’SATO’--South Atlantic Treaty Organization

    Science.gov (United States)

    1982-05-10

    p. 704. 21Ibid. 2 2 Ibid. 2 3A.R.A. Nicolas Piccaluga, Control Del Trafico Maritimo en el Atlantic Sud. Politics para la Defensa v Seguridad de las...1981. Piccaluga, A. R. A, Nicolis. Control Del Trafico Maritimo En El Atlantico Sud. Politicar, Para La Defensa Y Seguridad De Las Rutas Maritimas En

  12. Quelques noddees sur l’ecologie de la vedetation des dunes et sur la fonction de l’enraciment dans l’edification de dunes a la Cote Mediterraneenne de la France. I

    NARCIS (Netherlands)

    Boterenbrood, A.J.; Donsellaar-Ten Bokkel Huinink, van W.A.E.; Donselaar, van J.

    1956-01-01

    Dans la végétation des dunes du Languedoc J. BRAUN-BLANQUET (1952) distingue trois associations, à savoir; 1) l’Agropyretum mediterraneum parmi et sur les premières dunes basses; 2) l’Ammophiletum arundinaceae sur les dunes plus hautes; et 3) le Crucianelletum maritimae dans les dépressions et en

  13. Chromatographic fingerprint analysis of Pycnogenol® dietary supplements

    Science.gov (United States)

    French maritime bark (Pinus maritima) has been widely used as an herbal remedy for various degenerative diseases. A standardized bark extract is available that complies with its USP monograph and is derived from Pinus pinaster, Ait. (Pycnogenol®, Horphag Research Ltd., UK). The method specified in...

  14. Klaasikunstnikud puhuvad ja valavad klaasi taas Haapsalus / Maris Sander

    Index Scriptorium Estoniae

    Sander, Maris

    2004-01-01

    Haapsalu 2. klaasipäevad "Maritima" (lad. keeles mereäärne) Evald Okase majamuuseumis. Osa võtavad kunstnikud Eestist, Koreast, Jaapanist ja Kanadast. Korraldaja Kai Koppel. Kommentaar Kati Kerstna. Valminud tööd pannakse näitusele

  15. 1. VI avati Haapsalus taas Evald Okase muuseum

    Index Scriptorium Estoniae

    2004-01-01

    Väljas on Evald Okase graafika ja joonistute uuendatud ekspositsioon, kunstniku 1930. aastate maalid. Programmis: fotonäitus "Väikeste majade Haapsalu", rahvusvaheline kuumklaasi workshop ja klaasinäitus "Maritima", kunsti suvekool, Eesti Maalikunstnike Liidu näitus "Ma olen olnud Haapsalus" jm.

  16. Antifungal Activities of Extracts from Selected Lebanese Wild Plants against Plant Pathogenic Fungi

    Directory of Open Access Journals (Sweden)

    Y. Abou-Jawdah

    2004-12-01

    Full Text Available Extracts of nine plant species growing wild in Lebanon were tested for their efficacy against seven plant pathogenic fungi: Botrytis cinerea, Alternaria solani, Penicillium sp., Cladosporium sp., Fusarium oxysporum f. sp. melonis, Rhizoctonia solani and Sphaerotheca cucurbitae. Extracts of three of the plants, Origanum syriacum, Micromeria nervosa and Plumbago maritima, showed the highest levels of in vitro activity against spore germination and mycelial growth of the fungi tested. Inula viscosa showed high activity against spore germination but only moderate activity against mycelial growth. The other five plant species tested Calamintha origanifolia, Micromeria juliana, Ruta sp., Sideritis pullulans and Urginea maritima showed only moderate to low activity against these fungi. Preventive sprays with extracts of O. syriacum, M. nervosa, P. maritima and I. viscosa, applied at concentrations ranging between 4 and 8% to squash and cucumber seedlings, gave efficient protection against gray mold caused by B. cinerea and powdery mildew caused by S. cucurbitae. However, these extracts did not control green mold of citrus fruits caused by Penicillium sp. Thin layer chromatography revealed three inhibitory bands in extracts of O. syriacum, two in I. viscosa and only one in each of the other plants tested: M. nervosa, P. maritima, C. origanifolia and Ruta sp.

  17. Mereäärset maad meenutavad nukk, klotsid ja nokats / Anu Jürisson

    Index Scriptorium Estoniae

    Jürisson, Anu, 1977-

    2009-01-01

    Matsalu piirkonna meenekonkursi võidutöödest. Esikoha sai Pilvi Kangro meisterdatud nokats, 2. koha Tiit Kaljuste fotodega mänguklotsid ja 3. koha Mirje Simsi rahvarõivais nukk. Selgitusi jagab Terra Maritima juht Nele Sõber. Töid saab näha Lihula käsitööpoes

  18. Long-term trends in abundance of cladocerans in the Central Baltic Sea

    DEFF Research Database (Denmark)

    Möllmann, C.; Köster, Fritz; Kornilovs, G.

    2002-01-01

    on the abundance of cladoceran species was investigated. A clear affinity to higher temperature was found for B. coregoni maritima in summer as well as for E. nordmanni and Podon spp. in spring. In addition to temperature, association tests with salinity revealed besides species-specific preferences, regional...

  19. Impact of soil nematodes on salt-marsh plants : a pilot experiment

    NARCIS (Netherlands)

    Dormann, CF; van der Wal, R

    2001-01-01

    We tested whether the removal of nematodes by means of nematicide application changed plant performance or influenced plant competition. The study involved the two common plant species Artemisia maritima and Festuca rubra growing in intact sods collected from a temperate salt marsh. Half of the sods

  20. Effects of ocean acidification on single and mixed seagrass species meadows in estuarine waters of the Northern Gulf of Mexico

    Science.gov (United States)

    In an outdoor mesocosm, we tested the hypothesis that OA would benefit seagrasses in mesohaline waters of the Northern Gulf of Mexico in homo- and hetero- specific seagrass beds of Halodule wrightii and Ruppia maritima. In this estuarine environment, short-term increases in CO2 ...

  1. Free radical scavenging and cytotoxic activity of five commercial ...

    African Journals Online (AJOL)

    Polygonum cuspidatum), and pomegranate (Punica granatum). It shows radical scavenging activity in the following order, according to their median effective concentration (ECmaritima 7 µg/ml, ...

  2. Florae Malesianae Praecursores LX. The Oleaceae of Malesia. II. The genus Olea

    NARCIS (Netherlands)

    Kiew, Ruth

    1979-01-01

    Olea comprises six species in Malesia: two from Malaya. O. brachiata (Lour.) Merrill (formerly O. maritima Wall. ex G. Don) and O. dentata Wall. ex G. Don (formerly O. penangiana Ridley); two from Borneo, O. borneensis Boerl. and O. decussata (Heine) Kiew and two from Java, O. javanica (Bl.) Knobl.

  3. Ecophysiological responses of the salt marsh grass Spartina ...

    African Journals Online (AJOL)

    The effects of salinity on growth and productivity of Spartina maritima (Curtis) Fernald were investigated in glasshouse and field experiments in 2008. In the glasshouse study, plants were subjected to 2%, 10%, 20%, 40% and 80% sea water, with tidal simulation, for 10 months. Increase in salinity from 2% to 20% sea water ...

  4. 4. VII kell 17 avatakse Haapsalu Evald Okase muuseumis rahvusvaheline klaasinäitus...

    Index Scriptorium Estoniae

    2004-01-01

    Ühtlasi lõpevad II Haapsalu klaasipäevad "Maritima". Korraldaja Kai Koppel. Klaasi puhuvad Rait Parts, Robert Tannahill Kanadast, Sun Hwan Hong Koreast, Kai Koppel. Näitusel näeb eesti klaasikunstnike Mare Saare, Ivo Lille, Rait Präätsa jt. töid

  5. Maritime Factors Affecting Iberian Security,

    Science.gov (United States)

    1980-10-01

    afirme/ que para las sovie’ticas una campaiia en gran escala dirigida contra las comunicaciones marftimas del Atlantica Norte durante el perfada inicial...defender sus l ’neas de comunicacion maritima; y cuarta -- y a mi parecer la consideracion mas importante -- las otras tareas que la marina de guerra

  6. The intricate nomenclatural questions around Plantago holosteum (Plantaginaceae)

    DEFF Research Database (Denmark)

    Iamonico, Duilio; Hassemer, Gustavo; Rønsted, Nina

    2017-01-01

    on illustration by Bauhin & Cherler, P. wulfenii by Willdenow on a specimen preserved at B while P. maritima var. apennina was neotypified using a specimen deposited at RO. For P. holosteum, an accepted and widely used name both in the floristic and the vegetation literature of SE-Europe, an epitype is designated...

  7. 78 FR 12703 - Fisheries of the Caribbean, Gulf of Mexico, and South Atlantic; Amendment to the Corals and Reef...

    Science.gov (United States)

    2013-02-25

    ... Associated Plants and Invertebrates Fishery Management Plan of Puerto Rico and the U.S. Virgin Islands AGENCY... Reef Associated Plants and Invertebrates of Puerto Rico and the U.S. Virgin Islands (USVI) (Coral FMP... maritima), and one group of species, the sea vines (Halophila spp., including H. decipiens, H. baillonis, H...

  8. Evidence of molecular adaptation to extreme environments and applicability to space environments

    Directory of Open Access Journals (Sweden)

    Filipović M.

    2008-01-01

    Full Text Available This is initial investigation of gene signatures responsible for adapting microscopic life to the extreme Earth environments. We present preliminary results on identification of the clusters of orthologous groups (COGs common to several hyperthermophiles and exclusion of those common to a mesophile (non-hyperthermophile: Escherichia coli (E. coli K12, will yield a group of proteins possibly involved in adaptation to life under extreme temperatures. Comparative genome analyses represent a powerful tool in discovery of novel genes responsible for adaptation to specific extreme environments. Methanogens stand out as the only group of organisms that have species capable of growth at 0ºC (Metarhizium frigidum (M. frigidum and Methanococcoides burtonii (M. burtonii and 110ºC (Methanopyrus kandleri (M. kandleri. Although not all the components of heat adaptation can be attributed to novel genes, the chaperones known as heat shock proteins stabilize the enzymes under elevated temperature. However, highly conserved chaperons found in bacteria and eukaryots are not present in hyperthermophilic Archea, rather, they have a unique chaperone TF55. Our aim was to use software which we specifically developed for extremophile genome comparative analyses in order to search for additional novel genes involved in hyperthermophile adaptation. The following hyperthermophile genomes incorporated in this software were used for these studies: Methanocaldococcus jannaschii (M. jannaschii, M. kandleri, Archaeoglobus fulgidus (A. fulgidus and three species of Pyrococcus. Common genes were annotated and grouped according to their roles in cellular processes where such information was available and proteins not previously implicated in the heat-adaptation of hyperthermophiles were identified. Additional experimental data are needed in order to learn more about these proteins. To address non-gene based components of thermal adaptation, all sequenced extremophiles were

  9. Remarkable morphological diversity of viruses and virus-like particles in hot terrestrial environments.

    Science.gov (United States)

    Rachel, R; Bettstetter, M; Hedlund, B P; Häring, M; Kessler, A; Stetter, K O; Prangishvili, D

    2002-12-01

    Electron microscopic studies of the viruses in two hot springs (85 degrees C, pH 1.5-2.0, and 75-93 degrees C, pH 6.5) in Yellowstone National Park revealed particles with twelve different morphotypes. This diversity encompassed known viruses of hyperthermophilic archaea, filamentous Lipothrixviridae, rod-shaped Rudiviridae, and spindle-shaped Fuselloviridae, and novel morphotypes previously not observed in nature. Two virus types resembled head-and-tail bacteriophages from the families Siphoviridae and Podoviridae, and constituted the first observation of these viruses in a hydrothermal environment. Viral hosts in the acidic spring were members of the hyperthermophilic archaeal genus Acidianus.

  10. Hyperthermostable cellulolytic and hemicellulolytic enzymes and their biotechnological applications

    Directory of Open Access Journals (Sweden)

    Tipparat Hongpattarakere

    2002-07-01

    Full Text Available Hyperthermal cellulases and hemicellulases have been intensively studied due to their highly potential applications at extreme temperatures, which mimic industrial processes involving cellulose and hemicellulose degradation. More than 50 species of hyperthermophiles have been isolated, many of which possess hyperthermal enzymes required for hydrolyzing cellulose and hemicelluloses. Endoglucanases, exoglucanases, cellobiohydrolases, xylanases, β-glucosidase and β-galactosidase, which are produced by the hyperthermophiles, are resistant to boiling temperature. The characteristics of these enzymes and the ability to maintain their functional integrity at high temperature as well as their biotechnological application are discussed.

  11. AcEST: DK951745 [AcEST

    Lifescience Database Archive (English)

    Full Text Available somal protein S5 OS=Thermotoga sp. (s... 33 10.0 tr|B7RFH6|B7RFH6_9THEM Ribosomal protein S5 OS=Marinitoga piezo...6_9THEM Ribosomal protein S5 OS=Marinitoga piezophila KA3 GN=rpsE PE=4 SV=1 Length = 178 Score = 33.5 bits (

  12. Exploring the reductive capacity of Pyrococcus furiosus : the reduction of carboxylic acids and pyridine nucleotides

    NARCIS (Netherlands)

    Ban, van den E.C.D.

    2001-01-01

    This Ph.D. project started in 1997 and its main goal was to obtain insight in the reductive capacity of the hyperthermophilic archaeon Pyrococcus furiosus . The research was focused on the biocatalytic reduction of carboxylic

  13. Stress management skills in the subsurface: H2 stress on thermophilic heterotrophs and methanogens

    Science.gov (United States)

    Topcuoglu, B. D.; Holden, J. F.

    2017-12-01

    Marine hyperthermophilic heterotrophs and methanogens belonging to the Thermococcales and Methanococcales are often found in subsurface environments such as coal and shale beds, marine sediments, and oil reservoirs where they encounter H2 stress conditions. It is important to study the H2 stress survival strategies of these organisms and their cooperation with one another for survival to better understand their biogeochemical impact in hot subsurface environments. In this study, we have shown that H2 inhibition changed the growth kinetics and the transcriptome of Thermococcus paralvinellae. We observed a significant decrease in batch phase growth rates and cell concentrations with high H2 background. Produced metabolite production measurements, RNA-seq analyses of differentially expressed genes and in silico experiments we performed with the T. paralvinellae metabolic model showed that T. paralvinellae produces formate by a formate hydrogenlyase to survive H2 inhibition. We have also shown that H2 limitation caused a significant decrease in batch phase growth rates and methane production rates of the methanogen, Methanocaldococcus jannaschii. H2 stress of both organisms can be ameliorated by syntrophic growth. H2 syntrophy was demonstrated in microcosm incubations for a natural assemblage of Thermococcus and hyperthermophilic methanogens present in hydrothermal fluid samples. This project aims to describe how a hyperthermophilic heterotroph and a hyperthermophilic methanogen eliminate H2 stress and explore cooperation among thermophiles in the hot subsurface.

  14. Tertiary structure in 7.9 M guanidinium chloride: the role of Glu-53 and Asp-287 in Pyrococcus furiosus endo-beta-1,3-glucanase

    NARCIS (Netherlands)

    Chiaraluce, R.; Florio, R.; Angelaccio, S.; Gianese, G.; Lieshout, van J.F.T.; Oost, van der J.; Consalvi, V.

    2007-01-01

    The thermodynamic stability of family 16 endo-ß-1,3-glucanase (EC 3.2.1.39) from the hyperthermophilic archaeon Pyrococcus furiosus is decreased upon single (D287A, E53A) and double (E53A/D287A) mutation of Asp287 and Glu53. In accordance with the homology model prediction, both carboxylic acids are

  15. Conditions for gene disruption by homologous recombination of exogenous DNA into the Sulfolobus solfataricus genome

    NARCIS (Netherlands)

    Albers, Sonja-Verena; Driessen, Arnold J.M.

    2008-01-01

    The construction of directed gene deletion mutants is an essential tool in molecular biology that allows functional studies on the role of genes in their natural environment. For hyperthermophilic archaea, it has been difficult to obtain a reliable system to construct such mutants. However, during

  16. Conditions for gene disruption by homologous recombination of exogenous DNA into the Sulfolobus solfataricus genome

    OpenAIRE

    Albers, Sonja-Verena; Driessen, Arnold J. M.

    2008-01-01

    The construction of directed gene deletion mutants is an essential tool in molecular biology that allows functional studies on the role of genes in their natural environment. For hyperthermophilic archaea, it has been difficult to obtain a reliable system to construct such mutants. However, during the past years, systems have been developed for Thermococcus kodakarensis and two Sulfolobus species, S. ac...

  17. Draft Genome Sequence of a Highly Flagellated, Fast-Swimming Archaeon, Methanocaldococcus villosus Strain KIN24-T80 (DSM 22612)

    KAUST Repository

    Thennarasu, Sugumar

    2013-07-11

    We report the draft genome sequence of a hyperthermophilic Methanocaldococcus villosus strain, KIN24-T80. The gene associated with its heavy flagellum formation was annotated in the 1.2-Mb draft genome sequence, and this strain may be a good model system to study the extensive functional role of flagella and their fast motor activity.

  18. Draft Genome Sequence of a Highly Flagellated, Fast-Swimming Archaeon, Methanocaldococcus villosus Strain KIN24-T80 (DSM 22612)

    KAUST Repository

    Thennarasu, Sugumar; Poli Reddy, Dinesh Reddy; Antony, Aju; Yada, Madhava; Alqarawi, Sami; Neelamegam, Sivakumar

    2013-01-01

    We report the draft genome sequence of a hyperthermophilic Methanocaldococcus villosus strain, KIN24-T80. The gene associated with its heavy flagellum formation was annotated in the 1.2-Mb draft genome sequence, and this strain may be a good model system to study the extensive functional role of flagella and their fast motor activity.

  19. Characterization of different crystal forms of the alpha-glucosidase MalA from Sulfolobus solfataricus

    DEFF Research Database (Denmark)

    Ernst, Heidi Asschenfeldt; Willemoës, Martin; Lo Leggio, Leila

    2005-01-01

    MalA is an alpha-glucosidase from the hyperthermophilic archaeon Sulfolobus solfataricus. It belongs to glycoside hydrolase family 31, which includes several medically interesting alpha-glucosidases. MalA and its selenomethionine derivative have been overproduced in Escherichia coli...

  20. ADP-dependent Phosphofructokinases in Mesophilic and Thermophilic Methanogenic Archaea

    NARCIS (Netherlands)

    Verhees, C.H.; Tuininga, J.E.; Kengen, S.W.M.; Stams, A.J.M.; Oost, van der J.; Vos, de W.M.

    2001-01-01

    Phosphofructokinase (PFK) is a key enzyme of the glycolytic pathway in all domains of life. Two related PFKs, ATP-dependent and PPi-dependent PFK, have been distinguished in bacteria and eucarya, as well as in some archaea. Hyperthermophilic archaea of the order Thermococcales, including Pyrococcus

  1. Molecular analysis of the UV-inducible pili operon from Sulfolobus acidocaldarius

    NARCIS (Netherlands)

    Wolferen, Marleen van; Ajon, Małgorzata; Driessen, Arnold J.M.; Albers, Sonja-Verena

    2013-01-01

    Upon ultraviolet (UV) stress, hyperthermophilic Sulfolobus species show a highly induced transcription of a gene cluster responsible for pili biogenesis: the UV-inducible pili operon (ups operon). This operon is involved in UV-induced pili assembly, cellular aggregation, and subsequent DNA exchange

  2. Crenarchaeal Viruses: Morphotypes and Genomes,

    DEFF Research Database (Denmark)

    Prangishvili, P.; Basta, P.; Garrett, Roger Antony

    2008-01-01

    In this article we present our current knowledge about double-stranded (dsDNA) viruses infecting hyperthermophilic Crenarchaeaota, the organisms which predominate in hot terrestrial springs with temperatures over 80 °C. These viruses exhibit extraordinary diversity of morphotypes most of which have...

  3. Four newly isolated fuselloviruses from extreme geothermal environments reveal unusual morphologies and a possible interviral recombination mechanism

    DEFF Research Database (Denmark)

    Redder, Peter; Peng, Xu; Brügger, Kim

    2009-01-01

    Spindle-shaped virus-like particles are abundant in extreme geothermal environments, from which five spindle-shaped viral species have been isolated to date. They infect members of the hyperthermophilic archaeal genus Sulfolobus, and constitute the Fuselloviridae, a family of double-stranded DNA...

  4. Dynamic fluorescence studies of beta-glycosidase mutants from Sulfolobus solfataricus: effects of single mutations on protein thermostability.

    Science.gov (United States)

    Bismuto, Ettore; Febbraio, Ferdinando; Limongelli, Simona; Briante, Raffaella; Nucci, Roberto

    2003-04-01

    Multiple sequence alignment on 73 proteins belonging to glycosyl hydrolase family 1 reveals the occurrence of a segment (83-124) in the enzyme sequences from hyperthermophilic archaea bacteria, which is absent in all the mesophilic members of the family. The alignment of the known three-dimensional structures of hyperthermophilic glycosidases with the known ones from mesophilic organisms shows a similar spatial organizations of beta-glycosidases except for this sequence segment whose structure is located on the external surface of each of four identical subunits, where it overlaps two alpha-helices. Site-directed mutagenesis substituting N97 or S101 with a cysteine residue in the sequence of beta-glycosidase from hyperthermophilic archaeon Sulfolobus solfataricus caused some changes in the structural and dynamic properties as observed by circular dichroism in far- and near-UV light, as well as by frequency domain fluorometry, with a simultaneous loss of thermostability. The results led us to hypothesize an important role of the sequence segment present only in hyperthermophilic beta-glycosidases, in the thermal adaptation of archaea beta-glycosidases. The thermostabilization mechanism could occur as a consequence of numerous favorable ionic interactions of the 83-124 sequence with the other part of protein matrix that becomes more rigid and less accessible to the insult of thermal-activated solvent molecules. Copyright 2003 Wiley-Liss, Inc.

  5. Viruses of the Archaea

    DEFF Research Database (Denmark)

    Basta, T.; Garrett, Roger Antony; Prangishvili,, David

    2009-01-01

    Double-stranded deoxyribonucleic acid (DNA) viruses that infect members of the third domain of life, the Archaea, are diverse and exceptional in both their morphotypes and their genomic properties. The majority of characterized species infect hyperthermophilic hosts and carry morphological featur...

  6. Viruses of the Archaea

    DEFF Research Database (Denmark)

    Prangishvili,, David; Basta, Tamara; Garrett, Roger Antony

    2016-01-01

    Viruses infecting members of Archaea, the third domain of life, constitute an integral, yet unique part of the virosphere. Many of these viruses, specifically the species that infect hyperthermophilic hosts, display morphotypes – for example, bottle shaped, spindle shaped, droplet shaped, coil sh...

  7. Structure and genome organization of AFV2, a novel archaeal lipothrixvirus with unusual terminal and core structures

    DEFF Research Database (Denmark)

    Häring, Monika; Vestergaard, Gisle Alberg; Brügger, Kim

    2005-01-01

    A novel filamentous virus, AFV2, from the hyperthermophilic archaeal genus Acidianus shows structural similarity to lipothrixviruses but differs from them in its unusual terminal and core structures. The double-stranded DNA genome contains 31,787 bp and carries eight open reading frames homologous...

  8. Production and characterization of a thermostable alcohol dehydrogenase that belongs to the aldo-keto reductase superfamily

    NARCIS (Netherlands)

    Machielsen, M.P.; Uria, A.R.; Kengen, S.W.M.; Oost, van der J.

    2006-01-01

    The gene encoding a novel alcohol dehydrogenase that belongs to the aldo-keto reductase superfamily has been identified in the hyperthermophilic archaeon Pyrococcus furiosus. The gene, referred to as adhD, was functionally expressed in Escherichia coli and subsequently purified to homogeneity. The

  9. A site-specific endonuclease encoded by a typical archaeal intron

    DEFF Research Database (Denmark)

    Dalgaard, Jacob; Garrett, Roger Antony; Belfort, Malene

    1993-01-01

    The protein encoded by the archaeal intron in the 23S rRNA gene of the hyperthermophile Desulfurococcus mobilis is a double-strand DNase that, like group I intron homing endonucleases, is capable of cleaving an intronless allele of the gene. This enzyme, I-Dmo I, is unusual among the intron...

  10. Laboratory Directed Research and Development Program, FY 1995

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1995-12-31

    This program provides the resources for Berkeley Lab scientists to make rapid and significant contributions to critical nation science and technology problems: accelerators and fusion, chemical sciences, earth sciences, energy and environment, engineering, life sciences, materials, nuclear science, physics, and structural biology (hyperthermophilic microorganisms).

  11. Virology: Independent virus development outside a host

    DEFF Research Database (Denmark)

    Häring, M.; Vestergaard, Gisle Alberg; Rachel, R.

    2005-01-01

    Viruses are thought to be functionally inactive once they are outside and independent of their host cell 1 . Here we describe an exceptional property of a newly discovered virus that infects a hyperthermophilic archaeon growing in acidic hot springs: the lemon-shaped viral particle develops a very...

  12. Amylomaltase of Pyrobaculum aerophilum IM2 produces thermoreversible starch gels

    NARCIS (Netherlands)

    Kaper, T.; Talik, B.; Ettema, T.J.; Bos, H.; Maarel, M.J.E.C. van der; Dijkhuizen, L.

    2005-01-01

    Amylomaltases are 4-α-glucanotransferases (EC 2.4.1.25) of glycoside hydrolase family 77 that transfer α-1,4-linked glucans to another acceptor, which can be the 4-OH group of an α-1,4-linked glucan or glucose. The amylomaltase-encoding gene (PAE1209) from the hyperthermophilic archaeon Pyrobaculum

  13. Diversity of bacteria and archaea from two shallow marine hydrothermal vents from Vulcano Island.

    Science.gov (United States)

    Antranikian, Garabed; Suleiman, Marcel; Schäfers, Christian; Adams, Michael W W; Bartolucci, Simonetta; Blamey, Jenny M; Birkeland, Nils-Kåre; Bonch-Osmolovskaya, Elizaveta; da Costa, Milton S; Cowan, Don; Danson, Michael; Forterre, Patrick; Kelly, Robert; Ishino, Yoshizumi; Littlechild, Jennifer; Moracci, Marco; Noll, Kenneth; Oshima, Tairo; Robb, Frank; Rossi, Mosè; Santos, Helena; Schönheit, Peter; Sterner, Reinhard; Thauer, Rudolf; Thomm, Michael; Wiegel, Jürgen; Stetter, Karl Otto

    2017-07-01

    To obtain new insights into community compositions of hyperthermophilic microorganisms, defined as having optimal growth temperatures of 80 °C and above, sediment and water samples were taken from two shallow marine hydrothermal vents (I and II) with temperatures of 100 °C at Vulcano Island, Italy. A combinatorial approach of denaturant gradient gel electrophoresis (DGGE) and metagenomic sequencing was used for microbial community analyses of the samples. In addition, enrichment cultures, growing anaerobically on selected polysaccharides such as starch and cellulose, were also analyzed by the combinatorial approach. Our results showed a high abundance of hyperthermophilic archaea, especially in sample II, and a comparable diverse archaeal community composition in both samples. In particular, the strains of the hyperthermophilic anaerobic genera Staphylothermus and Thermococcus, and strains of the aerobic hyperthermophilic genus Aeropyrum, were abundant. Regarding the bacterial community, ε-Proteobacteria, especially the genera Sulfurimonas and Sulfurovum, were highly abundant. The microbial diversity of the enrichment cultures changed significantly by showing a high dominance of archaea, particularly the genera Thermococcus and Palaeococcus, depending on the carbon source and the selected temperature.

  14. Laboratory Directed Research and Development Program, FY 1995

    International Nuclear Information System (INIS)

    1995-01-01

    This program provides the resources for Berkeley Lab scientists to make rapid and significant contributions to critical nation science and technology problems: accelerators and fusion, chemical sciences, earth sciences, energy and environment, engineering, life sciences, materials, nuclear science, physics, and structural biology (hyperthermophilic microorganisms)

  15. The biochemical diversity of life near and above 100°C in marine environments.

    Science.gov (United States)

    Adams, M W

    1998-12-01

    Hyperthermophilic micro-organisms grow at temperatures above 90 °C with a current upper limit of 113 °C. They are a recent discovery in the microbial world and have been isolated mainly from marine geothermal environments, which include both shallow and deep sea hydrothermal vents. By 16S rRNA analyses they are the most slowly evolving of all extant life forms, and all but two of the nearly 20 known genera are classified as Archaea (formerly Archaebacteria). Almost all hyperthermophiles are strict anaerobes. They include species of methanogens, iron-oxidizers and sulphate reducers, but the majority are obligate heterotrophs that depend upon the reduction of elemental sulphur (S°) to hydrogen sulphide for significant growth. The heterotrophs utilize proteinaceous materials as carbon and energy sources, although a few species are also saccharolytic. A scheme for electron flow during the oxidation of carbohydrates and peptides and the reduction of S° has been proposed. Two S°-reducing enzymes have been purified from the cytoplasm of one hyperthermophile (T(opt) 100 °C) that is able to grow either with and without S°. However, the mechanisms by which S° reduction is coupled to energy conservation in this organism and in obligate S°-reducing hyperthermophiles is not known. In the heterotrophs, sugar fermentation is achieved by a novel glycolytic pathway involving unusual ADP-dependent kinases and ATP synthetases, and novel oxidoreductases that are ferredoxin- rather than NAD(P)-linked. Similarly, peptide fermentation involves several unusual ferredoxin-linked oxidoreductases not found in mesophilic organisms. Several of these oxido-reductases contain tungsten, an element that is rarely used in biological systems. Tungsten is present in exceedingly low concentrations in normal sea water, but hydrothermal systems contain much higher tungsten concentrations, more than sufficient to support hyperthermophilic life. 1998 Society of Applied Microbiology.

  16. Evidence of Molecular Adaptation to Extreme Environments and Applicability to Space Environments

    Directory of Open Access Journals (Sweden)

    Filipović, M. D.

    2008-06-01

    Full Text Available This is initial investigation of gene signatures responsible for adapting microscopic life to the extreme Earth environments. We present preliminary results on identification of the clusters of orthologous groups (COGs common to several hyperthermophiles and exclusion of those common to a mesophile (non-hyperthermophile: {it Escherichia coli (E. coli K12}, will yield a group of proteins possibly involved in adaptation to life under extreme temperatures. Comparative genome analyses represent a powerful tool in discovery of novel genes responsible for adaptation to specific extreme environments. Methanogens stand out as the only group of organisms that have species capable of growth at 0D C ({it Metarhizium frigidum (M.~frigidum} and {it Methanococcoides burtonii (M.~burtonii} and 110D C ({it Methanopyrus kandleri (M.~kandleri}. Although not all the components of heat adaptation can be attributed to novel genes, the {it chaperones} known as heat shock proteins stabilize the enzymes under elevated temperature. However, highly conserved {it chaperons} found in bacteria and eukaryots are not present in hyperthermophilic Archea, rather, they have a unique {it chaperone TF55}. Our aim was to use software which we specifically developed for extremophile genome comparative analyses in order to search for additional novel genes involved in hyperthermophile adaptation. The followinghyperthermophile genomes incorporated in this software were used forthese studies: {it Methanocaldococcus jannaschii (M.~jannaschii, M.~kandleri, Archaeoglobus fulgidus (A.~fulgidus} and threespecies of {it Pyrococcus}. Common genes were annotated and groupedaccording to their roles in cellular processes where such informationwas available and proteins not previously implicated in theheat-adaptation of hyperthermophiles were identified. Additionalexperimental data are needed in order to learn more about theseproteins. To address non-gene based components of thermaladaptation

  17. Toxicological, chemical and antibacterial evaluation of squill vinegar, a useful product in Persian Traditional Medicine

    OpenAIRE

    M. Bozorgi; G.R. Amin; S.N. Ostad; N. Samadi; E. Nazem; M. Shekarchi

    2017-01-01

    Background and objectives:  Squill [Drimia maritima (L.) Stearn] is an important medicinal plant that has been used for medicinal purposes such as cardiovascular diseases and asthma since ancient times. Bufadienolides are the main compounds of this plant and are responsible for some reported adverse effects. In order to reduce adverse effects, different methods like boiling with vinegar were applied by traditional practitioners. In the present study, the acute oral toxicity, cytotoxic effects...

  18. Thermostable cellulases, and mutants thereof, capable of hydrolyzing cellulose in ionic liquid

    Science.gov (United States)

    Sapra, Rajat; Datta, Supratim; Chen, Zhiwei; Holmes, Bradley M.; Simmons, Blake A.; Blanch, Harvey W.

    2016-04-26

    The present invention provides for a composition comprising an ionic liquid and a thermostable cellulose, and a method of hydrolyzing a cellulose, comprising: (a) providing a composition comprising a solution comprising an ionic liquid and a cellulose, and (b) introducing a thermostable cellulase to the solution, such that the cellulose is hydrolyzed by the cellulase. The present invention also provides for a Thermatoga maritima thermostable cellulase mutant with increased cellulase activity.

  19. Agronomic aspects of strip intercropping lettuce with alyssum for biological control of aphids

    OpenAIRE

    Brennan, Eric B.

    2013-01-01

    Organic lettuce growers in California typically use insectary strips of alyssum (Lobularia maritima (L.) Desv.) to attract hoverflies (Syrphidae) that provide biological control of aphids. A two year study with transplanted organic romaine lettuce in Salinas, California investigated agronomic aspects of lettuce monoculture and lettuce-alyssum strip intercropping on beds in replacement intercropping treatments where alyssum transplants replaced 2 to 8% of the lettuce transplants, and in additi...

  20. Final Environmental Impact Statement Evolved Expendable Launch Vehicle Program

    Science.gov (United States)

    1998-04-01

    including increased blood pressure and higher levels of available glucose and corticosteroids in the bloodstream. Continued disturbances and prolonged...treefrogs. Besides the common American alligator , reptiles observed include the Florida box turtle, the gopher tortoise, the Florida softshell, the...star Remirea maritima - E Reptiles and Amphibians Gopher frog Rana capito C SSC American alligator Alligator mississippiensis T(S/A) SSC Eastern Indigo

  1. The relationship between habitat complexity and nursery provision for an estuarine-dependent fish species in a permanently open South African Estuary

    Science.gov (United States)

    Leslie, Timothy; James, Nicola C.; Potts, Warren M.; Rajkaran, Anusha

    2017-11-01

    Estuarine-dependent marine fish species rely on shallow, sheltered and food rich habitats for protection from predators, growth and ultimately recruitment to adult populations. Hence, habitats within estuaries function as critical nursery areas for an abundance of fish species. However, these habitats vary in the degree of nursery function they provide and few studies have quantitatively assessed the relative nursery value of different habitat types within estuaries, particularly in the context of habitat complexity. This study aimed to assess the nursery value of the dominant vegetated habitats, namely the submergent Zostera capensis (Setch.) (seagrass) beds and emergent Spartina maritima (Curtis) Fernald (salt marsh) beds in the Bushmans Estuary, South Africa. Biomass and stem density were sampled seasonally in order to gain insight into the vegetation dynamics of seagrass and salt marsh beds. Aerial cover, canopy height and underwater camera imagery were used to develop multiple complexity indices for prioritizing habitat complexity. The relatively consistent results of the dimensionless indices (interstitial space indices and fractal geometry) suggest that Z. capensis exhibits an overall greater degree of complexity than S. maritima, and hence it can be expected that fish abundance is likely to be higher in Z. capensis beds than in S. maritima habitats. Underwater video cameras were deployed in seagrass, salt marsh and sand flat habitats to assess the relative abundance and behaviour of the estuarine-dependent sparid Rhabosargus holubi (Steindachner 1881) in different habitats. The relative abundance of R. holubi was significantly higher in Z. capensis seagrass than S. maritima salt marsh and sand flats, whilst the behaviour of R. holubi indicated a high degree of habitat use in structured habitats (both Z. capensis and S. martima) and a low degree of habitat use in unstructured sand flat habitats.

  2. Halophyte vegetation influences in salt marsh retention capacity for heavy metals

    International Nuclear Information System (INIS)

    Reboreda, Rosa; Cacador, Isabel

    2007-01-01

    We analysed concentrations of Cu, Cd and Pb in above and belowground tissues of the halophyte species Halimione portulacoides and Spartina maritima, as well as in sediments and pore water between the roots in a Tagus estuary salt marsh (Portugal). From these results we calculated the pools of metals in the compartments mentioned above. Relative percentages of accumulation in each pool were also determined. Our aim was to determine how the type of vegetation in the salt marsh affects overall metal retention capacity of the system. It was concluded that areas colonised by H. portulacoides are potential sources of Cu, Cd and Pb to the marsh ecosystem, whereas areas colonised by S. maritima are more effective sinks at least for Cu and Cd. Consequently, S. maritima seems to contribute more effectively to the stabilisation of metals in salt marsh sediments, reducing their availability to the estuarine system. - The type of vegetal cover can affect the overall retention capacity of a salt marsh as well as the functioning of the salt marsh as a sink or source of metals to the estuarine system

  3. Utilization of keratin-containing biowaste to produce biohydrogen

    Energy Technology Data Exchange (ETDEWEB)

    Balint, B.; Rakhely, G.; Kovacs, K.L. [Szeged Univ. (Hungary). Dept. of Biotechnology; Hungarian Academy of Sciences, Szeged (Hungary). Inst. of Biophysics; Bagi, Z.; Perei, K. [Szeged Univ. (Hungary). Dept. of Biotechnology; Toth, A. [Hungarian Academy of Sciences, Szeged (Hungary). Inst. of Biophysics

    2005-12-01

    A two-stage fermentation system was constructed to test and demonstrate the feasibility of biohydrogen generation from keratin-rich biowaste. We isolated a novel aerobic Bacillus strain (Bacillus licheniformis KK1) that displays outstanding keratinolytic activity. The isolated strain was employed to convert keratin-containing biowaste into a fermentation product that is rich in amino acids and peptides. The process was optimized for the second fermentation step, in which the product of keratin fermentation-supplemented with essential minerals-was metabolized by Thermococcus litoralis, an anaerobic hyperthermophilic archaeon. T. litoralis grew on the keratin hydrolysate and produced hydrogen gas as a physiological fermentation byproduct. Hyperthermophilic cells utilized the keratin hydrolysate in a similar way as their standard nutrient, i.e., bacto-peptone. The generalization of the findings to protein-rich waste treatment and production of biohydrogen is discussed and possible means of further improvements are listed. (orig.)

  4. Crystallization and preliminary X-ray characterization of a PaaX-like protein from Sulfolobus solfataricus P2

    International Nuclear Information System (INIS)

    Cao, Yi; Lou, Zhiyong; Sun, Yuna; Xue, Fei; Feng, Changzeng; Gong, Xiaocui; Yang, Dongmei; Bartlam, Mark; Meng, Zhaohui; Zhang, Keqin

    2009-01-01

    In this study, the PaaX-like protein from the hyperthermophilic archaeon Sulfolobus solfataricus P2 was successfully crystallized by the hanging-drop vapour-diffusion method using ammonium sulfate as a precipitant. PaaX is a global regulator of the phenylacetyl-coenzyme A catabolon that adjusts the expression of different operons to that of the paa-encoded central pathway. In this study, the PaaX-like protein from the hyperthermophilic archaeon Sulfolobus solfataricus P2 was successfully crystallized by the hanging-drop vapour-diffusion method using ammonium sulfate as a precipitant. Diffraction data were obtained to a resolution of 3.0 Å using synchrotron radiation at the Photon Factory. The crystal belonged to space group P321, with unit-cell parameters a = 86.4, b = 86.4, c = 105.5 Å

  5. Biochemical properties and base excision repair complex formation of apurinic/apyrimidinic endonuclease from Pyrococcus furiosus

    OpenAIRE

    Kiyonari, Shinichi; Tahara, Saki; Shirai, Tsuyoshi; Iwai, Shigenori; Ishino, Sonoko; Ishino, Yoshizumi

    2009-01-01

    Apurinic/apyrimidinic (AP) sites are the most frequently found mutagenic lesions in DNA, and they arise mainly from spontaneous base loss or modified base removal by damage-specific DNA glycosylases. AP sites are cleaved by AP endonucleases, and the resultant gaps in the DNA are repaired by DNA polymerase/DNA ligase reactions. We identified the gene product that is responsible for the AP endonuclease activity in the hyperthermophilic euryarchaeon, Pyrococcus furiosus. Furthermore, we detected...

  6. Biochemical characterization of a new nicotinamidase from an unclassified bacterium thriving in a geothermal water stream microbial mat community

    OpenAIRE

    Zapata-P?rez, Rub?n; Mart?nez-Mo?ino, Ana-Bel?n; Garc?a-Saura, Antonio-Gin?s; Cabanes, Juana; Takami, Hideto; S?nchez-Ferrer, ?lvaro

    2017-01-01

    Nicotinamidases are amidohydrolases that convert nicotinamide into nicotinic acid, contributing to NAD+ homeostasis in most organisms. In order to increase the number of nicotinamidases described to date, this manuscript characterizes a nicotinamidase obtained from a metagenomic library fosmid clone (JFF054_F02) obtained from a geothermal water stream microbial mat community in a Japanese epithermal mine. The enzyme showed an optimum temperature of 90?C, making it the first hyperthermophilic ...

  7. Widespread distribution of archaeal reverse gyrase in thermophilic bacteria suggests a complex history of vertical inheritance and lateral gene transfers

    Directory of Open Access Journals (Sweden)

    Céline Brochier-Armanet

    2006-01-01

    Full Text Available Reverse gyrase, an enzyme of uncertain funtion, is present in all hyperthermophilic archaea and bacteria. Previous phylogenetic studies have suggested that the gene for reverse gyrase has an archaeal origin and was transferred laterally (LGT to the ancestors of the two bacterial hyperthermophilic phyla, Thermotogales and Aquificales. Here, we performed an in-depth analysis of the evolutionary history of reverse gyrase in light of genomic progress. We found genes coding for reverse gyrase in the genomes of several thermophilic bacteria that belong to phyla other than Aquificales and Thermotogales. Several of these bacteria are not, strictly speaking, hyperthermophiles because their reported optimal growth temperatures are below 80 °C. Furthermore, we detected a reverse gyrase gene in the sequence of the large plasmid of Thermus thermophilus strain HB8, suggesting a possible mechanism of transfer to the T. thermophilus strain HB8 involving plasmids and transposases. The archaeal part of the reverse gyrase tree is congruent with recent phylogenies of the archaeal domain based on ribosomal proteins or RNA polymerase subunits. Although poorly resolved, the complete reverse gyrase phylogeny suggests an ancient acquisition of the gene by bacteria via one or two LGT events, followed by its secondary distribution by LGT within bacteria. Finally, several genes of archaeal origin located in proximity to the reverse gyrase gene in bacterial genomes have bacterial homologues mostly in thermophiles or hyperthermophiles, raising the possibility that they were co-transferred with the reverse gyrase gene. Our new analysis of the reverse gyrase history strengthens the hypothesis that the acquisition of reverse gyrase may have been a crucial evolutionary step in the adaptation of bacteria to high-temperature environments. However, it also questions the role of this enzyme in thermophilic bacteria and the selective advantage its presence could provide.

  8. Widespread distribution of archaeal reverse gyrase in thermophilic bacteria suggests a complex history of vertical inheritance and lateral gene transfers.

    Science.gov (United States)

    Brochier-Armanet, Céline; Forterre, Patrick

    2007-05-01

    Reverse gyrase, an enzyme of uncertain funtion, is present in all hyperthermophilic archaea and bacteria. Previous phylogenetic studies have suggested that the gene for reverse gyrase has an archaeal origin and was transferred laterally (LGT) to the ancestors of the two bacterial hyperthermophilic phyla, Thermotogales and Aquificales. Here, we performed an in-depth analysis of the evolutionary history of reverse gyrase in light of genomic progress. We found genes coding for reverse gyrase in the genomes of several thermophilic bacteria that belong to phyla other than Aquificales and Thermotogales. Several of these bacteria are not, strictly speaking, hyperthermophiles because their reported optimal growth temperatures are below 80 degrees C. Furthermore, we detected a reverse gyrase gene in the sequence of the large plasmid of Thermus thermophilus strain HB8, suggesting a possible mechanism of transfer to the T. thermophilus strain HB8 involving plasmids and transposases. The archaeal part of the reverse gyrase tree is congruent with recent phylogenies of the archaeal domain based on ribosomal proteins or RNA polymerase subunits. Although poorly resolved, the complete reverse gyrase phylogeny suggests an ancient acquisition of the gene by bacteria via one or two LGT events, followed by its secondary distribution by LGT within bacteria. Finally, several genes of archaeal origin located in proximity to the reverse gyrase gene in bacterial genomes have bacterial homologues mostly in thermophiles or hyperthermophiles, raising the possibility that they were co-transferred with the reverse gyrase gene. Our new analysis of the reverse gyrase history strengthens the hypothesis that the acquisition of reverse gyrase may have been a crucial evolutionary step in the adaptation of bacteria to high-temperature environments. However, it also questions the role of this enzyme in thermophilic bacteria and the selective advantage its presence could provide.

  9. Evolutionary genomics of archaeal viruses: unique viral genomes in the third domain of life

    DEFF Research Database (Denmark)

    Prangishvili, D.; Garrett, R. A.; Koonin, E.

    2006-01-01

    In terms of virion morphology, the known viruses of archaea fall into two distinct classes: viruses of mesophilic and moderately thermophilic Eueryarchaeota closely resemble head-and-tail bacteriophages whereas viruses of hyperthermophilic Crenarchaeota show a variety of unique morphotypes...... of bacteriophages. The proteins encoded by the genes belonging to this pool include predicted transcription regulators, ATPases implicated in viral DNA replication and packaging, enzymes of DNA precursor metabolism, RNA modification enzymes, and glycosylases. In addition, each of the crenarchaeal viruses encodes...

  10. PCOGR: Phylogenetic COG ranking as an online tool to judge the specificity of COGs with respect to freely definable groups of organisms

    OpenAIRE

    Meereis, Florian; Kaufmann, Michael

    2004-01-01

    Abstract Background The rapidly increasing number of completely sequenced genomes led to the establishment of the COG-database which, based on sequence homologies, assigns similar proteins from different organisms to clusters of orthologous groups (COGs). There are several bioinformatic studies that made use of this database to determine (hyper)thermophile-specific proteins by searching for COGs containing (almost) exclusively proteins from (hyper)thermophilic genomes. However, public softwar...

  11. Measurements of Protein Crystal Face Growth Rates

    Science.gov (United States)

    Gorti, S.

    2014-01-01

    Protein crystal growth rates will be determined for several hyperthermophile proteins.; The growth rates will be assessed using available theoretical models, including kinetic roughening.; If/when kinetic roughening supersaturations are established, determinations of protein crystal quality over a range of supersaturations will also be assessed.; The results of our ground based effort may well address the existence of a correlation between fundamental growth mechanisms and protein crystal quality.

  12. Genome stability: recent insights in the topoisomerase reverse gyrase and thermophilic DNA alkyltransferase.

    Science.gov (United States)

    Vettone, Antonella; Perugino, Giuseppe; Rossi, Mosè; Valenti, Anna; Ciaramella, Maria

    2014-09-01

    Repair and defence of genome integrity from endogenous and environmental hazard is a primary need for all organisms. Natural selection has driven the evolution of multiple cell pathways to deal with different DNA damaging agents. Failure of such processes can hamper cell functions and induce inheritable mutations, which in humans may cause cancerogenicity or certain genetic syndromes, and ultimately cell death. A special case is that of hyperthermophilic bacteria and archaea, flourishing at temperatures higher than 80 °C, conditions that favor genome instability and thus call for specific, highly efficient or peculiar mechanisms to keep their genome intact and functional. Over the last few years, numerous studies have been performed on the activity, function, regulation, physical and functional interaction of enzymes and proteins from hyperthermophilic microorganisms that are able to bind, repair, bypass damaged DNA, or modify its structure or conformation. The present review is focused on two enzymes that act on DNA catalyzing unique reactions: reverse gyrase and DNA alkyltransferase. Although both enzymes belong to evolutionary highly conserved protein families present in organisms of the three domains (Eucarya, Bacteria and Archaea), recently characterized members from hyperthermophilic archaea show both common and peculiar features.

  13. Looking for the most ``primitive'' organism(s) on Earth today: the state of the art

    Science.gov (United States)

    Forterre, Patrick

    1995-02-01

    Molecular phylogenetic studies have revealed a tripartite division of the living world into two procaryotic groups, Bacteria and Archaea, and one eucaryotic group, Eucarya. Which group is the most "primitive"? Which groups are sister? The answer to these questions would help to delineate the characters of the last common ancestor to all living beings, as a first step to reconstruct the earliest periods of biological evolution on Earth. The current "Procaryotic dogma" claims that procaryotes are primitive. Since the ancestor of Archaea was most probably a hyperthermophile, and since bacteria too might have originated from hyperthermophiles, the procaryotic dogma has been recently connected to the hot origin of life hypothesis. However, the notion that present-day hyperthermophiles are primitive has been challenged by recent findings, in these unique microorganisms, of very elaborate adaptative devices for life at high temperature. Accordingly, I discuss here alternative hypotheses that challenge the procaryotic dogma, such as the idea of a universal ancestor with molecular features in between those of eucaryotes and procaryotes, or the origin of procaryotes via thermophilic adaptation. Clearly, major evolutionary questions about early cellular evolution on Earth remain to be settled before we can speculate with confidence about which kinds of life might have appeared on other planets.

  14. The evolution of lipids

    Science.gov (United States)

    Itoh, Y. H.; Sugai, A.; Uda, I.; Itoh, T.

    2001-01-01

    Living organisms on the Earth which are divided into three major domains - Archaea, Bacteria, and Eucarya, probably came from a common ancestral cell. Because there are many thermophilic microorganisms near the root of the universal phylogenetic tree, the common ancestral cell should be considered to be a thermophilic microorganism. The existence of a cell is necessary for the living organisms; the cell membrane is the essential structural component of a cell, so its amphiphilic property is vital for the molecule of lipids for cell membranes. Tetraether type glycerophospholipids with C 40 isoprenoid chains are major membrane lipids widely distributed in archaeal cells. Cyclization number of C 40 isoprenoid chains in thermophilic archaea influences the fluidity of lipids whereas the number of carbons and degree of unsaturation in fatty acids do so in bacteria and eucarya. In addition to the cyclization of the tetraether lipids, covalent bonding of two C 40 isoprenoid chains was found in hyperthermophiles. These characteristic structures of the lipids seem to contribute to their fundamental physiological roles in hyperthermophiles. Stereochemical differences between G-1-P archaeal lipids and G-3-P bacterial and eucaryal lipids might have occured by the function of some proteins long after the first cell was developed by the reactions of small organic molecules. We propose that the structure of lipids of the common ancestral cell may have been similar to those of hyperthermophilic archaea.

  15. Differences in substrate specificity of C(5)-substituted or C(5)-unsubstituted pyrimidine nucleotides by DNA polymerases from thermophilic bacteria, archaea, and phages.

    Science.gov (United States)

    Sawai, Hiroaki; Nagashima, Junichi; Kuwahara, Msayasu; Kitagata, Rina; Tamura, Takehiro; Matsui, Ikuo

    2007-09-01

    The pyrimidine bases of RNA are uracil (U) and cytosine (C), while thymine (T) and C are used for DNA. The C(5) position of C and U is unsubstituted, whereas the C(5) of T is substituted with a Me group. Miller et al. hypothesized that various C(5)-substituted uracil derivatives were formed during chemical evolution, and that C(5)-substituted U derivatives may have played important roles in the transition from an 'RNA world' to a 'DNA-RNA-protein world'. Hyperthermophilic bacteria and archaea are considered to be primitive organisms that are evolutionarily close to the universal ancestor of all life on earth. Thus, we examined the substrate specificity of several C(5)-substituted or C(5)-unsubstituted dUTP and dCTP analogs for several DNA polymerases from hyperthermophilic bacteria, hyperthermophilic archaea, and viruses during PCR or primer extension reaction. The substrate specificity of the C(5)-substituted or C(5)-unsubstituted pyrimidine nucleotides varied greatly depending on the type of DNA polymerase. The significance of this difference in substrate specificity in terms of the origin and evolution of the DNA replication system is discussed briefly.

  16. Genetic and functional properties of uncultivated thermophilic crenarchaeotes from a subsurface gold mine as revealed by analysis of genome fragments.

    Science.gov (United States)

    Nunoura, Takuro; Hirayama, Hisako; Takami, Hideto; Oida, Hanako; Nishi, Shinro; Shimamura, Shigeru; Suzuki, Yohey; Inagaki, Fumio; Takai, Ken; Nealson, Kenneth H; Horikoshi, Koki

    2005-12-01

    Within a phylum Crenarchaeota, only some members of the hyperthermophilic class Thermoprotei, have been cultivated and characterized. In this study, we have constructed a metagenomic library from a microbial mat formation in a subsurface hot water stream of the Hishikari gold mine, Japan, and sequenced genome fragments of two different phylogroups of uncultivated thermophilic Crenarchaeota: (i) hot water crenarchaeotic group (HWCG) I (41.2 kb), and (ii) HWCG III (49.3 kb). The genome fragment of HWCG I contained a 16S rRNA gene, two tRNA genes and 35 genes encoding proteins but no 23S rRNA gene. Among the genes encoding proteins, several genes for putative aerobic-type carbon monoxide dehydrogenase represented a potential clue with regard to the yet unknown metabolism of HWCG I Archaea. The genome fragment of HWCG III contained a 16S/23S rRNA operon and 44 genes encoding proteins. In the 23S rRNA gene, we detected a homing-endonuclease encoding a group I intron similar to those detected in hyperthermophilic Crenarchaeota and Bacteria, as well as eukaryotic organelles. The reconstructed phylogenetic tree based on the 23S rRNA gene sequence reinforced the intermediate phylogenetic affiliation of HWCG III bridging the hyperthermophilic and non-thermophilic uncultivated Crenarchaeota.

  17. Trace metal chemistry and silicification of microorganisms in geothermal sinter, Taupo Volcanic Zone, New Zealand

    Energy Technology Data Exchange (ETDEWEB)

    McKenzie, E.J.; Brown, K.L.; Campbell, K.A. [University of Auckland (New Zealand). Dept. of Geology; Cady, S.L. [Portland State University, Portland, OR (United States). Dept. of Geology

    2001-08-01

    As part of a pilot study investigating the role of microorganisms in the immobilisation ol As, Sb, B, Tl and Ug, the inorganic geochemistry of seven different active sinter deposits and their contact fluids were characterised. A comprehensive series of sequential extractions for a suite of trace elements was carried out on siliceous sinter and a mixed silica-carbonate sinter. The extractions showed whether metals were loosely exchangeable or bound to carbonate, oxide, organic or crystalline fractions. Hyperthermophilic microbial communities associated with sinters deposited from high temperature (92-94{sup o}C) fluids at a variety of geothermal sources were investigated using SEM. The rapidity and style of silicification of the hyperthermophiles can be correlated with the dissolved silica content of the fluid. Although high concentrations of Hg and TI were found associated with the organic fraction of the sinters, there was no evidence to suggest that any of the heavy metals were associated preferentially with the hyperthermophiles at the high temperature (92-94{sup o}C) ends of the terrestrial thermal spring ecosystems studied. (author)

  18. Extremely thermophilic microorganisms for biomass conversion: status and prospects.

    Science.gov (United States)

    Blumer-Schuette, Sara E; Kataeva, Irina; Westpheling, Janet; Adams, Michael Ww; Kelly, Robert M

    2008-06-01

    Many microorganisms that grow at elevated temperatures are able to utilize a variety of carbohydrates pertinent to the conversion of lignocellulosic biomass to bioenergy. The range of substrates utilized depends on growth temperature optimum and biotope. Hyperthermophilic marine archaea (T(opt)>or=80 degrees C) utilize alpha- and beta-linked glucans, such as starch, barley glucan, laminarin, and chitin, while hyperthermophilic marine bacteria (T(opt)>or=80 degrees C) utilize the same glucans as well as hemicellulose, such as xylans and mannans. However, none of these organisms are able to efficiently utilize crystalline cellulose. Among the thermophiles, this ability is limited to a few terrestrial bacteria with upper temperature limits for growth near 75 degrees C. Deconstruction of crystalline cellulose by these extreme thermophiles is achieved by 'free' primary cellulases, which are distinct from those typically associated with large multi-enzyme complexes known as cellulosomes. These primary cellulases also differ from the endoglucanases (referred to here as 'secondary cellulases') reported from marine hyperthermophiles that show only weak activity toward cellulose. Many extremely thermophilic enzymes implicated in the deconstruction of lignocellulose can be identified in genome sequences, and many more promising biocatalysts probably remain annotated as 'hypothetical proteins'. Characterization of these enzymes will require intensive effort but is likely to generate new opportunities for the use of renewable resources as biofuels.

  19. Extremophiles: developments of their special functions and potential resources.

    Science.gov (United States)

    Fujiwara, Shinsuke

    2002-01-01

    Extremophilles are valuable resources in biotechnology. Enzymes from extremophiles are expected to fill the gap between biological and chemical processes due to their unusual properties. Especially enzymes from hyperthermophiles that can grow at above 90 degrees C were devoted owing to its extraordinary thermostability and denaturant tolerance. Screening trials of hyperthermophilic microorganisms were performed by a number of microbiologists and various unique strains were isolated from natural environments. One of the most successful uses of thermostable enzymes was DNA polymerase in the polymerase chain reaction (PCR). Thermostable enzymes are used in the chemical, food, pharmaceutical, paper and textile industries. Recombinant forms of thermostable enzymes that have been expressed in Escherichia coli are commonly utilized in industrial applications however their enzymatic characteristics and tertiary structure are different from the native ones produced in the original strains. In vitro heat treatment induces a structural conversion of the recombinant protein to its natural form. High temperature itself plays an important role in determining the specific characteristics and tertiary structure of the enzyme. Recent studies have revealed that hyperthermophiles can grow under numerous conditions not only in geothermal or deep-sea thermal environments. Technological advances have allowed DNA to be isolated from natural environments. Now genes could be isolated from microorganisms that have not been cultured. In this review, innovative approaches to hunt genes from natural environments without pure culturing of microorganisms are also discussed.

  20. Multiple Syntrophic Interactions in a Terephthalate-Degrading Methanogenic Consortium

    Energy Technology Data Exchange (ETDEWEB)

    Lykidis, Athanasios; Chen, Chia-Lung; Tringe, Susannah G.; McHardy, Alice C.; Copeland, Alex 5; Kyrpides, Nikos C.; Hugenholtz, Philip; Liu, Wen-Tso

    2010-08-05

    Terephthalate (TA) is one of the top 50 chemicals produced worldwide. Its production results in a TA-containing wastewater that is treated by anaerobic processes through a poorly understood methanogenic syntrophy. Using metagenomics, we characterized the methanogenic consortium tinside a hyper-mesophilic (i.e., between mesophilic and thermophilic), TA-degrading bioreactor. We identified genes belonging to dominant Pelotomaculum species presumably involved in TA degradation through decarboxylation, dearomatization, and modified ?-oxidation to H{sub 2}/CO{sub 2} and acetate. These intermediates are converted to CH{sub 4}/CO{sub 2} by three novel hyper-mesophilic methanogens. Additional secondary syntrophic interactions were predicted in Thermotogae, Syntrophus and candidate phyla OP5 and WWE1 populations. The OP5 encodes genes capable of anaerobic autotrophic butyrate production and Thermotogae, Syntrophus and WWE1 have the genetic potential to oxidize butyrate to COsub 2}/H{sub 2} and acetate. These observations suggest that the TA-degrading consortium consists of additional syntrophic interactions beyond the standard H{sub 2}-producing syntroph ? methanogen partnership that may serve to improve community stability.

  1. Regeneração e riqueza da formação arbustiva de Palmae em uma cronoseqüência pós-fogo na Restinga da Marambaia, Rio de Janeiro, RJ, Brasil The structure and diversity of three areas of shrubby restinga vegetation were analyzed 3, 12 and 84 months after fire in the Marambaia Restinga, Rio de Janeiro State, Brazil

    Directory of Open Access Journals (Sweden)

    Luis Fernando Tavares de Menezes

    2004-12-01

    Full Text Available A estrutura e a riqueza da formação arbustiva de Palmae foram analisadas em três sítios numa cronoseqüência de regeneração (3, 12 e 84 meses após a última queimada na Restinga da Marambaia, registrando-se a presença de 29, 41 e 64 táxons, respectivamente. No sítio com maior tempo de regeneração, Allagoptera arenaria (Gomes Kuntze representou 79% da dominância relativa (DoR, seguida das nanofanerófitas Inga maritima Benth. e Manilkara subsericea (Mart. Dubard. Na área queimada há 12 meses, A. arenaria representou 88% da DoR, seguida de Inga maritima, Setaria setosa (Sw. P. Beauv. e Paspalum arenarium Schrad. No sítio com três meses de regeneração, a DoR de A. arenaria foi de 82%, acompanhada de Clitoria sp., Inga maritima e Portulaca mucronata Link. Nos três sítios estudados, a forma de vida mais importante foi geófita rizomatosa, devido à dominância de A. arenaria. Caméfita herbácea escaposa foi a forma de vida que apresentou maior número de espécies nos sítios com três e 12 meses de regeneração e no sítio queimado há 84 meses, as nanofanerófitas acompanharam as caméfitas herbáceas escaposas em número de espécies. Das 29 espécies registradas no sítio com três meses de regeneração, só Portulaca mucronata e Sebastiania corniculata (Vahl Müll. Arg. originaramse a partir de sementes, sendo que as demais rebrotaram ou se regeneraram. Sete dias após a queimada A. arenaria apresentou, em média, 8cm de sua parte vegetativa regenerada e com 180 dias apresentou as primeiras inflorescências.A total of 29, 41 and 64 taxa, respectively, were found. In the area sampled 84 months after fire, relative dominance of Allagoptera arenaria (Gomes Kuntze was 79%, followed by the nanophanerophytes Inga maritima Benth. and Manilkara subsericea (Mart. Dubard. In the area sampled 12 months after fire, relative dominance of A. arenaria was 88%, followed by Inga maritima, Setaria setosa (Sw. P. Beauv and Paspalum

  2. Effects of migratory geese on plant communities of an Alaskan salt marsh

    Science.gov (United States)

    Zacheis, Amy B.; Hupp, Jerry W.; Ruess, Roger W.

    2001-01-01

    1. We studied the effects of lesser snow geese (Anser caerulescens caerulescens) and Canada geese (Branta canadensis) on two salt marsh plant communities in Cook Inlet, Alaska, a stopover area used during spring migration. From 1995 to 1997 we compared plant species composition and biomass on plots where geese were excluded from feeding with paired plots where foraging could occur. 2. Foraging intensity was low (650-1930 goose-days km-2) compared to other goose-grazing systems. 3. Canada geese fed mainly on above-ground shoots of Triglochin maritimum, Puccinellia spp. and Carex ramenskii, whereas the majority of the snow goose diet consisted of below-ground tissues of Plantago maritima and Triglochin maritimum. 4. Plant communities responded differently to goose herbivory. In the sedge meadow community, where feeding was primarily on above-ground shoots, there was no effect of grazing on the dominant species Carex ramenskii and Triglochin maritimum. In the herb meadow community, where snow geese fed on Plantago maritima roots and other below-ground tissues, there was a difference in the relative abundance of plant species between treatments. Biomass of Plantago maritima and Potentilla egedii was lower on grazed plots compared with exclosed, whereas biomass of Carex ramenskii was greater on grazed plots. There was no effect of herbivory on total standing crop biomass in either community. The variable effect of herbivory on Carex ramenskii between communities suggests that plant neighbours and competitive interactions are important factors in a species' response to herbivory. In addition, the type of herbivory (above- or below-ground) was important in determining plant community response to herbivory. 5. Litter accumulation was reduced in grazed areas compared with exclosed in both communities. Trampling of the previous year's litter into the soil surface by geese incorporated more litter into soils in grazed areas. 6. This study illustrates that even light herbivore

  3. AcEST: DK948593 [AcEST

    Lifescience Database Archive (English)

    Full Text Available DKLSHFNYVVDVLIPYPIHLEI 163 >sp|A4QKR2|MATK_CRUWA Maturase K OS=Crucihimalaya wallichii GN=matK PE=3 SV=2 Len..._LOBMA Maturase K OS=Lobularia maritima GN=matK PE... 32 2.3 sp|A4QKR2|MATK_CRUWA Maturase K OS=Crucihimal...aya wallichii GN=ma... 32 3.0 sp|Q9GF51|MATK_ARAHA Maturase K OS=Arabidopsis haller

  4. Ability of salt marsh plants for TBT remediation in sediments.

    Science.gov (United States)

    Carvalho, Pedro N; Basto, M Clara P; Silva, Manuela F G M; Machado, Ana; Bordalo, A A; Vasconcelos, M Teresa S D

    2010-07-01

    The capability of Halimione portulacoides, Spartina maritima, and Sarcocornia fruticosa (halophytes very commonly found in salt marshes from Mediterranean areas) for enhancing remediation of tributyltin (TBT) from estuarine sediments was investigated, using different experimental conditions. The influence of H. portulacoides on degradation of the butyltin compounds was assessed in two different ways: (1) a 9-month ex situ study carried out in a site of Sado River estuary, center of Portugal, which used polluted sediments collected at other nonvegetated site from the same estuary; and (2) a 12-month laboratorial study, using both plant and sediment collected at a relatively clean site of Cávado River estuary, north of Portugal, the sediment being doped with TBT, DBT, and MBT at the beginning of the experiment. The role of both S. fruticosa and S. maritima on TBT remediation in sediments was evaluated in situ, in salt marshes from Marim channel of Ria Formosa lagoon, south of Portugal, which has large areas colonized by each one of these two plants. For estimation of microbial abundance, total cell counts of sediment samples were enumerated by the DAPI direct count method. Butyltin analyses in sediment were performed using a method previously validated, which consisted of headspace solid-phase micro-extraction combined with gas chromatography-mass spectrometry after in situ ethylation (with tetraethylborate). Sediments colonized both ex situ and at lab by H. portulacoides displayed TBT levels about 30% lower than those for nonvegetated sediments with identical initial composition, after 9-12 months of plant exposure. In addition, H. portulacoides showed to be able of stimulating bacterial growth in the plant rhizosphere, which probably included degraders of TBT. In the in situ study, which compared the levels of TBT, DBT, and MBT in nonvegetated sediment and in sediments colonized by either S. maritima or S. fruticosa from the same area, TBT and DBT were only

  5. Expansion of southern distributional range of Ucides occidentalis (Decapoda: Ucididae and Cardisoma crassum (Decapoda: Gecarcinidae

    Directory of Open Access Journals (Sweden)

    Solange Alemán

    2017-04-01

    Full Text Available Is recorded the species of crabs brachyuran Ucides occidentalis (mangrove crab and Cardisoma crassum (Blue crab or without mouth in the mangroves of San Pedro (Piura, expanding its geographical distribution south of Tumbes, which was the known limit. The habitat of these species is characterized by the presence of two varieties of mangrove trees, Jeli white (Laguncularia racemosa and salty Jeli (Avicenia germinans and halophytic shrub called glass (Batis maritima, it observing that the depth of the burrows is shallow (< 60 cm. Biometric information and some biological aspects of the collected specimens are also presented.

  6. Screening of selected ethnomedicinal plants from South Africa for larvicidal activity against the mosquito Anopheles arabiensis

    CSIR Research Space (South Africa)

    Maharaj, R

    2012-09-01

    Full Text Available through effects of hor- mone regulation with subsequent disruption of instar development of Anopheles stephensi, Culex quinquefas- ciatus and Aedes aegypti [34]. Tiwari et al. [35] found that the essential oil obtained from seeds of Zanthoxy- lum.... Geerts S, Van Blerk K, Triest L: Effect of Ambrosia maritima on Anopheles stephensi and Aedes aegypti. J Ethnopharmacol 1994, 42:7?11. 20. Evans DA, Raj RK: Extracts of Indian plants as mosquito larvicides. Indian J Med Res 1988, 88:38?41. 21...

  7. Antioxidant activity of raw, cooked and Rhizopus oligosporus fermented beans of Canavalia of coastal sand dunes of Southwest India.

    Science.gov (United States)

    Niveditha, Vedavyas R; Sridhar, Kandikere R

    2014-11-01

    The raw and processed (cooked and cooked + solid-state fermented with Rhizopus oligosporus) split beans of two landraces of coastal sand dune wild legumes (Canavalia cathartica and Canavalia maritima) of the southwest coast of India were examined for bioactive compounds (total phenolics, tannins and vitamin C) and antioxidant potential (total antioxidant activity, ferrous-ion chelating capacity, DPPH free radical-scavenging activity and reducing activity). One-way ANOVA revealed significant elevation of bioactive compounds as well as antioxidant activities in fermented beans compared to raw and cooked beans in both legumes (p beans of both legumes were significantly lowest compared to raw and cooked beans (p beans of C. cathartica, while total antioxidant and free radical-scavenging activities of fermented beans of C. maritima were clustered. The present study demonstrated that split beans of coastal sand dune Canavalia fermented by R. oligosporus endowed with high bioactive principles as well as antioxidant potential and thus serve as future nutraceutical source.

  8. Feeding ecology of Rhabdosargus holubi (family Sparidae) in multiple vegetated refugia of selected warm temperate estuaries in South Africa

    Science.gov (United States)

    Nel, L.; Strydom, N. A.; Perissinotto, R.; Adams, J. B.; Lemley, D. A.

    2017-10-01

    Estuarine marine-dependent species, such as Rhabdosargus holubi, depend greatly on structured sheltered environments and important feeding areas provided by estuaries. In this study, we investigate the ecological feeding niches of the estuarine marine-dependent sparid, R. holubi, by using conventional stomach contents and stable isotope methods (δ13C and δ15N signatures). The study has been carried out in five temperate estuaries in order to understand how fish feed in multiple intertidal vegetated habitats. These habitats included the submerged seagrass, Zostera capensis, and both previously unexplored small intertidal cord grass, Spartina maritima, and the common reed, Phragmites australis. The diet varied amongst habitats, estuaries and fish sizes and data consistently confirmed their omnivorous diet relating to ontogenetic niche shifts. Stomach contents revealed the importance of benthic prey within both the S. maritima and P. australis habitats in the absence of large intertidal vegetation, available during low tides. Similarly, isotopic mixing models showed that R. holubi from these habitats have a greater isotopic niche compared to the Z. capensis habitat, due to their limited availability during the falling tide, suggesting migration between available habitats. Stable isotopes confirmed that R. holubi actively feeds on the epiphytic algae (especially diatoms) covering the leaves and stalks of plant matter, as supported by Bayesian mixing models. These findings add to the current knowledge regarding habitat partitioning in multiple aquatic vegetation types critical to fish ecology and the effective management and conservation of estuaries.

  9. Impact of solid waste burning air pollution on some physio-anatomical characteristics of some plants

    International Nuclear Information System (INIS)

    Laghari, S.K.; Zaidi, M.A.

    2015-01-01

    Present study evaluated the effect of solid waste burning pollution on carbohydrate, stomata and chlorophyll contents of seven different plant species. Leaf samples of Artemisia maritima L., Fraxinus excelsior L., Amaranthus viridis L., Cynodon dactylon L., Chenopodium album L., Robinia pseudoacacia L., and Sophora mollis (Royle) Baker, growing in the (1m, 500m and 1000m distance) vicinity of burning points at residential colony, University of Baluchistan Quetta were collected. Results revealed that the carbohydrate, chlorophyll a and b and total chlorophyll contents in the leaves of selected plant species were found to be significantly low at 1m distance, but as the distance from the source of pollution increased (500m and 1000m) these contents increased accordingly. Generally the percentage of completely and partially clogged stomata was found higher near the pollution source (1m distance). The percentage of open stomata in all investigated plant species was noticed lower near the pollution source (1m distance), while with the increase of distance (500m-1000m) the percentage of open stomata increased accordingly. As regard to carbohydrate and chlorophyll contents, the Artemisia maritima L., were found most sensitive to air pollution in all four directions at 1m distances as compared to the other species. While plant species, Cynodon dactylon L. showed more resistant to air pollution effect as regard to carbohydrate contents and high percentage of open stomata at 1m distances with respect to other species. (author)

  10. Evaluation of a Florida coastal golf complex as a local and watershed source of bioavailable contaminants

    International Nuclear Information System (INIS)

    Lewis, Michael A.; Quarles, Robert L.; Dantin, Darrin D.; Moore, James C.

    2004-01-01

    Contaminant fate in coastal areas impacted by golf course runoff is not well understood. This report summarizes trace metal, pesticide and PCB residues for colonized periphyton, Ruppia maritima (widgeon grass), Callinectes sapidus Rathbun (blue crabs) and Crassostrea virginica Gemlin (Eastern oyster) collected from areas adjacent to a Florida golf course complex which receive runoff containing reclaimed municipal wastewater. Concentrations of 19 chlorinated pesticides and 18 PCB congeners were usually below detection in the biota. In contrast, 8 trace metals were commonly detected although concentrations were not usually significantly different for biota collected from reference and non-reference coastal areas. Residue concentrations in decreasing order were typically: zinc, arsenic, copper, chromium, lead, nickel, cadmium and mercury. Mean BCF values for the eight trace metals ranged between 160-57 000 (periphyton), 79-11 033 (R. maritima), 87-162 625 (C. virginica) and 12-9800 (C. sapidus). Most trace metal residues in periphyton colonized adjacent to the golf complex, were either similar to or significantly less than those reported for periphyton colonized in nearby coastal areas impacted by urban stormwater runoff and treated municipal and industrial wastewater discharges. Consequently, the recreational complex does not appear to be a major source of bioavailable contaminants locally nor in the immediate watershed based on results for the selected biota

  11. The weed species composition in a reed canary grass (Phalaris arundinacea L. plantation for energy purposes depending on its age

    Directory of Open Access Journals (Sweden)

    Tomasz R. Sekutowski

    2014-12-01

    Full Text Available The present experiment, carried out in nine production fields of reed canary grass (Phalaris arundinacea grown for energy purposes, evaluated the effect of plantation age on the occurrence and species composition of weeds. The selected plantations were divided into 3 groups that were conventionally called “young” (1–2 years old, “middle-aged” (3–5 years old, and “older” plantations (6–8 years old. Regardless of plantation age, altogether 43 species were found in the experimental fields. Moreover, 6 species were common for all the plantations and were found in them regardless of plantation age. The least species, only 18, were found on the “young” plantations, almost twice more on the “older” ones (30 species, whereas the largest spectrum of species was found in the “middle-aged” plantations (33 species. In the “young” plantations, annual weeds were the most common, with the highest constancy and coverage index found for Chenopodium album, Matricaria maritima ssp. inodora and Echinochloa crus-galli. The greatest variation in species was found in the “middle-aged” plantations. However, only 4 species achieved the highest constancy and coverage index: Matricaria maritima ssp. inodora, Cirsium arvense, Poa trivialis and Taraxacum officinale. Furthermore, perennial weeds were found to be dominant in the “older” plantations. Within this group, Poa trivialis, Taraxacum officinale, Urtica dioica, Plantago maior, and Cirsium arvense had the highest constancy and coverage index.

  12. Fish farming in «Baetica». The «piscine» of the halieutic site at Trafalgar Cape (Cádiz

    Directory of Open Access Journals (Sweden)

    Darío BERNAL CASASOLA

    2011-10-01

    Full Text Available The present paper presents a structure of Roman date, cut into bedrock, and possibly used for fish farming purposes. So far, this sort of evidence, which is well attested in Italian villae maritimae dating to the Late Republic and the Early Empire, had only been found in the Iberian Peninsula in the southern Tarraconense (coast of Alicante. The above mentioned structure is, therefore, the first of its kind found in the Baetica. Interpretation must combine archaeological evidence for Roman fish farming in Andalusia (ostrearum vivaria at Traducta, current Algeciras and geoarchaeological features; the structure is located in the intertidal zone and enjoys fresh water supply from the nearby halieutic site of Trafalgar Cape (Barbate, Cádiz. This paper aims at the re-interpretation of this coastal site, previously interpreted as a salted products factory, or cetaria, but the topographical and architectural features of which (vats with inner steps, cisterns, terraced structures, etc. are rather suggestive of a villa maritima or a complex centre for the exploitation of marine resources.

  13. [Different NaCl-dependence of the circadian CO2-gas-exchange of some halophil growing coastal plants].

    Science.gov (United States)

    Treichel, Siegfried; Bauer, Peter

    1974-03-01

    CO 2 -exchange, diurnal changes in malate- and ion concentrations of the halophytes Carpobrotus edulis, Crithmum maritimum, Mesembryanthemum nodiflorum, Salicornia fruticosa, Suaeda maritima, and Trifolium fragiferum were investigated after culture at different NaCl concentrations. In Carp. edulis and Mes. nodiflorum the diurnal rhythm of CO 2 -exchange is in accordance with that of crassulacean acid metabolism (CAM), in Sal. fruticosa, Crithm. maritimum, Suaeda maritima, and Trif. fragiferum with that of Benson-Calvin metabolism (C 3 ). Malate concentration and CO 2 uptake in the sap latter group are not influenced. On the other hand, Carp. edulis and Mes. nodiflorum show an accumulation of malate during the night, which can be interpreted as a further indication of CAM.The two species most resistant to NaCl, Carp. edulis and Sal. fruticosa, greatly differ very much in their NaCl content. NaCl concentration in Salicornia is four times higher than in Carpobrotus.The different metabolic properties studied might be of ecological importance for the plants in their natural habitats. The effect of NaCl on metabolic processes is discussed.

  14. The effects of soil amendments on heavy metal bioavailability in two contaminated Mediterranean soils

    Energy Technology Data Exchange (ETDEWEB)

    Walker, D.J.; Clemente, Rafael; Roig, Asuncion; Bernal, M.P

    2003-04-01

    The effects of organic amendments on metal bioavailability were not always related to their degree of humification. - Two heavy metal contaminated calcareous soils from the Mediterranean region of Spain were studied. One soil, from the province of Murcia, was characterised by very high total levels of Pb (1572 mg kg{sup -1}) and Zn (2602 mg kg{sup -1}), whilst the second, from Valencia, had elevated concentrations of Cu (72 mg kg{sup -1}) and Pb (190 mg kg{sup -1}). The effects of two contrasting organic amendments (fresh manure and mature compost) and the chelate ethylenediaminetetraacetic acid (EDTA) on soil fractionation of Cu, Fe, Mn, Pb and Zn, their uptake by plants and plant growth were determined. For Murcia soil, Brassica juncea (L.) Czern. was grown first, followed by radish (Raphanus sativus L.). For Valencia soil, Beta maritima L. was followed by radish. Bioavailability of metals was expressed in terms of concentrations extractable with 0.1 M CaCl{sub 2} or diethylenetriaminepentaacetic acid (DTPA). In the Murcia soil, heavy metal bioavailability was decreased more greatly by manure than by the highly-humified compost. EDTA (2 mmol kg{sup -1} soil) had only a limited effect on metal uptake by plants. The metal-solubilising effect of EDTA was shorter-lived in the less contaminated, more highly calcareous Valencia soil. When correlation coefficients were calculated for plant tissue and bioavailable metals, the clearest relationships were for Beta maritima and radish.

  15. Accumulation and biological cycling of heavy metal in four salt marsh species, from Tagus estuary (Portugal)

    Energy Technology Data Exchange (ETDEWEB)

    Duarte, B., E-mail: baduarte@fc.ul.p [Centro de Oceanografia, Instituto de Oceanografia, Campo Grande, 1749-1016 Lisboa (Portugal); Caetano, M. [INRB/IPIMAR - Instituto Nacional de Recursos Biologicos, Av. Brasilia, 1449-006 Lisboa (Portugal); Almeida, P.R. [Centro de Oceanografia, Instituto de Oceanografia, Campo Grande, 1749-1016 Lisboa (Portugal); Departamento de Biologia, Universidade de Evora, Largo dos Colegiais 2, 7004-516 Evora (Portugal); Vale, C. [INRB/IPIMAR - Instituto Nacional de Recursos Biologicos, Av. Brasilia, 1449-006 Lisboa (Portugal); Cacador, I. [Centro de Oceanografia, Instituto de Oceanografia, Campo Grande, 1749-1016 Lisboa (Portugal)

    2010-05-15

    Pools of Zn, Cu, Cd and Co in leaf, stem and root tissues of Sarcocornia fruticosa, Sarcocornia perennis, Halimione portulacoides and Spartina maritima were analyzed on a bimonthly basis, in a Tagus estuary salt marsh. All the major concentrations were found in the root tissues, being the concentrations in the aboveground organs neglectable for sediment budget proposes, as seen by the low root-aboveground translocation. Metal annual accumulation, root turnovers and cycling coefficients were also assessed. S. maritima showed the higher root turnovers and cycling coefficients for most of the analyzed metals, making this a phytostabilizer specie. By contrast the low root turnover, cycling coefficient and low root necromass generation makes S. perennis the most suitable specie for phytoremediation processes. Although the high amounts of metal return to the sediments, due to root senescence, salt marshes can still be considered sinks of heavy metals, cycling heavy metals mostly between sediment and root. - The efficiency of the phytoremediative processes and metal budgets are greatly influenced by the turnover periods and necromass generation.

  16. Structural comparison of tRNA m1A58 methyltransferases revealed different molecular strategies to maintain their oligomeric architecture under extreme conditions

    Directory of Open Access Journals (Sweden)

    Guelorget Amandine

    2011-12-01

    Full Text Available Abstract Background tRNA m1A58 methyltransferases (TrmI catalyze the transfer of a methyl group from S-adenosyl-L-methionine to nitrogen 1 of adenine 58 in the T-loop of tRNAs from all three domains of life. The m1A58 modification has been shown to be essential for cell growth in yeast and for adaptation to high temperatures in thermophilic organisms. These enzymes were shown to be active as tetramers. The crystal structures of five TrmIs from hyperthermophilic archaea and thermophilic or mesophilic bacteria have previously been determined, the optimal growth temperature of these organisms ranging from 37°C to 100°C. All TrmIs are assembled as tetramers formed by dimers of tightly assembled dimers. Results In this study, we present a comparative structural analysis of these TrmIs, which highlights factors that allow them to function over a large range of temperature. The monomers of the five enzymes are structurally highly similar, but the inter-monomer contacts differ strongly. Our analysis shows that bacterial enzymes from thermophilic organisms display additional intermolecular ionic interactions across the dimer interfaces, whereas hyperthermophilic enzymes present additional hydrophobic contacts. Moreover, as an alternative to two bidentate ionic interactions that stabilize the tetrameric interface in all other TrmI proteins, the tetramer of the archaeal P. abyssi enzyme is strengthened by four intersubunit disulfide bridges. Conclusions The availability of crystal structures of TrmIs from mesophilic, thermophilic or hyperthermophilic organisms allows a detailed analysis of the architecture of this protein family. Our structural comparisons provide insight into the different molecular strategies used to achieve the tetrameric organization in order to maintain the enzyme activity under extreme conditions.

  17. Intermolecular ion pairs maintain the toroidal structure of Pyrococcus furiosus PCNA

    OpenAIRE

    Matsumiya, Shigeki; Ishino, Sonoko; Ishino, Yoshizumi; Morikawa, Kosuke

    2003-01-01

    Two mutant proliferating cell nuclear antigens from the hyperthermophilic archaeon Pyrococcus furiosus, PfuPCNA(D143A) and PfuPCNA(D143A/D147A), were prepared by site-specific mutagenesis. The results from gel filtration showed that mutations at D143 and D147 drastically affect the stability of the trimeric structure of PfuPCNA. The PfuPCNA(D143A) still retained the activity to stimulate the DNA polymerase reaction, but PfuPCNA(D143A/D147A) lost the activity. Crystal structures of the mutant ...

  18. Sulfolobus Replication Factor C stimulates the activity of DNA Polymerase B1

    DEFF Research Database (Denmark)

    Xing, Xuanxuan; Zhang, Likui; Guo, Li

    2014-01-01

    the hyperthermophilic archaea of the genus Sulfolobus physically interacts with DNA polymerase B1 (PolB1) and enhances both the polymerase and 3'-5' exonuclease activities of PolB1 in an ATP-independent manner. Stimulation of the PolB1 activity by RFC is independent of the ability of RFC to bind DNA but is consistent...... with the ability of RFC to facilitate DNA binding by PolB1 through protein-protein interaction. These results suggest that Sulfolobus RFC may play a role in recruiting DNA polymerase for efficient primer extension, in addition to clamp loading, during DNA replication....

  19. Identifying members of the domain Archaea with rRNA-targeted oligonucleotide probes.

    Science.gov (United States)

    Burggraf, S; Mayer, T; Amann, R; Schadhauser, S; Woese, C R; Stetter, K O

    1994-09-01

    Two 16S rRNA-targeted oligonucleotide probes were designed for the archaeal kingdoms Euryachaeota and Crenarchaeota. Probe specificities were evaluated by nonradioactive dot blot hybridization against selected reference organisms. The successful application of fluorescent-probe derivatives for whole-cell hybridization required organism-specific optimizations of fixation and hybridization conditions to assure probe penetration and morphological integrity of the cells. The probes allowed preliminary grouping of three new hyperthermophilic isolates. Together with other group-specific rRNA-targeted oligonucleotide probes, these probes will facilitate rapid in situ monitoring of the populations present in hydrothermal systems and support cultivation attempts.

  20. Conditions for gene disruption by homologous recombination of exogenous DNA into the Sulfolobus solfataricus genome

    Directory of Open Access Journals (Sweden)

    Sonja-Verena Albers

    2008-01-01

    Full Text Available The construction of directed gene deletion mutants is an essential tool in molecular biology that allows functional studies on the role of genes in their natural environment. For hyperthermophilic archaea, it has been difficult to obtain a reliable system to construct such mutants. However, during the past years, systems have been developed for Thermococcus kodakarensis and two Sulfolobus species, S. acidocaldarius and derivatives of S. solfataricus 98/2. Here we describe an optimization of the method for integration of exogenous DNA into S. solfataricus PBL 2025, an S. solfataricus 98/2 derivative, based on lactose auxotrophy that now allows for routine gene inactivation.

  1. Conditions for gene disruption by homologous recombination of exogenous DNA into the Sulfolobus solfataricus genome.

    Science.gov (United States)

    Albers, Sonja-Verena; Driessen, Arnold J M

    2008-12-01

    The construction of directed gene deletion mutants is an essential tool in molecular biology that allows functional studies on the role of genes in their natural environment. For hyperthermophilic archaea, it has been difficult to obtain a reliable system to construct such mutants. However, during the past years, systems have been developed for Thermococcus kodakarensis and two Sulfolobus species, S. acidocaldarius and derivatives of S. solfataricus 98/2. Here we describe an optimization of the method for integration of exogenous DNA into S. solfataricus PBL 2025, an S. solfataricus 98/2 derivative, based on lactose auxotrophy that now allows for routine gene inactivation.

  2. Precambrian Surface Temperatures and Molecular Phylogeny

    Science.gov (United States)

    Schwartzman, David; Lineweaver, Charles H.

    2004-06-01

    The timing of emergence of major organismal groups is consistent with the climatic temperature being equal to their upper temperature limit of growth (T_{max}), implying a temperature constraint on the evolution of each group, with the climatic temperature inferred from the oxygen isotope record of marine cherts. Support for this constraint comes from the correlation of T_{max} with the rRNA molecular phylogenetic distance from the last common ancestor (LCA) for both thermophilic Archaea and Bacteria. In particular, this correlation for hyperthermophilic Archaea suggests a climatic temperature of about 120°C at the time of the LCA, likely in the Hadean.

  3. Phylogenetic and Physiological Diversity of Subseafloor Microbial Communities at Axial Seamount, Juan de Fuca Ridge: Summary of Results From the New Millenium Observatory (NeMO), 1998-2004

    Science.gov (United States)

    Baross, J. A.; Huber, J. A.; Mehta, M. P.; Opatkiewicz, A.; Bolton, S. A.; Butterfield, D. A.; Sogin, M. L.; Embley, R. W.

    2005-12-01

    Axial Seamount (45 ° 58' N; 130 ° 00' W) is an active submarine volcano located on the Juan de Fuca Ridge, approximately 300 miles off the coast of Oregon. Lying at the intersection of a seamount chain and a spreading axis, Axial is a unique study site from both the geological and biological perspective. In January of 1998, Axial experienced a week-long series of earthquakes, and subsequent water column and seafloor observations on the southeast portion of the caldera found temperature and chemical anomalies, extensive new seafloor lava flows, large "snow blower" type vents, and other characteristics commonly associated with diking-eruptive events. Due to its high activity and close proximity to shore, Axial was chosen as a site for a multi-year observatory (New Millenium Observatory, NeMO) to document changes and interactions between geology, chemistry, and biology on the mid-ocean ridge system. From 1998 through 2004, we extensively sampled diffuse vents at Axial Seamount to determine the physiological and phylogenetic diversity of subseafloor microbial communities and their relationship to the geochemical environment. Here we present a summary of those studies, including molecular-based phylogenetic surveys of bacteria, archaea, and potential nitrogen-fixing organisms, culturing results of thermophiles and hyperthermophiles from over 20 sites, and the distribution of one particular group of hyperthermophiles at diffuse vents throughout the caldera and how that distribution may be linked to the geochemical habitat. Results indicate that Axial supports a diverse subseafloor microbial community, including hydrogen and sulfur oxidizers, hyperthermophilic methane producers and heterotrophs, and many organisms with the potential to fix nitrogen. In addition, we find that the species composition of the microbial community changes in response to changes in the physical and chemical conditions at each vent site. The extent of seawater mixing with hydrothermal fluids

  4. Isolation of diverse members of the Aquificales from geothermal springs in Tengchong, China

    OpenAIRE

    Hedlund, Brian P.; Reysenbach, Anna-Louise; Huang, Liuquin; Ong, John C.; Liu, Zizhang; Dodsworth, Jeremy A.; Ahmed, Reham; Williams, Amanda J.; Briggs, Brandon R.; Liu, Yitai; Hou, Weiguo; Dong, Hailiang

    2015-01-01

    The order Aquificales (phylum Aquificae) consists of thermophilic and hyperthermophilic bacteria that are prominent in many geothermal systems, including those in Tengchong, Yunnan Province, China. However, Aquificales have not previously been isolated from Tengchong. We isolated five strains of Aquificales from diverse springs (temperature 45.2–83.3°C and pH 2.6–9.1) in the Rehai Geothermal Field from sites in which Aquificales were abundant. Phylogenetic analysis showed that four of the str...

  5. An abyssal mobilome: viruses, plasmids and vesicles from deep-sea hydrothermal vents.

    Science.gov (United States)

    Lossouarn, Julien; Dupont, Samuel; Gorlas, Aurore; Mercier, Coraline; Bienvenu, Nadege; Marguet, Evelyne; Forterre, Patrick; Geslin, Claire

    2015-12-01

    Mobile genetic elements (MGEs) such as viruses, plasmids, vesicles, gene transfer agents (GTAs), transposons and transpovirions, which collectively represent the mobilome, interact with cellular organisms from all three domains of life, including those thriving in the most extreme environments. While efforts have been made to better understand deep-sea vent microbial ecology, our knowledge of the mobilome associated with prokaryotes inhabiting deep-sea hydrothermal vents remains limited. Here we focus on the abyssal mobilome by reviewing accumulating data on viruses, plasmids and vesicles associated with thermophilic and hyperthermophilic Bacteria and Archaea present in deep-sea hydrothermal vents. Copyright © 2015 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  6. Extremely thermophilic microorganisms and their polymer-hidrolytic enzymes

    Directory of Open Access Journals (Sweden)

    Andrade Carolina M.M.C.

    1999-01-01

    Full Text Available Thermophilic and hyperthermophilic microorganisms are found as normal inhabitants of continental and submarine volcanic areas, geothermally heated sea-sediments and hydrothermal vents and thus are considered extremophiles. Several present or potential applications of extremophilic enzymes are reviewed, especially polymer-hydrolysing enzymes, such as amylolytic and hemicellulolytic enzymes. The purpose of this review is to present the range of morphological and metabolic features among those microorganisms growing from 70oC to 100°C and to indicate potential opportunities for useful applications derived from these features.

  7. C68 from the Sulfolobus islandicus plasmid-virus pSSVx is a novel member of the AbrB-like transcription factor family

    DEFF Research Database (Denmark)

    Contursi, Patrizia; D'Ambrosio, Katia; Pirone, Luciano

    2011-01-01

    The genetic element pSSVx from Sulfolobus islandicus, strain REY15/4, is a hybrid between a plasmid and a fusellovirus. This plasmid-virus hybrid infects several species of the hyperthermophilic acidophilic crenarchaeon Sulfolobus. The open reading frame orfc68 of pSSVx encodes a 7.7 kDa protein...... factors, such as AbrB from Bacillus subtilis. Nevertheless, C68 constitutes a novel representative of this family because it shows several peculiar structural and functional features....

  8. Bacterial Sulfate Reduction Above 100-Degrees-C in Deep-Sea Hydrothermal Vent Sediments

    DEFF Research Database (Denmark)

    JØRGENSEN, BB; ISAKSEN, MF; JANNASCH, HW

    1992-01-01

    -reducing bacteria was done in hot deep-sea sediments at the hydrothermal vents of the Guaymas Basin tectonic spreading center in the Gulf of California. Radiotracer studies revealed that sulfate reduction can occur at temperatures up to 110-degrees-C, with an optimum rate at 103-degrees to 106-degrees......-C. This observation expands the upper temperature limit of this process in deep-ocean sediments by 20-degrees-C and indicates the existence of an unknown groUp of hyperthermophilic bacteria with a potential importance for the biogeochemistry of sulfur above 100-degrees-C....

  9. Bacterial ecology of abattoir wastewater treated by an anaerobic digestor

    Directory of Open Access Journals (Sweden)

    Linda Jabari

    2016-03-01

    Full Text Available Abstract Wastewater from an anaerobic treatment plant at a slaughterhouse was analysed to determine the bacterial biodiversity present. Molecular analysis of the anaerobic sludge obtained from the treatment plant showed significant diversity, as 27 different phyla were identified. Firmicutes, Proteobacteria, Bacteroidetes, Thermotogae, Euryarchaeota (methanogens, and msbl6 (candidate division were the dominant phyla of the anaerobic treatment plant and represented 21.7%, 18.5%, 11.5%, 9.4%, 8.9%, and 8.8% of the total bacteria identified, respectively. The dominant bacteria isolated were Clostridium, Bacteroides, Desulfobulbus, Desulfomicrobium, Desulfovibrio and Desulfotomaculum. Our results revealed the presence of new species, genera and families of microorganisms. The most interesting strains were characterised. Three new bacteria involved in anaerobic digestion of abattoir wastewater were published.

  10. Bacterial ecology of abattoir wastewater treated by an anaerobic digestor.

    Science.gov (United States)

    Jabari, Linda; Gannoun, Hana; Khelifi, Eltaief; Cayol, Jean-Luc; Godon, Jean-Jacques; Hamdi, Moktar; Fardeau, Marie-Laure

    2016-01-01

    Wastewater from an anaerobic treatment plant at a slaughterhouse was analysed to determine the bacterial biodiversity present. Molecular analysis of the anaerobic sludge obtained from the treatment plant showed significant diversity, as 27 different phyla were identified. Firmicutes, Proteobacteria, Bacteroidetes, Thermotogae, Euryarchaeota (methanogens), and msbl6 (candidate division) were the dominant phyla of the anaerobic treatment plant and represented 21.7%, 18.5%, 11.5%, 9.4%, 8.9%, and 8.8% of the total bacteria identified, respectively. The dominant bacteria isolated were Clostridium, Bacteroides, Desulfobulbus, Desulfomicrobium, Desulfovibrio and Desulfotomaculum. Our results revealed the presence of new species, genera and families of microorganisms. The most interesting strains were characterised. Three new bacteria involved in anaerobic digestion of abattoir wastewater were published. Copyright © 2015 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  11. Biological hydrogen production by moderately thermophilic anaerobic bacteria

    International Nuclear Information System (INIS)

    HP Goorissen; AJM Stams

    2006-01-01

    This study focuses on the biological production of hydrogen at moderate temperatures (65-75 C) by anaerobic bacteria. A survey was made to select the best (moderate) thermophiles for hydrogen production from cellulolytic biomass. From this survey we selected Caldicellulosiruptor saccharolyticus (a gram-positive bacterium) and Thermotoga elfii (a gram-negative bacterium) as potential candidates for biological hydrogen production on mixtures of C 5 -C 6 sugars. Xylose and glucose were used as model substrates to describe growth and hydrogen production from hydrolyzed biomass. Mixed substrate utilization in batch cultures revealed differences in the sequence of substrate consumption and in catabolites repression of the two microorganisms. The regulatory mechanisms of catabolites repression in these microorganisms are not known yet. (authors)

  12. Extremophiles survival to simulated space conditions: an astrobiology model study.

    Science.gov (United States)

    Mastascusa, V; Romano, I; Di Donato, P; Poli, A; Della Corte, V; Rotundi, A; Bussoletti, E; Quarto, M; Pugliese, M; Nicolaus, B

    2014-09-01

    In this work we investigated the ability of four extremophilic bacteria from Archaea and Bacteria domains to resist to space environment by exposing them to extreme conditions of temperature, UV radiation, desiccation coupled to low pressure generated in a Mars' conditions simulator. All the investigated extremophilic strains (namely Sulfolobus solfataricus, Haloterrigena hispanica, Thermotoga neapolitana and Geobacillus thermantarcticus) showed a good resistance to the simulation of the temperature variation in the space; on the other hand irradiation with UV at 254 nm affected only slightly the growth of H. hispanica, G. thermantarcticus and S. solfataricus; finally exposition to Mars simulated condition showed that H. hispanica and G. thermantarcticus were resistant to desiccation and low pressure.

  13. Drivers of microbial community composition in mesophilic and thermophilic temperature-phased anaerobic digestion pre-treatment reactors.

    Science.gov (United States)

    Pervin, Hasina M; Dennis, Paul G; Lim, Hui J; Tyson, Gene W; Batstone, Damien J; Bond, Philip L

    2013-12-01

    Temperature-phased anaerobic digestion (TPAD) is an emerging technology that facilitates improved performance and pathogen destruction in anaerobic sewage sludge digestion by optimising conditions for 1) hydrolytic and acidogenic organisms in a first-stage/pre-treatment reactor and then 2) methogenic populations in a second stage reactor. Pre-treatment reactors are typically operated at 55-65 °C and as such select for thermophilic bacterial communities. However, details of key microbial populations in hydrolytic communities and links to functionality are very limited. In this study, experimental thermophilic pre-treatment (TP) and control mesophilic pre-treatment (MP) reactors were operated as first-stages of TPAD systems treating activated sludge for 340 days. The TP system was operated sequentially at 50, 60 and 65 °C, while the MP rector was held at 35 °C for the entire period. The composition of microbial communities associated with the MP and TP pre-treatment reactors was characterised weekly using terminal-restriction fragment length polymorphism (T-RFLP) supported by clone library sequencing of 16S rRNA gene amplicons. The outcomes of this approach were confirmed using 454 pyrosequencing of gene amplicons and fluorescence in-situ hybridisation (FISH). TP associated bacterial communities were dominated by populations affiliated to the Firmicutes, Thermotogae, Proteobacteria and Chloroflexi. In particular there was a progression from Thermotogae to Lutispora and Coprothermobacter and diversity decreased as temperature and hydrolysis performance increased. While change in the composition of TP associated bacterial communities was attributable to temperature, that of MP associated bacterial communities was related to the composition of the incoming feed. This study determined processes driving the dynamics of key microbial populations that are correlated with an enhanced hydrolytic functionality of the TPAD system. Copyright © 2013 Elsevier Ltd. All rights

  14. Solution structure of an archaeal DNA binding protein with an eukaryotic zinc finger fold.

    Directory of Open Access Journals (Sweden)

    Florence Guillière

    Full Text Available While the basal transcription machinery in archaea is eukaryal-like, transcription factors in archaea and their viruses are usually related to bacterial transcription factors. Nevertheless, some of these organisms show predicted classical zinc fingers motifs of the C2H2 type, which are almost exclusively found in proteins of eukaryotes and most often associated with transcription regulators. In this work, we focused on the protein AFV1p06 from the hyperthermophilic archaeal virus AFV1. The sequence of the protein consists of the classical eukaryotic C2H2 motif with the fourth histidine coordinating zinc missing, as well as of N- and C-terminal extensions. We showed that the protein AFV1p06 binds zinc and solved its solution structure by NMR. AFV1p06 displays a zinc finger fold with a novel structure extension and disordered N- and C-termini. Structure calculations show that a glutamic acid residue that coordinates zinc replaces the fourth histidine of the C2H2 motif. Electromobility gel shift assays indicate that the protein binds to DNA with different affinities depending on the DNA sequence. AFV1p06 is the first experimentally characterised archaeal zinc finger protein with a DNA binding activity. The AFV1p06 protein family has homologues in diverse viruses of hyperthermophilic archaea. A phylogenetic analysis points out a common origin of archaeal and eukaryotic C2H2 zinc fingers.

  15. Cloning, Sequencing, and Expression of the Gene Encoding Cyclic 2,3-Diphosphoglycerate Synthetase, the Key Enzyme of Cyclic 2,3-Diphosphoglycerate Metabolism in Methanothermus fervidus

    Science.gov (United States)

    Matussek, Karl; Moritz, Patrick; Brunner, Nina; Eckerskorn, Christoph; Hensel, Reinhard

    1998-01-01

    Cyclic 2,3-diphosphoglycerate synthetase (cDPGS) catalyzes the synthesis of cyclic 2,3-diphosphoglycerate (cDPG) by formation of an intramolecular phosphoanhydride bond in 2,3-diphosphoglycerate. cDPG is known to be accumulated to high intracellular concentrations (>300 mM) as a putative thermoadapter in some hyperthermophilic methanogens. For the first time, we have purified active cDPGS from a methanogen, the hyperthermophilic archaeon Methanothermus fervidus, sequenced the coding gene, and expressed it in Escherichia coli. cDPGS purification resulted in enzyme preparations containing two isoforms differing in their electrophoretic mobility under denaturing conditions. Since both polypeptides showed the same N-terminal amino acid sequence and Southern analyses indicate the presence of only one gene coding for cDPGS in M. fervidus, the two polypeptides originate from the same gene but differ by a not yet identified modification. The native cDPGS represents a dimer with an apparent molecular mass of 112 kDa and catalyzes the reversible formation of the intramolecular phosphoanhydride bond at the expense of ATP. The enzyme shows a clear preference for the synthetic reaction: the substrate affinity and the Vmax of the synthetic reaction are a factor of 8 to 10 higher than the corresponding values for the reverse reaction. Comparison with the kinetic properties of the electrophoretically homogeneous, apparently unmodified recombinant enzyme from E. coli revealed a twofold-higher Vmax of the enzyme from M. fervidus in the synthesizing direction. PMID:9811660

  16. Backbone and side-chain 1H, 15N and 13C resonance assignments of two Sac10b family members Mvo10b and Mth10bTQQA from archaea.

    Science.gov (United States)

    Xuan, Jinsong; Yao, Hongwei; Feng, Yingang; Wang, Jinfeng

    2017-10-01

    The Sac10b family proteins, also named as Alba, are small, basic, nucleic acid-binding proteins widely distributed in archaea. They possess divergent physiological functions such as binding to both DNA and RNA with a high affinity and involving in genomic DNA compaction, RNA transactions and transcriptional regulations. The structures of many Sac10b family proteins from hyperthermophilic archaea have been reported, while those from thermophilic and mesophilic archaea are largely unknown. As was pointed out, the homologous members from thermophilic and mesophilic archaea may have functions different from the hyperthermophilic members. Therefore, comparison of these homologous members can provide biophysical and structural insight into the functional diversity and thermal adaptation mechanism. The present work mainly focused on the NMR study of two Sac10b family members, Mvo10b and Mth10b, from the mesophilic and thermophilic archaea, respectively. To overcome the difficulties caused by the oligomerization and conformation heterogeneity of Mth10b, a M13T/L17Q/I20Q/P56A mutant Mth10b (Mth10bTQQA) was constructed and used together with Mvo10b for multi-dimensional NMR experiments. The resonance assignments of Mvo10b and Mth10bTQQA are reported for further structural determination which is a basis for understanding the functional diversity and their thermal adaption mechanisms.

  17. Expression, Purification, and Characterisation of Dehydroquinate Synthase from Pyrococcus furiosus

    Directory of Open Access Journals (Sweden)

    Leonardo Negron

    2011-01-01

    Full Text Available Dehydroquinate synthase (DHQS catalyses the second step of the shikimate pathway to aromatic compounds. DHQS from the archaeal hyperthermophile Pyrococcus furiosus was insoluble when expressed in Escherichia coli but was partially solubilised when KCl was included in the cell lysis buffer. A purification procedure was developed, involving lysis by sonication at 30∘C followed by a heat treatment at 70∘C and anion exchange chromatography. Purified recombinant P. furiosus DHQS is a dimer with a subunit Mr of 37,397 (determined by electrospray ionisation mass spectrometry and is active over broad pH and temperature ranges. The kinetic parameters are KM (3-deoxy-D-arabino-heptulosonate 7-phosphate 3.7 μM and kcat 3.0 sec-1 at 60∘C and pH 6.8. EDTA inactivates the enzyme, and enzyme activity is restored by several divalent metal ions including (in order of decreasing effectiveness Cd2+, Co2+, Zn2+, and Mn2+. High activity of a DHQS in the presence of Cd2+ has not been reported for enzymes from other sources, and may be related to the bioavailability of Cd2+ for P. furiosus. This study is the first biochemical characterisation of a DHQS from a thermophilic source. Furthermore, the characterisation of this hyperthermophilic enzyme was carried out at elevated temperatures using an enzyme-coupled assay.

  18. Crystallization and preliminary crystallographic analysis of mannosyl-3-phosphoglycerate synthase from Rubrobacter xylanophilus

    International Nuclear Information System (INIS)

    Sá-Moura, Bebiana; Albuquerque, Luciana; Empadinhas, Nuno; Costa, Milton S. da; Pereira, Pedro José Barbosa; Macedo-Ribeiro, Sandra

    2008-01-01

    The enzyme mannosyl-3-phosphoglycerate synthase from R. xylanophilus has been expressed, purified and crystallized. The crystals belong to the hexagonal space group P6 5 22 and diffract to 2.2 Å resolution. Rubrobacter xylanophilus is the only Gram-positive bacterium known to synthesize the compatible solute mannosylglycerate (MG), which is commonly found in hyperthermophilic archaea and some thermophilic bacteria. Unlike the salt-dependent pattern of accumulation observed in (hyper)thermophiles, in R. xylanophilus MG accumulates constitutively. The synthesis of MG in R. xylanophilus was tracked from GDP-mannose and 3-phosphoglycerate, but the genome sequence of the organism failed to reveal any of the genes known to be involved in this pathway. The native enzyme was purified and its N-terminal sequence was used to identify the corresponding gene (mpgS) in the genome of R. xylanophilus. The gene encodes a highly divergent mannosyl-3-phosphoglycerate synthase (MpgS) without relevant sequence homology to known mannosylphosphoglycerate synthases. In order to understand the specificity and enzymatic mechanism of this novel enzyme, it was expressed in Escherichia coli, purified and crystallized. The crystals thus obtained belonged to the hexagonal space group P6 5 22 and contained two protein molecules per asymmetric unit. The structure was solved by SIRAS using a mercury derivative

  19. Cloning, sequencing, and expression of the gene encoding cyclic 2, 3-diphosphoglycerate synthetase, the key enzyme of cyclic 2, 3-diphosphoglycerate metabolism in Methanothermus fervidus.

    Science.gov (United States)

    Matussek, K; Moritz, P; Brunner, N; Eckerskorn, C; Hensel, R

    1998-11-01

    Cyclic 2,3-diphosphoglycerate synthetase (cDPGS) catalyzes the synthesis of cyclic 2,3-diphosphoglycerate (cDPG) by formation of an intramolecular phosphoanhydride bond in 2,3-diphosphoglycerate. cDPG is known to be accumulated to high intracellular concentrations (>300 mM) as a putative thermoadapter in some hyperthermophilic methanogens. For the first time, we have purified active cDPGS from a methanogen, the hyperthermophilic archaeon Methanothermus fervidus, sequenced the coding gene, and expressed it in Escherichia coli. cDPGS purification resulted in enzyme preparations containing two isoforms differing in their electrophoretic mobility under denaturing conditions. Since both polypeptides showed the same N-terminal amino acid sequence and Southern analyses indicate the presence of only one gene coding for cDPGS in M. fervidus, the two polypeptides originate from the same gene but differ by a not yet identified modification. The native cDPGS represents a dimer with an apparent molecular mass of 112 kDa and catalyzes the reversible formation of the intramolecular phosphoanhydride bond at the expense of ATP. The enzyme shows a clear preference for the synthetic reaction: the substrate affinity and the Vmax of the synthetic reaction are a factor of 8 to 10 higher than the corresponding values for the reverse reaction. Comparison with the kinetic properties of the electrophoretically homogeneous, apparently unmodified recombinant enzyme from E. coli revealed a twofold-higher Vmax of the enzyme from M. fervidus in the synthesizing direction.

  20. Microbial and Mineral Descriptions of the Interior Habitable Zones of Active Hydrothermal Chimneys from the Endeavour Segment, Juan de Fuca Ridge

    Science.gov (United States)

    Holden, J. F.; Lin, T.; Ver Eecke, H. C.; Breves, E.; Dyar, M. D.; Jamieson, J. W.; Hannington, M. D.; Butterfield, D. A.; Bishop, J. L.; Lane, M. D.

    2013-12-01

    Actively venting hydrothermal chimneys and their associated hydrothermal fluids were collected from the Endeavour Segment, Juan de Fuca Ridge to determine the mineralogy, chemistry and microbial community composition of their interiors. To characterize the mineralogy, Mössbauer, FTIR, VNIR and thermal emission spectroscopies were used for the first time on this type of sample in addition to thin-section petrography, x-ray diffraction and elemental analyses. A chimney from the Bastille edifice was Fe-sulfide rich and composed primarily of chalcopyrite, marcasite-sphalerite, and pyrrhotite while chimneys from the Dante and Hot Harold edifices were Fe-sulfide poor and composed primarily of anhydrite. The bulk emissivity and reflectance spectroscopies corroborated well with the petrography and XRD analyses. The microbial community in the interior of Bastille was most closely related to mesophilic-to-thermophilic anaerobes of the deltaproteobacteria and hyperthermophilic archaea while those in the interiors of Dante and Hot Harold were most closely related to mesophilic-to-thermophilic aerobes of the beta-, gamma- and epsilonproteobacteria. The fluid temperatures (282-321°C) and chemistries of the three chimneys were very similar suggesting that differences in mineralogy and microbial community compositions were more dependent on fluid flow characteristics and paragenesis within the chimney. Thin-section petrography of the interior of another hydrothermal chimney collected from the Dante edifice (emitting 336°C fluid) shows a thin coat of Fe3+ oxide associated with amorphous silica on the exposed outer surfaces of pyrrhotite, sphalerite and chalcopyrite in pore spaces, along with anhydrite precipitation in the pores that is indicative of seawater ingress. The Fe-sulfide minerals were likely oxidized to ferrihydrite with increasing pH and Eh due to cooling and seawater exposure, providing reactants for bioreduction. Culture-based most-probable-number estimates of

  1. Borders of life: lessons from Microbiology of deep-sea hydrothermal vents

    Science.gov (United States)

    Prieur, D.

    Thirty years ago, the deep-sea was known as a low density biotope due to coldness, darkness and famine-like conditions. The discovery of deep-sea hydrothermal vents in the Eastern Pacific in 1977 and the associated black smokers in 1979 considerably changed our views about life on Earth. For the first time, an ecosystem almost independent (at least for tens of years) of solar nergy was discovered. Besides the spectacular and unexpected communities of invertebrates based on symbiotic associations with chemo-litho-autotrophic bacteria, prokaryotic communities associated with high temperature black smokers fascinated microbiologists of extreme environments. Within mineral structures where temperature gradients may fluctuate from ambient seawater temperatures (2°C) up to 350°C, thermophilic (optimal growth above 60°C) and hyperthermophilic (optimal growth above 80°C) microorganisms thrived under very severe conditions due to elevated hydrostatic pressure, toxic compounds or strong ionizing radiations. These organisms belong to both domains of Bacteria and Archaea and live aerobically but mostly anaerobically, using a variety of inorganic and organic carbon sources, and a variety of electron donnors and acceptors as well. The most thermophilic organism known on Earth was isolated from a mid-Atlantic-Ridge hydrotermal vent: Pyrolobus fumarii grows optimally at 110°c and its upper temperature limit for life is 113°C. Such an organism survived to autoclaving conditions currently used for sterilization procedures. Many other hyperthermophilic organisms were isolated and described, including fermenters, sulphate and sulphur reducers, hydrogen oxidizers, nitrate reducers, methanogens, etc. Although most of anaerobes are killed when exposed to oxygen, several deep-sea hyperthermophiles appeared to survive to both oxygen and starvation exposures, indicating that they probably can colonize rather distant environments Because of elevated hydrostatic pressure that exists at

  2. Gaps of maritime health research in Latin America – a literature review

    DEFF Research Database (Denmark)

    Jensen, Olaf Chresten; Andrioti, Despena; Canals, M. Luisa

    for research in this part of the world. Materials and Methods PubMed, Google Scholar, SciELO - Scientific Electronic Library Online, Pan American Journal of Public Health, Medicina Maritima and other relevant journals in Latin America in the Spanish and English languages were searched. Results 57 peer......-reviewed articles on fishermen´s health and safety and none for the seafarers were included. Brazil counted for the main part n =39, while each of the other countries had 0-4 studies. The study objectives include occupational injuries, divers disease, skin diseases, hearing loss and other issues. The cross......Background So far the maritime health and safety research for seafarers and fishermen mainly comes from the industrial developed countries with sparse contributions from the developing countries. The aim was to give an overview of the peer reviewed research in Latin America to point out the needs...

  3. Review of Maritime Health research gab in latin America

    DEFF Research Database (Denmark)

    Jensen, Olaf Chresten

    for research in this part of the world. Materials and Methods PubMed, Google Scholar, SciELO - Scientific Electronic Library Online, Pan American Journal of Public Health, Medicina Maritima and other relevant journals in Latin America in the Spanish and English languages were searched. Results 57 peer......-reviewed articles on fishermen´s health and safety and none for the seafarers were included. Brazil counted for the main part n =39, while each of the other countries had 0-4 studies. The study objectives include occupational injuries, divers disease, skin diseases, hearing loss and other issues. The cross......Background So far the maritime health and safety research for seafarers and fishermen mainly comes from the industrial developed countries with sparse contributions from the developing countries. The aim was to give an overview of the peer reviewed research in Latin America to point out the needs...

  4. Dispersion and establishment of the species of mangrove of the Rancheria River in the period of maximum fructification

    International Nuclear Information System (INIS)

    Lema Velez, Luisa Fernanda; Polania, Jaime; Urrego Giraldo, Ligia Estela

    2003-01-01

    Dispersion and establishment patterns of Rhizophora mangle, Avicennia germinans, and Laguncularia racemosa, at the Rancheria River were studied using marked propagules. These mangroves are small, and surrounded by subtropical dry forest, subtropical thorn forest and the city of Riohacha. Significant relationships were found between the number of propagules retained and species, time, distance to release site, and retention structure. Avicennia germinans and L. racemosa propagules left the ecosystem within two weeks while a portion of R. mangle propagules remained during the two months study and were able to settle. Goat predation affected mainly the propagules, especially of A. germinans and L. racemosa. Predation did not follow the current dominance-predation model. A pre-dispersion consumption of 30% of A. germinans propagules by Pyralidae larvae was also documented. Batis maritima plants were the most effective structures that retain propagules of the three species and R. mangle propagules remained in greater quantities and for longer periods in the ecosystem

  5. A redescription of Rhysida celeris (Humbert & Saussure, 1870), with a proposal of eight new synonyms (Scolopendromorpha, Scolopendridae, Otostigminae).

    Science.gov (United States)

    Chagas-Júnior, Amazonas

    2013-01-01

    Seven species of the genus Rhysida Wood, 1862 from Venezuela and one subspecies from Peru described by Manuel Angel González Sponga and Wolfgang Bücherl respectively, are revised. Rhysida caripensis González-Sponga, 2002, Rhysida neoespartana González-Sponga, 2002, Rhysida guayanica González-Sponga, 2002, Rhysida maritima González-Sponga, 2002, Rhysida monaguensis González-Sponga, 2002, Rhysida porlamarensis González-Sponga 2002, Rhysida sucupanensis González-Sponga, 2002 and Rhysida celeris andina Bücherl, 1953 are junior synonyms of Rhysida celeris (Humbert & Saussure, 1870), which is redescribed and illustrated for the first time. Its geographic distribution is updated and a map showing its distribution is presented.

  6. First record of the genus Schoettella and three new records of the family Hypogastruridae (Collembola, Hexapoda for fauna of Iran with an identification key for Mazandaran province

    Directory of Open Access Journals (Sweden)

    Elham Yoosefi Lafooraki

    2015-07-01

    Full Text Available The Hypogastruridae family belonging to the class Collembola (Springtails are among the most important and abundant soil arthropods. These animals play important roles in decomposition processes and nutrient cycling. However, their fauna have remained too much unknown in Iran. In order to study of Collembola fauna in the Mazandaran province, some sampling from soil, leaf litters and mosses were made from different regions of the province during 2012-2013 years. Then, the springtails of samples were separated using Berlese funnel and preserved in 75-85 % ethyl alcohol. During the investigation, some samples belonging to Hypogastruridae were collected and identified. The genus Schoettella and the three species S. unungiuculata, Hypogastrura purpurescens and Ceratophysella engadinensis are new records for fauna of Iran and the two species Xenylla maritima and C. stercoraria are recorded for the first time from Mazandaran province. In addition, an identification key for local genera and species of the family Hypogastruridae in Mazandaran is presented here.

  7. Assessment and mitigation of liquefaction

    International Nuclear Information System (INIS)

    Czelada, J. A.; Melentijevic, S.

    2014-01-01

    The simplified empirical procedure in its original form presented in Youd et al (2001) and some further developments given in Idriss and Boulanger (2006) for evaluating liquefaction resistance of soils is presented in this paper only for the criteria based on standard penetration test (SPT). Methods for estimating the ground improvement techniques by stone columns and dynamics compaction are presented. For stone columns Priebe method (1995) and homogenized method (equivalent parameters) are present. for dynamic compaction methods proposed by Recomendacion Geotecnica para las Obras Maritimas y/o Porturaria - ROM 0.5-05 (2005) and Nashed et al. (2009) are described. These analysis methods for each ground improvement technique are compared in two different case histories showing similar results in each one. (Author)

  8. Food habits of mute swans in the Chesapeake Bay

    Science.gov (United States)

    Perry, M.C.; Osenton, P.C.; Lohnes, E.J.R.; Perry, Matthew C.

    2004-01-01

    Unlike the tundra swan (Cygnus columbianus) that migrate to the Bay for the winter, the mute swan (Cygnus olor) is a year long resident and therefore has raised concerns among research managers over reports of conflicts with nesting native water birds and the consumption of submerged aquatic vegetation (SAV). Although data on the reduction of SAV by nesting mute swans and their offspring during the spring and summer are limited, food-habits data show that mute swans rely heavily on SAV during these months. Analyses of the gullet and gizzard of mute swans indicate that widgeon grass (Ruppia maritima) and eelgrass (Zostera marina) were the most important food items to mute swans during the winter and spring. Other organisms were eaten by mute swans, but represent small percentages of food. Corn (Zea mays) fed to the swans by Bay residents in late winter probably supplements their limited vegetative food resources at that time of year.

  9. Feeding habits of Carabidae (Coleoptera associated with herbaceous plants and the phenology of coloured cotton

    Directory of Open Access Journals (Sweden)

    Danilo Henrique da Matta

    2017-04-01

    Full Text Available The carabids (Coleoptera: Carabidae are recognized as polyphagous predators and important natural enemies of insect pests. However, little is known about the feeding habits of these beetles. In this work, we determine the types of food content in the digestive tracts of nine species of Carabidae associated with herbaceous plants and different growth stages of coloured cotton. The food contents were evaluated for beetles associated with the coloured cotton cv. BRS verde, Gossypium hirsutum L. latifolium Hutch., adjacent to weed plants and the flowering herbaceous plants (FHPs Lobularia maritima (L., Tagetes erecta L., and Fagopyrum esculentum Moench. The digestive tract analysis indicated various types of diets and related arthropods for Abaris basistriata, Galerita brasiliensis, Scarites sp., Selenophorus alternans, Selenophorus discopunctatus and Tetracha brasiliensis. The carabids were considered to be polyphagous predators, feeding on different types of prey.

  10. Effect of saline soil parameters on endo mycorrhizal colonisation of dominant halophytes in four Hungarian sites

    Energy Technology Data Exchange (ETDEWEB)

    Fuzy, A.; Biro, B.; Toth, T.

    2010-07-01

    Soil and root samples were collected from the rhizosphere of dominant halophytes (Artemisia santonicum, Aster tripolium, Festuca pseudovina, Lepidium crassifolium, Plantago maritima and Puccinellia limosa) at four locations with saline soils in Hungary. The correlations- between arbuscular mycorrhiza (AM) fungal colonisation parameters (% colonisation, % arbuscules) and soil physical, chemical and biological parameters were determined Endomycorrhiza colonisation was found to be negatively correlated with the electric conductivity of the soil paste, the salt-specific ion concentrations and the cation exchange capacity, showing the sensitivity of AM fungi at increasing salt concentrations, independently of the types of salt-specific anions. A positive correlation was detected between the mycorrhiza colonisation and the abundance of oligotroph bacteria known to be the less variable and more stable (k-strategist) group. This fact and the negative correlation found with the humus content underlines the importance of nutrient availability and the limitations of the symbiotic interactions in stressed saline or sodic soils. (Author) 29 refs.

  11. Assessment and mitigation of liquefaction; Evaluacion y mitigacion de la licuefaccion

    Energy Technology Data Exchange (ETDEWEB)

    Czelada, J. A.; Melentijevic, S.

    2014-07-01

    The simplified empirical procedure in its original form presented in Youd et al (2001) and some further developments given in Idriss and Boulanger (2006) for evaluating liquefaction resistance of soils is presented in this paper only for the criteria based on standard penetration test (SPT). Methods for estimating the ground improvement techniques by stone columns and dynamics compaction are presented. For stone columns Priebe method (1995) and homogenized method (equivalent parameters) are present. for dynamic compaction methods proposed by Recomendacion Geotecnica para las Obras Maritimas y/o Porturaria - ROM 0.5-05 (2005) and Nashed et al. (2009) are described. These analysis methods for each ground improvement technique are compared in two different case histories showing similar results in each one. (Author)

  12. Ecological correlates of variable organ sizes and fat loads in the most northerly-wintering shorebirds

    Science.gov (United States)

    Ruthrauff, Daniel R.; Dekinga, Anne; Gill, R.E.; Summers, R.W.; Piersma, Theunis

    2013-01-01

    Shorebirds at northern latitudes during the nonbreeding season typically carry relatively large lipid stores and exhibit an up-regulation of lean tissues associated with digestion and thermogenesis. Intraspecific variation in these tissues across sites primarily reflects differences in environmental conditions. Rock (Calidris ptilocnemis (Coues, 1873)) and Purple (Calidris maritima (Brünnich, 1764)) sandpipers are closely related species having the most northerly nonbreeding distributions among shorebirds, living at latitudes up to 61°N in Cook Inlet, Alaska, and up to 71°N in northern Norway, respectively. Cook Inlet is the coldest known site used by nonbreeding shorebirds, and the region’s mudflats annually experience extensive coverage of foraging sites by sea and shore-fast ice. Accordingly, Rock Sandpipers increase their fat stores to nearly 20% of body mass during winter. In contrast, Purple Sandpipers exploit predictably ice-free rocky intertidal foraging sites and maintain low (food resources.

  13. Feeding ecology of waterfowl wintering on evaporation ponds in California

    Science.gov (United States)

    Euliss, N.H.; Jarvis, R.L.; Gilmer, D.S.

    1991-01-01

    We examined the feeding ecology of Northern Pintails (Anas acuta), Northern Shovelers (A. clypeata), and Ruddy Ducks (Oxyura jamaicensis) wintering on drainwater evaporation ponds in California from 1982 through 1984. Pintails primarily consumed midges (Chironomidae) (39.3%) and widegeongrass (Ruppia maritima) nutlets (34.6%). Shovelers and Ruddy Ducks consumed 92.5% and 90.1% animal matter, respectively. Water boatmen (Corixidae) (51.6%), rotifers (Rotatoria) (20.4%), and copepods (Copepoda) (15.2%) were the most important Shoveler foods, and midges (49.7%) and water boatmen (36.0%) were the most important foods of Ruddy Ducks. All three species were opportunistic foragers, shifting their diets seasonally to the most abundant foods given their behavioral and morphological attributes.

  14. Relationships between the floral neighborhood and individual pollen limitation in two self-incompatible herbs.

    Science.gov (United States)

    Jakobsson, Anna; Lázaro, Amparo; Totland, Orjan

    2009-07-01

    Local flower density can affect pollen limitation and plant reproductive success through changes in pollinator visitation and availability of compatible pollen. Many studies have investigated the relationship between conspecific density and pollen limitation among populations, but less is known about within-population relationships and the effect of heterospecific flower density. In addition, few studies have explicitly assessed how the spatial scales at which flowers are monitored affect relationships. We investigated the effect of floral neighborhood on pollen limitation at four spatial scales in the self-incompatible herbs Armeria maritima spp. maritima and Ranunculus acris spp. acris. Moreover, we measured pollen deposition in Armeria and pollinator visits to Ranunculus. There was substantial variation in pollen limitation among Armeria individuals, and 25% of this variation was explained by the density of compatible and heterospecific flowers within a 3 m circle. Deposition of compatible pollen was affected by the density of compatible and incompatible inflorescences within a 0.5 m circle, and deposition of heterospecific pollen was affected by the density of heterospecific flowers within a 2 m circle. In Ranunculus, the number of pollinator visits was affected by both conspecific and heterospecific flower densities. This did not, however, result in effects of the floral neighborhood on pollen limitation, probably due to an absence of pollen limitation at the population level. Our study shows that considerable variation in pollen limitation may occur among individuals of a population, and that this variation is partly explained by floral neighborhood density. Such individual-based measures provide an important link between pollen limitation theory, which predicts ecological and evolutionary causes and consequences for individual plants, and studies of the effects of landscape fragmentation on plant species persistence. Our study also highlights the importance

  15. Impacts of oil sands process water on fen plants: Implications for plant selection in required reclamation projects

    International Nuclear Information System (INIS)

    Pouliot, Rémy; Rochefort, Line; Graf, Martha D.

    2012-01-01

    Fen plant growth in peat contaminated with groundwater discharges of oil sands process water (OSPW) was assessed in a greenhouse over two growing seasons. Three treatments (non-diluted OSPW, diluted OSPW and rainwater) were tested on five vascular plants and four mosses. All vascular plants tested can grow in salinity and naphthenic acids levels currently produced by oil sands activity in northwestern Canada. No stress sign was observed after both seasons. Because of plant characteristics, Carex species (C. atherodes and C. utriculata) and Triglochin maritima would be more useful for rapidly restoring vegetation and creating a new peat-accumulating system. Groundwater discharge of OSPW proved detrimental to mosses under dry conditions and ensuring adequate water levels would be crucial in fen creation following oil sands exploitation. Campylium stellatum would be the best choice to grow in contaminated areas and Bryum pseudotriquetrum might be interesting as it has spontaneously regenerated in all treatments. - Highlights: ► Fen plant growth was assessed under groundwater discharges of oil sands process water. ► Sedge and grass species were not stressed after two growing seasons in greenhouse. ► Carex species and Triglochin maritima would be helpful in created contaminated fens. ► In dry conditions, contaminated groundwater discharge was detrimental for mosses. ► Campylium stellatum would be the best choice in created fens with contaminated water. - Sedges and grasses tolerated the contact with oil sands process water and could probably grow well in contaminated created fens, but mosses were particularly affected under dry conditions.

  16. Chemical constituents of selected Sudanese medicinal and aromatic plants

    International Nuclear Information System (INIS)

    Burham, B.O.

    2007-11-01

    Sudanese medicinal and aromatic plants (Alternanthra repens, Ambrosia maritima, Citrus paradisi, Croton zambesicus, Lepidium sativum, Morettia phillaena, Nauclea latifolia, Plectranthus barbatus, Pluchea dioscorides, and Sphaeranthus suaveolens) were analyzed for their chemical composition, mineral contents and secondary constituents. The concentration of manganese, copper, iron, nickel, lead, zinc and potassium in plant samples was performed using x-ray fluorescence spectrometry. The trace elements found in the smallest amount of the investigated plant species are lead, nickel and copper, while high concentration was detected for potassium, iron and manganese. Mn was accumulated with high level in Alternanthra repens species. Potassium was abundant in S. suaveolens and Ambrosia maritima. The values of concentration obtained for all studied elements were compared with published values of reference material, trace elements in Hay (powder) by International Atomic Energy Agency. Phyto chemical analysis of investigated plants was performed for constituents: Flavonoids, saponins, tannins, alkaloids, amino acids and sugars. The methanolic extracts of P.barbatus, C.paradisi, A.repens, N.latifolia, L. sativum and C. zambesicus are found to contain alkaloids. Results of TLC analysis were shown as R f values for saponins, bitter principles, essential oils, flavonoids and alkaloids. Quantification of flavonoids and tannins showed that flavonoid content was highest in case of Alternanthera repens and Sphaeranthus suavertens, whereas the highest tannin content was in case of Nauclea latifolia and Sphaearanthus suavertens. The results suggest that the user of traditional Sudanese crude drugs should be warned of potential danger of heavy metal poisoning because their concentrations seem to be higher than maximum values allowed by health agencies in several countries. This study has provided some biochemical basis for the ethno medical use of extracts from different candidate

  17. A ESPACIALIDADE DO VILEGISTURISTA MARÍTIMO EM FORTALEZA, CEARÁ: práticas e transformações recentes

    Directory of Open Access Journals (Sweden)

    Antônio Tadeu Pinto Soares Júnior

    2010-01-01

    Full Text Available The relationship of the society with the sea is redesigned by the metropolis with the consolidation of the modern maritime practices and the valorization of the land close to the sea (swimming, walks, "vilegiatura", habitation and coastal tourism as a place for entertainment and living, observed in the capitals of the northeast like Fortaleza. The practice of the "vilegiatura maritima" became present in the city in the decade of 1930 in the Iracema beach with the "vilegiaturistas" belonging to the upper class of Fortaleza and of all the Province of Ceará. Later, with the coastal urbanization in the decade of 1970, the phenomenon reaches also the municipalities members of the metropolitan region of the city such as Caucaia and Aquiraz. This work has as a general goal to analyse the phenomenon of the "vilegiatura marítima" that takes place on the cost of Fortaleza, evidenced by the acquisition of secondary residences in order to live close to the beach, even in a seasonal way, in the period from 1991 to 2000. In order to understand the unfolding of theses social processes in their urban tissue, Fortaleza and how its coastal space presents the practice of "vilegiatura maritima" were analyzed, by the diverse social agents, named the "vilegiaturistas" (domestic and foreign, the State, the real state sector and society, responsible for the transformations caused by the occupation, investment practices and use of the coastal zone, be for entertainment, be for secondary residences in condos. In this study are analysed the neighbourhoods of Iracema beach, Meireles, and Future beach I and it showed that the "vilegiaturistas marítimos" are concentrated on the Central and East part of the city, and simultaneously their "dilution" in the metropolis generating multicultural spots, opposite to the fi rst hypothesis of the "vilegiatura maritíma" as "the scape from the city", once this phenomenon became predominantly urban.

  18. New crystal forms of Diocleinae lectins in the presence of different dimannosides

    International Nuclear Information System (INIS)

    Moreno, Frederico Bruno Mendes Batista; Bezerra, Gustavo Arruda; Oliveira, Taianá Maia de; Souza, Emmanuel Prata de; Rocha, Bruno Anderson Matias da; Benevides, Raquel Guimarães; Delatorre, Plínio; Cavada, Benildo Sousa; Azevedo, Walter Filgueira Jr de

    2006-01-01

    The crystallization and preliminary X-ray data of Canavalia gladiata lectin (CGL) and C. maritima lectin (CML) complexed with Man(α1-2)Man(α1)OMe, Man(α1-3)Man(α1)OMe and Man(α1-4)Man(α1)OMe in two crystal forms [the complexes with Man(α1-3)Man(α1)OMe and Man(α1-4)Man(α1)OMe crystallized in space group P3 2 and those with Man(α1-2)Man(α1)OMe crystallized in space group I222], which differed from those of the native proteins (P2 1 2 1 2 for CML and C222 for CGL), are reported. Studying the interactions between lectins and sugars is important in order to explain the differences observed in the biological activities presented by the highly similar proteins of the Diocleinae subtribe. Here, the crystallization and preliminary X-ray data of Canavalia gladiata lectin (CGL) and C. maritima lectin (CML) complexed with Man(α1-2)Man(α1)OMe, Man(α1-3)Man(α1)OMe and Man(α1-4)Man(α1)OMe in two crystal forms [the complexes with Man(α1-3)Man(α1)OMe and Man(α1-4)Man(α1)OMe crystallized in space group P3 2 and those with Man(α1-2)Man(α1)OMe crystallized in space group I222], which differed from those of the native proteins (P2 1 2 1 2 for CML and C222 for CGL), are reported. The crystal complexes of ConA-like lectins with Man(α1-4)Man(α1)OMe are reported here for the first time

  19. Chemical constituents of selected Sudanese medicinal and aromatic plants

    Energy Technology Data Exchange (ETDEWEB)

    Burham, B O [Atomic Energy Researches Coordination Council, Sudan Academy of Sciences, Khartoum (Sudan)

    2007-11-15

    Sudanese medicinal and aromatic plants (Alternanthra repens, Ambrosia maritima, Citrus paradisi, Croton zambesicus, Lepidium sativum, Morettia phillaena, Nauclea latifolia, Plectranthus barbatus, Pluchea dioscorides, and Sphaeranthus suaveolens) were analyzed for their chemical composition, mineral contents and secondary constituents. The concentration of manganese, copper, iron, nickel, lead, zinc and potassium in plant samples was performed using x-ray fluorescence spectrometry. The trace elements found in the smallest amount of the investigated plant species are lead, nickel and copper, while high concentration was detected for potassium, iron and manganese. Mn was accumulated with high level in Alternanthra repens species. Potassium was abundant in S. suaveolens and Ambrosia maritima. The values of concentration obtained for all studied elements were compared with published values of reference material, trace elements in Hay (powder) by International Atomic Energy Agency. Phyto chemical analysis of investigated plants was performed for constituents: Flavonoids, saponins, tannins, alkaloids, amino acids and sugars. The methanolic extracts of P.barbatus, C.paradisi, A.repens, N.latifolia, L. sativum and C. zambesicus are found to contain alkaloids. Results of TLC analysis were shown as R{sub f} values for saponins, bitter principles, essential oils, flavonoids and alkaloids. Quantification of flavonoids and tannins showed that flavonoid content was highest in case of Alternanthera repens and Sphaeranthus suavertens, whereas the highest tannin content was in case of Nauclea latifolia and Sphaearanthus suavertens. The results suggest that the user of traditional Sudanese crude drugs should be warned of potential danger of heavy metal poisoning because their concentrations seem to be higher than maximum values allowed by health agencies in several countries. This study has provided some biochemical basis for the ethno medical use of extracts from different candidate

  20. Biological Control Against the Cowpea Weevil (Callosobruchus Chinensis L., Coleoptera: Bruchidae Using Essential Oils of Some Medicinal Plants

    Directory of Open Access Journals (Sweden)

    Fatiha Righi Assia

    2014-07-01

    Full Text Available Chickpea (Cicer arietinum L. is a valuable foodstuff but unfortunately this legume is prone to insect attacks from the chick pea weevil (Callosobruchus chinensis L.. This serious pest damages the chickpea and causes decreases in the yield and in the nutritional quality. Biological control is being used to deal with this problem. We tried different doses of the essential oils of three new medicinal plants, namely Salvia verbenaca L., Scilla maritima L., and Artemisia herba-alba Asso to limit the damage of the chick pea weevil pest, and to protect consumer’s health. To determine the effect and efficiency of the oil, the tests were conducted using the different biological parameters of fertility, longevity, and fecundity, under controlled temperature and relative humidity (28°C and 75%. The effectiveness of organic oils was demonstrated. We tested these oils on the germination of seeds. The obtained results showed that the tested plant oils have a real organic insecticide effect. The essential oil of Artemisia proved most effective as a biocide; achieving a mortality rate of 100%. A significant reduction in longevity was observed under the effect of 30 μl of S. maritima (1.3 days and S. verbenaca (2.8, 4.6 days, respectively, for males and females compared to 8 and 15 days for the control. For fecundity, an inhibition of oviposition was obtained using 30 μl of Salvia and Scilla essential oils. The test on the seed germination using different essential oils, showed no damage to the germinating seeds. The germination rate was 99%. These findings suggest that the tested plants can be used as a bioinsecticide for control of the C. chinensis pest of stored products.

  1. Structure of the ribosomal interacting GTPase YjeQ from the enterobacterial species Salmonella typhimurium

    Energy Technology Data Exchange (ETDEWEB)

    Nichols, C. E. [Division of Structural Biology, The Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN (United Kingdom); Johnson, C.; Lamb, H. K. [Institute of Cell and Molecular Biosciences, Catherine Cookson Building, Medical School, Framlington Place, Newcastle University, Newcastle-upon-Tyne NE2 4HH (United Kingdom); Lockyer, M. [Arrow Therapeutics Ltd, Britannia House, Trinity Street, Borough, London SE1 1DA (United Kingdom); Charles, I. G. [The Wolfson Institute for Biomedical Research, The Cruciform Building, University College London, Gower Street, London WC1E 6BT (United Kingdom); Hawkins, A. R. [Institute of Cell and Molecular Biosciences, Catherine Cookson Building, Medical School, Framlington Place, Newcastle University, Newcastle-upon-Tyne NE2 4HH (United Kingdom); Stammers, D. K., E-mail: daves@strubi.ox.ac.uk [Division of Structural Biology, The Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN (United Kingdom)

    2007-11-01

    The X-ray crystal structure of the GTPase YjeQ from S. typhimurium is presented and compared with those of orthologues from T. maritima and B. subtilis. The YjeQ class of P-loop GTPases assist in ribosome biogenesis and also bind to the 30S subunit of mature ribosomes. YjeQ ribosomal binding is GTP-dependent and thought to specifically direct protein synthesis, although the nature of the upstream signal causing this event in vivo is as yet unknown. The attenuating effect of YjeQ mutants on bacterial growth in Escherichia coli makes it a potential target for novel antimicrobial agents. In order to further explore the structure and function of YjeQ, the isolation, crystallization and structure determination of YjeQ from the enterobacterial species Salmonella typhimurium (StYjeQ) is reported. Whilst the overall StYjeQ fold is similar to those of the previously reported Thematoga maritima and Bacillus subtilis orthologues, particularly the GTPase domain, there are larger differences in the three OB folds. Although the zinc-finger secondary structure is conserved, significant sequence differences alter the nature of the external surface in each case and may reflect varying signalling pathways. Therefore, it may be easier to develop YjeQ-specific inhibitors that target the N- and C-terminal regions, disrupting the metabolic connectivity rather than the GTPase activity. The availability of coordinates for StYjeQ will provide a significantly improved basis for threading Gram-negative orthologue sequences and in silico compound-screening studies, with the potential for the development of species-selective drugs.

  2. Structure of the ribosomal interacting GTPase YjeQ from the enterobacterial species Salmonella typhimurium

    International Nuclear Information System (INIS)

    Nichols, C. E.; Johnson, C.; Lamb, H. K.; Lockyer, M.; Charles, I. G.; Hawkins, A. R.; Stammers, D. K.

    2007-01-01

    The X-ray crystal structure of the GTPase YjeQ from S. typhimurium is presented and compared with those of orthologues from T. maritima and B. subtilis. The YjeQ class of P-loop GTPases assist in ribosome biogenesis and also bind to the 30S subunit of mature ribosomes. YjeQ ribosomal binding is GTP-dependent and thought to specifically direct protein synthesis, although the nature of the upstream signal causing this event in vivo is as yet unknown. The attenuating effect of YjeQ mutants on bacterial growth in Escherichia coli makes it a potential target for novel antimicrobial agents. In order to further explore the structure and function of YjeQ, the isolation, crystallization and structure determination of YjeQ from the enterobacterial species Salmonella typhimurium (StYjeQ) is reported. Whilst the overall StYjeQ fold is similar to those of the previously reported Thematoga maritima and Bacillus subtilis orthologues, particularly the GTPase domain, there are larger differences in the three OB folds. Although the zinc-finger secondary structure is conserved, significant sequence differences alter the nature of the external surface in each case and may reflect varying signalling pathways. Therefore, it may be easier to develop YjeQ-specific inhibitors that target the N- and C-terminal regions, disrupting the metabolic connectivity rather than the GTPase activity. The availability of coordinates for StYjeQ will provide a significantly improved basis for threading Gram-negative orthologue sequences and in silico compound-screening studies, with the potential for the development of species-selective drugs

  3. Survival of microbial cultures on mineral while passing dense layers of the atmosphere

    Science.gov (United States)

    Viacheslav, Ilyin; Novikova, Nataliya; Deshevaya, Elena; Polikarpov, Nikolay; Slobodkin, Alexander; Gavrilov, Sergey; Ionov, Viktor; Morozova, Julia

    The purpose of the experiment is to study the possibility of extremophilic microorganisms survival in meteorite-like mineral while passing through the dense layers of the atmosphere. For this purpose cultures of bacteria were placed into the holes made in basalt pieces fixed to the outer wall of the spacecraft Bion M1. Control: similar materials placed in the outer container, prevented from overheating in the dense layers of the atmosphere by lid. In the flight experiment five strains of thermophilic bacteria and 2 strains of hyperthermophilic archaea from the collection of the Institute of Microbiology, RAS were used. In addition, microorganisms were selected from the collection of the Institute of Biomedical Problems, isolated from the environment objects of ISS: 10 fungal cultures and a culture of bacteria Bacillus pumilus. For thermophiles and hyperthermophiles the ability to redox interactions with minerals is considered as a priority physiological property. Ability of thermophiles to anaerobic growth also meets the conditions of the experiment - testing cell survival of microorganisms in the conditions of extraterrestrial space and ancient anaerobic atmosphere of the Earth. After 30-days flight in orbit control all spore-forming microorganisms have been successfully survived. Hyperthermophilic archaea growth in all control was significantly less intensive. Meanwhile, in one experimental samples there was obtained signs of survival of spore forming bacteria culture Carboxydocella ferrireducens. However, the maximum concentration of cells was 2 orders of magnitude below the values characteristic of an actively growing culture of the microorganism. Due to damage of holes in the stone, this result was obtained only in one replicate and for final prove of survival of C. ferrireducens when returning through the dense layers of the atmosphere it is necessary to repeat the experiment It should be noted that an important indicator of the possibility of survival of C

  4. Marine Subsurface Microbial Communities Across a Hydrothermal Gradient in Okinawa Trough Sediments

    Science.gov (United States)

    Brandt, L. D.; Hser Wah Saw, J.; Ettema, T.; House, C. H.

    2015-12-01

    IODP Expedition 331 to the Okinawa backarc basin provided an opportunity to study the microbial stratigraphy within the sediments surrounding a hydrothermal vent. The Okinawa backarc basin is a sedimented region of the seafloor located on a continental margin, and also hosts a hydrothermal network within the subsurface. Site C0014 within the Iheya North hydrothermal field is located 450 m east of the active vent and has a surface temperature of 5°C with no evidence of hydrothermal alteration within the top 10 meters below sea floor (mbsf). Temperature increases with depth at an estimated rate of 3°C/m and transitions from non-hydrothermal margin sediments to a hydrothermally altered regime below 10 mbsf. In this study, we utilized deep 16S rRNA sequencing of DNA from IODP Expedition 331 Site C0014 sediment horizons in order to assess diversity throughout the sediment column as well as determine the potential limits of the biosphere. Analysis of the amplicon data shows a shift over 15 mbsf from a heterogeneous community of cosmopolitan marine subsurface taxa toward an archaeal-dominated community in the deepest horizons of the predicted biosphere. Notably, the phylum Chloroflexi represents a substantial taxon through most horizons, where it appears to be replaced below 10 mbsf by punctuations of thermophilic and methanotrophic Archaea and Miscellaneous Crenarchaeotic Group abundances. DNA from the aforementioned transition horizons was further analyzed using metagenomic sequencing. Preliminary taxonomic analysis of the metagenomic data agrees well with amplicon data in capturing the shift in relative abundance of Archaea increasing with depth. Additionally, reverse gyrase, a gene found exclusively in hyperthermophilic microorganisms, was recovered only in the metagenome of the deepest horizon. A BLAST search of this protein sequence against the GenBank non-redudnant protein database produced top hits with reverse gyrase from Thermococcus and Pyrococcus, which are

  5. Crystallization and preliminary X-ray analysis of PH1010 from Pyrococcus horikoshii OT3, a member of the archaeal DUF54 family of proteins

    International Nuclear Information System (INIS)

    Shirokane, Michio; Sawano, Yoriko; Miyazono, Ken-ichi; Nagata, Koji; Tanokura, Masaru

    2007-01-01

    PH1010, a DUF54-family protein from the hyperthermophilic archaeon P. horikoshii OT3, was crystallized and X-ray diffraction data were collected to 1.90 Å resolution. PH1010 from Pyrococcus horikoshii OT3, a member of the archaeal DUF54 family of proteins, was expressed, purified and crystallized. Crystallization was performed by the sitting-drop vapour-diffusion method using PEG 3350 as the precipitant. The crystal diffracted X-rays to 1.90 Å resolution using a synchrotron-radiation source. The space group of the crystal was determined to be P2 1 2 1 2 1 , with unit-cell parameters a = 46.9, b = 49.5, c = 132.7 Å. The crystal contained two PH1010 molecules in the asymmetric unit (V M = 2.4 Å 3 Da −1 ) and had a solvent content of 48%

  6. Crystallization and preliminary X-ray analysis of PH1010 from Pyrococcus horikoshii OT3, a member of the archaeal DUF54 family of proteins

    Energy Technology Data Exchange (ETDEWEB)

    Shirokane, Michio; Sawano, Yoriko; Miyazono, Ken-ichi; Nagata, Koji; Tanokura, Masaru, E-mail: amtanok@mail.ecc.u-tokyo.ac.jp [Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657 (Japan)

    2007-06-01

    PH1010, a DUF54-family protein from the hyperthermophilic archaeon P. horikoshii OT3, was crystallized and X-ray diffraction data were collected to 1.90 Å resolution. PH1010 from Pyrococcus horikoshii OT3, a member of the archaeal DUF54 family of proteins, was expressed, purified and crystallized. Crystallization was performed by the sitting-drop vapour-diffusion method using PEG 3350 as the precipitant. The crystal diffracted X-rays to 1.90 Å resolution using a synchrotron-radiation source. The space group of the crystal was determined to be P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 46.9, b = 49.5, c = 132.7 Å. The crystal contained two PH1010 molecules in the asymmetric unit (V{sub M} = 2.4 Å{sup 3} Da{sup −1}) and had a solvent content of 48%.

  7. Hydroxyurea-Mediated Cytotoxicity Without Inhibition of Ribonucleotide Reductase.

    Science.gov (United States)

    Liew, Li Phing; Lim, Zun Yi; Cohen, Matan; Kong, Ziqing; Marjavaara, Lisette; Chabes, Andrei; Bell, Stephen D

    2016-11-01

    In many organisms, hydroxyurea (HU) inhibits class I ribonucleotide reductase, leading to lowered cellular pools of deoxyribonucleoside triphosphates. The reduced levels for DNA precursors is believed to cause replication fork stalling. Upon treatment of the hyperthermophilic archaeon Sulfolobus solfataricus with HU, we observe dose-dependent cell cycle arrest, accumulation of DNA double-strand breaks, stalled replication forks, and elevated levels of recombination structures. However, Sulfolobus has a HU-insensitive class II ribonucleotide reductase, and we reveal that HU treatment does not significantly impact cellular DNA precursor pools. Profiling of protein and transcript levels reveals modulation of a specific subset of replication initiation and cell division genes. Notably, the selective loss of the regulatory subunit of the primase correlates with cessation of replication initiation and stalling of replication forks. Furthermore, we find evidence for a detoxification response induced by HU treatment. Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.

  8. Early steps of biosynthesis of ether lipids in archaebacteria; Eteru shishitsu seigosei no shoki dankai

    Energy Technology Data Exchange (ETDEWEB)

    Nishino, T. [Tohoku Univ., Sendai (Japan). Faculty of Engineering

    1997-05-20

    Membrane lipids in archaebacteria are different from those of eubacteria and eukaryote which are fatty acid esters of glycerol. Archaebacterial lipids are mainly ether-linked lipids composed of glycerol linked to two molecules of isoprenoid phytanyl groups or of ether-linked glycerol with phytanyl group. This structural feature is one of the origins of survival and growth of archaebacteria in extreme conditions of high temperature, strong acid or alkali. It is considered that geranylgeranyl phosphate (GGPP) is synthesized and attached to glycerol phosphate, followed by reduction of the double bond in the geranylgeranyl moieties to form the diether lipids while the head-to-heat condensation of the phytanyl groups produces the tetraether lipids. Aiming to elucidate the lipid biosynthesis mechanism in a hyperthermophilic archaebacterium, Sulfolobus acidocaldarius, the gene of GGPP synthase was cloned with the aid of carotenoid synthesis in phytopathogenic Erwinia uredovora and its sequence was studied. 29 refs., 9 figs.

  9. Cloning, characterization and sequence comparison of the gene coding for IMP dehydrogenase from Pyrococcus furiosus.

    Science.gov (United States)

    Collart, F R; Osipiuk, J; Trent, J; Olsen, G J; Huberman, E

    1996-10-03

    We have cloned and characterized the gene encoding inosine monophosphate dehydrogenase (IMPDH) from Pyrococcus furiosus (Pf), a hyperthermophillic archeon. Sequence analysis of the Pf gene indicated an open reading frame specifying a protein of 485 amino acids (aa) with a calculated M(r) of 52900. Canonical Archaea promoter elements, Box A and Box B, are located -49 and -17 nucleotides (nt), respectively, upstream of the putative start codon. The sequence of the putative active-site region conforms to the IMPDH signature motif and contains a putative active-site cysteine. Phylogenetic relationships derived by using all available IMPDH sequences are consistent with trees developed for other molecules; they do not precisely resolve the history of Pf IMPDH but indicate a close similarity to bacterial IMPDH proteins. The phylogenetic analysis indicates that a gene duplication occurred prior to the division between rodents and humans, accounting for the Type I and II isoforms identified in mice and humans.

  10. Structure of the acidianus filamentous virus 3 and comparative genomics of related archaeal lipothrixviruses

    DEFF Research Database (Denmark)

    Vestergaard, Gisle Alberg; Aramayo, Ricardo; Basta, Tamara

    2008-01-01

    Four novel filamentous viruses with double-stranded DNA genomes, namely, Acidianus filamentous virus 3 (AFV3), AFV6, AFV7, and AFV8, have been characterized from the hyperthermophilic archaeal genus Acidianus, and they are assigned to the Betalipothrixvirus genus of the family Lipothrixviridae....... The structures of the approximately 2-mum-long virions are similar, and one of them, AFV3, was studied in detail. It consists of a cylindrical envelope containing globular subunits arranged in a helical formation that is unique for any known double-stranded DNA virus. The envelope is 3.1 nm thick and encases...... structural proteins; (iii) multiple overlapping open reading frames, which may be indicative of gene recoding; (iv) putative 12-bp genetic elements; and (v) partial gene sequences corresponding closely to spacer sequences of chromosomal repeat clusters....

  11. Combined thermophilic aerobic process and conventional anaerobic digestion: effect on sludge biodegradation and methane production.

    Science.gov (United States)

    Dumas, C; Perez, S; Paul, E; Lefebvre, X

    2010-04-01

    The efficiency of hyper-thermophilic (65 degrees Celsius) aerobic process coupled with a mesophilic (35 degrees Celsius) digester was evaluated for the activated sludge degradation and was compared to a conventional mesophilic digester. For two Sludge Retention Time (SRT), 21 and 42 days, the Chemical Oxygen Demand (COD) solubilisation and biodegradation processes, the methanisation yield and the aerobic oxidation were investigated during 180 days. The best results were obtained at SRT of 44 days; the COD removal yield was 30% higher with the Mesophilic Anaerobic Digestion/Thermophilic Aerobic Reactor (MAD-TAR) co-treatment. An increase of the sludge intrinsic biodegradability is also observed (20-40%), showing that the unbiodegradable COD in mesophilic conditions becomes bioavailable. However, the methanisation yield was quite similar for both processes at a same SRT. Finally, such a process enables to divide by two the volume of digester with an equivalent efficiency. Copyright 2009 Elsevier Ltd. All rights reserved.

  12. High hydrostatic pressure adaptive strategies in an obligate piezophile Pyrococcus yayanosii

    KAUST Repository

    Michoud, Gregoire

    2016-06-02

    Pyrococcus yayanosii CH1, as the first and only obligate piezophilic hyperthermophilic microorganism discovered to date, extends the physical and chemical limits of life on Earth. It was isolated from the Ashadze hydrothermal vent at 4,100 m depth. Multi-omics analyses were performed to study the mechanisms used by the cell to cope with high hydrostatic pressure variations. In silico analyses showed that the P. yayanosii genome is highly adapted to its harsh environment, with a loss of aromatic amino acid biosynthesis pathways and the high constitutive expression of the energy metabolism compared with other non-obligate piezophilic Pyrococcus species. Differential proteomics and transcriptomics analyses identified key hydrostatic pressure-responsive genes involved in translation, chemotaxis, energy metabolism (hydrogenases and formate metabolism) and Clustered Regularly Interspaced Short Palindromic Repeats sequences associated with Cellular apoptosis susceptibility proteins.

  13. Development of in vitro transposon assisted signal sequence trapping and its use in screening Bacillus halodurans C125 and Sulfolobus solfataricus P2 gene libraries

    DEFF Research Database (Denmark)

    Becker, F.; Schnorr, K.; Wilting, R.

    2004-01-01

    To identify genes encoding extracytosolic proteins, a minitransposon, TnSig, containing a signal-less beta-lactamase ('bla) as reporter gene, was constructed and used for in vitro transposition of genomic libraries made in Escherichia coli. The 'bla gene was cloned into a bacteriophage MU...... minitransposon enabling translational fusions between 'bla and target genes. Fusion of TnSig in the correct reading frame to a protein carrying transmembrane domains or signal peptides resulted in ampicillin resistance of the corresponding clone. Prokaryotic gene libraries from the alkaliphilic bacterium...... Bacillus halodurans C125 and the hyperthermophilic archaeon Sulfolobus solfataricus P2 were tagged with TnSig. The genomic sequences, which are publicly available (EMBL BA000004 and EMBL AE006641), were used for rapid open reading frame (ORF) identification and prediction of protein localisation...

  14. Single-molecule fluorescence polarization study of conformational change in archaeal group II chaperonin.

    Directory of Open Access Journals (Sweden)

    Ryo Iizuka

    Full Text Available Group II chaperonins found in archaea and in eukaryotic cytosol mediate protein folding without a GroES-like cofactor. The function of the cofactor is substituted by the helical protrusion at the tip of the apical domain, which forms a built-in lid on the central cavity. Although many studies on the change in lid conformation coupled to the binding and hydrolysis of nucleotides have been conducted, the molecular mechanism of lid closure remains poorly understood. Here, we performed a single-molecule polarization modulation to probe the rotation of the helical protrusion of a chaperonin from a hyperthermophilic archaeum, Thermococcus sp. strain KS-1. We detected approximately 35° rotation of the helical protrusion immediately after photorelease of ATP. The result suggests that the conformational change from the open lid to the closed lid state is responsible for the approximately 35° rotation of the helical protrusion.

  15. Industrial relevance of thermophilic Archaea.

    Science.gov (United States)

    Egorova, Ksenia; Antranikian, Garabed

    2005-12-01

    The dramatic increase of newly isolated extremophilic microorganisms, analysis of their genomes and investigations of their enzymes by academic and industrial laboratories demonstrate the great potential of extremophiles in industrial (white) biotechnology. Enzymes derived from extremophiles (extremozymes) are superior to the traditional catalysts because they can perform industrial processes even under harsh conditions, under which conventional proteins are completely denatured. In particular, enzymes from thermophilic and hyperthermophilic Archaea have industrial relevance. Despite intensive investigations, our knowledge of the structure-function relationships of their enzymes is still limited. Information concerning the molecular properties of their enzymes and genes has to be obtained to be able to understand the mechanisms that are responsible for catalytic activity and stability at the boiling point of water.

  16. Characterization of technetium(vII) reduction by cell suspensions of thermophilic bacteria and archaea.

    Science.gov (United States)

    Chernyh, Nikolay A; Gavrilov, Sergei N; Sorokin, Vladimir V; German, Konstantin E; Sergeant, Claire; Simonoff, Monique; Robb, Frank; Slobodkin, Alexander I

    2007-08-01

    Washed cell suspensions of the anaerobic hyperthermophilic archaea Thermococcus pacificus and Thermoproteus uzoniensis and the anaerobic thermophilic gram-positive bacteria Thermoterrabacterium ferrireducens and Tepidibacter thalassicus reduced technetium [(99)Tc(VII)], supplied as soluble pertechnetate with molecular hydrogen as an electron donor, forming highly insoluble Tc(IV)-containing grayish-black precipitate. Apart from molecular hydrogen, T. ferrireducens reduced Tc(VII) with lactate, glycerol, and yeast extract as electron donors, and T. thalassicus reduced it with peptone. Scanning electron microscopy and X-ray microanalysis of cell suspensions of T. ferrireducens showed the presence of Tc-containing particles attached to the surfaces of non-lysed cells. This is the first report on the reduction in Tc(VII) by thermophilic microorganisms of the domain Bacteria and by archaea of the phylum Euryarchaeota.

  17. Structural and physicochemical properties of polar lipids from thermophilic archaea.

    Science.gov (United States)

    Ulrih, Natasa Poklar; Gmajner, Dejan; Raspor, Peter

    2009-08-01

    The essential general features required for lipid membranes of extremophilic archaea to fulfill biological functions are that they are in the liquid crystalline phase and have extremely low permeability of solutes that is much less temperature sensitive due to a lack of lipid-phase transition and highly branched isoprenoid chains. Many accumulated data indicate that the organism's response to extremely low pH is the opposite of that to high temperature. The high temperature adaptation does not require the tetraether lipids, while the adaptation of thermophiles to acidic environment requires the tetraether polar lipids. The presence of cyclopentane rings and the role of polar heads are not so straightforward regarding the correlations between fluidity and permeability of the lipid membrane. Due to the unique lipid structures and properties of archaeal lipids, they are a valuable resource in the development of novel biotechnological processes. This microreview focuses primarily on structural and physicochemical properties of polar lipids of (hyper)thermophilic archaea.

  18. Widespread Disulfide Bonding in Proteins from Thermophilic Archaea

    Directory of Open Access Journals (Sweden)

    Julien Jorda

    2011-01-01

    Full Text Available Disulfide bonds are generally not used to stabilize proteins in the cytosolic compartments of bacteria or eukaryotic cells, owing to the chemically reducing nature of those environments. In contrast, certain thermophilic archaea use disulfide bonding as a major mechanism for protein stabilization. Here, we provide a current survey of completely sequenced genomes, applying computational methods to estimate the use of disulfide bonding across the Archaea. Microbes belonging to the Crenarchaeal branch, which are essentially all hyperthermophilic, are universally rich in disulfide bonding while lesser degrees of disulfide bonding are found among the thermophilic Euryarchaea, excluding those that are methanogenic. The results help clarify which parts of the archaeal lineage are likely to yield more examples and additional specific data on protein disulfide bonding, as increasing genomic sequencing efforts are brought to bear.

  19. Chromatin structure and dynamics in hot environments: architectural proteins and DNA topoisomerases of thermophilic archaea.

    Science.gov (United States)

    Visone, Valeria; Vettone, Antonella; Serpe, Mario; Valenti, Anna; Perugino, Giuseppe; Rossi, Mosè; Ciaramella, Maria

    2014-09-25

    In all organisms of the three living domains (Bacteria, Archaea, Eucarya) chromosome-associated proteins play a key role in genome functional organization. They not only compact and shape the genome structure, but also regulate its dynamics, which is essential to allow complex genome functions. Elucidation of chromatin composition and regulation is a critical issue in biology, because of the intimate connection of chromatin with all the essential information processes (transcription, replication, recombination, and repair). Chromatin proteins include architectural proteins and DNA topoisomerases, which regulate genome structure and remodelling at two hierarchical levels. This review is focussed on architectural proteins and topoisomerases from hyperthermophilic Archaea. In these organisms, which live at high environmental temperature (>80 °C <113 °C), chromatin proteins and modulation of the DNA secondary structure are concerned with the problem of DNA stabilization against heat denaturation while maintaining its metabolic activity.

  20. Chromatin Structure and Dynamics in Hot Environments: Architectural Proteins and DNA Topoisomerases of Thermophilic Archaea

    Directory of Open Access Journals (Sweden)

    Valeria Visone

    2014-09-01

    Full Text Available In all organisms of the three living domains (Bacteria, Archaea, Eucarya chromosome-associated proteins play a key role in genome functional organization. They not only compact and shape the genome structure, but also regulate its dynamics, which is essential to allow complex genome functions. Elucidation of chromatin composition and regulation is a critical issue in biology, because of the intimate connection of chromatin with all the essential information processes (transcription, replication, recombination, and repair. Chromatin proteins include architectural proteins and DNA topoisomerases, which regulate genome structure and remodelling at two hierarchical levels. This review is focussed on architectural proteins and topoisomerases from hyperthermophilic Archaea. In these organisms, which live at high environmental temperature (>80 °C <113 °C, chromatin proteins and modulation of the DNA secondary structure are concerned with the problem of DNA stabilization against heat denaturation while maintaining its metabolic activity.

  1. Widespread disulfide bonding in proteins from thermophilic archaea.

    Science.gov (United States)

    Jorda, Julien; Yeates, Todd O

    2011-01-01

    Disulfide bonds are generally not used to stabilize proteins in the cytosolic compartments of bacteria or eukaryotic cells, owing to the chemically reducing nature of those environments. In contrast, certain thermophilic archaea use disulfide bonding as a major mechanism for protein stabilization. Here, we provide a current survey of completely sequenced genomes, applying computational methods to estimate the use of disulfide bonding across the Archaea. Microbes belonging to the Crenarchaeal branch, which are essentially all hyperthermophilic, are universally rich in disulfide bonding while lesser degrees of disulfide bonding are found among the thermophilic Euryarchaea, excluding those that are methanogenic. The results help clarify which parts of the archaeal lineage are likely to yield more examples and additional specific data on protein disulfide bonding, as increasing genomic sequencing efforts are brought to bear.

  2. Archaeal diversity in Icelandic hot springs

    DEFF Research Database (Denmark)

    Kvist, Thomas; Ahring, Birgitte Kiær; Westermann, Peter

    2007-01-01

    Whole-cell density gradient extractions from three solfataras (pH 2.5) ranging in temperature from 81 to 90 degrees C and one neutral hot spring (81 degrees C, pH 7) from the thermal active area of Hveragerethi (Iceland) were analysed for genetic diversity and local geographical variation...... of Archaea by analysis of amplified 16S rRNA genes. In addition to the three solfataras and the neutral hot spring, 10 soil samples in transects of the soil adjacent to the solfataras were analysed using terminal restriction fragment length polymorphism (t-RFLP). The sequence data from the clone libraries...... enzymes AluI and BsuRI. The sequenced clones from this solfatara belonged to Sulfolobales, Thermoproteales or were most closest related to sequences from uncultured Archaea. Sequences related to group I.1b were not found in the neutral hot spring or the hyperthermophilic solfatara (90 degrees C)....

  3. High hydrostatic pressure adaptive strategies in an obligate piezophile Pyrococcus yayanosii

    KAUST Repository

    Michoud, Gregoire; Jebbar, Mohamed

    2016-01-01

    Pyrococcus yayanosii CH1, as the first and only obligate piezophilic hyperthermophilic microorganism discovered to date, extends the physical and chemical limits of life on Earth. It was isolated from the Ashadze hydrothermal vent at 4,100 m depth. Multi-omics analyses were performed to study the mechanisms used by the cell to cope with high hydrostatic pressure variations. In silico analyses showed that the P. yayanosii genome is highly adapted to its harsh environment, with a loss of aromatic amino acid biosynthesis pathways and the high constitutive expression of the energy metabolism compared with other non-obligate piezophilic Pyrococcus species. Differential proteomics and transcriptomics analyses identified key hydrostatic pressure-responsive genes involved in translation, chemotaxis, energy metabolism (hydrogenases and formate metabolism) and Clustered Regularly Interspaced Short Palindromic Repeats sequences associated with Cellular apoptosis susceptibility proteins.

  4. Isolation of diverse members of the Aquificales from geothermal springs in Tengchong, China.

    Science.gov (United States)

    Hedlund, Brian P; Reysenbach, Anna-Louise; Huang, Liuquin; Ong, John C; Liu, Zizhang; Dodsworth, Jeremy A; Ahmed, Reham; Williams, Amanda J; Briggs, Brandon R; Liu, Yitai; Hou, Weiguo; Dong, Hailiang

    2015-01-01

    The order Aquificales (phylum Aquificae) consists of thermophilic and hyperthermophilic bacteria that are prominent in many geothermal systems, including those in Tengchong, Yunnan Province, China. However, Aquificales have not previously been isolated from Tengchong. We isolated five strains of Aquificales from diverse springs (temperature 45.2-83.3°C and pH 2.6-9.1) in the Rehai Geothermal Field from sites in which Aquificales were abundant. Phylogenetic analysis showed that four of the strains belong to the genera Hydrogenobacter, Hydrogenobaculum, and Sulfurihydrogenibium, including strains distant enough to likely justify new species of Hydrogenobacter and Hydrogenobaculum. The additional strain may represent a new genus in the Hydrogenothermaceae. All strains were capable of aerobic respiration under microaerophilic conditions; however, they had variable capacity for chemolithotrophic oxidation of hydrogen and sulfur compounds and nitrate reduction.

  5. Isolation of diverse members of the Aquificales from geothermal springs in Tengchong, China

    Directory of Open Access Journals (Sweden)

    Brian P. Hedlund

    2015-02-01

    Full Text Available The order Aquificales (phylum Aquificae consists of thermophilic and hyperthermophilic bacteria that are prominent in many geothermal systems, including those in Tengchong, Yunnan Province, China. However, Aquificales have not previously been isolated from Tengchong. We isolated five strains of Aquificales from diverse springs (temperature 60.0-82.9°C and pH 2.6-8.9 in the Rehai Geothermal Field from sites in which Aquificales were abundant. Phylogenetic analysis showed that four of the strains belong to the genera Hydrogenobacter, Hydrogenobaculum, and Sulfurihydrogenibium, including strains distant enough to likely justify new species of Hydrogenobacter and Hydrogenobaculum. The additional strain may represent a new genus in the Hydrogenothermaceae. All strains were capable of aerobic respiration under microaerophilic conditions; however, they had variable capacity for chemolithotrophic oxidation of hydrogen and sulfur compounds and nitrate reduction.

  6. The Effects of Temperature and Growth Phase on the Lipidomes of Sulfolobus islandicus and Sulfolobus tokodaii

    DEFF Research Database (Denmark)

    Jensen, Sara Munk; Neesgaard, Vinnie Lund; Skjoldbjerg, Sandra Landbo Nedergaard

    2015-01-01

    at three different temperatures, with samples withdrawn during lag, exponential, and stationary phases. Three abundant tetraether lipid classes and one diether lipid class were monitored. Beside the expected increase in the number of cyclopentane moieties with higher temperature in both archaea, we......The functionality of the plasma membrane is essential for all organisms. Adaption to high growth temperatures imposes challenges and Bacteria, Eukarya, and Archaea have developed several mechanisms to cope with these. Hyperthermophilic archaea have earlier been shown to synthesize tetraether...... membrane lipids with an increased number of cyclopentane moieties at higher growth temperatures. Here we used shotgun lipidomics to study this effect as well as the influence of growth phase on the lipidomes of Sulfolobus islandicus and Sulfolobus tokodaii for the first time. Both species were cultivated...

  7. TOPICAL REVIEW: Protein stability and enzyme activity at extreme biological temperatures

    Science.gov (United States)

    Feller, Georges

    2010-08-01

    Psychrophilic microorganisms thrive in permanently cold environments, even at subzero temperatures. To maintain metabolic rates compatible with sustained life, they have improved the dynamics of their protein structures, thereby enabling appropriate molecular motions required for biological activity at low temperatures. As a consequence of this structural flexibility, psychrophilic proteins are unstable and heat-labile. In the upper range of biological temperatures, thermophiles and hyperthermophiles grow at temperatures > 100 °C and synthesize ultra-stable proteins. However, thermophilic enzymes are nearly inactive at room temperature as a result of their compactness and rigidity. At the molecular level, both types of extremophilic proteins have adapted the same structural factors, but in opposite directions, to address either activity at low temperatures or stability in hot environments. A model based on folding funnels is proposed accounting for the stability-activity relationships in extremophilic proteins.

  8. Chemical and microbiological problems associated with research on the biodesulfurization of coal. A review

    Energy Technology Data Exchange (ETDEWEB)

    Olson, G J; Kelly, R M [National Institute of Standards and Technology, Gaithersburg, MD (USA). Polymer Division

    1991-04-01

    The study of microbial processes for the removal of organic and inorganic sulfur from coals is complicated by the lack of direct methods of measurement for organic sulfur content and the related incomplete understanding of the specific forms of organic sulfur in coal. In addition, the accessibility of specific chemical groups in the coal matrix to microorganisms and their enzymes is uncertain, raising questions about the nature and validity of model compound studies. Thus, interpretation of data from numerous efforts focussed on the microbial removal of inorganic and organic sulfur from coals remains controversial. The discussion here reviews recent developments in the chemical characterization of coal sulfur related to bioprocessing research and describes some recent efforts in involving sulfur transformation by hyperthermophilic archaebacteria. 26 refs., 4 figs., 1 tab.

  9. Protein stability and enzyme activity at extreme biological temperatures

    International Nuclear Information System (INIS)

    Feller, Georges

    2010-01-01

    Psychrophilic microorganisms thrive in permanently cold environments, even at subzero temperatures. To maintain metabolic rates compatible with sustained life, they have improved the dynamics of their protein structures, thereby enabling appropriate molecular motions required for biological activity at low temperatures. As a consequence of this structural flexibility, psychrophilic proteins are unstable and heat-labile. In the upper range of biological temperatures, thermophiles and hyperthermophiles grow at temperatures > 100 0 C and synthesize ultra-stable proteins. However, thermophilic enzymes are nearly inactive at room temperature as a result of their compactness and rigidity. At the molecular level, both types of extremophilic proteins have adapted the same structural factors, but in opposite directions, to address either activity at low temperatures or stability in hot environments. A model based on folding funnels is proposed accounting for the stability-activity relationships in extremophilic proteins. (topical review)

  10. Thermostability in rubredoxin and its relationship to mechanical rigidity

    Science.gov (United States)

    Rader, A. J.

    2010-03-01

    The source of increased stability in proteins from organisms that thrive in extreme thermal environments is not well understood. Previous experimental and theoretical studies have suggested many different features possibly responsible for such thermostability. Many of these thermostabilizing mechanisms can be accounted for in terms of structural rigidity. Thus a plausible hypothesis accounting for this remarkable stability in thermophilic enzymes states that these enzymes have enhanced conformational rigidity at temperatures below their native, functioning temperature. Experimental evidence exists to both support and contradict this supposition. We computationally investigate the relationship between thermostability and rigidity using rubredoxin as a case study. The mechanical rigidity is calculated using atomic models of homologous rubredoxin structures from the hyperthermophile Pyrococcus furiosus and mesophile Clostridium pasteurianum using the FIRST software. A global increase in structural rigidity (equivalently a decrease in flexibility) corresponds to an increase in thermostability. Locally, rigidity differences (between mesophilic and thermophilic structures) agree with differences in protection factors.

  11. Thermostability in rubredoxin and its relationship to mechanical rigidity

    International Nuclear Information System (INIS)

    Rader, A J

    2010-01-01

    The source of increased stability in proteins from organisms that thrive in extreme thermal environments is not well understood. Previous experimental and theoretical studies have suggested many different features possibly responsible for such thermostability. Many of these thermostabilizing mechanisms can be accounted for in terms of structural rigidity. Thus a plausible hypothesis accounting for this remarkable stability in thermophilic enzymes states that these enzymes have enhanced conformational rigidity at temperatures below their native, functioning temperature. Experimental evidence exists to both support and contradict this supposition. We computationally investigate the relationship between thermostability and rigidity using rubredoxin as a case study. The mechanical rigidity is calculated using atomic models of homologous rubredoxin structures from the hyperthermophile Pyrococcus furiosus and mesophile Clostridium pasteurianum using the FIRST software. A global increase in structural rigidity (equivalently a decrease in flexibility) corresponds to an increase in thermostability. Locally, rigidity differences (between mesophilic and thermophilic structures) agree with differences in protection factors

  12. An extensive phase space for the potential martian biosphere.

    Science.gov (United States)

    Jones, Eriita G; Lineweaver, Charles H; Clarke, Jonathan D

    2011-12-01

    We present a comprehensive model of martian pressure-temperature (P-T) phase space and compare it with that of Earth. Martian P-T conditions compatible with liquid water extend to a depth of ∼310 km. We use our phase space model of Mars and of terrestrial life to estimate the depths and extent of the water on Mars that is habitable for terrestrial life. We find an extensive overlap between inhabited terrestrial phase space and martian phase space. The lower martian surface temperatures and shallower martian geotherm suggest that, if there is a hot deep biosphere on Mars, it could extend 7 times deeper than the ∼5 km depth of the hot deep terrestrial biosphere in the crust inhabited by hyperthermophilic chemolithotrophs. This corresponds to ∼3.2% of the volume of present-day Mars being potentially habitable for terrestrial-like life.

  13. Phthalic acid esters found in municipal organic waste

    DEFF Research Database (Denmark)

    Hartmann, Hinrich; Ahring, Birgitte Kiær

    2003-01-01

    Contamination of the organic fraction of municipal solid waste (OFMSW) with xenobiotic compounds and their fate during anaerobic digestion was investigated. The phthalic acid ester di-(2- ethylhexyl)phthalate (DEHP) was identified as the main contaminant in OFMSW in concentrations more than half.......41-0.79 d(-1), which is much higher than in previous investigations. It can be concluded that the higher removal rates are due to the higher temperature and higher initial concentrations per kg dry matter. These results suggest that the limiting factor for DEHP degradation is the bioavailability, which...... is enhanced at higher temperature and higher degradation of solid organic matter, to which the highly hydrophobic DEHP is adsorbed. The investigated reactor configuration with a thermophilic and a hyper-thermophilic treatment is, therefore, a good option for CD combining high rate degradation of organic...

  14. 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)

  15. Perspectives on biotechnological applications of archaea

    Science.gov (United States)

    Schiraldi, Chiara; Giuliano, Mariateresa; De Rosa, Mario

    2002-01-01

    Many archaea colonize extreme environments. They include hyperthermophiles, sulfur-metabolizing thermophiles, extreme halophiles and methanogens. Because extremophilic microorganisms have unusual properties, they are a potentially valuable resource in the development of novel biotechnological processes. Despite extensive research, however, there are few existing industrial applications of either archaeal biomass or archaeal enzymes. This review summarizes current knowledge about the biotechnological uses of archaea and archaeal enzymes with special attention to potential applications that are the subject of current experimental evaluation. Topics covered include cultivation methods, recent achievements in genomics, which are of key importance for the development of new biotechnological tools, and the application of wild-type biomasses, engineered microorganisms, enzymes and specific metabolites in particular bioprocesses of industrial interest. PMID:15803645

  16. Circular Permutation of a Chaperonin Protein: Biophysics and Application to Nanotechnology

    Science.gov (United States)

    Paavola, Chad; Chan, Suzanne; Li, Yi-Fen; McMillan, R. Andrew; Trent, Jonathan

    2004-01-01

    We have designed five circular permutants of a chaperonin protein derived from the hyperthermophilic organism Sulfolobus shibatae. These permuted proteins were expressed in E. coli and are well-folded. Furthermore, all the permutants assemble into 18-mer double rings of the same form as the wild-type protein. We characterized the thermodynamics of folding for each permutant by both guanidine denaturation and differential scanning calorimetry. We also examined the assembly of chaperonin rings into higher order structures that may be used as nanoscale templates. The results show that circular permutation can be used to tune the thermodynamic properties of a protein template as well as facilitating the fusion of peptides, binding proteins or enzymes onto nanostructured templates.

  17. Earth's early biosphere

    Science.gov (United States)

    Des Marais, D. J.

    1998-01-01

    Understanding our own early biosphere is essential to our search for life elsewhere, because life arose on Earth very early and rocky planets shared similar early histories. The biosphere arose before 3.8 Ga ago, was exclusively unicellular and was dominated by hyperthermophiles that utilized chemical sources of energy and employed a range of metabolic pathways for CO2 assimilation. Photosynthesis also arose very early. Oxygenic photosynthesis arose later but still prior to 2.7 Ga. The transition toward the modern global environment was paced by a decline in volcanic and hydrothermal activity. These developments allowed atmospheric O2 levels to increase. The O2 increase created new niches for aerobic life, most notably the more advanced Eukarya that eventually spawned the megascopic fauna and flora of our modern biosphere.

  18. High hydrostatic pressure adaptive strategies in an obligate piezophile Pyrococcus yayanosii

    Science.gov (United States)

    Michoud, Grégoire; Jebbar, Mohamed

    2016-01-01

    Pyrococcus yayanosii CH1, as the first and only obligate piezophilic hyperthermophilic microorganism discovered to date, extends the physical and chemical limits of life on Earth. It was isolated from the Ashadze hydrothermal vent at 4,100 m depth. Multi-omics analyses were performed to study the mechanisms used by the cell to cope with high hydrostatic pressure variations. In silico analyses showed that the P. yayanosii genome is highly adapted to its harsh environment, with a loss of aromatic amino acid biosynthesis pathways and the high constitutive expression of the energy metabolism compared with other non-obligate piezophilic Pyrococcus species. Differential proteomics and transcriptomics analyses identified key hydrostatic pressure-responsive genes involved in translation, chemotaxis, energy metabolism (hydrogenases and formate metabolism) and Clustered Regularly Interspaced Short Palindromic Repeats sequences associated with Cellular apoptosis susceptibility proteins. PMID:27250364

  19. Crystal structure of Aquifex aeolicus gene product Aq1627: a putative phosphoglucosamine mutase reveals a unique C-terminal end-to-end disulfide linkage.

    Science.gov (United States)

    Sridharan, Upasana; Kuramitsu, Seiki; Yokoyama, Shigeyuki; Kumarevel, Thirumananseri; Ponnuraj, Karthe

    2017-06-27

    The Aq1627 gene from Aquifex aeolicus, a hyperthermophilic bacterium has been cloned and overexpressed in Escherichia coli. The protein was purified to homogeneity and its X-ray crystal structure was determined to 1.3 Å resolution using multiple wavelength anomalous dispersion phasing. The structural and sequence analysis of Aq1627 is suggestive of a putative phosphoglucosamine mutase. The structural features of Aq1627 further indicate that it could belong to a new subclass of the phosphoglucosamine mutase family. Aq1627 structure contains a unique C-terminal end-to-end disulfide bond, which links two monomers and this structural information can be used in protein engineering to make proteins more stable in different applications.

  20. Proteomic properties reveal phyloecological clusters of Archaea.

    Directory of Open Access Journals (Sweden)

    Nela Nikolic

    Full Text Available In this study, we propose a novel way to describe the variety of environmental adaptations of Archaea. We have clustered 57 Archaea by using a non-redundant set of proteomic features, and verified that the clusters correspond to environmental adaptations to the archaeal habitats. The first cluster consists dominantly of hyperthermophiles and hyperthermoacidophilic aerobes. The second cluster joins together halophilic and extremely halophilic Archaea, while the third cluster contains mesophilic (mostly methanogenic Archaea together with thermoacidophiles. The non-redundant subset of proteomic features was found to consist of five features: the ratio of charged residues to uncharged, average protein size, normalized frequency of beta-sheet, normalized frequency of extended structure and number of hydrogen bond donors. We propose this clustering to be termed phyloecological clustering. This approach could give additional insights into relationships among archaeal species that may be hidden by sole phylogenetic analysis.