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Sample records for chemolithoautotroph sulfurimonas denitrificans

  1. The Genome of the Epsilonproteobacterial Chemolithoautotroph Sulfurimonas dentrificans

    Energy Technology Data Exchange (ETDEWEB)

    USF Genomics Class; Sievert, Stefan M.; Scott, Kathleen M.; Klotz, Martin G.; Chain, Patrick S.G.; Hauser, Loren J.; Hemp, James; Hugler, Michael; Land, Miriam; Lapidus, Alla; Larimer, Frank W.; Lucas, Susan; Malfatti, Stephanie A.; Meyer, Folker; Paulsen, Ian T.; Ren, Qinghu; Simon, Jorg

    2007-08-08

    Sulfur-oxidizing epsilonproteobacteria are common in a variety of sulfidogenic environments. These autotrophic and mixotrophic sulfur-oxidizing bacteria are believed to contribute substantially to the oxidative portion of the global sulfur cycle. In order to better understand the ecology and roles of sulfur-oxidizing epsilonproteobacteria, in particular those of the widespread genus Sulfurimonas, in biogeochemical cycles, the genome of Sulfurimonas denitrificans DSM1251 was sequenced. This genome has many features, including a larger size (2.2 Mbp), that suggest a greater degree of metabolic versatility or responsiveness to the environment than seen for most of the other sequenced epsilonproteobacteria. A branched electron transport chain is apparent, with genes encoding complexes for the oxidation of hydrogen, reduced sulfur compounds, and formate and the reduction of nitrate and oxygen. Genes are present for a complete, autotrophic reductive citric acid cycle. Many genes are present that could facilitate growth in the spatially and temporally heterogeneous sediment habitat from where Sulfurimonas denitrificans was originally isolated. Many resistance-nodulation-development family transporter genes (10 total) are present; of these, several are predicted to encode heavy metal efflux transporters. An elaborate arsenal of sensory and regulatory protein-encoding genes is in place, as are genes necessary to prevent and respond to oxidative stress.

  2. Genetic manipulation of the obligate chemolithoautotrophic bacterium Thiobacillus denitrificans

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    Beller, H.R.; Legler, T.C.; Kane, S.R.

    2011-07-15

    Chemolithoautotrophic bacteria can be of industrial and environmental importance, but they present a challenge for systems biology studies, as their central metabolism deviates from that of model organisms and there is a much less extensive experimental basis for their gene annotation than for typical organoheterotrophs. For microbes with sequenced genomes but unconventional metabolism, the ability to create knockout mutations can be a powerful tool for functional genomics and thereby render an organism more amenable to systems biology approaches. In this chapter, we describe a genetic system for Thiobacillus denitrificans, with which insertion mutations can be introduced by homologous recombination and complemented in trans. Insertion mutations are generated by in vitro transposition, the mutated genes are amplified by the PCR, and the amplicons are introduced into T. denitrificans by electroporation. Use of a complementation vector, pTL2, based on the IncP plasmid pRR10 is also addressed.

  3. Anaerobic, Nitrate-Dependent Oxidation of U(IV) Oxide Minerals by the Chemolithoautotrophic Bacterium Thiobacillus denitrificans

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    Beller, H R

    2004-06-25

    Under anaerobic conditions and at circumneutral pH, cells of the widely-distributed, obligate chemolithoautotrophic bacterium Thiobacillus denitrificans oxidatively dissolved synthetic and biogenic U(IV) oxides (uraninite) in nitrate-dependent fashion: U(IV) oxidation required the presence of nitrate and was strongly correlated to nitrate consumption. This is the first report of anaerobic U(IV) oxidation by an autotrophic bacterium.

  4. Structural Stability, Transitions, and Interactions within SoxYZCD-Thiosulphate from Sulfurimonas denitrificans: An In Silico Molecular Outlook for Maintaining Environmental Sulphur Cycle

    Science.gov (United States)

    Banerjee, Arundhati

    2016-01-01

    Thiosulphate oxidation (an essential mechanism) serves to maintain the global sulphur cycle. Earlier experimental and computational studies dealt with environmental thiosulphate oxidation but none dealt with thiosulphate oxidation from deep ocean belts. Wet-laboratory experimental research shows that epsilon-proteobacteria Sulfurimonas denitrificans possess sox (sulphur-oxidizing) operon and perform thiosulphate oxidation efficiently underneath the oceans. From this specific sox operon, SoxCD complex recycles the thiosulphate-bound SoxY from SoxYZ complex to balance the environmental sulphur cycle. So, four chief proteins were variedly modeled and relevant simulated interactive structures were obtained. The final simulated tetraprotein complex (SoxYZCD) from docked SoxYZ and SoxCD complexes was disclosed to be a highly interactive one with predominant ionic residues. Free energy of folding, solvent accessibility, and conformational shifts (coil-like conformation to helices and sheets) were observed in SoxYZ complex after interacting with SoxCD. The stability of the complex (SoxYZCD) after simulation was also observed through the electrostatic surface potential values. These evaluations were rationalized via biostatistics. This aids SoxCD for recycling SoxY along with thiosulphate, which remains interconnected by four H-bonds with SoxY. Therefore, this novel exploration is endowed with the detailed molecular viewpoint for maintaining the sulphur cycle (globally) including the ocean belts. PMID:27777586

  5. Genome-enabled studies of anaerobic, nitrate-dependent iron oxidation in the chemolithoautotrophic bacterium Thiobacillus denitrificans

    Directory of Open Access Journals (Sweden)

    Harry R Beller

    2013-08-01

    Full Text Available Thiobacillus denitrificans is a chemolithoautotrophic bacterium capable of anaerobic, nitrate-dependent U(IV and Fe(II oxidation, both of which can strongly influence the long-term efficacy of in situ reductive immobilization of uranium in contaminated aquifers. We previously identified two c-type cytochromes involved in nitrate-dependent U(IV oxidation in T. denitrificans and hypothesized that c-type cytochromes would also catalyze Fe(II oxidation, as they have been found to play this role in anaerobic phototrophic Fe(II-oxidizing bacteria. Here we report on efforts to identify genes associated with nitrate-dependent Fe(II oxidation, namely (a whole-genome transcriptional studies [using FeCO3, Fe2+, and U(IV oxides as electron donors under denitrifying conditions], (b Fe(II oxidation assays performed with knockout mutants targeting primarily highly expressed or upregulated c-type cytochromes, and (c random transposon-mutagenesis studies with screening for Fe(II oxidation. Assays of mutants for 26 target genes, most of which were c-type cytochromes, indicated that none of the mutants tested were significantly defective in nitrate-dependent Fe(II oxidation. The non-defective mutants included the c1-cytochrome subunit of the cytochrome bc1 complex (complex III, which has relevance to a previously proposed role for this complex in nitrate-dependent Fe(II oxidation and to current concepts of reverse electron transfer. A transposon mutant with a disrupted gene associated with NADH:ubiquinone oxidoreductase (complex I was ~35% defective relative to the wild-type strain; this strain was similarly defective in nitrate reduction with thiosulfate as the electron donor. Overall, our results indicate that nitrate-dependent Fe(II oxidation in T. denitrificans is not catalyzed by the same c-type cytochromes involved in U(IV oxidation, nor have other c-type cytochromes yet been implicated in the process.

  6. Whole-Genome Transcriptional Analysis of Chemolithoautotrophic Thiosulfate Oxidation by Thiobacillus denitrificans Under Aerobic vs. Denitrifying Conditions

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    Beller, H R; Letain, T E; Chakicherla, A; Kane, S R; Legler, T C; Coleman, M A

    2006-04-22

    Thiobacillus denitrificans is one of the few known obligate chemolithoautotrophic bacteria capable of energetically coupling thiosulfate oxidation to denitrification as well as aerobic respiration. As very little is known about the differential expression of genes associated with ke chemolithoautotrophic functions (such as sulfur-compound oxidation and CO2 fixation) under aerobic versus denitrifying conditions, we conducted whole-genome, cDNA microarray studies to explore this topic systematically. The microarrays identified 277 genes (approximately ten percent of the genome) as differentially expressed using Robust Multi-array Average statistical analysis and a 2-fold cutoff. Genes upregulated (ca. 6- to 150-fold) under aerobic conditions included a cluster of genes associated with iron acquisition (e.g., siderophore-related genes), a cluster of cytochrome cbb3 oxidase genes, cbbL and cbbS (encoding the large and small subunits of form I ribulose 1,5-bisphosphate carboxylase/oxygenase, or RubisCO), and multiple molecular chaperone genes. Genes upregulated (ca. 4- to 95-fold) under denitrifying conditions included nar, nir, and nor genes (associated respectively with nitrate reductase, nitrite reductase, and nitric oxide reductase, which catalyze successive steps of denitrification), cbbM (encoding form II RubisCO), and genes involved with sulfur-compound oxidation (including two physically separated but highly similar copies of sulfide:quinone oxidoreductase and of dsrC, associated with dissimilatory sulfite reductase). Among genes associated with denitrification, relative expression levels (i.e., degree of upregulation with nitrate) tended to decrease in the order nar > nir > nor > nos. Reverse transcription, quantitative PCR analysis was used to validate these trends.

  7. Complete genome sequence of Sulfurimonas autotrophica type strain (OK10T)

    Energy Technology Data Exchange (ETDEWEB)

    Sikorski, Johannes [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Munk, Christine [U.S. Department of Energy, Joint Genome Institute; Lapidus, Alla L. [U.S. Department of Energy, Joint Genome Institute; Djao, Olivier Duplex [HZI - Helmholtz Centre for Infection Research, Braunschweig, Germany; Lucas, Susan [U.S. Department of Energy, Joint Genome Institute; Glavina Del Rio, Tijana [U.S. Department of Energy, Joint Genome Institute; Nolan, Matt [U.S. Department of Energy, Joint Genome Institute; Tice, Hope [U.S. Department of Energy, Joint Genome Institute; Han, Cliff [Los Alamos National Laboratory (LANL); Cheng, Jan-Fang [U.S. Department of Energy, Joint Genome Institute; Tapia, Roxanne [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; Ivanova, N [U.S. Department of Energy, Joint Genome Institute; Mavromatis, K [U.S. Department of Energy, Joint Genome Institute; Mikhailova, Natalia [U.S. Department of Energy, Joint Genome Institute; Pati, Amrita [U.S. Department of Energy, Joint Genome Institute; Sims, David [Los Alamos National Laboratory (LANL); Meincke, Linda [Los Alamos National Laboratory (LANL); Detter, J. Chris [U.S. Department of Energy, Joint Genome Institute; Caiazza, Nicky [Mascoma Corporation; 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; Chang, Yun-Juan [ORNL; Jeffries, Cynthia [Oak Ridge National Laboratory (ORNL); Rohde, Manfred [HZI - Helmholtz Centre for Infection Research, Braunschweig, Germany; Lang, Elke [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, 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

    2010-01-01

    Sulfurimonas autotrophica Inagaki et al. 2003 is the type species of the genus Sulfurimonas. This genus is of interest because of its significant contribution to the global sulfur cycle by oxidizing of sulfur compounds to sulfate and by its apparent occupation of deep-sea hydrothermal and marine sulfidic environments as potential ecological niche. Here we describe the features of this organism, together with the complete genome sequence and annotation. This is the second complete genome sequence of the genus Sulfurimonas and the duodenary in the family Helicobacteraceae. The 2,153,198 bp long genome with its 2,165 protein-coding and 55 RNA genes is a part of the Genomic Encyclopedia of Bacteria and Archaea project.

  8. The terminal oxidases of Paracoccus denitrificans

    OpenAIRE

    de Gier, Jan-Willem L.; Lübben, Mathias; Reijnders, Willem N.M.; Tipker, Corinne A.; Slotboom, Dirk-Jan; Van Spanning, Rob J. M.; Stouthamer, Adriaan H.; van der Oost, John

    1994-01-01

    Three distinct types of terminal oxidases participate in the aerobic respiratory pathways of Paracoccus denitrificans. Two alternative genes encoding subunit I of the aa3-type cytochrome c oxidase have been isolated before, namely ctaDI and ctaDII. Each of these genes can be expressed separately to complement a double mutant (ΔctaDI, ΔctaDII), indicating that they are isoforms of subunit I of the aa3-type oxidase. The genomic locus of a quinol oxidase has been isolated: cyoABC. This protohaem...

  9. Permanent draft genome of Thiobacillus thioparus DSM 505(T), an obligately chemolithoautotrophic member of the Betaproteobacteria.

    Science.gov (United States)

    Hutt, Lee P; Huntemann, Marcel; Clum, Alicia; Pillay, Manoj; Palaniappan, Krishnaveni; Varghese, Neha; Mikhailova, Natalia; Stamatis, Dimitrios; Reddy, Tatiparthi; Daum, Chris; Shapiro, Nicole; Ivanova, Natalia; Kyrpides, Nikos; Woyke, Tanja; Boden, Rich

    2017-01-01

    Thiobacillus thioparus DSM 505(T) is one of first two isolated strains of inorganic sulfur-oxidising Bacteria. The original strain of T. thioparus was lost almost 100 years ago and the working type strain is Culture C(T) (=DSM 505(T) = ATCC 8158(T)) isolated by Starkey in 1934 from agricultural soil at Rutgers University, New Jersey, USA. It is an obligate chemolithoautotroph that conserves energy from the oxidation of reduced inorganic sulfur compounds using the Kelly-Trudinger pathway and uses it to fix carbon dioxide It is not capable of heterotrophic or mixotrophic growth. The strain has a genome size of 3,201,518 bp. Here we report the genome sequence, annotation and characteristics. The genome contains 3,135 protein coding and 62 RNA coding genes. Genes encoding the transaldolase variant of the Calvin-Benson-Bassham cycle were also identified and an operon encoding carboxysomes, along with Smith's biosynthetic horseshoe in lieu of Krebs' cycle sensu stricto. Terminal oxidases were identified, viz. cytochrome c oxidase (cbb3, EC 1.9.3.1) and ubiquinol oxidase (bd, EC 1.10.3.10). There is a partial sox operon of the Kelly-Friedrich pathway of inorganic sulfur-oxidation that contains soxXYZAB genes but lacking soxCDEF, there is also a lack of the DUF302 gene previously noted in the sox operon of other members of the 'Proteobacteria' that can use trithionate as an energy source. In spite of apparently not growing anaerobically with denitrification, the nar, nir, nor and nos operons encoding enzymes of denitrification are found in the T. thioparus genome, in the same arrangements as in the true denitrifier T. denitrificans.

  10. [Comparative susceptibility of Ochrobactrum anthropi, Agrobacterium tumefaciens, Alcaligenes faecalis, Alcaligenes denitrificans subsp. denitrificans, Alcaligenes denitrificans subsp. xylosidans and Bordetella bronchiseptica against 35 antibiotics including 17 beta-lactams].

    Science.gov (United States)

    Bizet, C; Bizet, J

    1995-04-01

    Ochrobactrum anthropi, formerly known as "Achromobacter sp." or CDC group Vd has been isolated from water, hospital environment (antiseptic solutions, dialysis fluids ... ). O. anthropi is a Gram negative, motile, strictly aerobic, oxydase positive and non-fermentative bacteria with a strong urease activity. The susceptibility of 13 strains of O. anthropi was determined by agar diffusion method and compared to those of type strains of Agrobacterium tumefaciens, Alcaligenes faecalis, Alcaligenes denitrificans subsp. denitrificans, Alcaligenes denitrificans subsp. xylosoxydans and Bordetella bronchiseptica. The MICs of 20 antimicrobial agents confirmed the distinct phenotype susceptibility of O. anthropi. All the strains of O. anthropi are sensitive to imipenem, amikacin, gentamicin, netilmicin, nalidixic acid, pefloxacin, ciprofloxacin, tetracyclin, colistin, sulphonamides and rifampicin and resistant to ampicillin, amoxycillin + clavulanic acid, ticarcillin, mezlocillin, cefuroxime, cefamandol, cefoxitin, cefotaxime, cefoperazon, ceftazidime, cefsulodin, aztreonam, streptomycin, kanamycin, pipemidic acid, chloramphenicol, erythromicin, pristinamycin, trimethoprim and fosfomycin. O. anthropi is implicated in nosocomial infections. O. anthropi was the species with the greatest resistance to beta-lactamins.

  11. Carbohydrate metabolism and carbon fixation in Roseobacter denitrificans OCh114.

    Directory of Open Access Journals (Sweden)

    Kuo-Hsiang Tang

    Full Text Available The Roseobacter clade of aerobic marine proteobacteria, which compose 10-25% of the total marine bacterial community, has been reported to fix CO(2, although it has not been determined what pathway is involved. In this study, we report the first metabolic studies on carbohydrate utilization, CO(2 assimilation, and amino acid biosynthesis in the phototrophic Roseobacter clade bacterium Roseobacter denitrificans OCh114. We develop a new minimal medium containing defined carbon source(s, in which the requirements of yeast extract reported previously for the growth of R. denitrificans can be replaced by vitamin B(12 (cyanocobalamin. Tracer experiments were carried out in R. denitrificans grown in a newly developed minimal medium containing isotopically labeled pyruvate, glucose or bicarbonate as a single carbon source or in combination. Through measurements of (13C-isotopomer labeling patterns in protein-derived amino acids, gene expression profiles, and enzymatic activity assays, we report that: (1 R. denitrificans uses the anaplerotic pathways mainly via the malic enzyme to fix 10-15% of protein carbon from CO(2; (2 R. denitrificans employs the Entner-Doudoroff (ED pathway for carbohydrate metabolism and the non-oxidative pentose phosphate pathway for the biosynthesis of histidine, ATP, and coenzymes; (3 the Embden-Meyerhof-Parnas (EMP, glycolysis pathway is not active and the enzymatic activity of 6-phosphofructokinase (PFK cannot be detected in R. denitrificans; and (4 isoleucine can be synthesized from both threonine-dependent (20% total flux and citramalate-dependent (80% total flux pathways using pyruvate as the sole carbon source.

  12. Halobacterium denitrificans sp. nov., an extremely halophilic denitrifying bacterium

    Science.gov (United States)

    Tomlinson, G. A.; Jahnke, L. L.; Hochstein, L. I.

    1986-01-01

    Halobacterium denitrificans was one of several carbohydrate-utilizing, denitrifying, extremely halophilic bacteria isolated by anaerobic enrichment in the presence of nitrate. Anaerobic growth took place only when nitrate (or nitrite) was present and was accompanied by the production of dinitrogen. In the presence of high concentrations of nitrate (i.e., 0.5 percent), nitrous oxide and nitrite were also detected. When grown aerobically in a mineral-salts medium containing 0.005 percent yeast extract, H. denitrificans utilized a variety of carbohydrates as sources of carbon and energy. In every case, carbohydrate utilization was accompanied by acid production.

  13. Halobacterium denitrificans sp. nov. - An extremely halophilic denitrifying bacterium

    Science.gov (United States)

    Tomlinson, G. A.; Jahnke, L. L.; Hochstein, L. I.

    1986-01-01

    Halobacterium denitrificans was one of several carbohydrate-utilizing, denitrifying, extremely halophilic bacteria isolated by anaerobic enrichment in the presence of nitrate. Anaerobic growth took place only when nitrate (or nitrite) was present and was accompanied by the production of dinitrogen. In the presence of high concentrations of nitrate (i.e., 0.5 percent), nitrous oxide and nitrite were also detected. When grown aerobically in a mineral-salts medium containing 0.005 percent yeast extract, H. denitrificans utilized a variety of carbohydrates as sources of carbon and energy. In every case, carbohydrate utilization was accompanied by acid production.

  14. Novel cyanide-hydrolyzing enzyme from Alcaligenes xylosoxidans subsp. denitrificans

    Energy Technology Data Exchange (ETDEWEB)

    Ingvorsen, K.; Hojer-Pederson, B.; Godtfredsen, S.E. (Novo Nordisk A/S, Bagsvaerd (Denmark))

    1991-06-01

    A cyanide-metabolizing bacterium, strain DF3, isolated from soil was identified as Alcaligenes xylosoxidans subsp. denitrificans. Whole cells and cell extracts of strain DF3 catalyzed hydrolysis of cyanide to formate and ammonia (HCN + 2H{sub 2}O {r arrow} HCOOH + NH{sub 3}) without forming formamide as a free intermediate. The cyanide-hydrolyzing activity was inducibly produced in cells during growth in cyanide-containing media. Cyanate (OCN{sup {minus}}) and a wide range of aliphatic and aromatic nitriles were not hydrolyzed by intact cells of A. xylosoxidans subsp. denitrificans DF3. Strain DF3 hydrolyzed cyanide with great efficacy. Thus, by using resting induced cells at a concentration of 11.3 mg (dry weight) per ml, the cyanide concentration could be reduced from 0.97 M (approximately 25,220 ppm) to less than 77 nM (approximately 0.002 ppm) in 55 h. Enzyme purification established that cyanide hydrolysis by A. xylosoxidans subsp. denitrificans DF3 was due to a single intracellular enzyme. The molecular mass of the active enzyme (purity, {gt}97% as determined by amino acid sequencing) was estimated to be {gt}300,000 Da. The cyanide-hydrolyzing enzyme of A. xylosoxidans subsp. denitrificans DF3 was tentatively named cyanidase to distinguish it from known nitrilases (EC 3.5.5.1) which act on organic nitriles.

  15. Permanent draft genome of Thermithiobaclillus tepidarius DSM 3134(T), a moderately thermophilic, obligately chemolithoautotrophic member of the Acidithiobacillia.

    Science.gov (United States)

    Boden, Rich; Hutt, Lee P; Huntemann, Marcel; Clum, Alicia; Pillay, Manoj; Palaniappan, Krishnaveni; Varghese, Neha; Mikhailova, Natalia; Stamatis, Dimitrios; Reddy, Tatiparthi; Ngan, Chew Yee; Daum, Chris; Shapiro, Nicole; Markowitz, Victor; Ivanova, Natalia; Woyke, Tanja; Kyrpides, Nikos

    2016-01-01

    Thermithiobacillus tepidarius DSM 3134(T) was originally isolated (1983) from the waters of a sulfidic spring entering the Roman Baths (Temple of Sulis-Minerva) at Bath, United Kingdom and is an obligate chemolithoautotroph growing at the expense of reduced sulfur species. This strain has a genome size of 2,958,498 bp. Here we report the genome sequence, annotation and characteristics. The genome comprises 2,902 protein coding and 66 RNA coding genes. Genes responsible for the transaldolase variant of the Calvin-Benson-Bassham cycle were identified along with a biosynthetic horseshoe in lieu of Krebs' cycle sensu stricto. Terminal oxidases were identified, viz. cytochrome c oxidase (cbb3, EC 1.9.3.1) and ubiquinol oxidase (bd, EC 1.10.3.10). Metalloresistance genes involved in pathways of arsenic and cadmium resistance were found. Evidence of horizontal gene transfer accounting for 5.9 % of the protein-coding genes was found, including transfer from Thiobacillus spp. and Methylococcus capsulatus Bath, isolated from the same spring. A sox gene cluster was found, similar in structure to those from other Acidithiobacillia - by comparison with Thiobacillus thioparus and Paracoccus denitrificans, an additional gene between soxA and soxB was found, annotated as a DUF302-family protein of unknown function. As the Kelly-Friedrich pathway of thiosulfate oxidation (encoded by sox) is not used in Thermithiobacillus spp., the role of the operon (if any) in this species remains unknown. We speculate that DUF302 and sox genes may have a role in periplasmic trithionate oxidation.

  16. Enhanced fatty acid production in engineered chemolithoautotrophic bacteria using reduced sulfur compounds as energy sources

    DEFF Research Database (Denmark)

    Beller, Harry R.; Zhou, Peng; Jewell, Talia N.M.;

    2016-01-01

    Chemolithoautotrophic bacteria that oxidize reduced sulfur compounds, such as H2S, while fixing CO2 are an untapped source of renewable bioproducts from sulfide-laden waste, such as municipal wastewater. In this study, we report engineering of the chemolithoautotrophic bacterium Thiobacillus...

  17. Genome sequences of Alicycliphilus denitrificans strains BC and K601T

    NARCIS (Netherlands)

    Oosterkamp, M.J.; Veuskens, T.; Plugge, C.M.; Langenhoff, A.A.M.; Gerritsen, J.; Berkel, van W.J.H.; Junca, H.; Smidt, H.; Stams, A.J.M.

    2011-01-01

    Alicycliphilus denitrificans strain BC and A. denitrificans strain K601(T) degrade cyclic hydrocarbons. These strains have been isolated from a mixture of wastewater treatment plant material and benzene-polluted soil and from a wastewater treatment plant, respectively, suggesting their role in biore

  18. Proteomic analysis of nitrate-dependent acetone degradation by Alicycliphilus denitrificans strain BC

    NARCIS (Netherlands)

    Oosterkamp, M.J.; Boeren, S.; Atashgahi, S.; Plugge, C.M.; Schaap, P.J.; Stams, A.J.M.

    2015-01-01

    Alicycliphilus denitrificans strain BC grows anaerobically on acetone with nitrate as electron acceptor. Comparative proteomics of cultures of A. denitrificans strain BC grown on either acetone or acetate with nitrate was performed to study the enzymes involved in the acetone degradation pathway. In

  19. The Impact of Nitrite on Aerobic Growth of Paracoccus denitrificans PD1222

    OpenAIRE

    Hartop, Katherine

    2014-01-01

    The effect of nitrite stress induced in Paracoccus denitrificans PD1222 was examined using additions of sodium nitrite to an aerobic bacterial culture. Nitrite generates a strong stress response in P. denitrificans, causing growth inhibition. This is dependent on both the concentration of nitrite present and the pH. The pH dependent effect of nitrite growth inhibition is likely a result of nitrite and free nitrous acid (FNA; pKa = 3.16) and subsequent reactive nitrogen oxides, ...

  20. Substrate uptake and subcellular compartmentation of anoxic cholesterol catabolism in Sterolibacterium denitrificans.

    Science.gov (United States)

    Lin, Ching-Wen; Wang, Po-Hsiang; Ismail, Wael; Tsai, Yu-Wen; El Nayal, Ashraf; Yang, Chia-Ying; Yang, Fu-Chun; Wang, Chia-Hsiang; Chiang, Yin-Ru

    2015-01-09

    Cholesterol catabolism by actinobacteria has been extensively studied. In contrast, the uptake and catabolism of cholesterol by Gram-negative species are poorly understood. Here, we investigated microbial cholesterol catabolism at the subcellular level. (13)C metabolomic analysis revealed that anaerobically grown Sterolibacterium denitrificans, a β-proteobacterium, adopts an oxygenase-independent pathway to degrade cholesterol. S. denitrificans cells did not produce biosurfactants upon growth on cholesterol and exhibited high cell surface hydrophobicity. Moreover, S. denitrificans did not produce extracellular catabolic enzymes to transform cholesterol. Accordingly, S. denitrificans accessed cholesterol by direction adhesion. Cholesterol is imported through the outer membrane via a putative FadL-like transport system, which is induced by neutral sterols. The outer membrane steroid transporter is able to selectively import various C27 sterols into the periplasm. S. denitrificans spheroplasts exhibited a significantly higher efficiency in cholest-4-en-3-one-26-oic acid uptake than in cholesterol uptake. We separated S. denitrificans proteins into four fractions, namely the outer membrane, periplasm, inner membrane, and cytoplasm, and we observed the individual catabolic reactions within them. Our data indicated that, in the periplasm, various periplasmic and peripheral membrane enzymes transform cholesterol into cholest-4-en-3-one-26-oic acid. The C27 acidic steroid is then transported into the cytoplasm, in which side-chain degradation and the subsequent sterane cleavage occur. This study sheds light into microbial cholesterol metabolism under anoxic conditions.

  1. Microbiological Denitrification and Denitrifying Activity of Paracoccus Denitrificans

    Institute of Scientific and Technical Information of China (English)

    万曦; 万国江; 等

    2000-01-01

    With rapidly industrial and agricultural development,more and more fertilizers,chemicals and heavy ions will be discharged into lakes and rivers,which would cause lake eutrophication and quality deterioration in drinking water sources.Therefore,denitrification is essential for controlling the amounts of nitrogen,During the transformation process from nitrate to the end products-nitrogen and several intermediated[e.g.nitrite(NO2-),nitrous oxide(N2O) and nitric oxide(NO)]may be accumulated,which have more toxic influences on the environment.in This study,the denitrification effect of Paracoccus Denitrificans was examined on the changes between oxic and anoxic conditions at varying pH.At pH=7.5,denitrification proceeded well after 3 switches from oxic to anoxic conditions and vice versa,Production of N2 was constant and the amounts of NO2-,N2O and NO were extremely low.How ever,at pH=6.8,denitrification activity was inhitied and there large amounts of the intermaediates.The denitrifying bacteria decreased violently in dry weight and were washed out.

  2. Engineering the iron-oxidizing chemolithoautotroph Acidithiobacillus ferrooxidans for biochemical production.

    Science.gov (United States)

    Kernan, Timothy; Majumdar, Sudipta; Li, Xiaozheng; Guan, Jingyang; West, Alan C; Banta, Scott

    2016-01-01

    There is growing interest in developing non-photosynthetic routes for the conversion of CO2 to fuels and chemicals. One underexplored approach is the transfer of energy to the metabolism of genetically modified chemolithoautotrophic bacteria. Acidithiobacillus ferrooxidans is an obligate chemolithoautotroph that derives its metabolic energy from the oxidation of iron or sulfur at low pH. Two heterologous biosynthetic pathways have been expressed in A. ferrooxidans to produce either isobutyric acid or heptadecane from CO2 and the oxidation of Fe(2+). A sevenfold improvement in productivity of isobutyric acid was obtained through improved media formulations in batch cultures. Steady-state efficiencies were lower in continuous cultures, likely due to ferric inhibition. If coupled to solar panels, the photon-to-fuel efficiency of this proof-of-principle process approaches estimates for agriculture-derived biofuels. These efforts lay the foundation for the utilization of this organism in the exploitation of electrical energy for biochemical synthesis.

  3. The metabolic impact of extracellular nitrite on aerobic metabolism of Paracoccus denitrificans.

    Science.gov (United States)

    Hartop, K R; Sullivan, M J; Giannopoulos, G; Gates, A J; Bond, P L; Yuan, Z; Clarke, T A; Rowley, G; Richardson, D J

    2017-02-07

    Nitrite, in equilibrium with free nitrous acid (FNA), can inhibit both aerobic and anaerobic growth of microbial communities through bactericidal activities that have considerable potential for control of microbial growth in a range of water systems. There has been much focus on the effect of nitrite/FNA on anaerobic metabolism and so, to enhance understanding of the metabolic impact of nitrite/FNA on aerobic metabolism, a study was undertaken with a model denitrifying bacterium Paracoccus denitrificans PD1222. Extracellular nitrite inhibits aerobic growth of P. denitrificans in a pH-dependent manner that is likely to be a result of both nitrite and free nitrous acid (pKa = 3.25) and subsequent reactive nitrogen oxides generated from the intracellular passage of FNA into P. denitrificans. Increased expression of a gene encoding a flavohemoglobin protein (Fhp) (Pden_1689) was observed in response to extracellular nitrite. Construction and analysis of a deletion mutant established Fhp to be involved in endowing nitrite/FNA resistance at high extracellular nitrite concentrations. Global transcriptional analysis confirmed nitrite-dependent expression of fhp and indicated that P. denitrificans expressed a number of stress response systems associated with protein, DNA and lipid repair. It is therefore suggested that nitrite causes a pH-dependent stress response that is due to the production of associated reactive nitrogen species, such as nitric oxide from the internalisation of FNA.

  4. Differentiation of Kingella denitrificans from Neisseria gonorrhoeae by growth on a semisolid medium and sensitivity to amylase.

    Science.gov (United States)

    Odugbemi, T; Arko, R J

    1983-01-01

    All 239 strains of Neisseria gonorrhoeae tested for sensitivity to amylase-impregnated disks showed zones of growth inhibition ranging from 15 to 30 mm in diameter. None of 52 strains of Kingella denitrificans were inhibited by the same amylase test. In addition, all N. gonorrhoeae strains grew well on a gonococcal base medium supplemented with either IsoVitaleX (BBL Microbiology Systems) or Supplement B (Difco Laboratories), whereas K. denitrificans grew adequately only on the medium with IsoVitaleX. The addition of L-cysteine or cystine to the medium with Supplement B enhanced the growth of K. denitrificans. Images PMID:6187768

  5. NCBI nr-aa BLAST: CBRC-MDOM-08-0115 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-08-0115 ref|YP_393083.1| putative integral membrane protein [Sulfurimonas... denitrificans DSM 1251] gb|ABB43848.1| putative integral membrane protein [Sulfurimonas denitrificans DSM 1251] YP_393083.1 0.12 25% ...

  6. NCBI nr-aa BLAST: CBRC-MDOM-05-0049 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-05-0049 ref|YP_393083.1| putative integral membrane protein [Sulfurimonas... denitrificans DSM 1251] gb|ABB43848.1| putative integral membrane protein [Sulfurimonas denitrificans DSM 1251] YP_393083.1 0.47 25% ...

  7. NCBI nr-aa BLAST: CBRC-MDOM-06-0011 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-06-0011 ref|YP_393083.1| putative integral membrane protein [Sulfurimonas... denitrificans DSM 1251] gb|ABB43848.1| putative integral membrane protein [Sulfurimonas denitrificans DSM 1251] YP_393083.1 0.11 23% ...

  8. NCBI nr-aa BLAST: CBRC-MDOM-02-0387 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-02-0387 ref|YP_393083.1| putative integral membrane protein [Sulfurimonas... denitrificans DSM 1251] gb|ABB43848.1| putative integral membrane protein [Sulfurimonas denitrificans DSM 1251] YP_393083.1 0.033 24% ...

  9. NCBI nr-aa BLAST: CBRC-MDOM-07-0060 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-07-0060 ref|YP_393083.1| putative integral membrane protein [Sulfurimonas... denitrificans DSM 1251] gb|ABB43848.1| putative integral membrane protein [Sulfurimonas denitrificans DSM 1251] YP_393083.1 0.51 26% ...

  10. The genome of deep-sea vent chemolithoautotroph Thiomicrospira crunogena XCL-2.

    Directory of Open Access Journals (Sweden)

    Kathleen M Scott

    2006-11-01

    Full Text Available Presented here is the complete genome sequence of Thiomicrospira crunogena XCL-2, representative of ubiquitous chemolithoautotrophic sulfur-oxidizing bacteria isolated from deep-sea hydrothermal vents. This gammaproteobacterium has a single chromosome (2,427,734 base pairs, and its genome illustrates many of the adaptations that have enabled it to thrive at vents globally. It has 14 methyl-accepting chemotaxis protein genes, including four that may assist in positioning it in the redoxcline. A relative abundance of coding sequences (CDSs encoding regulatory proteins likely control the expression of genes encoding carboxysomes, multiple dissolved inorganic nitrogen and phosphate transporters, as well as a phosphonate operon, which provide this species with a variety of options for acquiring these substrates from the environment. Thiom. crunogena XCL-2 is unusual among obligate sulfur-oxidizing bacteria in relying on the Sox system for the oxidation of reduced sulfur compounds. The genome has characteristics consistent with an obligately chemolithoautotrophic lifestyle, including few transporters predicted to have organic allocrits, and Calvin-Benson-Bassham cycle CDSs scattered throughout the genome.

  11. The Genome of Deep-Sea Vent Chemolithoautotroph Thiomicrospira crunogena XCL-2

    Energy Technology Data Exchange (ETDEWEB)

    Scott, K M; Sievert, S M; Abril, F N; Ball, L A; Barrett, C J; Blake, R A; Boller, A J; Chain, P G; Clark, J A; Davis, C R; Detter, C; Do, K F; Dobrinski, K P; Faza, B I; Fitzpatrick, K A; Freyermuth, S K; Harmer, T L; Hauser, L J; Hugler, M; Kerfeld, C A; Klotz, M G; Kong, W W; Land, M; Lapidus, A; Larimer, F W; Longo, D L; Lucas, S; Malfatti, S A; Massey, S E; Martin, D D; McCuddin, Z; Meyer, F; Moore, J L; Ocampo Jr., L H; Paul, J H; Paulsen, I T; Reep, D K; Ren, Q; Ross, R L; Sato, P Y; Thomas, P; Tinkham, L E; Zerugh, G T

    2007-01-10

    Presented here is the complete genome sequence of Thiomicrospira crunogena XCL-2, representative of ubiquitous chemolithoautotrophic sulfur-oxidizing bacteria isolated from deep-sea hydrothermal vents. This gammaproteobacterium has a single chromosome (2,427,734 bp), and its genome illustrates many of the adaptations that have enabled it to thrive at vents globally. It has 14 methyl-accepting chemotaxis protein genes, including four that may assist in positioning it in the redoxcline. A relative abundance of CDSs encoding regulatory proteins likely control the expression of genes encoding carboxysomes, multiple dissolved inorganic nitrogen and phosphate transporters, as well as a phosphonate operon, which provide this species with a variety of options for acquiring these substrates from the environment. T. crunogena XCL-2 is unusual among obligate sulfur oxidizing bacteria in relying on the Sox system for the oxidation of reduced sulfur compounds. A 38 kb prophage is present, and a high level of prophage induction was observed, which may play a role in keeping competing populations of close relatives in check. The genome has characteristics consistent with an obligately chemolithoautotrophic lifestyle, including few transporters predicted to have organic allocrits, and Calvin-Benson-Bassham cycle CDSs scattered throughout the genome.

  12. Dynamic Water Networks in Cytochrome c Oxidase from Paracoccus denitrificans Investigated by Molecular Dynamics Simulations

    OpenAIRE

    Olkhova, Elena; Hutter, Michael C; Lill, Markus A.; Helms, Volkhard; Michel, Hartmut

    2004-01-01

    We present a molecular dynamics study of cytochrome c oxidase from Paracoccus denitrificans in the fully oxidized state, embedded in a fully hydrated dimyristoylphosphatidylcholine lipid bilayer membrane. Parallel simulations with different levels of protein hydration, 1.125 ns each in length, were carried out under conditions of constant temperature and pressure using three-dimensional periodic boundary conditions and full electrostatics to investigate the distribution and dynamics of water ...

  13. Biodegradation of isopropanol by a solvent-tolerant Paracoccus denitrificans strain.

    Science.gov (United States)

    Geng, Yucong; Deng, Yuanjie; Chen, Feilong; Jin, Hong; Tao, Ke; Hou, Taiping

    2015-01-01

    The biodegradation of high concentration isopropanol (2-propanol, IPA) at 16 g/L was investigated by a solvent-tolerant strain of bacteria identified as Paracoccus denitrificans for the first time by 16S rDNA gene sequencing. The strain P. denitrificans GH3 was able to utilize the high concentration of IPA as the sole carbon source within a minimal salts medium with a cell density of 1.5×10(8) cells/mL. The optimal conditions were found as follows: initial pH 7.0, incubation temperature 30°C, with IPA concentration 8 g/L. Under the optimal conditions, strain GH3 utilized 90.3% of IPA in 7 days. Acetone, the major intermediate of aerobic IPA biodegradation, was also monitored as an indicator of microbial IPA utilization. Both IPA and acetone were completely removed from the medium following 216 hr and 240 hr, respectively. The growth of strain GH3 on IPA as a sole carbon and energy source was well described by the Andrews model with a maximum growth rate (μmax)=0.0277/hr, a saturation constant (KS)=0.7333 g/L, and an inhibition concentration (Ki)=8.9887 g/L. Paracoccus denitrificans GH3 is considered to be well used in degrading IPA in wastewater.

  14. Novel Castellaniella denitrificans SA13P as a Potent Malachite Green Decolorizing Strain

    Directory of Open Access Journals (Sweden)

    Ankita Chawla

    2014-01-01

    Full Text Available Triphenylmethane dyes represent a major group of dyes causing serious environmental hazards. Malachite Green is one of the commonly and extensively used triphenylmethane dyes although it is carcinogenic and mutagenic in nature. Various physicochemical methods have been employed for its elimination but are highly expensive, coupled with the formation of huge amount of sludge. Hence, biological methods being ecofriendly are good alternatives. In the present study, the novel bacterial isolate SA13P was isolated from UASB tank of tannery effluent treatment plant. Phylogenetic characterization of 1470 bp fragment of SA13P has revealed its similarity with Castellaniella denitrificans. This strain has been found to decolorize the dye (malachite green at a concentration of 100 mg L−1 (80.29%. Decolorization was done by living bacterial cells rather than adsorption. Growth conditions have also been optimized for the decolorization. Maximum decolorization was observed at a temperature of 37°C and pH 8.0. Also, it has been found that bacterization of seeds of Vigna radiata with Castellaniella denitrificans SA13P increases germination rate. We have reported for the first time that Castellaniella denitrificans SA13P may be used as a novel strain for dye decolorization (malachite green and biological treatment of tannery effluent.

  15. Interactive performances of betaine on the metabolic processes of Pseudomonas denitrificans.

    Science.gov (United States)

    Xia, Wei; Peng, Wei-fu; Chen, Wei; Li, Kun-tai

    2015-02-01

    The performances of betaine on the metabolic processes of vitamin B12-producing Pseudomonas denitrificans were investigated in this paper. The results showed that betaine was an indispensable methyl-group donor for vitamin B12 biosynthesis, but large amounts of the extracellular glycine accompanied by betaine metabolism would impose a severe restriction on the cell growth of P. denitrificans. By further using a comparative metabolomics approach coupled with intracellular free amino acids analysis for the fermentation processes with betaine addition (10 g/l) or not, it was found that betaine could highly strengthen the formation of some key precursors and intermediates facilitating vitamin B12 biosynthesis, such as δ-aminolevulinic acid (ALA, the first precursor of vitamin B12), glutamate (an intermediate of ALA via C5 pathway), glycine (an intermediate of ALA via C4 pathway), and methionine (directly participating in the methylation reaction involved in vitamin B12 biosynthetic pathway). Therefore, the performances of betaine on P. denitrificans metabolic processes were not only serving as a decisive methyl-group donor for vitamin B12 biosynthesis, but also playing a powerfully promoting role in the generation of vitamin B12 precursors and intermediates.

  16. Isolation and analysis of the genes for cytochrome c oxidase in Paracoccus denitrificans

    OpenAIRE

    Raitio, Mirja; Jalli, Tuulikki; Saraste, Matti

    1987-01-01

    Synthetic oligonucleotide probes were used to clone two loci from the chromosomal DNA of Paracoccus denitrificans that contain the genes for cytochrome c oxidase (cytochrome aa3). One locus seems to contain four or five genes probably forming an operon. Two of these code for the oxidase subunits II and III. Three open reading frames are found between the COII and COIII genes. The other locus codes for the subunit I. A short open reading frame is found upstream of this gene. All three subunits...

  17. Achromobactor denitrificans SP1 produces pharmaceutically active 25C prodigiosin upon utilizing hazardous di(2-ethylhexyl)phthalate

    Science.gov (United States)

    Achromobacter denitrificans SP1 isolated from soil sludge heavily contaminated with plastic waste produced a novel pharmaceutically-active 25C prodigiosin analog during growth in a simple mineral salt medium supplemented with hazardous di(2-ethylhexyl)phthalate (DEHP) blended PVC plastics (in situ) ...

  18. Co-Cultures of Pseudomonas aeruginosa and Roseobacter denitrificans Reveal Shifts in Gene Expression Levels Compared to Solo Cultures

    Directory of Open Access Journals (Sweden)

    Crystal A. Conway

    2012-01-01

    Full Text Available Consistent biosynthesis of desired secondary metabolites (SMs from pure microbial cultures is often unreliable. In a proof-of-principle study to induce SM gene expression and production, we describe mixed “co-culturing” conditions and monitoring of messages via quantitative real-time PCR (qPCR. Gene expression of model bacterial strains (Pseudomonas aeruginosa PAO1 and Roseobacter denitrificans Och114 was analyzed in pure solo and mixed cocultures to infer the effects of interspecies interactions on gene expression in vitro, Two P. aeruginosa genes (PhzH coding for portions of the phenazine antibiotic pathway leading to pyocyanin (PCN and the RhdA gene for thiosulfate: cyanide sulfurtransferase (Rhodanese and two R. denitrificans genes (BetaLact for metallo-beta-lactamase and the DMSP gene for dimethylpropiothetin dethiomethylase were assessed for differential expression. Results showed that R. denitrificans DMSP and BetaLact gene expression became elevated in a mixed culture. In contrast, P. aeruginosa co-cultures with R. denitrificans or a third species did not increase target gene expression above control levels. This paper provides insight for better control of target SM gene expression in vitro and bypass complex genetic engineering manipulations.

  19. Bacterial community succession during the enrichment of chemolithoautotrophic arsenite oxidizing bacteria at high arsenic concentrations

    Institute of Scientific and Technical Information of China (English)

    Nguyen Ai Le; Akiko Sato; Daisuke Inoue; Kazunari Sei; Satoshi Soda; Michihiko Ike

    2012-01-01

    To generate cost-effective technologies for the removal of arsenic from water,we developed an enrichment culture of chemolithoautotrophic arsenite oxidizing bacteria (CAOs) that could effectively oxidize widely ranging concentrations of As(Ⅲ) to As(Ⅴ).In addition,we attempted to elucidate the enrichment process and characterize the microbial composition of the enrichment culture.A CAOs enrichment culture capable of stably oxidizing As(Ⅲ) to As(Ⅴ) was successfully constructed through repeated batch cultivation for more than 700 days,during which time the initial As(Ⅲ) concentrations were increased in a stepwise manner from l to 10-12 mmol/L.As(Ⅲ) oxidation activity of the enrichment culture gradually improved,and 10-12 mmol/L As(Ⅲ) was almost completely oxidized within four days.Terminal restriction fragment length polymorphism analysis showed that the dominant bacteria in the enrichment culture varied drastically during the enrichment process depending on the As(Ⅲ) concentration.Isolation and characterization of bacteria in the enrichment culture revealed that the presence of multiple CAOs with various As(Ⅲ) oxidation abilities enabled the culture to adapt to a wide range of As(Ⅲ) concentrations.The CAOs enrichment culture constructed here may he useful for pretreatment of water from which arsenic is being removed.

  20. Paracoccus denitrificans possesses two BioR homologs having a role in regulation of biotin metabolism.

    Science.gov (United States)

    Feng, Youjun; Kumar, Ritesh; Ravcheev, Dmitry A; Zhang, Huimin

    2015-08-01

    Recently, we determined that BioR, the GntR family of transcription factor, acts as a repressor for biotin metabolism exclusively distributed in certain species of α-proteobacteria, including the zoonotic agent Brucella melitensis and the plant pathogen Agrobacterium tumefaciens. However, the scenario is unusual in Paracoccus denitrificans, another closely related member of the same phylum α-proteobacteria featuring with denitrification. Not only does it encode two BioR homologs Pden_1431 and Pden_2922 (designated as BioR1 and BioR2, respectively), but also has six predictive BioR-recognizable sites (the two bioR homolog each has one site, whereas the two bio operons (bioBFDAGC and bioYB) each contains two tandem BioR boxes). It raised the possibility that unexpected complexity is present in BioR-mediated biotin regulation. Here we report that this is the case. The identity of the purified BioR proteins (BioR1 and BioR2) was confirmed with LC-QToF-MS. Phylogenetic analyses combined with GC percentage raised a possibility that the bioR2 gene might be acquired by horizontal gene transfer. Gel shift assays revealed that the predicted BioR-binding sites are functional for the two BioR homologs, in much similarity to the scenario seen with the BioR site of A. tumefaciens bioBFDAZ. Using the A. tumefaciens reporter system carrying a plasmid-borne LacZ fusion, we revealed that the two homologs of P. denitrificans BioR are functional repressors for biotin metabolism. As anticipated, not only does the addition of exogenous biotin stimulate efficiently the expression of bioYB operon encoding biotin transport/uptake system BioY, but also inhibits the transcription of the bioBFDAGC operon resembling the de novo biotin synthetic pathway. EMSA-based screening failed to demonstrate that the biotin-related metabolite is involved in BioR-DNA interplay, which is consistent with our former observation with Brucella BioR. Our finding defined a complex regulatory network for biotin

  1. Genome sequence of the chemolithoautotrophic nitrite-oxidizing bacterium Nitrobacter winogradskyi Nb-255

    Energy Technology Data Exchange (ETDEWEB)

    Hauser, Loren John [ORNL; Land, Miriam L [ORNL; Larimer, Frank W [ORNL; Arp, D J [Oregon State University; Hickey, W J [University of Wisconsin, Madison

    2006-03-01

    The alphaproteobacterium Nitrobacter winogradskyi (ATCC 25391) is a gram-negative facultative chemolithoautotroph capable of extracting energy from the oxidation of nitrite to nitrate. Sequencing and analysis of its genome revealed a single circular chromosome of 3,402,093 bp encoding 3,143 predicted proteins. There were extensive similarities to genes in two alphaproteobacteria, Bradyrhizobium japonicum USDA110 (1,300 genes) and Rhodopseudomonas palustris CGA009 CG (815 genes). Genes encoding pathways for known modes of chemolithotrophic and chemoorganotrophic growth were identified. Genes encoding multiple enzymes involved in anapleurotic reactions centered on C2 to C4 metabolism, including a glyoxylate bypass, were annotated. The inability of N. winogradskyi to grow on C6 molecules is consistent with the genome sequence, which lacks genes for complete Embden-Meyerhof and Entner-Doudoroff pathways, and active uptake of sugars. Two gene copies of the nitrite oxidoreductase, type I ribulose-1,5-bisphosphate carboxylase/oxygenase, cytochrome c oxidase, and gene homologs encoding an aerobic-type carbon monoxide dehydrogenase were present. Similarity of nitrite oxidoreductases to respiratory nitrate reductases was confirmed. Approximately 10% of the N. winogradskyi genome codes for genes involved in transport and secretion, including the presence of transporters for various organic-nitrogen molecules. The N. winogradskyi genome provides new insight into the phylogenetic identity and physiological capabilities of nitrite-oxidizing bacteria. The genome will serve as a model to study the cellular and molecular processes that control nitrite oxidation and its interaction with other nitrogen-cycling processes.

  2. Complete Genome Sequence of the Marine, Chemolithoautotrophic, Ammonia-Oxidizing Bacterium Nitrosococcus oceani ATCC 19707

    Energy Technology Data Exchange (ETDEWEB)

    Klots, Martin G. [University of Louisville, Louisville; Arp, D J [Oregon State University; Chain, Patrick S [ORNL; El-Sheikh, Amal F. [University of Louisville, Louisville; Hauser, Loren John [ORNL; Hommes, Norman G. [Oregon State University; Larimer, Frank W [ORNL; Malfatti, Stephanie [Lawrence Livermore National Laboratory (LLNL); Norton, Jeanette M. [Utah State University (USU); Poret-Peterson, Amisha T. [University of Louisville, Louisville; Vergez, Lisa [Lawrence Livermore National Laboratory (LLNL); Ward, Bess B. [Princeton University

    2006-01-01

    The gammaproteobacterium Nitrosococcus oceani (ATCC 19707) is a gram-negative obligate chemolithoautotroph capable of extracting energy and reducing power from the oxidation of ammonia to nitrite. Sequencing and annotation of the genome revealed a single circular chromosome (3,481,691 bp; G+C content of 50.4%) and a plasmid (40,420 bp) that contain 3,052 and 41 candidate protein-encoding genes, respectively. The genes encoding proteins necessary for the function of known modes of lithotrophy and autotrophy were identified. Contrary to betaproteobacterial nitrifier genomes, the N. oceani genome contained two complete rrn operons. In contrast, only one copy of the genes needed to synthesize functional ammonia monooxygenase and hydroxylamine oxidoreductase, as well as the proteins that relay the extracted electrons to a terminal electron acceptor, were identified. The N. oceani genome contained genes for 13 complete two-component systems. The genome also contained all the genes needed to reconstruct complete central pathways, the tricarboxylic acid cycle, and the Embden-Meyerhof-Parnass and pentose phosphate pathways. The N. oceani genome contains the genes required to store and utilize energy from glycogen inclusion bodies and sucrose. Polyphosphate and pyrophosphate appear to be integrated in this bacterium's energy metabolism, stress tolerance, and ability to assimilate carbon via gluconeogenesis. One set of genes for type I ribulose-1,5-bisphosphate carboxylase/oxygenase was identified, while genes necessary for methanotrophy and for carboxysome formation were not identified. The N. oceani genome contains two copies each of the genes or operons necessary to assemble functional complexes I and IV as well as ATP synthase (one H+-dependent F0F1 type, one Na+-dependent V type).

  3. The Complete Genome Sequence of the Marine, Chemolithoautotrophic, Ammonia-Oxidizing Bacterium Nitrosococcus oceani ATCC19707

    Energy Technology Data Exchange (ETDEWEB)

    Klotz, M G; Arp, D J; Chain, P S; El-Sheikh, A F; Hauser, L J; Hommes, N G; Larimer, F W; Malfatti, S A; Norton, J M; Poret-Peterson, A T; Vergez, L M; Ward, B B

    2006-08-03

    The Gammaproteobacterium, Nitrosococcus oceani (ATCC 19707), is a Gram-negative obligate chemolithoautotroph capable of extracting energy and reducing power from the oxidation of ammonia to nitrite. Sequencing and annotation of the genome revealed a single circular chromosome (3,481,691 bp; 50.4% G+C) and a plasmid (40,420 bp) that contain 3052 and 41 candidate protein-encoding genes, respectively. The genes encoding proteins necessary for the function of known modes of lithotrophy and autotrophy were identified. In contrast to betaproteobacterial nitrifier genomes, the N. oceani genome contained two complete rrn operons. In contrast, only one copy of the genes needed to synthesize functional ammonia monooxygenase and hydroxylamine oxidoreductase, as well as the proteins that relay the extracted electrons to a terminal electron acceptor were identified. The N. oceani genome contained genes for 13 complete two-component systems. The genome also contained all the genes needed to reconstruct complete central pathways, the tricarboxylic acid cycle and the Embden-Meyerhof-Parnass and pentose phosphate pathways. The N. oceani genome contains the genes required to store and utilize energy from glycogen inclusion bodies and sucrose. Polyphosphate and pyrophosphate appear to be integrated in this bacterium's energy metabolism, stress tolerance and the ability to assimilate carbon via gluconeogenesis. One set of genes for type I RuBisCO was identified, while genes necessary for methanotrophy and for carboxysome formation were not identified. The N. oceani genome contains two copies each of the genes or operons necessary to assemble functional complexes I and IV as well as ATP synthase (one H{sup +}-dependent F{sub 0}F{sub 1}-type, one Na{sup +}-dependent V-type).

  4. Electronic and vibrational spectroscopy of the cytochrome c:cytochrome c oxidase complexes from bovine and Paracoccus denitrificans.

    OpenAIRE

    Lynch, S. R.; Copeland, R. A.

    1992-01-01

    The 1:1 complex between horse heart cytochrome c and bovine cytochrome c oxidase, and between yeast cytochrome c and Paracoccus denitrificans cytochrome c oxidase have been studied by a combination of second derivative absorption, circular dichroism (CD), and resonance Raman spectroscopy. The second derivative absorption and CD spectra reveal changes in the electronic transitions of cytochrome a upon complex formation. These results could reflect changes in ground state heme structure or chan...

  5. Peroxidase activity and involvement in the oxidative stress response of roseobacter denitrificans truncated hemoglobin.

    Directory of Open Access Journals (Sweden)

    Yaya Wang

    Full Text Available Roseobacter denitrificans is a member of the widespread marine Roseobacter genus. We report the first characterization of a truncated hemoglobin from R. denitrificans (Rd. trHb that was purified in the heme-bound form from heterologous expression of the protein in Escherichia coli. Rd. trHb exhibits predominantly alpha-helical secondary structure and absorbs light at 412, 538 and 572 nm. The phylogenetic classification suggests that Rd. trHb falls into group II trHbs, whereas sequence alignments indicate that it shares certain important heme pocket residues with group I trHbs in addition to those of group II trHbs. The resonance Raman spectra indicate that the isolated Rd. trHb contains a ferric heme that is mostly 6-coordinate low-spin and that the heme of the ferrous form displays a mixture of 5- and 6-coordinate states. Two Fe-His stretching modes were detected, notably one at 248 cm-1, which has been reported in peroxidases and some flavohemoglobins that contain an Fe-His-Asp (or Glu catalytic triad, but was never reported before in a trHb. We show that Rd. trHb exhibits a significant peroxidase activity with a (kcat/Km value three orders of magnitude higher than that of bovine Hb and only one order lower than that of horseradish peroxidase. This enzymatic activity is pH-dependent with a pKa value ~6.8. Homology modeling suggests that residues known to be important for interactions with heme-bound ligands in group II trHbs from Mycobacterium tuberculosis and Bacillus subtilis are pointing toward to heme in Rd. trHb. Genomic organization and gene expression profiles imply possible functions for detoxification of reactive oxygen and nitrogen species in vivo. Altogether, Rd. trHb exhibits some distinctive features and appears equipped to help the bacterium to cope with reactive oxygen/nitrogen species and/or to operate redox biochemistry.

  6. Complete Genome Sequence of the Unclassified Iron-Oxidizing, Chemolithoautotrophic Burkholderiales Bacterium GJ-E10, Isolated from an Acidic River.

    Science.gov (United States)

    Fukushima, Jun; Tojo, Fuyumi; Asano, Ryoki; Kobayashi, Yayoi; Shimura, Yoichiro; Okano, Kunihiro; Miyata, Naoyuki

    2015-02-05

    Burkholderiales bacterium GJ-E10, isolated from the Tamagawa River in Akita Prefecture, Japan, is an unclassified, iron-oxidizing chemolithoautotrophic bacterium. Its single circular genome, consisting of 3,276,549 bp, was sequenced by using three types of next-generation sequencers and the sequences were then confirmed by PCR-based Sanger sequencing.

  7. Studies on the Glutathione-Dependent Formaldehyde-Activating Enzyme from Paracoccus denitrificans

    Science.gov (United States)

    Hopkinson, Richard J.; Leung, Ivanhoe K. H.; Smart, Tristan J.; Rose, Nathan R.; Henry, Luc; Claridge, Timothy D. W.; Schofield, Christopher J.

    2015-01-01

    Formaldehyde is a toxin and carcinogen that is both an environmental pollutant and an endogenous metabolite. Formaldehyde metabolism, which is probably essential for all aerobic cells, likely proceeds via multiple mechanisms, including via a glutathione-dependent pathway that is widely conserved in bacteria, plants and animals. However, it is unclear whether the first step in the glutathione-dependent pathway (i.e. formation of S-hydroxymethylglutathione (HMG)) is enzyme-catalysed. We report studies on glutathione-dependent formaldehyde-activating enzyme (GFA) from Paracoccus denitrificans, which has been proposed to catalyse HMG formation from glutathione and formaldehyde on the basis of studies using NMR exchange spectroscopy (EXSY). Although we were able to replicate the EXSY results, time course experiments unexpectedly imply that GFA does not catalyse HMG formation under standard conditions. However, GFA was observed to bind glutathione using NMR and mass spectrometry. Overall, the results reveal that GFA binds glutathione but does not directly catalyse HMG formation under standard conditions. Thus, it is possible that GFA acts as a glutathione carrier that acts to co-localise glutathione and formaldehyde in a cellular context. PMID:26675168

  8. Studies on the Glutathione-Dependent Formaldehyde-Activating Enzyme from Paracoccus denitrificans.

    Directory of Open Access Journals (Sweden)

    Richard J Hopkinson

    Full Text Available Formaldehyde is a toxin and carcinogen that is both an environmental pollutant and an endogenous metabolite. Formaldehyde metabolism, which is probably essential for all aerobic cells, likely proceeds via multiple mechanisms, including via a glutathione-dependent pathway that is widely conserved in bacteria, plants and animals. However, it is unclear whether the first step in the glutathione-dependent pathway (i.e. formation of S-hydroxymethylglutathione (HMG is enzyme-catalysed. We report studies on glutathione-dependent formaldehyde-activating enzyme (GFA from Paracoccus denitrificans, which has been proposed to catalyse HMG formation from glutathione and formaldehyde on the basis of studies using NMR exchange spectroscopy (EXSY. Although we were able to replicate the EXSY results, time course experiments unexpectedly imply that GFA does not catalyse HMG formation under standard conditions. However, GFA was observed to bind glutathione using NMR and mass spectrometry. Overall, the results reveal that GFA binds glutathione but does not directly catalyse HMG formation under standard conditions. Thus, it is possible that GFA acts as a glutathione carrier that acts to co-localise glutathione and formaldehyde in a cellular context.

  9. Genome analysis and physiological comparison of Alicycliphilus denitrificans strains BC and K601T

    Energy Technology Data Exchange (ETDEWEB)

    Oosterkamp, Margreet J. [Wageningen University and Research Centre, The Netherlands; Veuskens, Teun [Wageningen University and Research Centre, The Netherlands; Saia, Flavia Talarico [Wageningen University and Research Centre, The Netherlands; Weelink, Sander A.B. [Wageningen University and Research Centre, The Netherlands; Goodwin, Lynne A. [Los Alamos National Laboratory (LANL); Daligault, Hajnalka E. [Los Alamos National Laboratory (LANL); Bruce, David [Los Alamos National Laboratory (LANL); Detter, J. Chris [U.S. Department of Energy, Joint Genome Institute; Tapia, Roxanne [Los Alamos National Laboratory (LANL); Han, Cliff [Los Alamos National Laboratory (LANL); Land, Miriam L [ORNL; Hauser, Loren John [ORNL; Langenhoff, A. M. [Deltares, The Netherlands; Gerritse, Jan [Deltares, The Netherlands; Van Berkel, Willem J. H. [Wageningen University and Research Centre, The Netherlands; Pieper, Dietmar [HZI - Helmholtz Centre for Infection Research, Braunschweig, Germany; Junca, Howard [CorpoGen, Bogota Colombia; Smidt, Hauke [Wageningen University and Research Centre, The Netherlands; Schraa, Gosse [Wageningen University and Research Centre, The Netherlands; Davids, Mark [Wageningen University and Research Centre, The Netherlands; Schaap, Peter J [Wageningen University and Research Centre, The Netherlands; Plugge, Caroline M. [Wageningen University and Research Centre, The Netherlands; Stams, Alfons J. M. [Wageningen University and Research Centre, The Netherlands

    2013-01-01

    The genomes of the Betaproteobacteria Alicycliphilus denitrificans strains BC and K601T have been sequenced to get insight into the physiology of the two strains. Strain BC degrades benzene with chlorate as electron acceptor. The cyclohexanol-degrading denitrifying strain K601T is not able to use chlorate as electron acceptor, while strain BC cannot degrade cyclohexanol. The 16S rRNA sequences of strains BC and K601T are identical and the fatty acid methyl ester patterns of the strains are similar. Basic Local Alignment Search Tool (BLAST) analysis of predicted open reading frames of both strains showed most hits with Acidovorax sp. JS42, a bacterium that degrades nitro-aromatics. The genomes include strain-specific plasmids (pAlide201 in strain K601T and pAlide01 and pAlide02 in strain BC). Key genes of chlorate reduction in strain BC were located on a 120 kb megaplasmid (pAlide01), which was absent in strain K601T. Genes involved in cyclohexanol degradation were only found in strain K601T. Benzene and toluene are degraded via oxygenase-mediated pathways in both strains. Genes involved in the meta-cleavage pathway of catechol are present in the genomes of both strains. Strain BC also contains all genes of the ortho-cleavage pathway. The large number of mono- and dioxygenase genes in the genomes suggests that the two strains have a broader substrate range than known thus far.

  10. Phospholipid Fatty Acids as Physiological Indicators of Paracoccus denitrificans Encapsulated in Silica Sol-Gel Hydrogels

    Directory of Open Access Journals (Sweden)

    Josef Trögl

    2015-02-01

    Full Text Available The phospholipid fatty acid (PLFA content was determined in samples of Paracoccus denitrificans encapsulated in silica hydrogel films prepared from prepolymerized tetramethoxysilane (TMOS. Immediately after encapsulation the total PLFA concentration was linearly proportional to the optical density (600 nm of the input microbial suspension (R2 = 0.99. After 7 days this relationship remained linear, but with significantly decreased slope, indicating a higher extinction of bacteria in suspensions of input concentration 108 cells/mL and higher. trans-Fatty acids, indicators of cytoplasmatic membrane disturbances, were below the detection limit. The cy/pre ratio (i.e., ratio of cyclopropylated fatty acids (cy17:0 + cy19:0 to their metabolic precursors (16:1ω7 + 18:1ω7, an indicator of the transition of the culture to a stationary growth-phase, decreased depending on co-immobilization of nutrients in the order phosphate buffer > mineral medium > Luria Broth rich medium. The ratio, too, was logarithmically proportional to cell concentration. These results confirm the applicability of total PLFA as an indicator for the determination of living biomass and cy/pre ratio for determination of nutrient limitation of microorganisms encapsulated in sol-gel matrices. This may be of interest for monitoring of sol-gel encapsulated bacteria proposed as optical recognition elements in biosensor construction, as well as other biotechnological applications.

  11. Investigating the nitrification and denitrification kinetics under aerobic and anaerobic conditions by Paracoccus denitrificans ISTOD1.

    Science.gov (United States)

    Medhi, Kristina; Singhal, Anjali; Chauhan, D K; Thakur, Indu Shekhar

    2017-03-16

    Municipal wastewater contains multiple nitrogen contaminants such as ammonia, nitrate and nitrite. Two heterotrophic nitrifier and aerobic denitrifiers, bacterial isolates ISTOD1 and ISTVD1 were isolated from domestic wastewater. On the basis of removal efficiency of ammonia, nitrate and nitrite under both aerobic and anaerobic conditions, ISTOD1 was selected and identified as Paracoccus denitrificans. Aerobically, NH4(+)-N had maximum specific nitrogen removal rate (Rxi) of 7.6g/gDCW/h and anaerobically, NO3(-)N showed Rxi of 2.5*10(-1)g/g DCW/h. Monod equation described the bioprocess kinetic coefficients, µmax and Ks, obtained by regression. Error functions were calculated to validate the Monod equation experimental data. Aerobic NO3(-)N showed the highest YW of 0.372mg DCW/mg NO3(-)N among the five conditions. ISTOD1 serves as a potential candidate for treating nitrogen rich wastewater using simultaneous nitrification and aerobic denitrification. It can be used in bioaugmentation studies under varied condition.

  12. The Inhibitory Mechanism of the ζ Subunit of the F1FO-ATPase Nanomotor of Paracoccus denitrificans and Related α-Proteobacteria*

    OpenAIRE

    García-Trejo, José J.; Zarco-Zavala, Mariel; Mendoza-Hoffmann, Francisco; Hernández-Luna, Eduardo; Ortega, Raquel; Mendoza-Hernández, Guillermo

    2015-01-01

    The ζ subunit is a novel inhibitor of the F1FO-ATPase of Paracoccus denitrificans and related α-proteobacteria. It is different from the bacterial (ϵ) and mitochondrial (IF1) inhibitors. The N terminus of ζ blocks rotation of the γ subunit of the F1-ATPase of P. denitrificans (Zarco-Zavala, M., Morales-Ríos, E., Mendoza-Hernández, G., Ramírez-Silva, L., Pérez-Hernández, G., and García-Trejo, J. J. (2014) FASEB J. 24, 599–608) by a hitherto unknown quaternary structure that was first modeled h...

  13. Respiratory Complex I in Bos taurus and Paracoccus denitrificans Pumps Four Protons across the Membrane for Every NADH Oxidized.

    Science.gov (United States)

    Jones, Andrew J Y; Blaza, James N; Varghese, Febin; Hirst, Judy

    2017-03-24

    Respiratory complex I couples electron transfer between NADH and ubiquinone to proton translocation across an energy-transducing membrane to support the proton-motive force that drives ATP synthesis. The proton-pumping stoichiometry of complex I (i.e. the number of protons pumped for each two electrons transferred) underpins all mechanistic proposals. However, it remains controversial and has not been determined for any of the bacterial enzymes that are exploited as model systems for the mammalian enzyme. Here, we describe a simple method for determining the proton-pumping stoichiometry of complex I in inverted membrane vesicles under steady-state ADP-phosphorylating conditions. Our method exploits the rate of ATP synthesis, driven by oxidation of NADH or succinate with different sections of the respiratory chain engaged in catalysis as a proxy for the rate of proton translocation and determines the stoichiometry of complex I by reference to the known stoichiometries of complexes III and IV. Using vesicles prepared from mammalian mitochondria (from Bos taurus) and from the bacterium Paracoccus denitrificans, we show that four protons are pumped for every two electrons transferred in both cases. By confirming the four-proton stoichiometry for mammalian complex I and, for the first time, demonstrating the same value for a bacterial complex, we establish the utility of P. denitrificans complex I as a model system for the mammalian enzyme. P. denitrificans is the first system described in which mutagenesis in any complex I core subunit may be combined with quantitative proton-pumping measurements for mechanistic studies.

  14. The nitric-oxide reductase from Paracoccus denitrificans uses a single specific proton pathway.

    Science.gov (United States)

    ter Beek, Josy; Krause, Nils; Reimann, Joachim; Lachmann, Peter; Ädelroth, Pia

    2013-10-18

    The NO reductase from Paracoccus denitrificans reduces NO to N2O (2NO + 2H(+) + 2e(-) → N2O + H2O) with electrons donated by periplasmic cytochrome c (cytochrome c-dependent NO reductase; cNOR). cNORs are members of the heme-copper oxidase superfamily of integral membrane proteins, comprising the O2-reducing, proton-pumping respiratory enzymes. In contrast, although NO reduction is as exergonic as O2 reduction, there are no protons pumped in cNOR, and in addition, protons needed for NO reduction are derived from the periplasmic solution (no contribution to the electrochemical gradient is made). cNOR thus only needs to transport protons from the periplasm into the active site without the requirement to control the timing of opening and closing (gating) of proton pathways as is needed in a proton pump. Based on the crystal structure of a closely related cNOR and molecular dynamics simulations, several proton transfer pathways were suggested, and in principle, these could all be functional. In this work, we show that residues in one of the suggested pathways (denoted pathway 1) are sensitive to site-directed mutation, whereas residues in the other proposed pathways (pathways 2 and 3) could be exchanged without severe effects on turnover activity with either NO or O2. We further show that electron transfer during single-turnover reduction of O2 is limited by proton transfer and can thus be used to study alterations in proton transfer rates. The exchange of residues along pathway 1 showed specific slowing of this proton-coupled electron transfer as well as changes in its pH dependence. Our results indicate that only pathway 1 is used to transfer protons in cNOR.

  15. Cloning, expression and characterization of D-aminoacylase from Achromobacter xylosoxidans subsp. denitrificans ATCC 15173.

    Science.gov (United States)

    Wang, Wei; Xi, Huange; Bi, Qirui; Hu, Ying; Zhang, Yang; Ni, Mengxiang

    2013-07-19

    D-Aminoacylase catalyzes the conversion of N-acyl-D-amino acids to d-amino acids and fatty acids. The aim of this study was to identify the D-aminoacylase gene from Achromobacter xylosoxidans subsp. denitrificans ATCC 15173 and investigate the biochemical characterization of the enzyme. A previously uncharacterized D-aminoacylase gene (ADdan) from this organism was cloned and sequenced. The open reading frame (ORF) of ADdan was 1467 bp in size encoding a 488-amino acid polypeptide. ADdan, with a high amino acid similarity to N-acyl-D-aspartate amidohydrolase from Alcaligenes A6, showed relatively low sequence similarities to other characterized D-aminoacylases. The recombinant ADdan protein was expressed in Escherichia coli BL21 (DE3) using pET-28a with a T7 promoter. The enzyme was purified in a single chromatographic step using nickel affinity gel column. The molecular mass of the expressed protein, calculated by SDS-PAGE, was about 52 kDa. The purified ADdan showed optimal activity at pH 8.0 and 50°C, and was stable at pH 6.0-8.0 and up to 45°C. Its activity was inhibited by Cu(2+), Fe(2+), Ca(2+), Mn(2+), Ni(2+), Zn(2+) and Hg(2+), whereas Mg(2+) had no significant influence on this recombinant D-aminoacylase. This is the first report on the characterization of D-aminoacylase with activity towards both N-acyl derivatives of neutral D-amino acids and N-acyl-D-aspartate. The characteristics of ADdan could prove to be of interest in industrial production of D-amino acids.

  16. Tightly bound nucleotides of the energy-transducing ATPase, and their role in oxidative phosphorylation. I. The Paracoccus denitrificans system.

    Science.gov (United States)

    Harris, D A; John, P; Radda, G K

    1977-03-11

    1. The coupling ATPase of Paracoccus denitrificans can be removed from the membrane by washing coupled membrane fragments at low salt concentrations. 2. This ATPase resembles coupling ATPases of mitochondria, chloroplasts and other bacteria. It is a negatively charged protein of molecular weight about 300,000. An inhibitor protein in bound tightly to the ATPase in vivo, and can be destroyed by trypsin treatment. 3. ATP and ADP are found tightly bound to the coupling ATPase of P. denitrificans, both in its membrane-bound and isolated state. The ATP/ADP ratio on the enzyme is greater than one. 4. Under de-energised condtions, the bound nucleotides are not available to the suspending medium. When the membrane is energised however, the bound nucleotides can exchange with added nucleotides and incorporate 32Pi. 32Ppi is incorporated into the beta and gamma positions of the bound nucleotides, but beta-labelling probably does not occur on the coupling ATPase. 5. Uncouplers inhibit the exchange of the free nucleotides or 32Pi into the bound nucleotides, while venturicidin (an energy transfer inhibitor) and aurovertin stimulate the exchange. 6. The response of the bound nucleotides to energisation is consistent with their being involved directly in the mechanism of oxidative phosphorylation.

  17. Assembly of respiratory complexes I, III, and IV into NADH oxidase supercomplex stabilizes complex I in Paracoccus denitrificans.

    Science.gov (United States)

    Stroh, Anke; Anderka, Oliver; Pfeiffer, Kathy; Yagi, Takao; Finel, Moshe; Ludwig, Bernd; Schägger, Hermann

    2004-02-01

    Stable supercomplexes of bacterial respiratory chain complexes III (ubiquinol:cytochrome c oxidoreductase) and IV (cytochrome c oxidase) have been isolated as early as 1985 (Berry, E. A., and Trumpower, B. L. (1985) J. Biol. Chem. 260, 2458-2467). However, these assemblies did not comprise complex I (NADH:ubiquinone oxidoreductase). Using the mild detergent digitonin for solubilization of Paracoccus denitrificans membranes we could isolate NADH oxidase, assembled from complexes I, III, and IV in a 1:4:4 stoichiometry. This is the first chromatographic isolation of a complete "respirasome." Inactivation of the gene for tightly bound cytochrome c552 did not prevent formation of this supercomplex, indicating that this electron carrier protein is not essential for structurally linking complexes III and IV. Complex I activity was also found in the membranes of mutant strains lacking complexes III or IV. However, no assembled complex I but only dissociated subunits were observed following the same protocols used for electrophoretic separation or chromatographic isolation of the supercomplex from the wild-type strain. This indicates that the P. denitrificans complex I is stabilized by assembly into the NADH oxidase supercomplex. In addition to substrate channeling, structural stabilization of a membrane protein complex thus appears as one of the major functions of respiratory chain supercomplexes.

  18. Genes and pathways for CO2 fixation in the obligate, chemolithoautotrophic acidophile, Acidithiobacillus ferrooxidans, Carbon fixation in A. ferrooxidans

    Directory of Open Access Journals (Sweden)

    Esparza Mario

    2010-08-01

    Full Text Available Abstract Background Acidithiobacillus ferrooxidans is chemolithoautotrophic γ-proteobacterium that thrives at extremely low pH (pH 1-2. Although a substantial amount of information is available regarding CO2 uptake and fixation in a variety of facultative autotrophs, less is known about the processes in obligate autotrophs, especially those living in extremely acidic conditions, prompting the present study. Results Four gene clusters (termed cbb1-4 in the A. ferrooxidans genome are predicted to encode enzymes and structural proteins involved in carbon assimilation via the Calvin-Benson-Bassham (CBB cycle including form I of ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO, EC 4.1.1.39 and the CO2-concentrating carboxysomes. RT-PCR experiments demonstrated that each gene cluster is a single transcriptional unit and thus is an operon. Operon cbb1 is divergently transcribed from a gene, cbbR, encoding the LysR-type transcriptional regulator CbbR that has been shown in many organisms to regulate the expression of RubisCO genes. Sigma70-like -10 and -35 promoter boxes and potential CbbR-binding sites (T-N11-A/TNA-N7TNA were predicted in the upstream regions of the four operons. Electrophoretic mobility shift assays (EMSAs confirmed that purified CbbR is able to bind to the upstream regions of the cbb1, cbb2 and cbb3 operons, demonstrating that the predicted CbbR-binding sites are functional in vitro. However, CbbR failed to bind the upstream region of the cbb4 operon that contains cbbP, encoding phosphoribulokinase (EC 2.7.1.19. Thus, other factors not present in the assay may be required for binding or the region lacks a functional CbbR-binding site. The cbb3 operon contains genes predicted to encode anthranilate synthase components I and II, catalyzing the formation of anthranilate and pyruvate from chorismate. This suggests a novel regulatory connection between CO2 fixation and tryptophan biosynthesis. The presence of a form II Rubis

  19. Structure of apo-azurin from Alcaligenes denitrificans at 1.8 A resolution.

    Science.gov (United States)

    Shepard, W E; Kingston, R L; Anderson, B F; Baker, E N

    1993-05-01

    The structure of apo-azurin from Alcaligenes denitrificans has been determined at high resolution by X-ray crystallography. Two separate structure analyses have been carried out, (i) on crystals obtained from solutions of apo-azurin and (ii) on crystals obtained by removal of copper from previously formed crystals of holo-azurin. Data to 1.8 A resolution were collected from the apo-azurin crystals, by Weissenberg photography (with image plates) using synchrotron radiation and by diffractometry, and the structure was refined by restrained least-squares methods to a final R value of 0.160 for all data in the range 10.0-1.8 A. The final model of 1954 protein atoms, 246 water molecules (66 half-weighted), four SO(4)(2-) ions, and two low-occupancy (0.13 and 0.15) Cu atoms has r.m.s. deviations of 0.012, 0.045 and 0.013 A from standard bond lengths, angle distances and planar groups. For copper-removed azurin, data to 2.2 A were collected by diffractometry and the structure refined by restrained least squares to a final R value of 0.158 for all data in the range 10.0-2.2 A. The final model of 1954 protein atoms, 264 water molecules, two SO(4)(2-) ions, two low occupancy (0.18 and 0.22) metal atoms and one unidentified atom (modelled as S) has r.m.s. deviations of 0.013, 0.047 and 0.012 A from standard bond lengths, angle distances and planar groups. The two structures are essentially identical to each other and show no significant differences from the oxidized and reduced holo-azurin structures. The ligand side chains move slightly closer together following the removal of copper, with the radius of the cavity between the three strongly binding ligands, His 46, His 117 and Cys 112, shrinking from 1.31 A in reduced azurin to 1.24 A in oxidized azurin and 1.16 A in apo-azurin. There is a suggestion of increased flexibility in one of the copper-binding loops but the structure supports the view that the copper site found in holo-azurin is a stable structure, defined by the

  20. Thiobacter subterraneus gen. nov., sp. nov., an obligately chemolithoautotrophic, thermophilic, sulfur-oxidizing bacterium from a subsurface hot aquifer.

    Science.gov (United States)

    Hirayama, Hisako; Takai, Ken; Inagaki, Fumio; Nealson, Kenneth H; Horikoshi, Koki

    2005-01-01

    A novel, thermophilic, obligately chemolithoautotrophic, sulfur/thiosulfate-oxidizing bacterium was isolated from subsurface geothermal aquifer water (temperature approximately 70 degrees C) in the Hishikari gold mine, Japan. Cells of the isolate, designated strain C55T, were motile, straight rods with a single polar flagellum. Growth was observed at temperatures between 35 and 62 degrees C (optimum 50-55 degrees C; 60 min doubling time) and pH between 5.2 and 7.7 (optimum pH 6.5-7.0). High growth rate of strain C55T was observed on either thiosulfate or elemental sulfur as a sole energy source, with molecular oxygen as the only electron acceptor. None of the organic compounds tested supported or stimulated growth of strain C55T. The G+C content of the genomic DNA was 66.9 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain C55T was affiliated to the beta-Proteobacteria, but was distantly related to recognized genera. On the basis of its physiological and molecular properties, strain C55T (=JCM12421T=DSM 16629T=ATCC BAA-941T) is proposed as the type strain of Thiobacter subterraneus gen. nov., sp. nov.

  1. Complete genome sequence of the facultatively chemolithoautotrophic and methylotrophic alpha Proteobacterium Starkeya novella type strain (ATCC 8093T)

    Energy Technology Data Exchange (ETDEWEB)

    Kappler, Ulrike [University of Queensland, The, Brisbane, Queensland, Australia; Davenport, Karen W. [Los Alamos National Laboratory (LANL); Beatson, Scott [University of Queensland, The, Brisbane, Queensland, Australia; Lucas, Susan [U.S. Department of Energy, Joint Genome Institute; Lapidus, Alla L. [U.S. Department of Energy, Joint Genome Institute; Copeland, A [U.S. Department of Energy, Joint Genome Institute; Berry, Kerrie W. [U.S. Department of Energy, Joint Genome Institute; Glavina Del Rio, Tijana [U.S. Department of Energy, Joint Genome Institute; Hammon, Nancy [U.S. Department of Energy, Joint Genome Institute; Dalin, Eileen [U.S. Department of Energy, Joint Genome Institute; Tice, Hope [U.S. Department of Energy, Joint Genome Institute; Pitluck, Sam [U.S. Department of Energy, Joint Genome Institute; Richardson, P M [U.S. Department of Energy, Joint Genome Institute; Bruce, David [Los Alamos National Laboratory (LANL); Goodwin, Lynne A. [Los Alamos National Laboratory (LANL); Han, Cliff [Los Alamos National Laboratory (LANL); Tapia, Roxanne [Los Alamos National Laboratory (LANL); Detter, J. Chris [U.S. Department of Energy, Joint Genome Institute; Chang, Yun-Juan [ORNL; Jeffries, Cynthia [Oak Ridge National Laboratory (ORNL); Land, Miriam L [ORNL; Hauser, Loren John [ORNL; Kyrpides, Nikos C [U.S. Department of Energy, Joint Genome Institute; Goker, Markus [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Ivanova, N [U.S. Department of Energy, Joint Genome Institute; Klenk, Hans-Peter [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Woyke, Tanja [U.S. Department of Energy, Joint Genome Institute

    2012-01-01

    Starkeya novella (Starkey 1934) Kelly et al. 2000 is a member of the family Xanthobacteraceae in the order Rhizobiales , which is thus far poorly characterized at the genome level. Cultures from this spe- cies are most interesting due to their facultatively chemolithoautotrophic lifestyle, which allows them to both consume carbon dioxide and to produce it. This feature makes S. novella an interesting model or- ganism for studying the genomic basis of regulatory networks required for the switch between con- sumption and production of carbon dioxide, a key component of the global carbon cycle. In addition, S. novella is of interest for its ability to grow on various inorganic sulfur compounds and several C1- compounds such as methanol. Besides Azorhizobium caulinodans, S. novella is only the second spe- cies in the family Xanthobacteraceae with a completely sequenced genome of a type strain. The cur- rent taxonomic classification of this group is in significant conflict with the 16S rRNA data. The ge- nomic data indicate that the physiological capabilities of the organism might have been underestimat- ed. The 4,765,023 bp long chromosome with its 4,511 protein-coding and 52 RNA genes was se- quenced as part of the DOE Joint Genome Institute Community Sequencing Program (CSP) 2008.

  2. Thiocyanate hydrolase, the primary enzyme initiating thiocyanate degradation in the novel obligately chemolithoautotrophic halophilic sulfur-oxidizing bacterium Thiohalophilus thiocyanoxidans.

    Science.gov (United States)

    Bezsudnova, Ekaterina Yu; Sorokin, Dimitry Yu; Tikhonova, Tamara V; Popov, Vladimir O

    2007-12-01

    Thiohalophilus thiocyanoxidans is a first halophilic sulfur-oxidizing chemolithoautotrophic bacterium capable of growth with thiocyanate as an electron donor at salinity up to 4 M NaCl. The cells, grown with thiocyanate, but not with thiosulfate, contained an enzyme complex hydrolyzing thiocyanate to sulfide and ammonia under anaerobic conditions with carbonyl sulfide as an intermediate. Despite the fact of utilization of the , high cyanase activity was also detected in thiocyanate-induced cells. Three-stage column chromotography resulted in a highly purified thiocyanate-hydrolyzing protein with an apparent molecular mass of 140 kDa that consists of three subunits with masses 17, 19 and 29 kDa. The enzyme is a Co,Fe-containing protein resembling on its function and subunit composition the enzyme thiocyanate hydrolase from the Betaproteobacterium Thiobacillus thioparus. Cyanase, copurified with thiocyanate hydrolase, is a bisubstrate multisubunit enzyme with an apparent subunit molecular mass of 14 kDa. A possible role of cyanase in thiocyanate degradation by T. thiocyanoxidans is discussed.

  3. Electrochemistry suggests proton access from the exit site to the binuclear center in Paracoccus denitrificans cytochrome c oxidase pathway variants.

    Science.gov (United States)

    Meyer, Thomas; Melin, Frédéric; Richter, Oliver-M H; Ludwig, Bernd; Kannt, Aimo; Müller, Hanne; Michel, Hartmut; Hellwig, Petra

    2015-02-27

    Two different pathways through which protons access cytochrome c oxidase operate during oxygen reduction from the mitochondrial matrix, or the bacterial cytoplasm. Here, we use electrocatalytic current measurements to follow oxygen reduction coupled to proton uptake in cytochrome c oxidase isolated from Paracoccus denitrificans. Wild type enzyme and site-specific variants with defects in both proton uptake pathways (K354M, D124N and K354M/D124N) were immobilized on gold nanoparticles, and oxygen reduction was probed electrochemically in the presence of varying concentrations of Zn(2+) ions, which are known to inhibit both the entry and the exit proton pathways in the enzyme. Our data suggest that under these conditions substrate protons gain access to the oxygen reduction site via the exit pathway.

  4. Protoplast fusion technology for improved production of coenzyme Q10 using Paracoccus denitrificans ATCC 19367 mutant strains

    Directory of Open Access Journals (Sweden)

    Pradipta Tokdar

    2014-01-01

    Full Text Available Normal 0 false false false EN-US X-NONE X-NONE MicrosoftInternetExplorer4 Induced mutants generated from Paracoccus denitrificans ATCC 19367 having antibiotic resistant markers, were used as parent strains to carry out protoplast fusion. The generated fusants were screened using standardized protocol for CoQ10 production. Among the generated fusants, one fusant namely PF-P1 showed 1.73 folds enhancements in specific CoQ10 content than wild type strain. Fusant PF-P1 was characterized by biochemical and molecular approaches where it showed differences than wild type strain. The fusant was further identified by 16S rRNA gene sequence analysis that showed eight nucleotide base pair mutation on conserved region and 99% homology with Paracoccus denitrificans strains. /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:minor-bidi;}

  5. Catabolic and anabolic energy for chemolithoautotrophs in deep-sea hydrothermal systems hosted in different rock types

    Science.gov (United States)

    Amend, Jan P.; McCollom, Thomas M.; Hentscher, Michael; Bach, Wolfgang

    2011-10-01

    Active deep-sea hydrothermal vents are hosted by a range of different rock types, including basalt, peridotite, and felsic rocks. The associated hydrothermal fluids exhibit substantial chemical variability, which is largely attributable to compositional differences among the underlying host rocks. Numerical models were used to evaluate the energetics of seven inorganic redox reactions (potential catabolisms of chemolithoautotrophs) and numerous biomolecule synthesis reactions (anabolism) in a representative sampling of these systems, where chemical gradients are established by mixing hydrothermal fluid with seawater. The wide ranging fluid compositions dictate demonstrable differences in Gibbs energies (Δ G r) of these catabolic and anabolic reactions in three peridotite-hosted, six basalt-hosted, one troctolite-basalt hybrid, and two felsic rock-hosted systems. In peridotite-hosted systems at low to moderate temperatures (10), hydrogen oxidation yields the most catabolic energy, but the oxidation of methane, ferrous iron, and sulfide can also be moderately exergonic. At higher temperatures, and consequent SW:HF mixing ratios catabolic energy source at all temperatures (and SW:HF ratios) considered. The energetics of catabolism at the troctolite-basalt hybrid system were intermediate to these extremes. Reaction energetics for anabolism in chemolithoautotrophs—represented here by the synthesis of amino acids, nucleotides, fatty acids, saccharides, and amines—were generally most favorable at moderate temperatures (22-32 °C) and corresponding SW:HF mixing ratios (˜15). In peridotite-hosted and the troctolite-basalt hybrid systems, Δ G r for primary biomass synthesis yielded up to ˜900 J per g dry cell mass. The energetics of anabolism in basalt- and felsic rock-hosted systems were far less favorable. The results suggest that in peridotite-hosted (and troctolite-basalt hybrid) systems, compared with their basalt (and felsic rock) counterparts, microbial

  6. Structural and functional analysis of aa3-type and cbb3-type cytochrome c oxidases of Paracoccus denitrificans reveals significant differences in proton-pump design

    OpenAIRE

    de Gier, Jan-Willem L.; Schepper, Mike; Reijnders, Willem N.M.; Dyck, Stef J. van; Slotboom, Dirk Jan; Warne, Antony; Saraste, Matti; Krab, Klaas; Finel, Moshe; Stouthamer, Adriaan H.; Van Spanning, Rob J. M.; van der Oost, John

    1996-01-01

    In Paracoccus denitrificans the aa3-type cytochrome c oxidase and the bb3-type quinol oxidase have previously been characterized in detail, both biochemically and genetically. Here we report on the isolation of a genomic locus that harbours the gene cluster ccoNOQP, and demonstrate that it encodes an alternative cbb3-type cytochrome c oxidase. This oxidase has previously been shown to be specifically induced at low oxygen tensions, suggesting that its expression is controlled by an oxygen-sen...

  7. Cytochromes c550, c552, and c1 in the Electron Transport Network of Paracoccus denitrificans: Redundant or Subtly Different in Function?

    OpenAIRE

    Otten, M.F.; Oost, van der, J.; Reijnders, W. N. M.; Westerhoff, H.V.; Ludwig, B.; Spanning, van, R.J.M.

    2001-01-01

    Paracoccus denitrificans strains with mutations in the genes encoding the cytochrome c550, c552, or c1 and in combinations of these genes were constructed, and their growth characteristics were determined. Each mutant was able to grow heterotrophically with succinate as the carbon and free-energy source, although their specific growth rates and maximum cell numbers fell variably behind those of the wild type. Maximum cell numbers and rates of growth were also reduced when these strains were g...

  8. Structure of a catalytic dimer of the α- and β-subunits of the F-ATPase from Paracoccus denitrificans at 2.3 Å resolution

    Energy Technology Data Exchange (ETDEWEB)

    Morales-Ríos, Edgar; Montgomery, Martin G. [The Medical Research Council Mitochondrial Biology Unit, Cambridge Biomedical Campus, Hills Road, Cambridge CB2 0XY (United Kingdom); Leslie, Andrew G. W. [The Medical Research Council Laboratory of Molecular Biology, Cambridge Biomedical Campus, Francis Crick Avenue, Cambridge CB2 0QH (United Kingdom); García-Trejo, José J. [Universidad Nacional Autónoma de México, Mexico City (Mexico); Walker, John E., E-mail: walker@mrc-mbu.cam.ac.uk [The Medical Research Council Mitochondrial Biology Unit, Cambridge Biomedical Campus, Hills Road, Cambridge CB2 0XY (United Kingdom)

    2015-09-23

    The structure of the αβ heterodimer of the F-ATPase from the α-proteobacterium P. denitrificans has been determined at 2.3 Å resolution. It corresponds to the ‘open’ or ‘empty’ catalytic interface found in other F-ATPases. The structures of F-ATPases have predominantly been determined from mitochondrial enzymes, and those of the enzymes in eubacteria have been less studied. Paracoccus denitrificans is a member of the α-proteobacteria and is related to the extinct protomitochondrion that became engulfed by the ancestor of eukaryotic cells. The P. denitrificans F-ATPase is an example of a eubacterial F-ATPase that can carry out ATP synthesis only, whereas many others can catalyse both the synthesis and the hydrolysis of ATP. Inhibition of the ATP hydrolytic activity of the P. denitrificans F-ATPase involves the ζ inhibitor protein, an α-helical protein that binds to the catalytic F{sub 1} domain of the enzyme. This domain is a complex of three α-subunits and three β-subunits, and one copy of each of the γ-, δ- and ∊-subunits. Attempts to crystallize the F{sub 1}–ζ inhibitor complex yielded crystals of a subcomplex of the catalytic domain containing the α- and β-subunits only. Its structure was determined to 2.3 Å resolution and consists of a heterodimer of one α-subunit and one β-subunit. It has no bound nucleotides, and it corresponds to the ‘open’ or ‘empty’ catalytic interface found in other F-ATPases. The main significance of this structure is that it aids in the determination of the structure of the intact membrane-bound F-ATPase, which has been crystallized.

  9. Structure of the two-domain hexameric APS kinase from Thiobacillus denitrificans: structural basis for the absence of ATP sulfurylase activity

    Energy Technology Data Exchange (ETDEWEB)

    Gay, Sean C. [Department of Chemistry, University of California, One Shields Avenue, Davis, CA 95616 (United States); Segel, Irwin H. [Section of Molecular and Cellular Biology, University of California, One Shields Avenue, Davis, CA 95616 (United States); Fisher, Andrew J., E-mail: fisher@chem.ucdavis.edu [Department of Chemistry, University of California, One Shields Avenue, Davis, CA 95616 (United States); Section of Molecular and Cellular Biology, University of California, One Shields Avenue, Davis, CA 95616 (United States)

    2009-10-01

    APS kinase from Thiobacillus denitrificans contains an inactive N-terminal ATP sulfurylase domain. The structure presented unveils the first hexameric assembly for an APS kinase, and reveals that structural changes in the N-terminal domain disrupt the ATP sulfurylase active site thus prohibiting activity. The Tbd-0210 gene of the chemolithotrophic bacterium Thiobacillus denitrificans is annotated to encode a 60.5 kDa bifunctional enzyme with ATP sulfurylase and APS kinase activity. This putative bifunctional enzyme was cloned, expressed and structurally characterized. The 2.95 Å resolution X-ray crystal structure reported here revealed a hexameric assembly with D{sub 3} symmetry. Each subunit contains a large N-terminal sulfurylase-like domain and a C-terminal APS kinase domain reminiscent of the two-domain fungal ATP sulfurylases of Penicillium chrysogenum and Saccharomyces cerevisiae, which also exhibit a hexameric assembly. However, the T. denitrificans enzyme exhibits numerous structural and sequence differences in the N-terminal domain that render it inactive with respect to ATP sulfurylase activity. Surprisingly, the C-terminal domain does indeed display APS kinase activity, indicating that this gene product is a true APS kinase. Therefore, these results provide the first structural insights into a unique hexameric APS kinase that contains a nonfunctional ATP sulfurylase-like domain of unknown function.

  10. From Chemolithoautotrophs to Electrolithoautotrophs: CO2 Fixation by Fe(II-Oxidizing Bacteria Coupled with Direct Uptake of Electrons from Solid Electron Sources

    Directory of Open Access Journals (Sweden)

    Takumi eIshii

    2015-09-01

    Full Text Available At deep-sea vent systems, hydrothermal emissions rich in reductive chemicals replace solar energy as fuels to support microbial carbon assimilation. Until recently, all the microbial components at vent systems have been assumed to be fostered by the primary production of chemolithoautotrophs; however, both the laboratory and on-site studies demonstrated electrical current generation at vent systems and have suggested that a portion of microbial carbon assimilation is stimulated by the direct uptake of electrons from electrically conductive minerals. Here we show that chemolithoautotrophic Fe(II-oxidizing bacterium, Acidithiobacillus ferrooxidans, switches the electron source for carbon assimilation from diffusible Fe2+ ions to an electrode under the condition that electrical current is the only source of energy and electrons. Site-specific marking of a cytochrome aa3 complex (aa3 complex and a cytochrome bc1 complex (bc1 complex in viable cells demonstrated that the electrons taken directly from an electrode are used for O2 reduction via a down-hill pathway, which generates proton motive force that is used for pushing the electrons to NAD+ through a bc1 complex. Activation of carbon dioxide fixation by a direct electron uptake was also confirmed by the clear potential dependency of cell growth. These results reveal a previously unknown bioenergetic versatility of Fe(II-oxidizing bacteria to use solid electron sources and will help with understanding carbon assimilation of microbial components living in electronically conductive chimney habitats.

  11. AztD, a Periplasmic Zinc Metallochaperone to an ATP-binding Cassette (ABC) Transporter System in Paracoccus denitrificans.

    Science.gov (United States)

    Handali, Melody; Roychowdhury, Hridindu; Neupane, Durga P; Yukl, Erik T

    2015-12-11

    Bacterial ATP-binding cassette (ABC) transporters of transition metals are essential for acquisition of necessary elements from the environment. A large number of Gram-negative bacteria, including human pathogens, have a fourth conserved gene of unknown function adjacent to the canonical permease, ATPase, and solute-binding protein (SBP) genes of the AztABC zinc transporter system. To assess the function of this putative accessory factor (AztD) from Paracoccus denitrificans, we have analyzed its transcriptional regulation, metal binding properties, and interaction with the SBP (AztC). Transcription of the aztD gene is significantly up-regulated under conditions of zinc starvation. Recombinantly expressed AztD purifies with slightly substoichiometric zinc from the periplasm of Escherichia coli and is capable of binding up to three zinc ions with high affinity. Size exclusion chromatography and a simple intrinsic fluorescence assay were used to determine that AztD as isolated is able to transfer bound zinc nearly quantitatively to apo-AztC. Transfer occurs through a direct, associative mechanism that prevents loss of metal to the solvent. These results indicate that AztD is a zinc chaperone to AztC and likely functions to maintain zinc homeostasis through interaction with the AztABC system. This work extends our understanding of periplasmic zinc trafficking and the function of chaperones in this process.

  12. Influence of salts and pH on growth and activity of a novel facultatively alkaliphilic, extremely salt-tolerant, obligately chemolithoautotrophic sufur-oxidizing Gammaproteobacterium Thioalkalibacter halophilus gen. nov., sp. nov. from South-Western Siberian soda lakes

    NARCIS (Netherlands)

    Banciu, H.L.; Sorokin, D.Y.; Tourova, T.P.; Galinski, E.A.; Muntyan, M.S.; Kuenen, J.G.; Muyzer, G.

    2008-01-01

    A chemolithoautotrophic sulfur-oxidizing bacterium (SOB) strain ALCO 1 capable of growing at both near-neutral and extremely alkaline pH was isolated from hypersaline soda lakes in S-W Siberia (Altai, Russia). Strain ALCO 1 represents a novel separate branch within the halothiobacilli in the Gammapr

  13. 脱氮硫杆菌对废水中硫氮的脱除%Removal of Nitrate and Sulfide from Wastewater by Thiobacillus denitrificans

    Institute of Scientific and Technical Information of China (English)

    武鑫; 赵晋忠

    2013-01-01

    The growth curve of Thiobacillus denitrificans and pH of the culture media during the domestication process were determined to make sure the optimum condition for the removal of nitrate and sulfide in highly-concentrated wastewater. Then the ratio of sulfide to nitrate and the removing rate of NO3- were also examined in this paper. The results showed that when the S2- concentration in wastewater was 400 mg/L and the ratio of sulfide to nitrate was 5:3, the removing rate of NO3- could reach 63.19%, and S2- could simultaneously be converted into sulfate or elementary sulfur. These results indicated that Thiobacillus denitrificans had application potential in pollution control for nitrate and sulfide in wastewater. Fig 6, Tab 1, Ref 16%为脱除废水中高浓度的硫氮,首先对脱氮硫杆菌(Thiobacillus denitrificans)的生长曲线和驯化期间营养液pH 的变化进行了测定,确定了脱氮硫杆菌合适的使用条件.然后检测了接菌后废水中的S2-浓度控制在400 mg/L时合适的硫氮比以及脱氮硫杆菌对NO3的去除率.结果表明,处理富含硫氮的废水时,当硫氮比控制在5:3、S2-浓度控制在400mg/L时,脱氮硫杆菌对NO3的去除率可达63.19%,同时S2-转化成硫酸盐或可回收的单质硫,使废水的硫氮污染得到了有效的控制.

  14. ω-Amino Acid:Pyruvate Transaminase from Alcaligenes denitrificans Y2k-2: a New Catalyst for Kinetic Resolution of β-Amino Acids and Amines

    OpenAIRE

    Yun, Hyungdon; Lim, Seongyop; Cho, Byung-Kwan; Kim, Byung-Gee

    2004-01-01

    Alcaligenes denitrificans Y2k-2 was obtained by selective enrichment followed by screening from soil samples, which showed ω-amino acid:pyruvate transaminase activity, to kinetically resolve aliphatic β-amino acid, and the corresponding structural gene (aptA) was cloned. The gene was functionally expressed in Escherichia coli BL21 by using an isopropyl-β-d-thiogalactopyranoside (IPTG)-inducible pET expression system (9.6 U/mg), and the recombinant AptA was purified to show a specific activity...

  15. Effects of ionic strength on the coordination of Eu(III) and Cm(III) to a Gram-negative bacterium, Paracoccus denitrificans

    Energy Technology Data Exchange (ETDEWEB)

    Ozaki, T.; Ohnuki, T. [Advanced Science Research Center, Japan Atomic Energy Agency, Tokai, Ibaraki (Japan); Kimura, T. [Nuclear Science and Engineering Directorate, Japan Atomic Energy Agency, Tokai, Ibaraki (Japan); Francis, A.J. [Environmental Sciences Dept., Brookhaven National Lab., Upton, NY (United States)

    2006-07-01

    We studied the effect of ionic strength on the interactions of Europium(III) and Curium(III) with a Gram-negative bacterium Paracoccus denitrificans. Bacterial cells grown in 0.5-, 3.5-, and 5.0% NaCl were used in adsorption experiments and laser experiments that were performed at the same ionic strengths as those in the original growth media. The distribution ratio (log K{sub d}) for Eu(III) and Cm(III) was determined at pHs 3-5. To elucidate the coordination environment of Eu(III) adsorbed on P. denitrificans, we estimated the number of water molecules in the inner sphere and strength of the ligand field by time-resolved laser-induced fluorescence spectroscopy (TRLFS) at pHs 4-6. The log K{sub d} of Eu(III) and Cm(III) increased with an increase of pH at all ionic strengths because there was less competition for ligands in cells with H{sup +} at higher pHs, wherein less H{sup +} was present in solution: cation adsorption generally occurs through an exchange with H{sup +} on the functional groups of coordination sites. No significant differences were observed in the log K{sub d} of Eu(III) and Cm(III) at each pH in 0.5-, 3.5-, and 5.0% NaCl solutions, though competition for ligands with Na{sup +} would be expected to increase at higher NaCl concentrations. The log K{sub d} of Eu(III) was almost equivalent to that of Cm(III) under all the experimental conditions. TRLFS showed that the coordination environments of Eu(III) did not differ from each other at 0.5-, 3.5-, and 5.0% NaCl at pHs 4-6. TRLFS also showed that the characteristic of the coordination environment of Eu(III) on P. denitrificans was similar to that on a halophile, Nesterenkonia halobia, while it significantly differed from that on a non-halophile, Pseudomonas putida. These findings indicate that the number of coordination sites for Eu(III) on P. denitrificans, whose cell surface may have similar structures to that of halophiles, increased with increasing ionic strength, though their structure

  16. The structural and functional basis of catalysis mediated by NAD(PH:acceptor Oxidoreductase (FerB of Paracoccus denitrificans.

    Directory of Open Access Journals (Sweden)

    Vojtěch Sedláček

    Full Text Available FerB from Paracoccus denitrificans is a soluble cytoplasmic flavoprotein that accepts redox equivalents from NADH or NADPH and transfers them to various acceptors such as quinones, ferric complexes and chromate. The crystal structure and small-angle X-ray scattering measurements in solution reported here reveal a head-to-tail dimer with two flavin mononucleotide groups bound at the opposite sides of the subunit interface. The dimers tend to self-associate to a tetrameric form at higher protein concentrations. Amino acid residues important for the binding of FMN and NADH and for the catalytic activity are identified and verified by site-directed mutagenesis. In particular, we show that Glu77 anchors a conserved water molecule in close proximity to the O2 of FMN, with the probable role of facilitating flavin reduction. Hydride transfer is shown to occur from the 4-pro-S position of NADH to the solvent-accessible si side of the flavin ring. When using deuterated NADH, this process exhibits a kinetic isotope effect of about 6 just as does the NADH-dependent quinone reductase activity of FerB; the first, reductive half-reaction of flavin cofactor is thus rate-limiting. Replacing the bulky Arg95 in the vicinity of the active site with alanine substantially enhances the activity towards external flavins that obeys the standard bi-bi ping-pong reaction mechanism. The new evidence for a cryptic flavin reductase activity of FerB justifies the previous inclusion of this enzyme in the protein family of NADPH-dependent FMN reductases.

  17. Thiomicrospira hydrogeniphila sp. nov., an aerobic, hydrogen- and sulfur-oxidizing chemolithoautotroph isolated from a seawater tank containing a block of beef tallow.

    Science.gov (United States)

    Watsuji, Tomo-O; Hada, Emi; Miyazaki, Masayuki; Ichimura, Masako; Takai, Ken

    2016-09-01

    A moderately psychrophilic, aerobic, hydrogen- and sulfur-oxidizing bacterium, designated strain MAS2T, was isolated from a tank containing coastal seawater from Tokyo Bay and a block of beef tallow added as organic material. Growth occurred under aerobic chemolithoautotrophic conditions in the presence of molecular hydrogen, thiosulfate, tetrathionate, elemental sulfur or sulfide as the sole energy source and bicarbonate as a carbon source. The isolate represented a Gram-staining-negative rod with a single polar flagellum and grew in artificial seawater medium with thiosulfate at 2-40 °C (optimum 30 °C). The isolate grew in media with thiosulfate at Na+ concentrations between 30 and 1380 mM (optimum 270 mM). MAS2T possessed C16 : 0, C16 : 1 and C18 : 1 as the major fatty acids. The G+C content of the genomic DNA was 39.6 mol%. The 16S rRNA gene sequence similarity analysis showed that the isolate represented a member of the genus Thiomicrospira within the class Gammaproteobacteria and was most closely related to Thiomicrospira frisia JB-A2T. On the basis of phenotypic and molecular properties, the isolate represents a novel species of the genus Thiomicrospira, for which the name Thiomicrospira hydrogeniphila sp. nov. is proposed (type strain, MAS2T=JCM 30760T=DSM 100274T).

  18. The structure of RdDddP from Roseobacter denitrificans reveals that DMSP lyases in the DddP-family are metalloenzymes.

    Directory of Open Access Journals (Sweden)

    Jan-Hendrik Hehemann

    Full Text Available Marine microbes degrade dimethylsulfoniopropionate (DMSP, which is produced in large quantities by marine algae and plants, with DMSP lyases into acrylate and the gas dimethyl sulfide (DMS. Approximately 10% of the DMS vents from the sea into the atmosphere and this emission returns sulfur, which arrives in the sea through rivers and runoff, back to terrestrial systems via clouds and rain. Despite their key role in this sulfur cycle DMSP lyases are poorly understood at the molecular level. Here we report the first X-ray crystal structure of the putative DMSP lyase RdDddP from Roseobacter denitrificans, which belongs to the abundant DddP family. This structure, determined to 2.15 Å resolution, shows that RdDddP is a homodimeric metalloprotein with a binuclear center of two metal ions located 2.7 Å apart in the active site of the enzyme. Consistent with the crystallographic data, inductively coupled plasma mass spectrometry (ICP-MS and total reflection X-ray fluorescence (TRXF revealed the bound metal species to be primarily iron. A 3D structure guided analysis of environmental DddP lyase sequences elucidated the critical residues for metal binding are invariant, suggesting all proteins in the DddP family are metalloenzymes.

  19. The Online Morphology Control and Dynamic Studies on Improving Vitamin B12 Production by Pseudomonas denitrificans with Online Capacitance and Specific Oxygen Consumption Rate.

    Science.gov (United States)

    Wang, Ze-Jian; Shi, Hui-Lin; Wang, Ping

    2016-07-01

    The relationship between the morphological character of Pseudomonas denitrificans and vitamin B12 synthesis based on real-time capacitance measurement and online specific oxygen consumption rate (Q O2) control was established for enhancing vitamin B12 production. Results demonstrated that the threshold Q O2 value lower than 2.0 mmol/gDCW/l would greatly stimulate the state transfer from the cell number growth phase to the cell elongation phase and promote rapid vitamin B12 biosynthesis, while the vitamin B12 biosynthesis rate could also be inhibited when the rate of cell's length-to-width ratio (ratio-LW) was higher than 10:1. Furthermore, the optimal morphology controlling strategy was achieved based on online Q O2 control, which increases the appropriate active cell numbers at the former phase, and then control the elongation of ratio-LW no more than 10:1 at the vitamin B12 biosynthesis phase. The maximal vitamin B12 production reached 239.7 mg/l at 168 h, which was improved by 14.7 % compared with the control (208 mg/l). This online controlling strategy would be effectively applied for improving industrial vitamin B12 fermentation.

  20. 养殖池塘系统脱氮硫杆菌(Thiobacillus denitrificans)的分离、生长特性及脱氮特征研究%Isolation and Culture of Thiobacillus denitrificans from Different Area of the Intensive Pond System and Their Capacities of Removing Nitrate

    Institute of Scientific and Technical Information of China (English)

    范立民; 裘丽萍; 陈家长; 宋超; 胡庚东; 瞿建宏; 孟顺龙; 吴伟

    2013-01-01

    分别采集集约化养殖吉富罗非鱼(Oreochromis niloticus)的池塘底泥、表层水、塘边裸露土壤样品,采用脱氮硫杆菌选择培养基富集培养、分离得到3株细菌,通过菌落特征、形态学特征、生理生化检验和16S rDNA序列分析等手段,鉴定为脱氮硫杆菌(Thiobacillus denitrificans),命名为TD1、TD2和TD3.在30℃下,采用静置、厌氧培养实验研究这3株脱氮硫杆菌的生长繁殖特征、对硝酸盐氮的去除速率特征、硫酸根产生状况、亚硝酸盐产生情况、培养液pH值和氧化还原电位变化特征等方面的差异进行研究,探索养殖环境修复中应用的可能性.结果表明:在一定的时间内菌株生长速率、介质pH降低速率和氧化还原电位上升幅度均表现为TD1>TD2>TD3;菌株硝态氮脱除效率和硫酸根浓度升高速率均表现为TD1>TD2>TD3,亚硝酸盐产生情况则表现为TD1<TD2<TD3;TD1更适合异位修复,TD2更适合原位修复.%In this study, an intensive pond in which GIFT tilapia( Oreochromis niloticus ) were cultured was used. And sampling were done in different areas of the pond system, including sediments of the pond, the waters 50 centimeters below the surface and the bare soil at the edge of the pond. After that, enrichment and elective cultures were prepared anaerobically on these samples in 30 centigrade by incubating glass-stoppered bottles with the medium for Thiobacillus denitrificans full filled. After the culture, by using the spread-plate method, three strains were got. Through the appearance of the colonies, the shape of the strains, the detection of physiological and biochemical indicators, and the analysis of the sequences of 16S rDNA, the strains was identified as Thiobacillus denitrificans, and they were named as TD1, TD2 and TD3. Through the studies on the characteristics of growth and reproduction, of the removing rates of the nitrate, of the production of the sulfate and nitrite, and of

  1. Proteome-wide dataset generated by iTRAQ-3DLCMS/MS technique for studying the role of FerB protein in oxidative stress in Paracoccus denitrificans

    Directory of Open Access Journals (Sweden)

    Vendula Pernikářová

    2015-09-01

    Full Text Available 3DLC protein- and peptide-fractionation technique combined with iTRAQ-peptide labeling and Orbitrap mass spectrometry was employed to quantitate Paracoccus dentirificans total proteome with maximal coverage. This resulted in identification of 24,948 peptides representing 2627 proteins (FDR<0.01 in P. dentirificans wild type and ferB mutant strains grown in the presence or absence of methyl viologen as an oxidative stressor. The data were generated for assessment of FerB protein role in oxidative stress as published by Pernikářová et al.; proteomic responses to a methyl viologen-induced oxidative stress in the wild type and FerB mutant strains of P. denitrificans, J. Proteomics 2015;125:68–75. Dataset is supplied in the article.

  2. 脱氮硫杆菌生长特性及其对SRB生长的影响%CHARACTERISTICS OF THIOBACILLUS DENITRIFICANS AND THE EFFECT ON THE GROWTH OF SRB

    Institute of Scientific and Technical Information of China (English)

    刘宏芳; 汪梅芳; 许立铭

    2003-01-01

    由土壤中分离得到一株自养型的脱氮硫杆菌(Thiobacillus denitrificans,硫杆菌属,硫杆菌科,革兰氏阴性化能自养细菌),该菌株的最佳生长pH为7.0.将此菌株与硫酸盐还原菌(Sulfate Reducing Bacteria, SRB,脱硫弧菌属,革兰氏阴性厌氧细菌)混合培养,测定SRB的菌量变化,结果表明,脱氮硫杆菌的生长抑制了硫酸盐还原菌的生长,降低了SRB的腐蚀性的代谢产物硫化物的浓度,腐蚀速率降低,有利于防治SRB引起的微生物腐蚀.

  3. Isolation and characterization of the moxJ, moxG, moxI, and moxR genes of Paracoccus denitrificans: inactivation of moxJ, moxG, and moxR and the resultant effect on methylotrophic growth.

    OpenAIRE

    Van Spanning, R J; Wansell, C W; de Boer, T.; Hazelaar, M J; Anazawa, H; Harms, N.; Oltmann, L F; Stouthamer, A H

    1991-01-01

    By using the moxF gene encoding the large fragment of methanol dehydrogenase as a probe, a downstream linked chromosomal fragment was isolated from a genomic bank of Paracoccus denitrificans. The nucleotide sequence of the fragment was determined and revealed the 3' part of moxF, four additional open reading frames, and the 5' part of a sixth one. The organization and deduced amino acid sequences of the first three frames downstream from moxF were found to be largely homologous to the moxJ, m...

  4. 脱氮硫杆菌利用不同硫源去除地下水硝酸盐的实验研究%The Thiobacillus Denitrificans Use of Different Sulfur Source to Remove the Nitrate of Groundwater

    Institute of Scientific and Technical Information of China (English)

    孙莹

    2012-01-01

    从斛兵塘底土壤中分离得到一株自养反硝化菌。该菌株经过生理生化实验,初步判定为脱氮硫杆菌。该菌株可以分别利用黄铁矿、硫代硫酸钠、硫单质作电子供体,进行反硝化代谢。实验表明,在黄铁矿存在下该菌种对硝酸盐氮去除率最高,同时也有较商的硫酸根生成率。为了去除硫酸根新污染,尝试在培养基中加入石灰石。经实验证明.在培养基中加入石灰石几乎不影响反硝化代谢,并且有效的去除硫酸根离子,达到去除硝酸盐的目的。%A strain of Thiobacillus was isolated from the soil of the bottom of the Hubing Pool. Based on the analysis of physiological and biochemical measurements, this stain was identified with Thiobacillus denitrificans. The stain was employed to reduce Nitrate nitrogen by using of pyrite, thiosulfate and brimstone as electron donor. Experiments show that, the T.D get the highest nitrate nitrogen removal rate by using of pyrite, and also higher sulfate formation rate. In order to remove the sulfate pollution, try to add limestone in the pyrite medium, The pyrite medium by adding limestone make the nitrate nitrogen removal rate Almost no decreased, and effective removal of sulfate produced from denitrification, and ultimately achieve the purpose of remove the nitrate.

  5. Repair Effect of Thiobacillus Denitrificans and Denitrifying Bacterium on Pollution in Tianjin Waste Landfill Sites%脱氮硫杆菌和反硝化菌对天津市垃圾填埋场污染的修复研究

    Institute of Scientific and Technical Information of China (English)

    季彦; 郝金山; 薛祈

    2013-01-01

    Aiming at waste leachate samples from Tianjin waste landfill sites,the situation of waste with nitrifying bacteria and sulfate reducting bacteria in laboratory was discussed.The results showed that the metabolizable action of Thiobacillus denitrificans and denitrifying bacterium had strong function for nitrogen and organics removal.Under favorable conditions,it also could remove some metal pollutants.Through strengthening metabolizable action of some microorganism,the pollution system could be controlled and repaired.%针对天津市各大垃圾填埋场的渗沥液试样,论述试验室对垃圾掺入氮化物转化的硝化菌、硫化物转化的硫酸盐还原菌情况.结果表明:脱氮硫杆菌及反硝化菌的代谢作用具有较强的除氮及除去有机物的功能,有利条件下,还去除某些金属污染物质;通过强化某些微生物的代谢作用,可以对污染系统进行调控和修复.

  6. Isolation of Achromobacter xylosoxidans subsp. denitrificans and it's degradation of phenanthrene%木糖氧化无色杆菌反硝化亚种细菌的分离鉴定及其菲降解特性研究

    Institute of Scientific and Technical Information of China (English)

    武凤霞; 范丙全; 刘建玲

    2007-01-01

    利用选择性富集培养及升华法,从石油污染的土壤中分离到2株菲降解细菌,它们在以菲为唯一碳源的培养基上生长良好.应用B10LOG细菌鉴定系统和分子生物学方法对两株细菌进行鉴定,两株菌分别为坚强芽孢杆菌(Bacillus firmus)和木糖氧化无色杆菌反硝化亚种(Achromobacter xylosoxidans subsp.denitrificans),两株菌均具有邻苯二酚氧化酶活性.两株细菌在液体培养条件下都表现较强降解菲的能力,液体培养60 h约90%的加入菲被降解.通过测定液体培养基中菲浓度和菌体密度变化发现,菌株降解菲的量与其生长密度相关;随着菌体浓度(吸光度)的增加,代谢底物菲的浓度明显降低,两株菌混合使用能够大幅度提高降解菲的能力.

  7. Regulatory effect of the typical methyl-group donors on the oxygen metabolism in Pseudomonas denitrificans%典型性甲基供体对脱氮假单胞杆菌代谢过程的氧调控作用

    Institute of Scientific and Technical Information of China (English)

    周佳; 程新; 彭志远; 彭卫福; 李昆太

    2013-01-01

    In this paper,the fermentation processes of Pseudomonas denitrificans were investigated in 7L fermenter by using betaine and choline chloride as the typical methyl - group donors for vitamin B12 biosynthesis, respectively. When using choline chloride as the methyl - group donor, the dissolved oxygen ( DO) and pH respectively presented an obviously zooming and dropping phenomena, resulting in a slow cell growth and low vitamin B12 biosynthesis.However,the metabolism capability under betaine as methyl-group donor was significantly better than that of choline chloride, and the pH and DO could be maintained at a stable and appropriate level during the whole fermentation processes. As a result, the final cell growth and vitamin B12 production were significantly higher than those obtained in choline chloride.%本文在7L发酵罐中分别以甜菜碱和氯化胆碱作为合成维生素B12的甲基供体,分别考察了二者对脱氮假单胞杆菌代谢过程的影响.结果表明:使用氯化胆碱的罐批出现了溶氧过早回升、pH剧烈下降等异常的代谢现象,致使发酵过程的菌体生长缓慢且维生素B12合成量低;而使用甜菜碱作为甲基供体时,其菌体代谢活力要明显优于使用氯化胆碱的罐批,整个发酵过程的pH和溶氧均能保持在稳定且合适的水平,最终的菌体量和维生素B12产量也更高.

  8. Iron meteorites can support the growth of acidophilic chemolithoautotrophic microorganisms.

    Science.gov (United States)

    González-Toril, Elena; Martínez-Frías, Jesús; Gómez Gómez, José María; Rull, Fernando; Amils, Ricardo

    2005-06-01

    Chemolithoautotrophy based on reduced inorganic minerals is considered a primitive energy transduction system. Evidence that a high number of meteorites crashed into the planet during the early period of Earth history led us to test the ability of iron-oxidizing bacteria to grow using iron meteorites as their source of energy. Here we report the growth of two acidophilic iron-oxidizing bacteria, Leptospirillum ferrooxidans and Acidithiobacillus ferrooxidans, on a piece of the Toluca meteorite as the only source of energy. The alteration of the surface of the exposed piece of meteorite, the solubilization of its oxidized metal constituents, mainly ferric iron, and the formation of goethite precipitates all clearly indicate that iron-meteorite-based chemolithotrophic metabolism is viable.

  9. Metagenome of a Versatile Chemolithoautotroph from Expanding Oceanic Dead Zones

    Energy Technology Data Exchange (ETDEWEB)

    Walsh, David A.; Zaikova, Elena; Howes, Charles L.; Song, Young; Wright, Jody; Tringe, Susannah G.; Tortell, Philippe D.; Hallam, Steven J.

    2009-07-15

    Oxygen minimum zones (OMZs), also known as oceanic"dead zones", are widespread oceanographic features currently expanding due to global warming and coastal eutrophication. Although inhospitable to metazoan life, OMZs support a thriving but cryptic microbiota whose combined metabolic activity is intimately connected to nutrient and trace gas cycling within the global ocean. Here we report time-resolved metagenomic analyses of a ubiquitous and abundant but uncultivated OMZ microbe (SUP05) closely related to chemoautotrophic gill symbionts of deep-sea clams and mussels. The SUP05 metagenome harbors a versatile repertoire of genes mediating autotrophic carbon assimilation, sulfur-oxidation and nitrate respiration responsive to a wide range of water column redox states. Thus, SUP05 plays integral roles in shaping nutrient and energy flow within oxygen-deficient oceanic waters via carbon sequestration, sulfide detoxification and biological nitrogen loss with important implications for marine productivity and atmospheric greenhouse control.

  10. Improvement of the production of vitamin B12 by Pseudomonas denitrifican through increasing the fermenter pressure%增大罐压提高脱氮假单胞菌发酵生产维生素B12的产量

    Institute of Scientific and Technical Information of China (English)

    刘玉伟; 王建; 王泽建; 黄明志

    2012-01-01

    [Objective] The pressure of fermenter was enhanced in order to increase the concentration of vitamin B12. [Methods] Metabolic flux analysis (MFA) was used to analyse the fermentation process of vitamin B12 production by Pseudomonas denitrifwan. [Results] The results indicated that the metabolic flux of oxaloacetic acid from phosphoenolpyruvate car-boxylation was increased when the synthesis rate of vitamin B12 increased, in order to meet the demand for precursor supply to vitamin B12 production. A novel control strategy was designedas following. The pressure of fermenter was enhanced in order to increase the solubility of carbon dioxide in aqueous solution at steady phase of fermentation, so more carbon dioxide was carboxylated to oxaloacetic acid. The result showed that the concentration of vitamin B12 was 176 mg/L, 19.7% higher than the control batch (147 mg/L) at 160 hours. [Conclusion] the production of vitamin B12 by Pseudomonas denitrifican was improved by increasing the fermenter pressure.%[目的]提高发酵罐的罐压,增加维生素B12的产率.[方法]利用常规代谢通量分析(MFA)方法,对脱氮假单胞菌生产维生素B12的发酵过程进行研究.[结果]发现随着VB12合成速率的加快,磷酸烯醇式丙酮酸(PEP)羧化生成草酰乙酸(OAA)的通量明显加大,以满足维生素B12合成对前体的需求.根据该分析结果,对发酵工艺进行了改进,即在脱氮假单胞菌进入合成维生素B12阶段时,提高发酵罐的罐压,增加发酵液中二氧化碳的溶解度,从而强化了羧化回补途径.维生素B12的产率明显增加,发酵160 h的产物浓度为176 mg/L,比对照批次终浓度147 mg/L高出了19.7%.[结论]通过增大罐压提高了脱氮假单胞菌进入合成维生素B12的产量.

  11. The Genome of Deep-Sea Vent Chemolithoautotroph Thiomicrospiracrunogena XCL-2

    Energy Technology Data Exchange (ETDEWEB)

    Scott, Kathleen M.; Sievert, Stefan M.; Abril, Fereniki N.; Ball,Lois A.; Barrett, Chantell J.; Blake, Rodrigo A.; Boller, Amanda J.; Chain, Patrick S.G.; Clark, Justine A.; Davis, Carisa R.; Detter, Chris; Do, Kimberly F.; Dobrinski, Kimberly P.; Faza, BrandonI.; Fitzpatrick,Kelly A.; Freyermuth, Sharyn K.; Harmer, Tara L.; Hauser, Loren J.; Hugler, Michael; Kerfeld, Cheryl A.; Klotz, Martin G.; Kong, William W.; Land, Miriam; Lapidus, Alla; Larimer, Frank W.; Longo, Dana L.; Lucas,Susan; Malfatti, Stephanie A.; Massey, Steven E.; Martin, Darlene D.; McCuddin, Zoe; Meyer, Folker; Moore, Jessica L.; Ocampo, Luis H.; Paul,John H.; Paulsen, Ian T.; Reep, Douglas K.; Ren, Qinghu; Ross, Rachel L.; Sato, Priscila Y.; Thomas, Phaedra; Tinkham, Lance E.; Zeruth, Gary T.

    2006-08-23

    Presented here is the complete genome sequence ofThiomicrospira crunogena XCL-2, representative of ubiquitouschemolithoautotrophic sulfur-oxidizing bacteria isolated from deep-seahydrothermal vents. This gammaproteobacterium has a single chromosome(2,427,734 bp), and its genome illustrates many of the adaptations thathave enabled it to thrive at vents globally. It has 14 methyl-acceptingchemotaxis protein genes, including four that may assist in positioningit in the redoxcline. A relative abundance of CDSs encoding regulatoryproteins likely control the expression of genes encoding carboxysomes,multiple dissolved inorganic nitrogen and phosphate transporters, as wellas a phosphonate operon, which provide this species with a variety ofoptions for acquiring these substrates from the environment. T. crunogenaXCL-2 is unusual among obligate sulfur oxidizing bacteria in relying onthe Sox system for the oxidation of reduced sulfur compounds. A 38 kbprophage is present, and a high level of prophage induction was observed,which may play a role in keeping competing populations of close relativesin check. The genome has characteristics consistent with an obligatelychemolithoautotrophic lifestyle, including few transporters predicted tohave organic allocrits, and Calvin-Benson-Bassham cycle CDSs scatteredthroughout the genome.

  12. Influence of increasing dissolved inorganic carbon concentrations and decreasing pH on chemolithoautrophic bacteria from oxic-sulfidic interfaces

    Directory of Open Access Journals (Sweden)

    K. Mammitzsch

    2012-12-01

    Full Text Available Increases in the dissolved inorganic carbon (DIC concentration are expected to cause a decrease in the pH of ocean waters, a process known as ocean acidification. In oxygen-deficient zones this will add to already increased DIC and decreased pH values. It is not known how this might affect microbial communities and microbially mediated processes. In this study, the potential effects of ocean acidification on chemolithoautotrophic prokaryotes of marine oxic-anoxic transition zones were investigated, using the chemoautotrophic denitrifying ε-proteobacterium "Sulfurimonas gotlandica" strain GD1 as a model organism. This and related taxa use reduced sulfur compounds, e.g. sulfide and thiosulfate, as electron donors and were previously shown to be responsible for nitrate removal and sulfide detoxification in redox zones of the Baltic Sea water column but occur also in other oxygen-deficient marine systems. Bacterial cell growth within a broad range of DIC concentrations and pH values was monitored and substrate utilization was determined. The results showed that the DIC saturation concentration for growth was already reached at 800 μM, which is well below in situ DIC levels. The pH optimum was between 6.6 and 8.0. Within a pH range of 6.6–7.1 there was no significant difference in substrate utilization; however, at lower pH values cell growth decreased sharply and cell-specific substrate consumption increased. These findings suggest that a direct effect of ocean acidification, with the predicted changes in pH and DIC, on chemolithoautotrophic bacteria such as "S. gotlandica" str. GD1 is generally not very probable.

  13. An integrated bioremediation process for petroleum hydrocarbons removal and odor mitigation from contaminated marine sediment.

    Science.gov (United States)

    Zhang, Zhen; Lo, Irene M C; Yan, Dickson Y S

    2015-10-15

    This study developed a novel integrated bioremediation process for the removal of petroleum hydrocarbons and the mitigation of odor induced by reduced sulfur from contaminated marine sediment. The bioremediation process consisted of two phases. In Phase I, acetate was dosed into the sediment as co-substrate to facilitate the sulfate reduction process. Meanwhile, akaganeite (β-FeOOH) was dosed in the surface layer of the sediment to prevent S(2-) release into the overlying seawater. In Phase II, NO3(-) was injected into the sediment as an electron acceptor to facilitate the denitrification process. After 20 weeks of treatment, the sequential integration of the sulfate reduction and denitrification processes led to effective biodegradation of total petroleum hydrocarbons (TPH), in which about 72% of TPH was removed. In Phase I, the release of S(2-) was effectively controlled by the addition of akaganeite. The oxidation of S(2-) by Fe(3+) and the precipitation of S(2-) by Fe(2+) were the main mechanisms for S(2-) removal. In Phase II, the injection of NO3(-) completely inhibited the sulfate reduction process. Most of residual AVS and S(0) were removed within 4 weeks after NO3(-) injection. The 16S rRNA clone library-based analysis revealed a distinct shift of bacterial community structure in the sediment over different treatment phases. The clones affiliated with Desulfobacterales and Desulfuromonadales were the most abundant in Phase I, while the clones related to Thioalkalivibrio sulfidophilus, Thiohalomonas nitratireducens and Sulfurimonas denitrificans predominated in Phase II.

  14. Post-eruption colonization and community succession of hydrothermal microbial mats

    Science.gov (United States)

    Moyer, C. L.; Hager, K. W.; Fullerton, H.

    2015-12-01

    T-RFLP fingerprint cluster analysis and qPCR of microbial mat communities from hydrothermal vent habitats among recent post-eruption sites exhibit similar communities containing Epsilonproteobacteria that are phylogenetically similar and capable of hydrogen-oxidation (e.g., Nitratiruptor, Caminibacter, Nautilia, Thioreductor, and/or Lebetimonas). This community is the first (Group I) of three community types that represent different stages in the transition from vapor-dominated to brine-dominated water-rock interactions (i.e., vent effluent geochemistry). We have now observed this similar transition from four hydrothermal regions from across the Pacific Ocean. The second type of mat community (Group II) that has been observed is characterized by the presence of another group of Epsilonproteobacteria; however, these are mostly sulfur-oxidizing phylotypes (e.g., Sulfurimonas, Sulfurovum, and/or Sulfuricurvum). Finally, once the transition from sulfur to iron is complete, then the third type (Group III) cluster together by the presence of Zetaproteobacteria, which are known to use iron-oxidation. Each of these community types are dominated by groups of microorganisms characterized by cultured isolates, all of which are strict chemolithoautotrophs capable of carbon fixation and are hypothesized as both ecosystem engineers and primary producers in these energy-rich ecosystems. We also consider the thermodynamic implications towards carbon fixation for each of the three groups of mat communities.

  15. Isolation and properties of obligately chemolithoautotrophic and extremely alkali-tolerant ammonia-oxidizing bacteria from Mongolian soda lakes.

    Science.gov (United States)

    Sorokin, D; Tourova, T; Schmid, M C; Wagner, M; Koops, H P; Kuenen, J G; Jetten, M

    2001-09-01

    Five mixed samples prepared from the surface sediments of 20 north-east Mongolian soda lakes with total salt contents from 5 to 360 g/l and pH values from 9.7 to 10.5 were used to enrich for alkaliphilic ammonia-oxidizing bacteria. Successful enrichments at pH 10 were achieved on carbonate mineral medium containing 0.6 M total Na(+) and < or =4 mM NH(4)Cl. Five isolates (ANs1-ANs5) of ammonia-oxidizing bacteria capable of growth at pH 10 were obtained from the colonies developed on bilayered gradient plates. The cells were motile and coccoid, with well-developed intracytoplasmic membranes (ICPM) and carboxysomes. At pH 10.0, ammonia was toxic for growth at concentrations higher than 5 mM NH(4)Cl. The bacteria were able to grow within the salinity range of 0.1-1.0 M of total Na+ (optimum 0.3 M). In media containing 0.3-0.6 M total Na(+), optimal growth in batch cultures occurred in the presence of a bicarbonate/carbonate buffer system within the pH range 8.5-9.5, with the highest pH limit at pH 10.5. At pH lower than 8.0, growth was slower, most probably due to decreasing free ammonia. The pH profile of the respiratory activity was broader, with limits at 6.5-7.0 and 11.0 and an optimum at 9.5-10.0. In pH-controlled, NH(3)-limited continuous culture, isolate ANs5 grew up to pH 11.3, which is the highest pH limit known for ammonia-oxidizing bacteria so far. This showed the existence of extremely alkali-tolerant ammonia-oxidizing bacteria in the soda lakes. Comparative 16S rDNA sequence analysis of the five isolates demonstrated that they possess identical 16S rDNA genes and that they are closely related to Nitrosomonas halophila (sequence similarity 99.3%), a member of the beta-subclass of the Proteobacteria. This affiliation was confirmed by comparative sequence analysis of the amoA gene, encoding the active-site subunit of the ammonia-monoxygenase, of one of the isolates. DNA-DNA hybridization data further supported that the soda lake isolates are very similar to each other and represent an alkali-tolerant subpopulation of N. halophila whose species description is herewith amended.

  16. Influence of medium composition on the characteristics of a denitrifying biofilm formed by Alcaligenes denitrificans in a fluidised bed reactor

    OpenAIRE

    Alves, C. F.; Melo, L. F.; Vieira, M. J.

    2002-01-01

    The influence of the ratio carbon/nitrogen and phosphorus concentration on the performance of a biofilm fluidised bed reactor used for denitrification and on the properties of the biofilm was studied. Although the removal efficiencies of C and N reached steady-state values, the thickness of the biofilm steadily increased. The dry density of the biofilm did not seem to be dependent on the loading conditions, although a denser biofilm was obtained when there was no nutrient limitation ...

  17. Sulfuriferula thiophila sp. nov., a chemolithoautotrophic sulfur-oxidizing bacterium, and correction of the name Sulfuriferula plumbophilusWatanabe, Kojima and Fukui 2015 to Sulfuriferula plumbiphila corrig.

    Science.gov (United States)

    Watanabe, Tomohiro; Kojima, Hisaya; Fukui, Manabu

    2016-05-01

    A novel sulfur-oxidizing bacterium designated strain mst6T was isolated from spring water of Masutomi hot spring in Japan. The cells were rod-shaped (1.2-4.0 × 0.5-0.7 μm) and Gram-stain-negative. The G+C content of genomic DNA was around 52.6 mol%. The isolate possessed summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), C16 : 0 and C12 : 0 as major cellular fatty acids. Strain mst6T grew by inorganic carbon fixation and oxidation of inorganic sulfur compounds with oxygen as an electron acceptor. The isolate grew over a temperature range of 5-34 °C, a NaCl concentration range of 0-110 mM and a pH range of 4.6-8.1. Optimum growth occurred at 32 °C, in the absence of NaCl and at pH 5.9-6.2. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain mst6T belongs to the family Sulfuricellaceae in the class Betaproteobacteria. The closest cultured relative was Sulfuriferula multivorans TTNT with a 16S rRNA gene sequence similarity of 97.0 %. On the basis of the data obtained in this study, strain mst6T represents a novel species of the genus Sulfuriferula, for which the name Sulfuriferula thiophila sp. nov. is proposed. The type strain is mst6T ( = NBRC 111150T = DSM 101871T). In addition, we propose correcting the name Sulfuriferula plumbophilus Watanabe, Kojima and Fukui 2015 to Sulfuriferula plumbiphila corrig. based on Rule 12c, Rule 61 and Appendix 9 of the International Code of Nomenclature of Prokaryotes.

  18. Nitrolancea hollandica gen. nov., sp. nov., a chemolithoautotrophic nitrite-oxidizing bacterium isolated from a bioreactor belonging to the phylum Chloroflexi

    NARCIS (Netherlands)

    Sorokin, D.Y.; Vejmelkova, D.; Lücker, S.; Streshinskaya, G.M.; Rijpstra, W.I.C.; Sinninghe Damste, J.S.; Kleerbezem, R.; van Loosdrecht, M.; Muyzer, G.; Daims, H.

    2014-01-01

    A novel nitrite-oxidizing bacterium (NOB), strain LbT, was isolated from a nitrifying bioreactor with a high loading of ammonium bicarbonate in a mineral medium with nitrite as the energy source. The cells were oval (lancet-shaped) rods with pointed edges, non-motile, Gram-positive (by staining and

  19. Defluviimonas denitrificans gen. nov., sp. nov., and Pararhodobacter aggregans gen. nov., sp. nov., non-phototrophic Rhodobacteraceae from the biofilter of a marine aquaculture

    DEFF Research Database (Denmark)

    Foesel, Bärbel U.; Drake, Harold L.; Schramm, Andreas

    2011-01-01

    Three Gram-negative bacterial strains were isolated from the biofilter of a recirculating marine aquaculture. They were non-pigmented rods, mesophiles, moderately halophilic, and showed chemoorganoheterotrophic growth on various sugars, fatty acids, and amino acids, with oxygen as electron acceptor...

  20. Genome Analysis of the Biotechnologically Relevant Acidophilic Iron Oxidising Strain JA12 Indicates Phylogenetic and Metabolic Diversity within the Novel Genus “Ferrovum”

    Science.gov (United States)

    Ullrich, Sophie R.; Poehlein, Anja; Tischler, Judith S.; González, Carolina; Ossandon, Francisco J.; Daniel, Rolf; Holmes, David S.; Schlömann, Michael; Mühling, Martin

    2016-01-01

    Background Members of the genus “Ferrovum” are ubiquitously distributed in acid mine drainage (AMD) waters which are characterised by their high metal and sulfate loads. So far isolation and microbiological characterisation have only been successful for the designated type strain “Ferrovum myxofaciens” P3G. Thus, knowledge about physiological characteristics and the phylogeny of the genus “Ferrovum” is extremely scarce. Objective In order to access the wider genetic pool of the genus “Ferrovum” we sequenced the genome of a “Ferrovum”-containing mixed culture and successfully assembled the almost complete genome sequence of the novel “Ferrovum” strain JA12. Phylogeny and Lifestyle The genome-based phylogenetic analysis indicates that strain JA12 and the type strain represent two distinct “Ferrovum” species. “Ferrovum” strain JA12 is characterised by an unusually small genome in comparison to the type strain and other iron oxidising bacteria. The prediction of nutrient assimilation pathways suggests that “Ferrovum” strain JA12 maintains a chemolithoautotrophic lifestyle utilising carbon dioxide and bicarbonate, ammonium and urea, sulfate, phosphate and ferrous iron as carbon, nitrogen, sulfur, phosphorous and energy sources, respectively. Unique Metabolic Features The potential utilisation of urea by “Ferrovum” strain JA12 is moreover remarkable since it may furthermore represent a strategy among extreme acidophiles to cope with the acidic environment. Unlike other acidophilic chemolithoautotrophs “Ferrovum” strain JA12 exhibits a complete tricarboxylic acid cycle, a metabolic feature shared with the closer related neutrophilic iron oxidisers among the Betaproteobacteria including Sideroxydans lithotrophicus and Thiobacillus denitrificans. Furthermore, the absence of characteristic redox proteins involved in iron oxidation in the well-studied acidophiles Acidithiobacillus ferrooxidans (rusticyanin) and Acidithiobacillus

  1. Unique and universal features of Epsilonproteobacterial origins of chromosome replication and DnaA-DnaA box interactions

    Directory of Open Access Journals (Sweden)

    Pawel Jaworski

    2016-09-01

    Full Text Available In bacteria, chromosome replication is initiated by the interaction of the initiator protein DnaA with a defined region of a chromosome at which DNA replication starts (oriC. While DnaA proteins share significant homology regardless of phylogeny, oriC regions exhibit more variable structures. The general architecture of oriCs is universal, i.e. they are composed of a cluster of DnaA binding sites, a DNA-unwinding element, and sequences that bind regulatory proteins. However, detailed structures of oriCs are shared by related species while being significantly different in unrelated bacteria. In this work, we characterised Epsilonproteobacterial oriC regions. Helicobacter pylori was the only species of the class for which oriC was characterised. A few unique features were found such as bipartite oriC structure, not encountered in any other Gram-negative species, and topology-sensitive DnaA-DNA interactions, which have not been found in any other bacterium. These unusual H. pylori oriC features raised questions of whether oriC structure and DnaA-DNA interactions are unique to this bacterium or they are common to related species. By in silico and in vitro analyses we identified putative oriCs in three Epsilonproteobacterial species: pathogenic Arcobacter butzleri, symbiotic Wolinella succinogenes and free-living Sulfurimonas denitrificans. We propose that oriCs typically co-localize with ruvC-dnaA-dnaN in Epsilonproteobacteria, with the exception of Helicobacteriaceae species. The clusters of DnaA boxes localize upstream (oriC1 and downstream (oriC2 of dnaA, and they likely constitute bipartite origins. In all cases, DNA unwinding was shown to occur in oriC2. Unlike the DnaA box pattern, which is not conserved in Epsilonproteobacterial oriCs, the consensus DnaA box sequences and the mode of DnaA-DnaA box interactions are common to the class. We propose that the typical Epsilonproteobacterial DnaA box consists of the core nucleotide sequence 5

  2. Unique and Universal Features of Epsilonproteobacterial Origins of Chromosome Replication and DnaA-DnaA Box Interactions

    Science.gov (United States)

    Jaworski, Pawel; Donczew, Rafal; Mielke, Thorsten; Thiel, Marcel; Oldziej, Stanislaw; Weigel, Christoph; Zawilak-Pawlik, Anna

    2016-01-01

    In bacteria, chromosome replication is initiated by the interaction of the initiator protein DnaA with a defined region of a chromosome at which DNA replication starts (oriC). While DnaA proteins share significant homology regardless of phylogeny, oriC regions exhibit more variable structures. The general architecture of oriCs is universal, i.e., they are composed of a cluster of DnaA binding sites, a DNA-unwinding element, and sequences that bind regulatory proteins. However, detailed structures of oriCs are shared by related species while being significantly different in unrelated bacteria. In this work, we characterized Epsilonproteobacterial oriC regions. Helicobacter pylori was the only species of the class for which oriC was characterized. A few unique features were found such as bipartite oriC structure, not encountered in any other Gram-negative species, and topology-sensitive DnaA-DNA interactions, which have not been found in any other bacterium. These unusual H. pylori oriC features raised questions of whether oriC structure and DnaA-DNA interactions are unique to this bacterium or whether they are common to related species. By in silico and in vitro analyses we identified putative oriCs in three Epsilonproteobacterial species: pathogenic Arcobacter butzleri, symbiotic Wolinella succinogenes, and free-living Sulfurimonas denitrificans. We propose that oriCs typically co-localize with ruvC-dnaA-dnaN in Epsilonproteobacteria, with the exception of Helicobacteriaceae species. The clusters of DnaA boxes localize upstream (oriC1) and downstream (oriC2) of dnaA, and they likely constitute bipartite origins. In all cases, DNA unwinding was shown to occur in oriC2. Unlike the DnaA box pattern, which is not conserved in Epsilonproteobacterial oriCs, the consensus DnaA box sequences and the mode of DnaA-DnaA box interactions are common to the class. We propose that the typical Epsilonproteobacterial DnaA box consists of the core nucleotide sequence 5′-TTCAC-3

  3. NCBI nr-aa BLAST: CBRC-MMUR-01-1403 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUR-01-1403 ref|YP_003160262.1| protein of unknown function DUF1212 [Jonesia ...denitrificans DSM 20603] gb|ACV07959.1| protein of unknown function DUF1212 [Jonesia denitrificans DSM 20603] YP_003160262.1 6.2 25% ...

  4. NCBI nr-aa BLAST: CBRC-LAFR-01-0953 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-LAFR-01-0953 ref|YP_316208.1| possible transmembrane protein [Thiobacillus den...itrificans ATCC 25259] gb|AAZ98403.1| possible transmembrane protein [Thiobacillus denitrificans ATCC 25259] YP_316208.1 0.005 24% ...

  5. NCBI nr-aa BLAST: CBRC-OCUN-01-0958 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OCUN-01-0958 ref|ZP_00629062.1| Major facilitator superfamily [Paracoccus deni...trificans PD1222] ref|YP_916821.1| major facilitator superfamily MFS_1 [Paracoccus denitrificans PD1222] gb|ABL71125.1| maj

  6. NCBI nr-aa BLAST: CBRC-DDIS-03-0142 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DDIS-03-0142 ref|YP_683041.1| 5-aminolevulinic acid synthase [Roseobacter deni...trificans OCh 114] gb|ABG32355.1| 5-aminolevulinic acid synthase [Roseobacter denitrificans OCh 114] YP_683041.1 1e-122 61% ...

  7. NCBI nr-aa BLAST: CBRC-DRER-26-0447 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DRER-26-0447 ref|YP_682607.1| hypothetical protein RD1_2345 [Roseobacter denit...rificans OCh 114] gb|ABG31921.1| conserved hypothetical protein [Roseobacter denitrificans OCh 114] YP_682607.1 8e-34 35% ...

  8. NCBI nr-aa BLAST: CBRC-DNOV-01-0806 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-0806 ref|YP_683172.1| sulfite reductase [Roseobacter denitrificans OCh... 114] gb|ABG32486.1| sulfite reductase [Roseobacter denitrificans OCh 114] YP_683172.1 2.3 25% ...

  9. NCBI nr-aa BLAST: CBRC-OSAT-10-0008 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OSAT-10-0008 ref|YP_683656.1| permease, putative [Roseobacter denitrificans OC...h 114] gb|ABG32970.1| permease, putative [Roseobacter denitrificans OCh 114] YP_683656.1 1.6 42% ...

  10. UniProt search blastx result: AK288277 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288277 J090017K16 Q160X3|RL11_ROSDO 50S ribosomal protein L11 - Roseobacter denit...rificans (strain ATCC 33942 / OCh 114) (Erythrobacter sp. (strain OCh 114)) (Roseobacter denitrificans) 7.00E-34 ...

  11. UniProt search blastx result: AK288771 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288771 J090068A13 Q160Y1|RS12_ROSDO 30S ribosomal protein S12 - Roseobacter denit...rificans (strain ATCC 33942 / OCh 114) (Erythrobacter sp. (strain OCh 114)) (Roseobacter denitrificans) 2.00E-37 ...

  12. NCBI nr-aa BLAST: CBRC-DRER-26-0529 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DRER-26-0529 ref|YP_683352.1| hypothetical protein RD1_3158 [Roseobacter denit...rificans OCh 114] gb|ABG32666.1| conserved hypothetical protein [Roseobacter denitrificans OCh 114] YP_683352.1 2.5 41% ...

  13. NCBI nr-aa BLAST: CBRC-DNOV-01-2878 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-2878 ref|YP_680769.1| murein endopeptidase [Roseobacter denitrificans ...OCh 114] gb|ABG30083.1| murein endopeptidase [Roseobacter denitrificans OCh 114] YP_680769.1 1.4 29% ...

  14. NCBI nr-aa BLAST: CBRC-OLAT-26-0028 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OLAT-26-0028 ref|YP_771762.1| iron-regulated protein frpC, putative [Roseobact...er denitrificans OCh 114] gb|ABI93354.1| iron-regulated protein frpC, putative [Roseobacter denitrificans OCh 114] YP_771762.1 3e-32 23% ...

  15. NCBI nr-aa BLAST: CBRC-OPRI-01-0485 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OPRI-01-0485 ref|YP_680544.1| magnesium chelatase subunit D [Roseobacter denit...rificans OCh 114] gb|ABG29858.1| magnesium-chelatase 60 kDa subunit [Roseobacter denitrificans OCh 114] YP_680544.1 0.003 28% ...

  16. UniProt search blastx result: AK288890 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288890 J090080B13 Q160Y1|RS12_ROSDO 30S ribosomal protein S12 - Roseobacter denit...rificans (strain ATCC 33942 / OCh 114) (Erythrobacter sp. (strain OCh 114)) (Roseobacter denitrificans) 2.00E-37 ...

  17. Kinetic enrichment of 34S during proteobacterial thiosulfate oxidation and the conserved role of SoxB in S-S bond breaking

    Digital Repository Service at National Institute of Oceanography (India)

    Alam, M.; Pyne, P.; Mazumdar, A.; Peketi, A.; Ghosh, W.

    During chemolithoautotrophic thiosulfate oxidation the phylogenetically-diverged proteobacteria Paracoccus pantotrophus, Tetrathiobacter kashmirensis and Thiomicrospira crunogena rendered steady enrichment of 34S in the end product...

  18. Interactions between nitrifying and denitrifying bacteria in gnotobiotic microcosms planted with the emergent macrophyte Glyceria maxima

    NARCIS (Netherlands)

    Bodelier, P.L.E.; Duyts, H.; Blom, C.W.P.M.; Laanbroek, H.J.

    1998-01-01

    The population dynamics of the chemolithoautotrophic nitrifiers Nitrosomonas europaea and Nitrobacter winogradskyi were studied in gnotobiotic microcosms fed with ammonium in response to the presence or absence of the emergent macrophyte Glyceria maxima and the heterotrophic denitrifying bacterium P

  19. Habitability Potential of the Subsurface Martian Crust Constrained by SNC Meteorites

    Science.gov (United States)

    Shivak, J. N.; Banerjee, N. R.; Flemming, R. L.

    2013-09-01

    The SNC meteorites represent potentially habitable substrates for chemolithoautotrophs on Mars, but the conditions in the rocky subsurface will be site specific and much more limiting to microbiota than the inorganic mineral components of the crust.

  20. Tentative Study on a New Way of Simultaneous Desulfurization and Denitrification%一种利用脱氮硫杆菌的同步脱硫反硝化新工艺研究

    Institute of Scientific and Technical Information of China (English)

    王爱杰; 杜大仲; 任南琪; 程翔; 刘春爽

    2005-01-01

    Thiobacillus denitrificans, a kind of autotrophic facultative bacteria, can oxidize sulfide into elemental sulfur or sulfate when nitrate was adopted as its electron accepter and carbon dioxide as its carbon resource under anoxic or anaerobic environment. In this way, nitrate is converted into nitrogen. In addition, Thiobacillus denitrificans can accumulate sulfur extracellularly. In this study, in a process of simultaneous desulfurization and denitrification, a strain of Thiobacillus denitrificans is employed as sulfur-producer in the treatment of wastewater containing sulfide and nitrate. The key factors affecting this process are investigated through batch tests. The experimental results indicate that the sulfide concentration and the ratio of sulfide to nitrate (S2-/NO-3) in the respectively, in order to achieve high conversion of sulfur.

  1. Tentative Study on a New Way of Simultaneous Desulfurization and Denitrification

    Institute of Scientific and Technical Information of China (English)

    王爱杰; 杜大仲; 任南琪; 程翔; 刘春爽

    2005-01-01

    Thiobacillus denitrificans, a kind of autotrophic facultative bacteria, can oxidize sulfide into elemental sulfur or sulfate when nitrate was adopted as its electron accepter and carbon dioxide as its carbon resource under anoxic or anaerobic environment. In this way, nitrate is converted into nitrogen. In addition, Thiobacillus denitrificans can accumulate sulfur extracellularly. In this study, in a process of simultaneous desulfurization and denitrification, a strain of Thiobacillus denitrificans is employed as sulfur-producer in the treatment of wastewater containing sulfide and nitrate. The key factors affecting this process are investigated through batch tests. The experimental results indicate that the sulfide concentration and the ratio of sulfide to nitrate (S2-/NO3-) in the influent are the key factors, and their suitable values are suggested to be 5/3 and no more than 300mg·L-1, respectively, in order to achieve high conversion of sulfur.

  2. Stimulation of autotrophic denitrification by intrusions of the Bosporus Plume into the anoxic Black Sea

    Directory of Open Access Journals (Sweden)

    Clara A. Fuchsman

    2012-07-01

    Full Text Available Autotrophic denitrification was measured in the southwestern coastal Black Sea, where the Bosporus Plume injects oxidized chemical species (especially O2 and NO3- into the oxic, suboxic and anoxic layers. Prominent oxygen intrusions caused an overlap of NOx- and sulfide at the same station where autotrophic denitrification activity was detected with incubation experiments. Several bacteria that have been proposed to oxidize sulfide in other low oxygen environments were found in the Black Sea including SUP05, Sulfurimonas, Arcobacter, and BS-GSO2. Comparison of TRFLP profiles from this mixing zone station and the Western Gyre (a station not affected by the Bosporus Plume indicate the greatest relative abundance of Sulfurimonas and Arcobacter at the appropriate depths at the mixing zone station. The autotrophic gammaproteobacterium BS-GSO2 correlated with ammonium fluxes rather than with sulfide fluxes and the maximum in SUP05 peak height was shallower than the depths where autotrophic denitrification was detected. Notably, anammox activity was not detected at the mixing zone station, though low levels of DNA from the anammox bacteria Candidatus Scalindua were present. These results provide evidence for a modified ecosystem with different N2 production pathways in the southwest coastal region compared to that found in the rest of the Black Sea. Moreover, the same Sulfurimonas phylotype (BS139 was previously detected on >30 μm particles in the suboxic zone of the Western Gyre along with DNA of potential sulfate reducers, so it is possible that particle-attached autotrophic denitrification may be an overlooked N2 production pathway in the central Black Sea as well.

  3. CRYSTAL-STRUCTURE OF AN ELECTRON-TRANSFER COMPLEX BETWEEN METHYLAMINE DEHYDROGENASE AND AMICYANIN

    NARCIS (Netherlands)

    CHEN, LY; DURLEY, R; POLIKS, BJ; HAMADA, K; CHEN, ZW; MATHEWS, FS; DAVIDSON, VL; SATOW, Y; HUIZINGA, E; VELLIEUX, FMD; HOL, WGJ

    1992-01-01

    The crystal structure of the complex between the quinoprotein methylamine dehydrogenase (MADH) and the type I blue copper protein amicyanin, both from Paracoccus denitrificans, has been determined at 2.5-angstrom resolution using molecular replacement. The search model was MADH from Thiobacillus ver

  4. Purification, characterization, and cloning of a bifunctional molybdoenzyme with hydratase and alcohol dehydrogenase activity

    NARCIS (Netherlands)

    Jin, J.; Straathof, A.J.J.; Pinkse, M.W.H.; Hanefeld, U.

    2010-01-01

    A bifunctional hydratase/alcohol dehydrogenase was isolated from the cyclohexanol degrading bacterium Alicycliphilus denitrificans DSMZ 14773. The enzyme catalyzes the addition of water to α,β-unsaturated carbonyl compounds and the subsequent alcohol oxidation. The purified enzyme showed three subun

  5. Michael hydratase alcohol dehydrogenase or just alcohol dehydrogenase?

    NARCIS (Netherlands)

    Resch, V.A.; Jin, J.; Chen, B.S.; Hanefeld, U.

    2014-01-01

    The Michael hydratase – alcohol dehydrogenase (MhyADH) from Alicycliphilus denitrificans was previously identified as a bi-functional enzyme performing a hydration of α,β-unsaturated ketones and subsequent oxidation of the formed alcohols. The investigations of the bi-functionality were based on a s

  6. Three-dimensional model of stellacyanin and its implications for electron transfer reactivity

    DEFF Research Database (Denmark)

    Wherland, S; Farver, O; Pecht, I

    1988-01-01

    . The structure also indicates that a carbonyl oxygen atom is near the copper, thus the site may have analogy to the Alcaligenes denitrificans azurin (Az) site, although the amino acid sequence is more homologous to that of Pc. The model indicates that aspartate 49, reductively labeled by Cr(III), is near...

  7. COMPETITION BETWEEN ANOXYGENIC PHOTOTROPHIC BACTERIA AND COLORLESS SULFUR BACTERIA IN A MICROBIAL MAT

    NARCIS (Netherlands)

    VISSCHER, PT; VANDENENDE, FP; SCHAUB, BEM; VANGEMERDEN, H

    1992-01-01

    The populations of chemolithoautotrophic (colorless) sulfur bacteria and anoxygenic phototrophic bacteria were enumerated in a marine microbial mat. The highest population densities were found in the 0-5 mm layer of the mat: 2.0 X 10(9) cells CM-3 sediment, and 4.0 X 10(7) cells cm-3 sediment for th

  8. RATES OF SULFATE REDUCTION AND THIOSULFATE CONSUMPTION IN A MARINE MICROBIAL MAT

    NARCIS (Netherlands)

    VISSCHER, PT; PRINS, RA; VANGEMERDEN, H

    1992-01-01

    The sulfur cycle in a microbial mat was studied by determining viable counts of sulfate-reducing bacteria, chemolithoautotrophic sulfur bacteria and anoxygenic phototrophic bacteria. All three functional groups of sulfur bacteria revealed a maximum population density in the uppermost 5 mm of the mat

  9. Physiology of Haloalkaliphilic Sulfur-oxidizing Bacteria

    NARCIS (Netherlands)

    Banciu, H.L.

    2004-01-01

    The inorganic sulfur oxidation by obligate haloalkaliphilic chemolithoautotrophs was only recently discovered and investigated. These autotrophic sulfur oxidizing bacteria (SOB), capable of oxidation of inorganic sulfur compounds at moderate to high salt concentration and at high pH, can be divided

  10. Assessment of the stoichiometry and efficiency of CO2 fixation coupled to reduced sulfur oxidation

    NARCIS (Netherlands)

    Klatt, Judith M.; Polerecky, Lubos

    2015-01-01

    Chemolithoautotrophic sulfur oxidizing bacteria (SOB) couple the oxidation of reduced sulfur compounds to the production of biomass. Their role in the cycling of carbon, sulfur, oxygen, and nitrogen is, however, difficult to quantify due to the complexity of sulfur oxidation pathways. We describe a

  11. Complete nitrification by Nitrospira bacteria

    DEFF Research Database (Denmark)

    Daims, Holger; Lebedeva, Elena V.; Pjevac, Petra

    2015-01-01

    Nitrification, the oxidation of ammonia via nitrite to nitrate, has always been considered to be a two-step process catalysed by chemolithoautotrophic microorganisms oxidizing either ammonia or nitrite. No known nitrifier carries out both steps, although complete nitrification should be energetic...

  12. Draft genome sequence of the extremely acidophilic biomining bacterium Acidithiobacillus thiooxidans ATCC 19377 provides insights into the evolution of the Acidithiobacillus genus.

    Science.gov (United States)

    Valdes, Jorge; Ossandon, Francisco; Quatrini, Raquel; Dopson, Mark; Holmes, David S

    2011-12-01

    Acidithiobacillus thiooxidans is a mesophilic, extremely acidophilic, chemolithoautotrophic gammaproteobacterium that derives energy from the oxidation of sulfur and inorganic sulfur compounds. Here we present the draft genome sequence of A. thiooxidans ATCC 19377, which has allowed the identification of genes for survival and colonization of extremely acidic environments.

  13. Draft Genome Sequence of the Extremely Acidophilic Biomining Bacterium Acidithiobacillus thiooxidans ATCC 19377 Provides Insights into the Evolution of the Acidithiobacillus Genus

    OpenAIRE

    Valdes, Jorge; Ossandon, Francisco; Quatrini, Raquel; Dopson, Mark; Holmes, David S

    2011-01-01

    Acidithiobacillus thiooxidans is a mesophilic, extremely acidophilic, chemolithoautotrophic gammaproteobacterium that derives energy from the oxidation of sulfur and inorganic sulfur compounds. Here we present the draft genome sequence of A. thiooxidans ATCC 19377, which has allowed the identification of genes for survival and colonization of extremely acidic environments.

  14. Complete Genome Sequence of Nitrosomonas cryotolerans ATCC 49181, a Phylogenetically Distinct Ammonia-Oxidizing Bacterium Isolated from Arctic Waters.

    Science.gov (United States)

    Rice, Marlen C; Norton, Jeanette M; Stein, Lisa Y; Kozlowski, Jessica; Bollmann, Annette; Klotz, Martin G; Sayavedra-Soto, Luis; Shapiro, Nicole; Goodwin, Lynne A; Huntemann, Marcel; Clum, Alicia; Pillay, Manoj; Varghese, Neha; Mikhailova, Natalia; Palaniappan, Krishna; Ivanova, Natalia; Mukherjee, Supratim; Reddy, T B K; Yee Ngan, Chew; Daum, Chris; Kyrpides, Nikos; Woyke, Tanja

    2017-03-16

    Nitrosomonas cryotolerans ATCC 49181 is a cold-tolerant marine ammonia-oxidizing bacterium isolated from seawater collected in the Gulf of Alaska. The high-quality complete genome contains a 2.87-Mbp chromosome and a 56.6-kbp plasmid. Chemolithoautotrophic modules encoding ammonia oxidation and CO2 fixation were identified.

  15. Complete genome sequence of Nitrosomonas sp. Is79, an ammonia oxidizing bacterium adapted to low ammonium concentrations

    NARCIS (Netherlands)

    Bollmann, A.; Sedlacek, C.J.; Norton, J.; Laanbroek, H.J.; Suwa, Y.; Stein, L.Y.; Klotz, M.G.; Arp, D.; Sayavedra-Soto, L.; Lu, M.; Bruce, D.; Detter, C.; Tapia, R.; Han, J.; Woyke, T.; Lucas, S.; Pitluck, S.; Pennacchio, L.; Nolan, M.; Land, M.L.; Huntemann, M.; Deshpande, S.; Han, C.; Chen, A.; Kyrpides, N.; Mavromatis, K.; Markowitz, V.; Szeto, E.; Ivanova, N.; Mikhailova, N.; Pagani, I.; Pati, A.; Peters, L.; Ovchinnikova, G.; Goodwin, L.

    2013-01-01

    Nitrosomonas sp. Is79 is a chemolithoautotrophic ammonia-oxidizing bacterium that belongs to the family Nitrosomonadaceae within the phylum Proteobacteria. Ammonia oxidation is the first step of nitrification, an important process in the global nitrogen cycle ultimately resulting in the production o

  16. Microbial control of the production of hydrogen sulfide by sulfate-reducing bacteria.

    Science.gov (United States)

    Montgomery, A D; McLnerney, M J; Sublette, K L

    1990-03-01

    A sulfide-resistant ctrain of Thiobacillus denitrificans, strain F, prevented the accumulation of sulfide by Desulfovibrio desulfuricans when both organisms were grown in liquid medium or in Berea sandstone cores. The wild-type strain of T. denitrificans did not prevent the accumulation of sulfide produced by D. desulfuricans. Strain F also prevented the accumulation of sulfide by a mixed population of sulfate-reducing bacteria enriched from an oil field brine. Fermentation balances showed that strain F stoichiometrically oxidized the sulfide produced by D. desulfuricans and the oil field brine enrichment to sulfate. These data suggest that strain F would be effective in controlling sulfide production in oil reservoirs and other environments.

  17. Oxidation/Biodegradation of Solid Propellants Used in Legacy Chemical Rounds

    Science.gov (United States)

    2007-08-01

    Bioreactor Sample Source Sample Number Similarity Index Genus Species ICB M28-1 Sample 1A 0.771 Kluyvera cryocrescenes 0.704 Enterobacter cloacae...0.678 Photorhabdus luminencent 0.676 Entrobacter aerogenes Sample 1B 0.901 Alcaligenes faecalis Sample 2 0.894 Pseudomonas stutzeri 0.807 Pseudomonas...et. al. 13 has also described the role of Enterobacter cloacae NADH in the degradation of nitro aromatic compounds. Paracoccus denitrificans, commonly

  18. Construction and characterization of an azurin analog for the purple copper site in cytochrome c oxidase.

    OpenAIRE

    Hay, M; Richards, J. H.; Lu, Y.

    1996-01-01

    A protein analog of a purple copper center has been constructed from a recombinant blue copper protein (Pseudomonas aeruginosa azurin) by replacing the loop containing the three ligands to the blue copper center with the corresponding loop of the CuA center in cytochrome c oxidase (COX) from Paracoccus denitrificans. The electronic absorption in the UV and visible region (UV-vis) and electron paramagnetic resonance (EPR) spectra of this analog are remarkably similar to those of the native CuA...

  19. A two-subunit cytochrome c oxidase (cytochrome aa3) from Paracoccus dentrificans.

    OpenAIRE

    Ludwig, B.; Schatz, G

    1980-01-01

    Cytochrome c oxidase (ferrocytochrome c: oxygen oxidoreductase, EC 1.9.3.1) was purified from the cytoplasmic membrane of the bacterium Paracoccus denitrificans. The enzyme contains two heme groups (a and a3) and two copper atoms per minimal unit, oxidizes mammalian cytochrome c at a high rate, and, when incorporated into liposomes, generates an electrochemical proton gradient during cytochrome c oxidation. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis reveals only two subunits of...

  20. Prediction and Biochemical Demonstration of a Catabolic Pathway for the Osmoprotectant Proline Betaine

    OpenAIRE

    Kumar, Ritesh; Zhao, Suwen; Vetting, Matthew W.; Wood, B. McKay; Sakai, Ayano; Cho, Kyuil; Solbiati, José; Steven C Almo; Jonathan V Sweedler; Matthew P Jacobson; Gerlt, John A.; Cronan, John E.

    2014-01-01

    ABSTRACT Through the use of genetic, enzymatic, metabolomic, and structural analyses, we have discovered the catabolic pathway for proline betaine, an osmoprotectant, in Paracoccus denitrificans and Rhodobacter sphaeroides. Genetic and enzymatic analyses showed that several of the key enzymes of the hydroxyproline betaine degradation pathway also function in proline betaine degradation. Metabolomic analyses detected each of the metabolic intermediates of the pathway. The proline betaine catab...

  1. 脱窒細菌を固定化する架橋poly (vinyl alcohol) ゲルマイクロカプセルの調製と脱窒能力の評価

    OpenAIRE

    大久保, 一心; 吉田, 昌弘; 上村, 芳三; 幡手, 泰雄; 畑中, 千秋; オオクボ, カズヒト; ヨシダ, マサヒロ; ウエムラ, ヨシミツ; ハタテ, ヤスオ; ハタナカ, チアキ; OHKUBO, Kazuhito; Yoshida, Masahiro; Uemura, Yoshimitsu; HATATE, Yasuo; HATANAKA, Chiaki

    2004-01-01

    Recently, nitrate-nitrogen concentration in groundwater increases gradually. Drinking water including nitrate-nitrogen is possible to cause several human-and animal-health damages. Therefore, efficient denitrification methods have been investigated from all angles. In this study, autotrophic denitrifying bacteria, Paracoccus denitrificans IFO 13301 using H_2 gas as a hydrogen donor was immobilized in microcapsules composed of cross-linked poly (vinyl alcohol) gel. The efficient microencapsula...

  2. Identify biosorption effects of Thiobacillus towards perfluorooctanoic acid (PFOA): Pilot study from field to laboratory.

    Science.gov (United States)

    Li, Lei; Wang, Tieyu; Sun, Yajun; Wang, Pei; Yvette, Baninla; Meng, Jing; Li, Qifeng; Zhou, Yunqiao

    2017-03-01

    The concentration of Perfluoroalkyl acids (PFAAs) and the bacterial community composition along the Xiaoqing River were explored with HPLC-MS/MS and Illumina high-throughput sequencing in present study. The results showed that perfluorooctanoic acid (PFOA) was the predominant PFAAs in all sediment samples, and high level of PFOA could lead to an evident increase in the abundance of Thiobacillus. Thiobacillus was identified with the survival ability in high concentrations of PFOA accordingly. Therefore, Thiobacillus thioparus and Thiobacillus denitrificans were selected as receptors to design indoor biosorption experiment. The growth curves under different PFOA concentrations and residual rates of PFOA in the processes of cultivation were analyzed. The results showed that upwards concentrations of PFOA below 5000 ng/L led to an obvious increase in the growth rate of T. thioparus. Whereas PFOA promoted the growth of T. denitrificans in a relatively limited range of concentration, and the effect was not obvious. The addition of different concentrations of PFOA had no apparent effects on pH values in the media of both T. thioparus and T. denitrificans. The concentrations of PFOA in liquid media reduced after the process of bacteria culturing. The removal rates of T. thioparus and T. denitrificans to PFOA were 21.1-26.8% and 13.5-18.4%, respectively. The current findings indicated that T. thioparus could play a significant role as potential biosorbent with the ability to eliminate PFOA effectively in aquatic environment, which would provide novel information for PFOA ecological decontamination and remediation.

  3. Associations of Europium(III) with gram-negative bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Ozaki, T.; Ohnuki, T. [Advanced Science Research Center, Japan Atomic Energy Research Institute, Tokai, Ibaraki 319-1195 (Japan); Kimura, T. [Department of Materials Science, Japan Atomic Energy Research Institute, Tokai, Ibaraki 319-1195 (Japan); Takahashi, Y. [Department of Earth and Planetary Systems Science, Hiroshima University, Hiroshima 739- 8526 (Japan); Francis, A.J. [Brookhaven National Laboratory, Upton, New York 11973 (United States)

    2005-07-01

    Full text of publication follows: Migration of radionuclides in the environment is greatly affected by bacteria. Gram-negative bacteria are ubiquitous in the environment and can preferentially bind radionuclides because of the presence of the cell envelop consisting of two membrane bilayers with an intervening thin peptidoglycan layer, where carboxyl and phosphate functional groups are mainly involved in metal cation adsorption. In this study, we investigated the association of Eu(III) with four Gram-negative bacteria Pseudomonas fluorescens, Alcaligenes faecalis, Shewanella putrefaciens, and Paracoccus denitrificans. Europium(III) is a good analogue of Am(III) and Cm(III). The association of Eu(III) with the bacteria were determined by time-resolved laser-induced fluorescence spectroscopy (TRLFS). The kinetics study showed that the Eu(III) adsorption on the bacteria proceeded rapidly. The Eu(III) adsorption on P. fluorescens at pH 3, A. faecalis and P. denitrificans at pHs 3, 4, and 5, and S. putrefaciens at pHs 4 and 5 reached a maximum within 5 minutes after contact. For P. denitrificans, the percent adsorption of Eu(III) decreased after the maximum percent adsorption was attained, which suggests the existence of exudates with an affinity with Eu(III). TRLFS showed that the coordination of Eu(III) on these bacteria is multi-dentate through an inner-spherical process. The ligand field of Eu(III) on P. denitrificans was as strong as the ones observed for halo-philic microorganisms, while that of P. fluorescens, A. faecalis, and S. putrefaciens was the typical one observed for non-halo-philic microorganisms. The coordination environment of Eu(III) on the bacteria differed from each other, though they are categorized as Gram-negative bacteria with the similar cell wall components. (authors)

  4. Electron transfer flavoprotein domain II orientation monitored using double electron-electron resonance between an enzymatically reduced, native FAD cofactor, and spin labels.

    Science.gov (United States)

    Swanson, Michael A; Kathirvelu, Velavan; Majtan, Tomas; Frerman, Frank E; Eaton, Gareth R; Eaton, Sandra S

    2011-03-01

    Human electron transfer flavoprotein (ETF) is a soluble mitochondrial heterodimeric flavoprotein that links fatty acid β-oxidation to the main respiratory chain. The crystal structure of human ETF bound to medium chain acyl-CoA dehydrogenase indicates that the flavin adenine dinucleotide (FAD) domain (αII) is mobile, which permits more rapid electron transfer with donors and acceptors by providing closer access to the flavin and allows ETF to accept electrons from at least 10 different flavoprotein dehydrogenases. Sequence homology is high and low-angle X-ray scattering is identical for Paracoccus denitrificans (P. denitrificans) and human ETF. To characterize the orientations of the αII domain of P. denitrificans ETF, distances between enzymatically reduced FAD and spin labels in the three structural domains were measured by double electron-electron resonance (DEER) at X- and Q-bands. An FAD to spin label distance of 2.8 ± 0.15 nm for the label in the FAD-containing αII domain (A210C) agreed with estimates from the crystal structure (3.0 nm), molecular dynamics simulations (2.7 nm), and rotamer library analysis (2.8 nm). Distances between the reduced FAD and labels in αI (A43C) were between 4.0 and 4.5 ± 0.35 nm and for βIII (A111C) the distance was 4.3 ± 0.15 nm. These values were intermediate between estimates from the crystal structure of P. denitrificans ETF and a homology model based on substrate-bound human ETF. These distances suggest that the αII domain adopts orientations in solution that are intermediate between those which are observed in the crystal structures of free ETF (closed) and ETF bound to a dehydrogenase (open).

  5. Respiratory arsenate reductase as a bidirectional enzyme

    Science.gov (United States)

    Richey, C.; Chovanec, P.; Hoeft, S.E.; Oremland, R.S.; Basu, P.; Stolz, J.F.

    2009-01-01

    The haloalkaliphilic bacterium Alkalilimnicola ehrlichii is capable of anaerobic chemolithoautotrophic growth by coupling the oxidation of arsenite (As(III)) to the reduction of nitrate and carbon dioxide. Analysis of its complete genome indicates that it lacks a conventional arsenite oxidase (Aox), but instead possesses two operons that each encode a putative respiratory arsenate reductase (Arr). Here we show that one homolog is expressed under chemolithoautotrophic conditions and exhibits both arsenite oxidase and arsenate reductase activity. We also demonstrate that Arr from two arsenate respiring bacteria, Alkaliphilus oremlandii and Shewanella sp. strain ANA-3, is also biochemically reversible. Thus Arr can function as a reductase or oxidase. Its physiological role in a specific organism, however, may depend on the electron potentials of the molybdenum center and [Fe–S] clusters, additional subunits, or constitution of the electron transfer chain. This versatility further underscores the ubiquity and antiquity of microbial arsenic metabolism.

  6. Construction and characterization of an azurin analog for the purple copper site in cytochrome c oxidase.

    Science.gov (United States)

    Hay, M; Richards, J H; Lu, Y

    1996-01-09

    A protein analog of a purple copper center has been constructed from a recombinant blue copper protein (Pseudomonas aeruginosa azurin) by replacing the loop containing the three ligands to the blue copper center with the corresponding loop of the CuA center in cytochrome c oxidase (COX) from Paracoccus denitrificans. The electronic absorption in the UV and visible region (UV-vis) and electron paramagnetic resonance (EPR) spectra of this analog are remarkably similar to those of the native CuA center in COX from Paracoccus denitrificans. The above spectra can be obtained upon addition of a mixture of Cu2+ and Cu+. Addition of Cu2+ only results in a UV-vis spectrum consisting of absorptions from both a purple copper center and a blue copper center. This spectrum can be converted to the spectrum of a pure purple copper by a prolonged incubation in the air, or by addition of excess ascorbate. The azurin mutant reported here is an example of an engineered purple copper center with the A480/A530 ratio greater than 1 and with no detectable hyperfines, similar to those of the CuA sites in COX of bovine heart and of Paracoccus denitrificans.

  7. EVELOPMENT OF AN ENVIRONMENTALLY BENIGN MICROBIAL INHIBITOR TO CONTROL INTERNAL PIPELINE CORROSION

    Energy Technology Data Exchange (ETDEWEB)

    Bill W. Bogan; Wendy R. Sullivan; Kristine M. H. Cruz; Kristine L. Lowe; John J. Kilbane II

    2004-04-30

    The overall program objective is to develop and evaluate environmentally benign agents or products that are effective in the prevention, inhibition, and mitigation of microbially influenced corrosion (MIC) in the internal surfaces of metallic natural gas pipelines. The goal is to develop one or more environmentally benign (a.k.a. ''green'') products that can be applied to maintain the structure and dependability of the natural gas infrastructure. Previous testing of pepper extracts resulted in preliminary data indicating that some pepper extracts inhibit the growth of some corrosion-associated microorganisms. This quarter additional tests were performed to more specifically investigate the ability of three pepper extracts to inhibit the growth, and to influence the metal corrosion caused by two microbial species: Desulfovibrio vulgaris, and Comomonas denitrificans. All three pepper extracts rapidly killed Desulfovibrio vulgaris, but did not appear to inhibit Comomonas denitrificans. While corrosion rates were at control levels in experiments with Desulfovibrio vulgaris that received pepper extract, corrosion rates were increased in the presence of Comomonas denitrificans plus pepper extract. Further testing with a wider range of pure bacterial cultures, and more importantly, with mixed bacterial cultures should be performed to determine the potential effectiveness of pepper extracts to inhibit MIC.

  8. Coenzyme B12 can be produced by engineered Escherichia coli under both anaerobic and aerobic conditions.

    Science.gov (United States)

    Ko, Yeounjoo; Ashok, Somasundar; Ainala, Satish Kumar; Sankaranarayanan, Mugesh; Chun, Ah Yeong; Jung, Gyoo Yeol; Park, Sunghoon

    2014-12-01

    Coenzyme B12 (Vitamin B12 ) is one of the most complex biomolecules and an essential cofactor required for the catalytic activity of many enzymes. Pseudomonas denitrificans synthesizes coenzyme B12 in an oxygen-dependent manner using a pathway encoded by more than 25 genes that are located in six different operons. Escherichia coli, a robust and suitable host for metabolic engineering was used to produce coenzyme B12 . These genes were cloned into three compatible plasmids and expressed heterologously in E. coli BL21 (DE3). Real-time PCR, SDS-PAGE analysis and bioassay showed that the recombinant E. coli expressed the coenzyme B12 synthetic genes and successfully produced coenzyme B12 . However, according to the quantitative determination by inductively coupled plasma-mass spectrometry, the amount of coenzyme B12 produced by the recombinant E. coli (0.21 ± 0.02 μg/g cdw) was approximately 13-fold lower than that by P. denitrificans (2.75 ± 0.22 μg/g cdw). Optimization of the culture conditions to improve the production of coenzyme B12 by the recombinant E. coli was successful, and the highest titer (0.65 ± 0.03 μg/g cdw) of coenzyme B12 was obtained. Interestingly, although the synthesis of coenzyme B12 in P. denitrificans is strictly oxygen-dependent, the recombinant E. coli could produce coenzyme B12 under anaerobic conditions.

  9. The Genome of Nitrospina gracilis Illuminates the Metabolism and Evolution of the Major Marine Nitrite Oxidizer

    OpenAIRE

    2013-01-01

    In marine systems, nitrate is the major reservoir of inorganic fixed nitrogen. The only known biological nitrate-forming reaction is nitrite oxidation, but despite its importance, our knowledge of the organisms catalyzing this key process in the marine N-cycle is very limited. The most frequently encountered marine NOB are related to Nitrospina gracilis, an aerobic chemolithoautotrophic bacterium isolated from ocean surface waters. To date, limited physiological and genomic data for this orga...

  10. The genome of Nitrospina gracilis illuminates the metabolism and evolution of the major marine nitrite oxidizer

    OpenAIRE

    2013-01-01

    In marine systems, nitrate is the major reservoir of inorganic fixed nitrogen. The only known biological nitrate-forming reaction is nitrite oxidation, but despite its importance, our knowledge of the organisms catalyzing this key process in the marine N-cycle is very limited. The most frequently encountered marine NOB are related to Nitrospina gracilis, an aerobic chemolithoautotrophic bacterium isolated from ocean surface waters. To date, limited physiological and genomic data for this orga...

  11. Carbon isotope fractionation by the marine ammonia-oxidizing archaeon Nitrosopumilus maritimus

    OpenAIRE

    Könneke, Martin; Lipp, Julius Sebastian; Hinrichs, Kai-Uwe

    2012-01-01

    Abstract Ammonia-oxidizing archaea (AOA) are abundant and widely distributed microorganisms in aquatic and terrestrial habitats. By catalyzing the first and rate limiting step in nitrification, these chemolithoautotrophs play a significant role in the global nitrogen cycle and contribute to primary production. Here, the carbon isotopic fractionation relative to inorganic carbon source was determined for bulk biomass, biphytanes and polar lipid bound sugars of a marine AOA pure culture. Bu...

  12. Giant hydrogen sulfide plume in the oxygen minimum zone off Peru supports chemolithoautotrophy.

    Directory of Open Access Journals (Sweden)

    Harald Schunck

    Full Text Available In Eastern Boundary Upwelling Systems nutrient-rich waters are transported to the ocean surface, fuelling high photoautotrophic primary production. Subsequent heterotrophic decomposition of the produced biomass increases the oxygen-depletion at intermediate water depths, which can result in the formation of oxygen minimum zones (OMZ. OMZs can sporadically accumulate hydrogen sulfide (H2S, which is toxic to most multicellular organisms and has been implicated in massive fish kills. During a cruise to the OMZ off Peru in January 2009 we found a sulfidic plume in continental shelf waters, covering an area >5500 km(2, which contained ∼2.2×10(4 tons of H2S. This was the first time that H2S was measured in the Peruvian OMZ and with ∼440 km(3 the largest plume ever reported for oceanic waters. We assessed the phylogenetic and functional diversity of the inhabiting microbial community by high-throughput sequencing of DNA and RNA, while its metabolic activity was determined with rate measurements of carbon fixation and nitrogen transformation processes. The waters were dominated by several distinct γ-, δ- and ε-proteobacterial taxa associated with either sulfur oxidation or sulfate reduction. Our results suggest that these chemolithoautotrophic bacteria utilized several oxidants (oxygen, nitrate, nitrite, nitric oxide and nitrous oxide to detoxify the sulfidic waters well below the oxic surface. The chemolithoautotrophic activity at our sampling site led to high rates of dark carbon fixation. Assuming that these chemolithoautotrophic rates were maintained throughout the sulfidic waters, they could be representing as much as ∼30% of the photoautotrophic carbon fixation. Postulated changes such as eutrophication and global warming, which lead to an expansion and intensification of OMZs, might also increase the frequency of sulfidic waters. We suggest that the chemolithoautotrophically fixed carbon may be involved in a negative feedback loop that

  13. The Diversity of Sulfide Oxidation and Sulfate Reduction Genes Expressed by the Bacterial Communities of the Cariaco Basin, Venezuela.

    Science.gov (United States)

    Rodriguez-Mora, Maria J; Edgcomb, Virginia P; Taylor, Craig; Scranton, Mary I; Taylor, Gordon T; Chistoserdov, Andrei Y

    2016-01-01

    Qualitative expression of dissimilative sulfite reductase (dsrA), a key gene in sulfate reduction, and sulfide:quinone oxidoreductase (sqr), a key gene in sulfide oxidation was investigated. Neither of the two could be amplified from mRNA retrieved with Niskin bottles but were amplified from mRNA retrieved by the Deep SID. The sqr and sqr-like genes retrieved from the Cariaco Basin were related to the sqr genes from a Bradyrhizobium sp., Methylomicrobium alcaliphilum, Sulfurovum sp. NBC37-1, Sulfurimonas autotrophica, Thiorhodospira sibirica and Chlorobium tepidum. The dsrA gene sequences obtained from the redoxcline of the Cariaco Basin belonged to chemoorganotrophic and chemoautotrophic sulfate and sulfur reducers belonging to the class Deltaproteobacteria (phylum Proteobacteria) and the order Clostridiales (phylum Firmicutes).

  14. Microbial reduction of SO[sub 2] and NO[sub x] as a means of by- product recovery/disposal from regenerable processes for the desulfurization of flue gas

    Energy Technology Data Exchange (ETDEWEB)

    Sublette, K.L.

    1992-01-01

    Based on the work described simultaneous SO[sub 2]/No[sub x] removal from flue gas based on direct contact of the gas with SRB and T. denitrificans co-cultures or cultures-in-series has been eliminated as a viable process concept at this time. The technical reasons are as follows: (1) NO inhibition of SO[sub 2] reduction by D. desulfuricans - Although the NO concentrations used in the experiments described above are somewhat higher than that found in a typical flue gas, it is quite possible that at lower NO concentrations (or partial pressures) the inhibiting effects will simply take longer to become apparent. (2) Nitrate suppression of NO removal - As noted previously, the cultivation of T. denitrificans in a microbial flue gas treatment system (either one or two stages) would require sulfide-limiting conditions. Therefore, the electron acceptor must be in excess, requiring nitrate in the T. denitrificans process culture. As shown in experiments described above, nitrate significantly suppresses the removal of NO from a feed gas making simultaneous SO[sub 2]/NO[sub x] removal impractical by microbial means. (3) O[sub 2] inhibition of SO[sub 2] and NO reduction - It has been demonstrated that D. desulfuricans working cultures are tolerant of up to 1.7% O[sub 2] in the feed gas. However, further increases in the O[sub 2] partial pressure in the feed gas resulted in O[sub 2] inhibition of SO[sub 2] reduction. These inhibiting levels of O[sub 2] are comparable to those concentrations found in flue gases (3). Therefore, in any process in which raw flue gas contacts a D. desulfuricans culture marginal stability at best can be expected.

  15. Microbial reduction of SO{sub 2} and NO{sub x} as a means of by- product recovery/disposal from regenerable processes for the desulfurization of flue gas. Technical progress report, June 11, 1992--September 11, 1992

    Energy Technology Data Exchange (ETDEWEB)

    Sublette, K.L.

    1992-12-31

    Based on the work described simultaneous SO{sub 2}/No{sub x} removal from flue gas based on direct contact of the gas with SRB and T. denitrificans co-cultures or cultures-in-series has been eliminated as a viable process concept at this time. The technical reasons are as follows: (1) NO inhibition of SO{sub 2} reduction by D. desulfuricans - Although the NO concentrations used in the experiments described above are somewhat higher than that found in a typical flue gas, it is quite possible that at lower NO concentrations (or partial pressures) the inhibiting effects will simply take longer to become apparent. (2) Nitrate suppression of NO removal - As noted previously, the cultivation of T. denitrificans in a microbial flue gas treatment system (either one or two stages) would require sulfide-limiting conditions. Therefore, the electron acceptor must be in excess, requiring nitrate in the T. denitrificans process culture. As shown in experiments described above, nitrate significantly suppresses the removal of NO from a feed gas making simultaneous SO{sub 2}/NO{sub x} removal impractical by microbial means. (3) O{sub 2} inhibition of SO{sub 2} and NO reduction - It has been demonstrated that D. desulfuricans working cultures are tolerant of up to 1.7% O{sub 2} in the feed gas. However, further increases in the O{sub 2} partial pressure in the feed gas resulted in O{sub 2} inhibition of SO{sub 2} reduction. These inhibiting levels of O{sub 2} are comparable to those concentrations found in flue gases (3). Therefore, in any process in which raw flue gas contacts a D. desulfuricans culture marginal stability at best can be expected.

  16. Microbial reduction of SO{sub 2} and NO{sub x} as a means of by-product recovery/disposal from regenerable processes for the desulfurization of flue gas. Technical progress report, September 11, 1992--December 11, 1992

    Energy Technology Data Exchange (ETDEWEB)

    Sublette, K.L.

    1992-12-31

    With the continual increase in the utilization of high sulfur and high nitrogen containing fossil fuels, the release of airborne pollutants into the environment has become a critical problem. The fuel sulfur is converted to SO{sub 2} during combustion. Fuel nitrogen and a fraction of the nitrogen from the combustion air are converted to nitric oxide and nitrogen dioxide, NO{sub x}. For the past five years Combustion Engineering (now Asea Brown Boveri or ABB) and, since 1986, the University of Tulsa (TU) have been investigating the oxidation of H{sub 2}S by the facultatively anaerobic and autotrophic bacterium Thiobacillus denitrificans and have developed a process, concept for the microbial removal of H{sub 2}S from a gas stream the simultaneous removal of SO{sub 2} and NO by D. desulfuricans and T. denitrificans co-cultures and cultures-in-series was demonstrated. These systems could not be sustained due to NO inhibition of D. desulfuricans. However, a preliminary economic analysis has shown that microbial reduction of SO{sub 2} to H{sub 2}S with subsequent conversion to elemental sulfur by the Claus process is both technically and economically feasible if a less expensive carbon and/or energy source can be found. It has also been demonstrated that T. denitrificans can be grown anaerobically on NO(g) as a terminal electron acceptor with reduction to elemental nitrogen. Microbial reduction of NO{sub x} is a viable process concept for the disposal of concentrated streams of NO{sub x} as may be produced by certain regenerable processes for the removal of SO{sub 2} and NO{sub x} from flue gas.

  17. Electron transfer among the CuA-, heme b- and a3-centers of Thermus thermophilus cytochrome ba3

    DEFF Research Database (Denmark)

    Farver, Ole; Chen, Ying; Fee, James A

    2006-01-01

    . and Pecht, I. (2006) Rates and equilibrium of CuA to heme a electron transfer in Paracoccus denitrificans cytochrome c oxidase. Biophys. J. 90, 2131-2137]. Investigating this process in the cytochrome ba(3) of Thermus thermophilus (Tt), we now show that MNA(*) also reduces Cu(A) with a subsequent ET...... to the heme b and then to heme a(3), with first-order rate constants 11200 s(-1), and 770 s(-1), respectively. The results provide clear evidence for ET among the three spectroscopically distinguishable centers and indicate that the binuclear a(3)-Cu(B) center can be reduced in molecules containing a single...

  18. In Situ Gene Expression Responsible for Sulfide Oxidation and CO2 Fixation of an Uncultured Large Sausage-Shaped Aquificae Bacterium in a Sulfidic Hot Spring.

    Science.gov (United States)

    Tamazawa, Satoshi; Yamamoto, Kyosuke; Takasaki, Kazuto; Mitani, Yasuo; Hanada, Satoshi; Kamagata, Yoichi; Tamaki, Hideyuki

    2016-06-25

    We investigated the in situ gene expression profile of sulfur-turf microbial mats dominated by an uncultured large sausage-shaped Aquificae bacterium, a key metabolic player in sulfur-turfs in sulfidic hot springs. A reverse transcription-PCR analysis revealed that the genes responsible for sulfide, sulfite, and thiosulfate oxidation and carbon fixation via the reductive TCA cycle were continuously expressed in sulfur-turf mats taken at different sampling points, seasons, and years. These results suggest that the uncultured large sausage-shaped bacterium has the ability to grow chemolithoautotrophically and plays key roles as a primary producer in the sulfidic hot spring ecosystem in situ.

  19. Complete genome sequence of Nitrosomonas sp. Is79, an ammonia oxidizing bacterium adapted to low ammonium concentrations

    Energy Technology Data Exchange (ETDEWEB)

    Bollmann, Annette [Miami University, Oxford, OH; Sedlacek, Christopher J [Miami University, Oxford, OH; Laanbroek, Hendrikus J [Netherlands Institute of Ecology (NIOO-KNAW); Suwa, Yuichi [Chuo University, Tokyo, Japan; Stein, Lisa Y [University of California, Riverside; Klotz, Martin G [University of Louisville, Louisville; Arp, D J [Oregon State University; Sayavedra-Soto, LA [Oregon State University; Lu, Megan [Los Alamos National Laboratory (LANL); Bruce, David [Los Alamos National Laboratory (LANL); Detter, J. Chris [U.S. Department of Energy, Joint Genome Institute; Tapia, Roxanne [Los Alamos National Laboratory (LANL); Han, James [U.S. Department of Energy, Joint Genome Institute; Woyke, Tanja [U.S. Department of Energy, Joint Genome Institute; Lucas, Susan [U.S. Department of Energy, Joint Genome Institute; Pitluck, Sam [U.S. Department of Energy, Joint Genome Institute; Pennacchio, Len [U.S. Department of Energy, Joint Genome Institute; Nolan, Matt [U.S. Department of Energy, Joint Genome Institute; Land, Miriam L [ORNL; Huntemann, Marcel [U.S. Department of Energy, Joint Genome Institute; Deshpande, Shweta [U.S. Department of Energy, Joint Genome Institute; Han, Cliff [Los Alamos National Laboratory (LANL); Chen, Amy [U.S. Department of Energy, Joint Genome Institute; Kyrpides, Nikos C [U.S. Department of Energy, Joint Genome Institute; Mavromatis, K [U.S. Department of Energy, Joint Genome Institute; Markowitz, Victor [U.S. Department of Energy, Joint Genome Institute; Szeto, Ernest [U.S. Department of Energy, Joint Genome Institute; Ivanova, N [U.S. Department of Energy, Joint Genome Institute; Mikhailova, Natalia [U.S. Department of Energy, Joint Genome Institute; Pagani, Ioanna [U.S. Department of Energy, Joint Genome Institute; Pati, Amrita [U.S. Department of Energy, Joint Genome Institute; Peters, Lin [U.S. Department of Energy, Joint Genome Institute; Ovchinnikova, Galina [U.S. Department of Energy, Joint Genome Institute; Goodwin, Lynne A. [Los Alamos National Laboratory (LANL)

    2013-01-01

    Nitrosomonas sp. Is79 is a chemolithoautotrophic ammonia-oxidizing bacterium that belongs to the family Nitrosomonadaceae within the phylum Proteobacteria. Ammonia oxidation is the first step of nitrification, an important process in the global nitrogen cycle ultimately resulting in the production of nitrate. Nitrosomonas sp. Is79 is an ammonia oxidizer of high interest because it is adapted to low ammonium and can be found in freshwater environments around the world. The 3,783,444-bp chromosome with a total of 3,553 protein coding genes and 44 RNA genes was sequenced by the DOE-Joint Genome Institute Program CSP 2006.

  20. Methanococcus igneus sp. nov., a novel hyperthermophilic methanogen from a shallow submarine hydrothermal system

    Science.gov (United States)

    Burggraf, S.; Fricke, H.; Neuner, A.; Kristjansson, J.; Rouvier, P.; Mandelco, L.; Woese, C. R.; Stetter, K. O.

    1990-01-01

    A novel hyperthermophilic strictly chemolithoautotrophic member of the genus Methanococcus was isolated from a shallow (depth: 106 m) submarine vent system at the Kolbeinsey ridge, Iceland. The isolate grew between 45 and 91 degrees C with an optimum around 88 degrees C (doubling time: 25 min). It differs from Methanococcus jannaschii in its 16S rRNA sequence, its non-hybridizing DNA, and its selenium-independent growth. Therefore, the isolate represents a new species which we name Methanococcus igneus. Type strain is isolate "Kol 5" (DSM 5666).

  1. Nitrate stimulation of indigenous nitrate-reducing, sulfide-oxidising bacterial community in wastewater anaerobic biofilms.

    Science.gov (United States)

    Garcia-de-Lomas, Juan; Corzo, Alfonso; Carmen Portillo, M; Gonzalez, Juan M; Andrades, Jose A; Saiz-Jimenez, Cesáreo; Garcia-Robledo, Emilio

    2007-07-01

    The role of the nitrate-reducing, sulfide-oxidising bacteria (NR-SOB) in the nitrate-mediated inhibition of sulfide net production by anaerobic wastewater biofilms was analyzed in two experimental bioreactors, continuously fed with the primary effluent of a wastewater treatment plant, one used as control (BRC) and the other one supplemented with nitrate (BRN). This study integrated information from H(2)S and pH microelectrodes, RNA-based molecular techniques, and the time course of biofilm growth and bioreactors water phase. Biofilms were a net source of sulfide for the water phase (2.01 micromol S(2-)(tot)m(-2)s(-1)) in the absence of nitrate dosing. Nitrate addition effectively led to the cessation of sulfide release from biofilms despite which a low rate of net sulfate reduction activity (0.26 micromol S(2-)(tot)m(-2)s(-1)) persisted at a deep layer within the biofilm. Indigenous NR-SOB including Thiomicrospira denitrificans, Arcobacter sp., and Thiobacillus denitrificans were stimulated by nitrate addition resulting in the elimination of most sulfide from the biofilms. Active sulfate reducing bacteria (SRB) represented comparable fractions of total metabolically active bacteria in the libraries obtained from BRN and BRC. However, we detected changes in the taxonomic composition of the SRB community suggesting its adaptation to a higher level of NR-SOB activity in the presence of nitrate.

  2. Effects of pentachlorophenol on the bacterial denitrification process

    Directory of Open Access Journals (Sweden)

    Bairen Yang

    2016-10-01

    Full Text Available The use of pentachlorophenol (PCP was banned or restricted in many countries worldwide because of its adverse influences on the ecological environment and humans. However, the potential disrupting effects of PCP on denitrifying microorganisms have warranted more analysis. In this study, the impacts of PCP on denitrification were investigated by using Paracoccus denitrificans as a model denitrifying bacterium. Compared with the control, the presences of 10 and 50 μM of PCP were found to significantly decrease the denitrification efficiencies from 98.5 to 87.2% and 68.7%, respectively. The mechanism studies showed that PCP induced the generation of reactive oxygen species, which decreased the vital enzymes activities related to glycolysis process, causing the disturbance of the metabolism of P. denitrificans utilizing carbon source (glucose and the growth of the cell, and subsequently the generation of electron donor (NADH for denitrification via NAD+ reduction was severely depressed. Further studies indicated that PCP also decreased the genes expression of several key enzymes responsible for denitrification, such as napA of nitrate reductase (NAR, nirS of nitrite reductase, norB of nitric oxide reductase, and nosZ of nitrous oxide reductase; however, there was only the enzyme activity of NAR was remarkably inhibited.

  3. Microbial growth at hyperaccelerations up to 403,627 x g.

    Science.gov (United States)

    Deguchi, Shigeru; Shimoshige, Hirokazu; Tsudome, Mikiko; Mukai, Sada-atsu; Corkery, Robert W; Ito, Susumu; Horikoshi, Koki

    2011-05-10

    It is well known that prokaryotic life can withstand extremes of temperature, pH, pressure, and radiation. Little is known about the proliferation of prokaryotic life under conditions of hyperacceleration attributable to extreme gravity, however. We found that living organisms can be surprisingly proliferative during hyperacceleration. In tests reported here, a variety of microorganisms, including Gram-negative Escherichia coli, Paracoccus denitrificans, and Shewanella amazonensis; Gram-positive Lactobacillus delbrueckii; and eukaryotic Saccharomyces cerevisiae, were cultured while being subjected to hyperaccelerative conditions. We observed and quantified robust cellular growth in these cultures across a wide range of hyperacceleration values. Most notably, the organisms P. denitrificans and E. coli were able to proliferate even at 403,627 × g. Analysis shows that the small size of prokaryotic cells is essential for their proliferation under conditions of hyperacceleration. Our results indicate that microorganisms cannot only survive during hyperacceleration but can display such robust proliferative behavior that the habitability of extraterrestrial environments must not be limited by gravity.

  4. Biological reduction of iron to the elemental state from ochre deposits of Skelton Beck in Northeast England

    Directory of Open Access Journals (Sweden)

    Pattanathu K S M Rahman

    2014-06-01

    Full Text Available Ochre, consequence of acid mine drainage, is iron oxides-rich soil pigments that can be found in the water drainage from historic base metal and coal mines. The anaerobic strains of Geobacter sulfurreducens and Shewanella denitrificans were used for the microbial reduction of iron from samples of ochre collected from Skelton Beck (Saltburn Orange River, NZ 66738 21588 in Northeast England. The aim of the research was to determine the ability of the two anaerobic bacteria to reduce the iron present in ochre and to determine the rate of the reduction process. The physico-chemical changes in the ochre sample after the microbial reduction process were observed by the production of zero-valent iron which was later confirmed by the detection of elemental Fe in XRD spectrum. The XRF results revealed that 69.16% and 84.82% of iron oxide can be reduced using G. sulfurreducens and S. denitrificans respectively after 8 days of incubation. These results could provide the basis for the development of a biohydrometallurgical process for the production of elemental iron from ochre sediments.

  5. Corrigenda: Epigean and hypogean Palaemonetes sp. (Decapoda, Palaemonidae from Edwards Aquifer: An examination of trophic structure and metabolism. Subterranean Biology 14: 79–102.

    Directory of Open Access Journals (Sweden)

    Renee Bishop

    2015-06-01

    Full Text Available This study addresses the causes of the metabolic depression observed when examining the metabolism of hypogean versus epigean organisms. We examined the two current hypotheses regarding the cause of metabolic cave adaptation, a paucity of food and low oxygen availability, both necessary for ATP production, by first determining if the hypogean environment examined, Edwards Aquifer, was resource limited. Stable isotope analyses indicate that there is extensive microbial chemolithoautotrophic production providing resources for the hypogean organisms. δ13C values ( ≤30‰ were well below that of terrestrial biome indicating that C in the aquifer originates from chemolithoautotrophic inorganic carbon fixation, not photosynthetically derived material resulting from terrigenous sources. Data suggest the artesian system is a complex geochemical ecosystem providing inorganic energy sources from both methane and sulfates. Metabolism, examined via key aerobic and anaerobic proxies, and organismal proximate composition indicated there was no difference between metabolic rates and energy storage of Palaemonetes antrorum (stygobitic and Palaemonetes kadiakensis (epigean. This indicates that resources within the oxic aquifer are not limited. We demonstrate that it is necessary for one, or both, of these selective pressures to be present for metabolic cave adaptation to occur.

  6. Metals and minerals as a biotechnology feedstock: engineering biomining microbiology for bioenergy applications.

    Science.gov (United States)

    Banerjee, Indrani; Burrell, Brittany; Reed, Cara; West, Alan C; Banta, Scott

    2017-03-31

    Developing new feedstocks for the efficient production of biochemicals and biofuels will be a critical challenge as we diversify away from petrochemicals. One possible opportunity is the utilization of sulfide-based minerals in the Earth's crust. Non-photosynthetic chemolithoautotrophic bacteria are starting to be developed to produce biochemicals from CO2 using energy obtained from the oxidation of inorganic feedstocks. Biomining of metals like gold and copper already exploit the native metabolism of these bacteria and these represent perhaps the largest-scale bioprocesses ever developed. The metabolic engineering of these bacteria could be a desirable alternative to classical heterotrophic bioproduction. In this review, we discuss biomining operations and the challenges and advances in the engineering of associated chemolithoautotrophic bacteria for biofuel production. The co-generation of biofuels integrated with mining operations is a largely unexplored opportunity that will require advances in fundamental microbiology and the development of new genetic tools and techniques for these organisms. Although this approach is presently in its infancy, the production of biochemicals using energy from non-petroleum mineral resources is an exciting new biotechnology opportunity.

  7. Sulphur-cycling bacteria and ciliated protozoans in a Beggiatoaceae mat covering organically enriched sediments beneath a salmon farm in a southern Chilean fjord.

    Science.gov (United States)

    Aranda, Carlos P; Valenzuela, Cristian; Matamala, Yessica; Godoy, Félix A; Aranda, Nicol

    2015-11-15

    The colourless mat covering organically enriched sediments underlying an intensive salmon farm in Estero Pichicolo, southern Chile, was surveyed by combined 454 PyroTag and conventional Sanger sequencing of 16S/18S ribosomal RNA genes for Bacteria and Eukarya. The mat was dominated by the sulphide-oxidizing bacteria (SOB) Candidatus Isobeggiatoa, Candidatus Parabeggiatoa and Arcobacter. By order of their abundances, sulphate-reducing bacteria (SRB) were represented by diverse deltaproteobacterial Desulfobacteraceae, but also within Desulfobulbaceae, Desulfuromonadaceae and Desulfovibrionaceae. The eukaryotic PyroTags were dominated by polychaetes, copepods and nematodes, however, ciliated protozoans were highly abundant in microscopy observations, and were represented by the genera Condylostoma, Loxophyllum and Peritromus. Finally, the abundant Sulfurimonas/Sulfurovum also suggest the occurrence of zero-valence sulphur oxidation, probably derived from Beggiatoaceae as a result of bacteriovorus infaunal activity or generated as free S(0) by the Arcobacter bacteria. The survey suggests an intense and complex sulphur cycle within the surface of salmon-farm impacted sediments.

  8. Active Microbial Communities Inhabit Sulphate-Methane Interphase in Deep Bedrock Fracture Fluids in Olkiluoto, Finland

    Directory of Open Access Journals (Sweden)

    Malin Bomberg

    2015-01-01

    Full Text Available Active microbial communities of deep crystalline bedrock fracture water were investigated from seven different boreholes in Olkiluoto (Western Finland using bacterial and archaeal 16S rRNA, dsrB, and mcrA gene transcript targeted 454 pyrosequencing. Over a depth range of 296–798 m below ground surface the microbial communities changed according to depth, salinity gradient, and sulphate and methane concentrations. The highest bacterial diversity was observed in the sulphate-methane mixing zone (SMMZ at 250–350 m depth, whereas archaeal diversity was highest in the lowest boundaries of the SMMZ. Sulphide-oxidizing ε-proteobacteria (Sulfurimonas sp. dominated in the SMMZ and γ-proteobacteria (Pseudomonas spp. below the SMMZ. The active archaeal communities consisted mostly of ANME-2D and Thermoplasmatales groups, although Methermicoccaceae, Methanobacteriaceae, and Thermoplasmatales (SAGMEG, TMG were more common at 415–559 m depth. Typical indicator microorganisms for sulphate-methane transition zones in marine sediments, such as ANME-1 archaea, α-, β- and δ-proteobacteria, JS1, Actinomycetes, Planctomycetes, Chloroflexi, and MBGB Crenarchaeota were detected at specific depths. DsrB genes were most numerous and most actively transcribed in the SMMZ while the mcrA gene concentration was highest in the deep methane rich groundwater. Our results demonstrate that active and highly diverse but sparse and stratified microbial communities inhabit the Fennoscandian deep bedrock ecosystems.

  9. Active Microbial Communities Inhabit Sulphate-Methane Interphase in Deep Bedrock Fracture Fluids in Olkiluoto, Finland

    Science.gov (United States)

    Bomberg, Malin; Nyyssönen, Mari; Pitkänen, Petteri; Lehtinen, Anne; Itävaara, Merja

    2015-01-01

    Active microbial communities of deep crystalline bedrock fracture water were investigated from seven different boreholes in Olkiluoto (Western Finland) using bacterial and archaeal 16S rRNA, dsrB, and mcrA gene transcript targeted 454 pyrosequencing. Over a depth range of 296–798 m below ground surface the microbial communities changed according to depth, salinity gradient, and sulphate and methane concentrations. The highest bacterial diversity was observed in the sulphate-methane mixing zone (SMMZ) at 250–350 m depth, whereas archaeal diversity was highest in the lowest boundaries of the SMMZ. Sulphide-oxidizing ε-proteobacteria (Sulfurimonas sp.) dominated in the SMMZ and γ-proteobacteria (Pseudomonas spp.) below the SMMZ. The active archaeal communities consisted mostly of ANME-2D and Thermoplasmatales groups, although Methermicoccaceae, Methanobacteriaceae, and Thermoplasmatales (SAGMEG, TMG) were more common at 415–559 m depth. Typical indicator microorganisms for sulphate-methane transition zones in marine sediments, such as ANME-1 archaea, α-, β- and δ-proteobacteria, JS1, Actinomycetes, Planctomycetes, Chloroflexi, and MBGB Crenarchaeota were detected at specific depths. DsrB genes were most numerous and most actively transcribed in the SMMZ while the mcrA gene concentration was highest in the deep methane rich groundwater. Our results demonstrate that active and highly diverse but sparse and stratified microbial communities inhabit the Fennoscandian deep bedrock ecosystems. PMID:26425566

  10. Sulfate reduction and sulfide oxidation in extremely steep salinity gradients formed by freshwater springs emerging into the Dead Sea.

    Science.gov (United States)

    Häusler, Stefan; Weber, Miriam; Siebert, Christian; Holtappels, Moritz; Noriega-Ortega, Beatriz E; De Beer, Dirk; Ionescu, Danny

    2014-12-01

    Abundant microbial mats, recently discovered in underwater freshwater springs in the hypersaline Dead Sea, are mostly dominated by sulfur-oxidizing bacteria. We investigated the source of sulfide and the activity of these communities. Isotopic analysis of sulfide and sulfate in the spring water showed a fractionation of 39-50‰ indicative of active sulfate reduction. Sulfate reduction rates (SRR) in the spring sediment (Dead Sea water are responsible for the abundant microbial biomass around the springs. The springs flow is highly variable and accordingly the local salinities. We speculate that the development of microbial mats dominated by either Sulfurimonas/Sulfurovum-like or Thiobacillus/Acidithiobacillus-like sulfide-oxidizing bacteria, results from different mean salinities in the microenvironment of the mats. SRR of up to 10 nmol cm(-3) day(-1) detected in the Dead Sea sediment are surprisingly higher than in the less saline springs. While this shows the presence of an extremely halophilic sulfate-reducing bacteria community in the Dead Sea sediments, it also suggests that extensive salinity fluctuations limit these communities in the springs due to increased energetic demands for osmoregulation.

  11. Sulfur oxidation to sulfate coupled with electron transfer to electrodes by Desulfuromonas strain TZ1

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, T; Bain, TS; Barlett, MA; Dar, SA; Snoeyenbos-West, OL; Nevin, KP; Lovley, DR

    2014-01-02

    Microbial oxidation of elemental sulfur with an electrode serving as the electron acceptor is of interest because this may play an important role in the recovery of electrons from sulfidic wastes and for current production in marine benthic microbial fuel cells. Enrichments initiated with a marine sediment inoculum, with elemental sulfur as the electron donor and a positively poised (+300 mV versus Ag/AgCl) anode as the electron acceptor, yielded an anode biofilm with a diversity of micro-organisms, including Thiobacillus, Sulfurimonas, Pseudomonas, Clostridium and Desulfuromonas species. Further enrichment of the anode biofilm inoculum in medium with elemental sulfur as the electron donor and Fe(III) oxide as the electron acceptor, followed by isolation in solidified sulfur/Fe(III) medium yielded a strain of Desulfuromonas, designated strain TZ1. Strain TZ1 effectively oxidized elemental sulfur to sulfate with an anode serving as the sole electron acceptor, at rates faster than Desulfobulbus propionicus, the only other organism in pure culture previously shown to oxidize S with current production. The abundance of Desulfuromonas species enriched on the anodes of marine benthic fuel cells has previously been interpreted as acetate oxidation driving current production, but the results presented here suggest that sulfur-driven current production is a likely alternative.

  12. Diversity of prokaryotes at a shallow submarine vent of Panarea Island (Italy by high-throughput sequencing

    Directory of Open Access Journals (Sweden)

    Teresa L. Maugeri

    2013-09-01

    Full Text Available To determine microbial community composition and possible key microbial processes in the shallow-sea hydrothermal vent system off Panarea Island (Italy, we examined bacterial and archaeal communities of sediment and fluid samples from a hot vent by 16S rDNA Illumina sequencing technique. Both high abundant (>1% of total sequences, low abundant (from 0.1 to <1% and rare (< 0.1% phylogenetic groups were responsible for the distinct prokaryotic communities characterizing the heated sediment and fluid. The bacterial and archaeal communities from sediment were dominated by sequences affiliated with Rhodovulum genus (Alphaproteobacteria, including phototrophic ferrous-iron-oxidizing purple bacteria, Thiohalospira and Thiomicrospira (Gammaproteobacteria, typically involved in the sulphur cycle, and Methanococcus (Euryarchaeota. Fluid communities were dominated by anoxygenic phototrophic members of Chlorobium, followed by Thiomicrospira (Gammaproteobacteria, Sulfurimonas, Arcobacter and Sulfurospirillum (Epsilonproteobacteria, and Methanosarcina (Euryarchaeota. Obtained sequences were affiliated with prokaryotes taking a key part in the carbon, iron and sulphur cycling at the shallow hydrothermal system off Panarea Island. Despite the huge sequencing efforts, a great number of Bacteria and Archaea still remains unaffiliated at genus level, indicating that Black Point vent represents a hotspot of prokaryotic diversity.

  13. Evaluation of several modifications of an ecometric technique for assessment of media performance.

    Science.gov (United States)

    Kornacki, Jeffrey L; Gurtler, Joshua B; Yan, Zhinong; Cooper, Chad M

    2003-09-01

    Recovery of Listeria monocytogenes 101M, Jonesia denitrificans, salmonellae, and Pediococcus sp. NRRL B-2354 across nine media was evaluated with three modified versions of an ecometric method. Two approaches involved the use of broth cultures (10(8) to 10(9) CFU/ml) of individual strains and either large (10-microl) or small (1-microl) presterilized plastic loops. The third approach involved precultured slants and the inoculation of media with presterilized plastic inoculating needles (10(4) CFU per needle). Absolute growth indices (AGIs) were compared. No significant differences (P agar supplemented with 0.6% yeast extract (TSAYE) was used for the recovery of L. monocytogenes, J. denitrificans, Pediococcus sp. NRRL B-2354, and Salmonella spp. However, the small loop-broth technique recovered significantly fewer Salmonella enterica Typhimurium DT104 and Salmonella Senftenberg 775W cells than the other two techniques did. The performance of each individual bacterial strain on each of nine media was assayed. The recovery of L. monocytogenes was excellent (AGI > 4.8) with TSAYE, PALCAM, modified Oxford medium (MOX), and Baird-Parker agar and slight with modified PRAB (AGI = 0.4) and deMan Rogosa Sharpe (MRS) agar (lysine iron agar [MLIA], xylose lysine desoxycholate [XLD] agar, and xylose lysine tergitol 4 [XLT4] agar). The recovery of J. denitrificans with TSAYE and MOX was excellent, significantly better than that achieved with PALCAM (AGI = 3.0), but the organism was not recovered with Baird-Parker agar or with the other media tested. The recovery of Pediococcus sp. NRRL B-2354 was excellent with TSAYE and modified PRAB medium > Baird-Parker agar > acidified MRS agar, but the organism was not recovered with any of the other media tested. The best recovery of S. enterica Typhimurium DT104 was achieved with TSAYE > MLIA > or = XLD agar > or = XLT4 agar > Baird-Parker > PALCAM, MOX, acidified MRS agar, modified PRAB, and MRS agar. The best recovery of Salmonella

  14. HACEK endocarditis: state-of-the-art.

    Science.gov (United States)

    Revest, Matthieu; Egmann, Gérald; Cattoir, Vincent; Tattevin, Pierre

    2016-01-01

    The HACEK group of bacteria - Haemophilus parainfluenzae, Aggregatibacter spp. (A. actinomycetemcomitans, A. aphrophilus, A. paraphrophilus, and A. segnis), Cardiobacterium spp. (C. hominis, C. valvarum), Eikenella corrodens, and Kingella spp. (K. kingae, K. denitrificans) - are fastidious gram-negative bacteria, part of the normal microbiota of oral and upper respiratory tract in humans. Although their pathogenicity is limited, they are responsible for 1-3% of all infective endocarditis. HACEK endocarditis mostly affect patients with underlying heart disease or prosthetic valves, and are characterized by an insidious course, with a mean diagnosis delay of 1 month (Haemophilus spp.) to 3 months (Aggregatibacter and Cardiobacterium spp.). The advent of continuously monitored blood culture systems with enriched media has erased the need for extended incubation for the diagnosis of HACEK endocarditis. Medical treatment relies on third-generation cephalosporin, with a favorable outcome in 80-90% of cases, with or without cardiac surgery.

  15. Biochemistry of Dissimilatory Sulfur Oxidation

    Energy Technology Data Exchange (ETDEWEB)

    Blake II, R.

    2003-05-30

    The long term goals of this research were to define the substrate oxidation pathways, the electron transport mechanisms, and the modes of energy conservation employed during the dissimilatory oxidation of sulfur practiced by various species of the thiobacilli. Specific adhesion of the thiobacilli to elemental sulfur was studied by electrical impedance, dynamic light scattering, laser Doppler velocimetry, and optical trapping methods. The conclusion is that the thiobacilli appear to express specific receptors that enable the bacteria to recognize and adhere to insoluble sulfur. The enzyme tetrathionate oxidase was purified from two species of the thiobacilli. Extensive structural and functional studies were conducted on adenosine 5'-phosphosulfate reductase purified from cell-free extracts of Thiobacillus denitrificans. The kinetic mechanism of rhodanese was studied.

  16. Denitrification characteristics of a sulfur autotrophic denitrification reactor

    Directory of Open Access Journals (Sweden)

    Chenxiao ZHANG

    2016-02-01

    Full Text Available The denitrification characteristics of a sulfur autotrophic denitrification reactor are investigated. The results show that domestication of sulfur autotrophic bacteria is completed within 15 days after biofilm formation in the reactor, which is shorter than other similar researches. The nitrogen removal rate remains over than 90%, and the denitrification rate reaches 18.5 mg N/(L·h with influent NO-3-N of 70 mg/L , influent pH of 8 and HRT of 4.3 h . Thiobacillus denitrificans are observed in the whole reactor when domestication finishes, while it is more abundant in the middle and lower part. The optimal influent NO-3-N concentration for the reactor is 50 mg/L, the optimal temperature is 30~35 ℃, the optimal influent pH is 7~8, and the nitrogen removal rate is over than 90%.

  17. Feasibility of an innovative integrated process of simultaneous desulfurization and denitrification for high strength wastewater

    Institute of Scientific and Technical Information of China (English)

    WANG Ai-jie; LIU Chun-shuang; REN Nan-qi; DENG Xu-liang; WAN Chun-li; YU Zhen-guo; XU Xin

    2008-01-01

    An anaerobic expanding-bed reactor was adopted to investigate the feasibility of an innovative inte-grated process of simultaneous desulfurization and denitrification (SDD) for high strength wastewater. In the re-actor, beterotrophic bacteria (including sulfate reducing bacterium and denitrifying bacteria) and autotrophic bacteria (including Thiobacillus denitrificans) cooperated together by incubating and enriching functional bac-teria on different carriers in the anaerobic activated sludge. Synthetic wastewater with high concentrations of sul-fate and nitrate was employed. The experimental results showed that the removal efficiency of sulfate and nitrate was above 85%, elemental sulfur was observed while nitrate was absent in effluent. The balance of sulfur, ni-trogen and electron was discussed respectively, which indicated that the integrated SDD process could be actual-ized. These results might provide a guidance to further investigate the key factors affecting the integrated SDD process and to improve the efficiency of desulfurization and denitrification in wastewater treatment.

  18. Final Report: Molecular mechanisms and kinetics of microbial anaerobic nitrate-dependent U(IV) and Fe(II) oxidation

    Energy Technology Data Exchange (ETDEWEB)

    O' Day, Peggy A. [Univ. of California, Merced, CA (United States); Asta, Maria P. [Univ. of California, Merced, CA (United States); Kanematsu, Masakazu [Univ. of California, Merced, CA (United States); Beller, Harry [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Zhou, Peng [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Steefel, Carl [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)

    2015-02-27

    In this project, we combined molecular genetic, spectroscopic, and microscopic techniques with kinetic and reactive transport studies to describe and quantify biotic and abiotic mechanisms underlying anaerobic, nitrate-dependent U(IV) and Fe(II) oxidation, which influences the long-term efficacy of in situ reductive immobilization of uranium at DOE sites. In these studies, Thiobacillus denitrificans, an autotrophic bacterium that catalyzes anaerobic U(IV) and Fe(II) oxidation, was used to examine coupled oxidation-reduction processes under either biotic (enzymatic) or abiotic conditions in batch and column experiments with biogenically produced UIVO2(s). Synthesis and quantitative analysis of coupled chemical and transport processes were done with the reactive transport modeling code Crunchflow. Research focused on identifying the primary redox proteins that catalyze metal oxidation, environmental factors that influence protein expression, and molecular-scale geochemical factors that control the rates of biotic and abiotic oxidation.

  19. Surface residues dynamically organize water bridges to enhance electron transfer between proteins

    CERN Document Server

    de la Lande, Aurélien; Řezáč, Jan; Sanders, Barry C; Salahub, Dennis R; 10.1073/pnas.0914457107

    2010-01-01

    Cellular energy production depends on electron transfer (ET) between proteins. In this theoretical study, we investigate the impact of structural and conformational variations on the electronic coupling between the redox proteins methylamine dehydrogenase and amicyanin from Paracoccus denitrificans. We used molecular dynamics simulations to generate configurations over a duration of 40ns (sampled at 100fs intervals) in conjunction with an ET pathway analysis to estimate the ET coupling strength of each configuration. In the wild type complex, we find that the most frequently occurring molecular configurations afford superior electronic coupling due to the consistent presence of a water molecule hydrogen-bonded between the donor and acceptor sites. We attribute the persistence of this water bridge to a "molecular breakwater" composed of several hydrophobic residues surrounding the acceptor site. The breakwater supports the function of nearby solvent-organizing residues by limiting the exchange of water molecul...

  20. [Major results of research and development of heterogenous biocatalysts for regenerated water clearing from harmful admixture].

    Science.gov (United States)

    2012-01-01

    The biological method of clearing atmospheric condensate in pressurized habitats exploits filters with a heterogenic biocatalyst produced by way of immobilizing harmless for human, animal and plant microoganisms on water-insoluble solid carrier--foam polyvinyl-formal (FPVF), and a hydrogen peroxide biofilter containing triacetate cellulose-immobilized catalase. Experience of forming an immobilized bacterial association as a polyenzyme system is particularly promising for development of advanced biotechnologies. Biocatalysts with expanded applicability can be manufactured using a FPVF-immobilized associative bacterial culture composed of Paracoccus denitrificans, Pseudomonas esterophilus and Methilopila capsulata. In aerobic condition at room temperature the heterogenic biocatalyst is capable to transform harmful organics in atmospheric condensate, e.g. methyl amine, ethyl acetate, acetic acid, ethanol and acetone into the end-products, i.e. carbon dioxide and water. Ammonia is consumed by 3 cultures as a source of nitrogen.

  1. Achromobacter species endocarditis: A Case Report and Literature Review

    Directory of Open Access Journals (Sweden)

    Catherine Derber

    2011-01-01

    Full Text Available Endocarditis due to Achromobacter species is a rare, yet serious, endovascular infection. Achromobacter species infective endocarditis is associated with underlying immunodeficiencies or prosthetic heart valves and devices. A case of prosthetic pulmonary valve endocarditis secondary to Achromobacter xylosoxidans subspecies denitrificans is described in the present report. This life-threatening infection was successfully treated with combined valve replacement and prolonged antibiotic therapy. A Medline/PubMed literature review of Achromobacter endocarditis was also performed. Achromobacter species are an uncommon, yet important, cause of nosocomial endocarditis. Given the significant associated morbidity and mortality, along with a high degree of intrinsic antibiotic resistance, Achromobacter species infective endocarditis remains a clinical treatment challenge.

  2. Structural Adaptation of a Thermostable Biotin-binding Protein in a Psychrophilic Environment

    Science.gov (United States)

    Meir, Amit; Bayer, Edward A.; Livnah, Oded

    2012-01-01

    Shwanavidin is an avidin-like protein from the marine proteobactrium Shewanella denitrificans, which exhibits an innate dimeric structure while maintaining high affinity toward biotin. A unique residue (Phe-43) from the L3,4 loop and a distinctive disulfide bridge were shown to account for the high affinity toward biotin. Phe-43 emulates the function and position of the critical intermonomeric Trp that characterizes the tetrameric avidins but is lacking in shwanavidin. The 18 copies of the apo-monomer revealed distinctive snapshots of L3,4 and Phe-43, providing rare insight into loop flexibility, binding site accessibility, and psychrophilic adaptation. Nevertheless, as in all avidins, shwanavidin also displays high thermostability properties. The unique features of shwanavidin may provide a platform for the design of a long sought after monovalent form of avidin, which would be ideal for novel types of biotechnological application. PMID:22493427

  3. Characterization of Pseudomonas Species Isolated from the Rhizosphere of Plants Grown in Serozem Soil, Semi-Arid Region of Uzbekistan

    Directory of Open Access Journals (Sweden)

    Dilfuza Egamberdiyeva

    2005-01-01

    Full Text Available Collections of native Pseudomonas spp. are kept at the NCAM of Uzbekistan. Some of those organisms were isolated from the rhizosphere of cotton, wheat, corn, melon, alfalfa, and tomato grown in field locations within a semi-arid region of Uzbekistan. Strains used for this study were Pseudomonas alcaligenes, P. aurantiaca, P. aureofaciens, P. denitrificans, P. mendocina, P. rathonis, and P. stutzeri. Some of the pseudomonads have been characterized in this report. These strains produced enzymes, phytohormone auxin (IAA, and were antagonist against plant pathogenic fungi in in vitro experiments. Most of the strains were salt tolerant and temperature resistant. Some of the Pseudomonas spp. isolated in this study have been found to increase the growth of wheat, corn, and tomato in pot experiments.

  4. Structure of a new azurin from the denitrifying bacterium Alcaligenes xylosoxidans at high resolution.

    Science.gov (United States)

    Dodd, F E; Hasnain, S S; Abraham, Z H; Eady, R R; Smith, B E

    1995-11-01

    It has been reported previously that Alcaligenes xylosoxidans (NC1MB 11015) grown under denitrifying conditions produces two azurins instead of the single previously identified azurin [Dodd, Hasnain, Hunter, Abraham, Debenham, Kanzler, Eldridge, Eady, Ambler & Smith (1995). Biochemistry. In the press]. The new azurin, called azurin II, has been crystallized as blue elongated rectangular prisms with the tetragonal space group P4(1)22 and unit-cell parameters a = b = 52.65, c = 100.63 A. X-ray crystallographic data extending to 1.9 A resolution were collected by the Weissenberg method using 200 x 400 mm image plates and synchrotron X-rays of wavelength 0.97 A. The three-dimensional structure of azurin II has been solved by the molecular-replacement method using the structure of azurin from Alcaligenes denitrificans NCTC 8582 with which this new azurin shows a close homology. The quality of the initial map was sufficient to predict a number of sequence differences. The model is currently refined to an R-factor of 18.8% with X-ray data between 8.5 and 1.9 A. The final model of 961 protein atoms, one Cu atom and 50 water molecules has r.m.s. deviations from ideality of 0.009 A for bond lengths and 1.7 degrees for bond angles. The overall structure is similar to that of the azurin from A. denitrificans NCTC 8582. It has a beta-barrel structure with the Cu atom located near the top end of the molecule. The Cu atom is coordinated to Ndelta of His46 and His117 at 2.02 A and to Sgamma of Cys112 at 2.12 A, while the carbonyl O atom of Gly45 and Sdelta atom of Met121 provide the additional interactions at 2.75 and 3.26 A, respectively.

  5. The complete genome sequence and analysis of the epsilonproteobacterium Arcobacter butzleri.

    Directory of Open Access Journals (Sweden)

    William G Miller

    Full Text Available BACKGROUND: Arcobacter butzleri is a member of the epsilon subdivision of the Proteobacteria and a close taxonomic relative of established pathogens, such as Campylobacter jejuni and Helicobacter pylori. Here we present the complete genome sequence of the human clinical isolate, A. butzleri strain RM4018. METHODOLOGY/PRINCIPAL FINDINGS: Arcobacter butzleri is a member of the Campylobacteraceae, but the majority of its proteome is most similar to those of Sulfuromonas denitrificans and Wolinella succinogenes, both members of the Helicobacteraceae, and those of the deep-sea vent Epsilonproteobacteria Sulfurovum and Nitratiruptor. In addition, many of the genes and pathways described here, e.g. those involved in signal transduction and sulfur metabolism, have been identified previously within the epsilon subdivision only in S. denitrificans, W. succinogenes, Sulfurovum, and/or Nitratiruptor, or are unique to the subdivision. In addition, the analyses indicated also that a substantial proportion of the A. butzleri genome is devoted to growth and survival under diverse environmental conditions, with a large number of respiration-associated proteins, signal transduction and chemotaxis proteins and proteins involved in DNA repair and adaptation. To investigate the genomic diversity of A. butzleri strains, we constructed an A. butzleri DNA microarray comprising 2238 genes from strain RM4018. Comparative genomic indexing analysis of 12 additional A. butzleri strains identified both the core genes of A. butzleri and intraspecies hypervariable regions, where <70% of the genes were present in at least two strains. CONCLUSION/SIGNIFICANCE: The presence of pathways and loci associated often with non-host-associated organisms, as well as genes associated with virulence, suggests that A. butzleri is a free-living, water-borne organism that might be classified rightfully as an emerging pathogen. The genome sequence and analyses presented in this study are an

  6. Biohydrometallurgy for nonsulfidic minerals - A review

    Energy Technology Data Exchange (ETDEWEB)

    Jain, N.; Sharma, D.K. [Indian Institute of Technology of Delhi, New Delhi (India). Center for Energy Studies

    2004-05-01

    Bioleaching is a technology applicable to metal extraction from low-grade ores, ore beneficiation, coal beneficiation, metal detoxification, and recovery of metals from waste materials. The technology is environmentally sound and it may lower operational cost and energy requirement. Whereas leaching of sulfidic minerals using chemolithoautotrophic bacteria is the most studied and commercially exploitable aspect of mineral biotechnology today, there is a dearth of literature on the dissolution of nonsulfidic minerals. Biohydrometallurgy of nonsulfidic minerals involves the action of heterotrophic microorganisms. Heterotrophic bacteria and fungi have the potential for producing acidic metabolites that are able to solubilize oxide, silicate, carbonate and hydroxide minerals by reduction, acidolysis and complexation mechanisms. It is an important aspect of biohydrometallugy that requires development to meet future needs.

  7. Reconstructing a hydrogen-driven microbial metabolic network in Opalinus Clay rock

    Science.gov (United States)

    Bagnoud, Alexandre; Chourey, Karuna; Hettich, Robert L.; de Bruijn, Ino; Andersson, Anders F.; Leupin, Olivier X.; Schwyn, Bernhard; Bernier-Latmani, Rizlan

    2016-10-01

    The Opalinus Clay formation will host geological nuclear waste repositories in Switzerland. It is expected that gas pressure will build-up due to hydrogen production from steel corrosion, jeopardizing the integrity of the engineered barriers. In an in situ experiment located in the Mont Terri Underground Rock Laboratory, we demonstrate that hydrogen is consumed by microorganisms, fuelling a microbial community. Metagenomic binning and metaproteomic analysis of this deep subsurface community reveals a carbon cycle driven by autotrophic hydrogen oxidizers belonging to novel genera. Necromass is then processed by fermenters, followed by complete oxidation to carbon dioxide by heterotrophic sulfate-reducing bacteria, which closes the cycle. This microbial metabolic web can be integrated in the design of geological repositories to reduce pressure build-up. This study shows that Opalinus Clay harbours the potential for chemolithoautotrophic-based system, and provides a model of microbial carbon cycle in deep subsurface environments where hydrogen and sulfate are present.

  8. The genome sequence of Desulfatibacillum alkenivorans AK-01: a blueprint for anaerobic alkane oxidation.

    Science.gov (United States)

    Callaghan, A V; Morris, B E L; Pereira, I A C; McInerney, M J; Austin, R N; Groves, J T; Kukor, J J; Suflita, J M; Young, L Y; Zylstra, G J; Wawrik, B

    2012-01-01

    Desulfatibacillum alkenivorans AK-01 serves as a model organism for anaerobic alkane biodegradation because of its distinctive biochemistry and metabolic versatility. The D. alkenivorans genome provides a blueprint for understanding the genetic systems involved in alkane metabolism including substrate activation, CoA ligation, carbon-skeleton rearrangement and decarboxylation. Genomic analysis suggested a route to regenerate the fumarate needed for alkane activation via methylmalonyl-CoA and predicted the capability for syntrophic alkane metabolism, which was experimentally verified. Pathways involved in the oxidation of alkanes, alcohols, organic acids and n-saturated fatty acids coupled to sulfate reduction and the ability to grow chemolithoautotrophically were predicted. A complement of genes for motility and oxygen detoxification suggests that D. alkenivorans may be physiologically adapted to a wide range of environmental conditions. The D. alkenivorans genome serves as a platform for further study of anaerobic, hydrocarbon-oxidizing microorganisms and their roles in bioremediation, energy recovery and global carbon cycling.

  9. Metal-containing residues from industry and in the environment: geobiotechnological urban mining.

    Science.gov (United States)

    Glombitza, Franz; Reichel, Susan

    2014-01-01

    This chapter explains the manifold geobiotechnological possibilities to separate industrial valuable metals from various industrial residues and stored waste products of the past. In addition to an overview of the different microbially catalyzed chemical reactions applicable for a separation of metals and details of published studies, results of many individual investigations from various research projects are described. These concern the separation of rare earth elements from phosphorous production slags, the attempts of tin leaching from mining flotation residues, the separation of metals from spent catalysts, or the treatment of ashes as valuable metal-containing material. The residues of environmental technologies are integrated into this overview as well. The description of the different known microbial processes offers starting points for suitable and new technologies. In addition to the application of chemolithoautotrophic microorganisms the use of heterotrophic microorganisms is explained.

  10. Bacterial and archaeal diversity in an iron-rich coastal hydrothermal field in Yamagawa, Kagoshima, Japan

    DEFF Research Database (Denmark)

    Kawaichi, Satoshi; Ito, Norihiro; Yoshida, Takashi;

    2013-01-01

    . The environmental settings of the coastal hydrothermal field were similar in some degree to those of deep-sea hydrothermal environments because of its emission of H2, CO2, and sulfide from the bottom of the hot spot. The results of clone analyses based on the 16S rRNA gene led us to speculate the presence...... of a chemo-synthetic microbial ecosystem, where chemolithoautotrophic thermophiles, primarily the bacterial order Aquificales, function as primary producers using H2 or sulfur compounds as their energy source and CO2 as their carbon source, and the organic compounds synthesized by them support the growth...... of chemoheterotrophic thermophiles, such as members of the order Thermales and the family Desulfurococcaceae. In addition, the dominance of members of the bacterial genus Herbaspirillum in the high temperature bottom layer led us to speculate the temporal formation of mesophilic zones where they can also function...

  11. Reconstructing a hydrogen-driven microbial metabolic network in Opalinus Clay rock

    Science.gov (United States)

    Bagnoud, Alexandre; Chourey, Karuna; Hettich, Robert L.; de Bruijn, Ino; Andersson, Anders F.; Leupin, Olivier X.; Schwyn, Bernhard; Bernier-Latmani, Rizlan

    2016-01-01

    The Opalinus Clay formation will host geological nuclear waste repositories in Switzerland. It is expected that gas pressure will build-up due to hydrogen production from steel corrosion, jeopardizing the integrity of the engineered barriers. In an in situ experiment located in the Mont Terri Underground Rock Laboratory, we demonstrate that hydrogen is consumed by microorganisms, fuelling a microbial community. Metagenomic binning and metaproteomic analysis of this deep subsurface community reveals a carbon cycle driven by autotrophic hydrogen oxidizers belonging to novel genera. Necromass is then processed by fermenters, followed by complete oxidation to carbon dioxide by heterotrophic sulfate-reducing bacteria, which closes the cycle. This microbial metabolic web can be integrated in the design of geological repositories to reduce pressure build-up. This study shows that Opalinus Clay harbours the potential for chemolithoautotrophic-based system, and provides a model of microbial carbon cycle in deep subsurface environments where hydrogen and sulfate are present. PMID:27739431

  12. Arsenic speciation in shrimp and mussel from the Mid-Atlantic hydrothermal vents

    DEFF Research Database (Denmark)

    Larsen, Erik Huusfeldt; Quetel, C. R.; Munoz, R.

    1997-01-01

    Specimens of shrimp (Rimicaris exoculata) and mussel (Bathymodiolus puteoserpentis) were collected 3500 m below the ocean surface at the hydrothermal vents of the mid-Atlantic Ridge (TAG and Snake Pit sites, respectively). Arsenic, a potentially toxic element, is among the substances emitted...... by the hydrothermal vents. The hydrothermal vent shrimp, which are known to be a primary consumer of the primary producing chemolithoautotrophic bacteria, contained arsenic at 13 mu g g(-1) almost exclusively as arsenobetaine (AsB). Arsenic was present in the soft:issues of the mussel at 40 mu g g(-1) and the major...... part of the extractable arsenic species in the adductor muscle/mantle tissues and in the gill were present as dimethylarsinylriboside-derivatives (arsenosugrars), while AsB was present at 16 and 3.6%, respectively, in these tissues. In spite of the absence of biosynthetically active algae, the pattern...

  13. Cultivation of a chemoautotroph from the SUP05 clade of marine bacteria that produces nitrite and consumes ammonium

    Science.gov (United States)

    Shah, Vega; Chang, Bonnie X; Morris, Robert M

    2017-01-01

    Marine oxygen minimum zones (OMZs) are expanding regions of intense nitrogen cycling. Up to half of the nitrogen available for marine organisms is removed from the ocean in these regions. Metagenomic studies have identified an abundant group of sulfur-oxidizing bacteria (SUP05) with the genetic potential for nitrogen cycling and loss in OMZs. However, SUP05 have defied cultivation and their physiology remains untested. We cultured, sequenced and tested the physiology of an isolate from the SUP05 clade. We describe a facultatively anaerobic sulfur-oxidizing chemolithoautotroph that produces nitrite and consumes ammonium under anaerobic conditions. Genetic evidence that closely related strains are abundant at nitrite maxima in OMZs suggests that sulfur-oxidizing chemoautotrophs from the SUP05 clade are a potential source of nitrite, fueling competing nitrogen removal processes in the ocean. PMID:27434424

  14. Active ammonia oxidizers in an acidic soil are phylogenetically closely related to neutrophilic archaeon.

    Science.gov (United States)

    Wang, Baozhan; Zheng, Yan; Huang, Rong; Zhou, Xue; Wang, Dongmei; He, Yuanqiu; Jia, Zhongjun

    2014-03-01

    All cultivated ammonia-oxidizing archaea (AOA) within the Nitrososphaera cluster (former soil group 1.1b) are neutrophilic. Molecular surveys also indicate the existence of Nitrososphaera-like phylotypes in acidic soil, but their ecological roles are poorly understood. In this study, we present molecular evidence for the chemolithoautotrophic growth of Nitrososphaera-like AOA in an acidic soil with pH 4.92 using DNA-based stable isotope probing (SIP). Soil microcosm incubations demonstrated that nitrification was stimulated by urea fertilization and accompanied by a significant increase in the abundance of AOA rather than ammonia-oxidizing bacteria (AOB). Real-time PCR analysis of amoA genes as a function of the buoyant density of the DNA gradient following the ultracentrifugation of the total DNA extracted from SIP microcosms indicated a substantial growth of soil AOA during nitrification. Pyrosequencing of the total 16S rRNA genes in the "heavy" DNA fractions suggested that archaeal communities were labeled to a much greater extent than soil AOB. Acetylene inhibition further showed that (13)CO2 assimilation by nitrifying communities depended solely on ammonia oxidation activity, suggesting a chemolithoautotrophic lifestyle. Phylogenetic analysis of both (13)C-labeled amoA and 16S rRNA genes revealed that most of the active AOA were phylogenetically closely related to the neutrophilic strains Nitrososphaera viennensis EN76 and JG1 within the Nitrososphaera cluster. Our results provide strong evidence for the adaptive growth of Nitrososphaera-like AOA in acidic soil, suggesting a greater metabolic versatility of soil AOA than previously appreciated.

  15. Fluid mixing and the deep biosphere of a fossil Lost City-type hydrothermal system at the Iberia Margin.

    Science.gov (United States)

    Klein, Frieder; Humphris, Susan E; Guo, Weifu; Schubotz, Florence; Schwarzenbach, Esther M; Orsi, William D

    2015-09-29

    Subseafloor mixing of reduced hydrothermal fluids with seawater is believed to provide the energy and substrates needed to support deep chemolithoautotrophic life in the hydrated oceanic mantle (i.e., serpentinite). However, geosphere-biosphere interactions in serpentinite-hosted subseafloor mixing zones remain poorly constrained. Here we examine fossil microbial communities and fluid mixing processes in the subseafloor of a Cretaceous Lost City-type hydrothermal system at the magma-poor passive Iberia Margin (Ocean Drilling Program Leg 149, Hole 897D). Brucite-calcite mineral assemblages precipitated from mixed fluids ca. 65 m below the Cretaceous paleo-seafloor at temperatures of 31.7 ± 4.3 °C within steep chemical gradients between weathered, carbonate-rich serpentinite breccia and serpentinite. Mixing of oxidized seawater and strongly reducing hydrothermal fluid at moderate temperatures created conditions capable of supporting microbial activity. Dense microbial colonies are fossilized in brucite-calcite veins that are strongly enriched in organic carbon (up to 0.5 wt.% of the total carbon) but depleted in (13)C (δ(13)C(TOC) = -19.4‰). We detected a combination of bacterial diether lipid biomarkers, archaeol, and archaeal tetraethers analogous to those found in carbonate chimneys at the active Lost City hydrothermal field. The exposure of mantle rocks to seawater during the breakup of Pangaea fueled chemolithoautotrophic microbial communities at the Iberia Margin, possibly before the onset of seafloor spreading. Lost City-type serpentinization systems have been discovered at midocean ridges, in forearc settings of subduction zones, and at continental margins. It appears that, wherever they occur, they can support microbial life, even in deep subseafloor environments.

  16. Submarine water-rock interactions and microbial life: a theoretical approach

    Science.gov (United States)

    Bach, W.; Hentscher, M.

    2009-04-01

    Mass and energy balances coupled with thermodynamic calculations indicate that a large amount of energy in the form of chemical work (about 100 Petajoule/yr) is transported to the seafloor by hydrothermal vent fluids. A similar amount of energy is tied to the affinity of reduced components in seafloor rocks and minerals for oxidation. Chemolithoautotrophic microorganisms harness an unknown fraction of that energy to produce primary biomass in the deep sea. The specific magmatic and fluid-rock interaction processes taking place within the geological system control what metabolic reactions can support chemolithoautotrophy-based microbial ecosystems at the seafloor. It turns out that basement composition, magmatic degassing, and subseafloor mineralization impose a first-order control on vent fluid chemistry. We used thermodynamic calculations to assess how much energy hot rocks can provide in different geotectonic setting to support biomass production by chemolithoautotrophic microorganisms. The dominant energy source varies greatly between vents in different submarine settings, from hydrogen sulfide in basalt-hosted systems to dihydrogen and methane in peridotite-hosted systems to Fe and S in felsic rock systesm in island arcs. The dihydrogen fluxes related to serpentinization are at least one order of magnitude greater than those related to global magmatism, and hydrogen consumption could be one of the most important catabolic reactions in deep-sea chemolithoautotrophy. In one example we show that peridotite-water interactions release quantities of hydrogen that are sufficient for methanogens and sulfate reducers to thrive under a range of temperature and fluid flux conditions. In contrast, hydrogen production within basaltic aquifers is barely enough under the best of circumstances to allow for growth of methanogens and sulfate reducers. This prediction appears to be corroborated by sulfur isotope compositions of hydrothermally altered peridotites and basalts

  17. Methane-carbon flow into the benthic food web at cold seeps--a case study from the Costa Rica subduction zone.

    Directory of Open Access Journals (Sweden)

    Helge Niemann

    Full Text Available Cold seep ecosystems can support enormous biomasses of free-living and symbiotic chemoautotrophic organisms that get their energy from the oxidation of methane or sulfide. Most of this biomass derives from animals that are associated with bacterial symbionts, which are able to metabolize the chemical resources provided by the seeping fluids. Often these systems also harbor dense accumulations of non-symbiotic megafauna, which can be relevant in exporting chemosynthetically fixed carbon from seeps to the surrounding deep sea. Here we investigated the carbon sources of lithodid crabs (Paralomis sp. feeding on thiotrophic bacterial mats at an active mud volcano at the Costa Rica subduction zone. To evaluate the dietary carbon source of the crabs, we compared the microbial community in stomach contents with surface sediments covered by microbial mats. The stomach content analyses revealed a dominance of epsilonproteobacterial 16S rRNA gene sequences related to the free-living and epibiotic sulfur oxidiser Sulfurovum sp. We also found Sulfurovum sp. as well as members of the genera Arcobacter and Sulfurimonas in mat-covered surface sediments where Epsilonproteobacteria were highly abundant constituting 10% of total cells. Furthermore, we detected substantial amounts of bacterial fatty acids such as i-C15∶0 and C17∶1ω6c with stable carbon isotope compositions as low as -53‰ in the stomach and muscle tissue. These results indicate that the white microbial mats at Mound 12 are comprised of Epsilonproteobacteria and that microbial mat-derived carbon provides an important contribution to the crab's nutrition. In addition, our lipid analyses also suggest that the crabs feed on other (13C-depleted organic matter sources, possibly symbiotic megafauna as well as on photosynthetic carbon sources such as sedimentary detritus.

  18. Microbiome Dynamics of a Polychlorobiphenyl (PCB) Historically Contaminated Marine Sediment under Conditions Promoting Reductive Dechlorination

    Science.gov (United States)

    Matturro, Bruna; Ubaldi, Carla; Rossetti, Simona

    2016-01-01

    The toxicity of polychlorinated biphenyls (PCB) can be efficiently reduced in contaminated marine sediments through the reductive dechlorination (RD) process lead by anaerobic organohalide bacteria. Although the process has been extensively investigated on PCB-spiked sediments, the knowledge on the identity and metabolic potential of PCB-dechlorinating microorganisms in real contaminated matrix is still limited. Aim of this study was to explore the composition and the dynamics of the microbial communities of the marine sediment collected from one of the largest Sites of National Interest (SIN) in Italy (Mar Piccolo, Taranto) under conditions promoting the PCBs RD. A long-term microcosm study revealed that autochthonous bacteria were able to sustain the PCB dechlorination at a high extent and the successive addition of an external fermentable organic substrate (lactate) caused the further depletion of the high-chlorinated PCBs (up to 70%). Next Generation Sequencing was used to describe the core microbiome of the marine sediment and to follow the changes caused by the treatments. OTUs affiliated to sulfur-oxidizing ε-proteobacteria, Sulfurovum, and Sulfurimonas, were predominant in the original sediment and increased up to 60% of total OTUs after lactate addition. Other OTUs detected in the sediment were affiliated to sulfate reducing (δ-proteobacteria) and to organohalide respiring bacteria within Chloroflexi phylum mainly belonging to Dehalococcoidia class. Among others, Dehalococcoides mccartyi was enriched during the treatments even though the screening of the specific reductive dehalogenase genes revealed the occurrence of undescribed strains, which deserve further investigations. Overall, this study highlighted the potential of members of Dehalococcoidia class in reducing the contamination level of the marine sediment from Mar Piccolo with relevant implications on the selection of sustainable bioremediation strategies to clean-up the site.

  19. Methane-Carbon Flow into the Benthic Food Web at Cold Seeps – A Case Study from the Costa Rica Subduction Zone

    Science.gov (United States)

    Niemann, Helge; Linke, Peter; Knittel, Katrin; MacPherson, Enrique; Boetius, Antje; Brückmann, Warner; Larvik, Gaute; Wallmann, Klaus; Schacht, Ulrike; Omoregie, Enoma; Hilton, David; Brown, Kevin; Rehder, Gregor

    2013-01-01

    Cold seep ecosystems can support enormous biomasses of free-living and symbiotic chemoautotrophic organisms that get their energy from the oxidation of methane or sulfide. Most of this biomass derives from animals that are associated with bacterial symbionts, which are able to metabolize the chemical resources provided by the seeping fluids. Often these systems also harbor dense accumulations of non-symbiotic megafauna, which can be relevant in exporting chemosynthetically fixed carbon from seeps to the surrounding deep sea. Here we investigated the carbon sources of lithodid crabs (Paralomis sp.) feeding on thiotrophic bacterial mats at an active mud volcano at the Costa Rica subduction zone. To evaluate the dietary carbon source of the crabs, we compared the microbial community in stomach contents with surface sediments covered by microbial mats. The stomach content analyses revealed a dominance of epsilonproteobacterial 16S rRNA gene sequences related to the free-living and epibiotic sulfur oxidiser Sulfurovum sp. We also found Sulfurovum sp. as well as members of the genera Arcobacter and Sulfurimonas in mat-covered surface sediments where Epsilonproteobacteria were highly abundant constituting 10% of total cells. Furthermore, we detected substantial amounts of bacterial fatty acids such as i-C15∶0 and C17∶1ω6c with stable carbon isotope compositions as low as −53‰ in the stomach and muscle tissue. These results indicate that the white microbial mats at Mound 12 are comprised of Epsilonproteobacteria and that microbial mat-derived carbon provides an important contribution to the crab's nutrition. In addition, our lipid analyses also suggest that the crabs feed on other 13C-depleted organic matter sources, possibly symbiotic megafauna as well as on photosynthetic carbon sources such as sedimentary detritus. PMID:24116017

  20. Biogeographic Congruency among Bacterial Communities from Terrestrial Sulfidic Springs

    Directory of Open Access Journals (Sweden)

    Brendan eHeadd

    2014-09-01

    Full Text Available Terrestrial sulfidic springs support diverse microbial communities by serving as stable conduits for geochemically diverse and nutrient-rich subsurface waters. Microorganisms that colonize terrestrial springs likely originate from groundwater, but may also be sourced from the surface. As such, the biogeographic distribution of microbial communities inhabiting sulfidic springs should be controlled by a combination of spring geochemistry and surface and subsurface transport mechanisms, and not necessarily geographic proximity to other springs. We examined the bacterial diversity of seven springs to test the hypothesis that occurrence of taxonomically similar microbes, important to the sulfur cycle, at each spring is controlled by geochemistry. Complementary Sanger sequencing and 454 pyrosequencing of 16S rRNA genes retrieved five proteobacterial classes, and Bacteroidetes, Chlorobi, Chloroflexi, and Firmicutes phyla from all springs, which suggested the potential for a core sulfidic spring microbiome. Among the putative sulfide-oxidizing groups (Epsilonproteobacteria and Gammaproteobacteria, up to 83% of the sequences from geochemically similar springs clustered together. Abundant populations of Hydrogenimonas-like or Sulfurovum-like spp. (Epsilonproteobacteria occurred with abundant Thiothrix and Thiofaba spp. (Gammaproteobacteria, but Arcobacter-like and Sulfurimonas spp. (Epsilonproteobacteria occurred with less abundant gammaproteobacterial populations. These distribution patterns confirmed that geochemistry rather than biogeography regulates bacterial dominance at each spring. Potential biogeographic controls were related to paleogeologic sedimentation patterns that could control long-term microbial transport mechanisms that link surface and subsurface environments. Knowing the composition of a core sulfidic spring microbial community could provide a way to monitor diversity changes if a system is threatened by anthropogenic processes or

  1. Microbiological production and ecological flux of northwestern subduction hydrothermal systems

    Science.gov (United States)

    Sunamura, M.; Okamura, K.; Noguchi, T.; Yamamoto, H.; Fukuba, T.; Yanagawa, K.

    2012-12-01

    Deep-sea hydrothermal system is one of the most important sources for heat and chemical flux from the oceanic crust to the global ocean. The rich biological community around the hydrothermal vent shows chemolithoautotrophic microbial production are important in deep sea ecosystems. More than 99% of microbiological available chemical components in hydrothermal vent fluid, e.g. sulfide, methane, hydrogen, Fe2+, and Mn2+, is released into surrounding seawater to construct hydrothermal plume, suggesting that the chemolithoautotrophic-microbial primary production in the hydrothermal plume is huge and important in the whole hydrothermal ecosystems. To understand the impact of hydrothermal plume to a microbial ecosystem and a connectivity with zooplankton, we targeted and investigated a total of 16 hydrothermal fileds (7 sites in Okinawa trough, 3 sites in Ogasawara arc, and 6 sites in Mariana arc and back arc) and investigated in several cruises under the TAIGA project in Japan. Hydrothermal fluids in the subduction system are rich in sulfide. The hydrothermal fluids in the Okinawa trough, Ogasawara arc. and Mariana trough are characterized by rich in methane, poor in other reduced chemicals, and rich in iron, respectively. The major microbial composition was a potential sulfur oxidizing microbes SUP05 in the plume ecosystems, while an aerobic methanotrophic bacteria was secondary major member in methane-rich hydrothermal systems in Okinawa trough. Microbial quantitative and spatial distribution analyses of each plume site showed that the microbial population size and community structures are influenced by original chemical components of hydrothermal fluid, e.g. sulfide, methane and iron concentration. Microbial quantitative data indicated the removal/sedimentation of microbial cells from the plume and effect of phase separation in a same vent field through construction of gas-rich or gas-poor plumes. After the correlation of plume mixing effect, we estimates that the

  2. Water-soluble, recombinant CuA-domain of the cytochrome ba3 subunit II from Thermus thermophilus.

    Science.gov (United States)

    Slutter, C E; Sanders, D; Wittung, P; Malmström, B G; Aasa, R; Richards, J H; Gray, H B; Fee, J A

    1996-03-19

    Recently, the genes of cytochrome ba3 from thermus thermophilus [Keightley, J.A., et al. (1995) J. Biol. Chem. 270, 20345-20358], a homolog of the heme-copper oxidase family, have been cloned. We report here expression of a truncated gene, encoding the copper A (CuA) domain of cytochrome ba3, that is regulated by a T7 RNA polymerase promoter in Escherichia coli. The CuA-containing domain is purified in high yields as a water-soluble, thermostable, purple-colored protein. Copper analysis by chemical assay, mass spectrometry, X-ray fluorescence, and EPR spin quantification show that this protein contains two copper ions bound in a mixed-valence state, indicating that the CuA site in cytochrome ba3, is a binuclear center. The absorption spectrum of the CuA site, free of the heme interference in cytochrome ba3, is similar to the spectra of other soluble fragments from the aa3-type oxidase of Parachccus denitrificans [Lappalainen, P., et al. (1993) J. Biol Chem. 268, 26416-26421] and the caa3-type oxidase of Bacillus subtilis [von Wachenfeldt, C. et al. (1994) FEBS Lett. 340, 109-113]. There are intense bands at 480 nm (3100 M(-1) cm(-1)) and 530 nm (3200 M(-1) cm(-1)), a band in the near -IR centered at 790 nm (1900 M(-1) cn(-1)), and a weaker band at 363 nm (1300M(-1) cm(-1)). The visible CD spectrum shows a positive-going band at 460 nm and a negative-going band at 527 nm, the opposite signs of which may result from the binuclear nature of the site. The secondary structure prediction from the far-UV CD spectrum indicates that this domain is predominantly beta-sheet, in agreement with the recent X-ray structure reported for the complete P. denitrificans cytochrome aa3 molecule [Iwata, S., et al. (1995) Nature 376, 660-669] and the engineered, purple CyoA protein [Wilmanns, M., et al. (1996) Proc. Natl Acad. Sci. U.S.A. 92, 11955-11959]. However, the thermostability of the fragment described here (Tm approximately 80 degrees C) and the stable binding of copper over a

  3. Microbial control of hydrogen sulfide production

    Energy Technology Data Exchange (ETDEWEB)

    Montgomery, A.D.; Bhupathiraju, V.K.; Wofford, N.; McInerney, M.J. [Univ. of Oklahoma, Tulsa, OK (United States)] [and others

    1995-12-31

    A sulfide-resistant strain of Thiobacillus denitrificans, strain F, prevented the accumulation of sulfide by Desulfovibrio desulfuricans when both organisms were grown in liquid medium. The wild-type strain of T. denitrificans did not prevent the accumulation of sulfide produced by D. desulfuricans. Strain F also prevented the accumulation of sulfide by a mixed population of sulfate-reducing bacteria enriched from an oil field brine. Fermentation balances showed that strain F stoichiometrically oxidized the sulfide produced by D. desulfuricans and the oil field brine enrichment to sulfate. The ability of a strain F to control sulfide production in an experimental system of cores and formation water from the Redfield, Iowa, natural gas storage facility was also investigated. A stable, sulfide-producing biofilm was established in two separate core systems, one of which was inoculated with strain F while the other core system (control) was treated in an identical manner, but was not inoculated with strain F. When formation water with 10 mM acetate and 5 mM nitrate was injected into both core systems, the effluent sulfide concentrations in the control core system ranged from 200 to 460 {mu}M. In the test core system inoculated with strain F, the effluent sulfide concentrations were lower, ranging from 70 to 110 {mu}M. In order to determine whether strain F could control sulfide production under optimal conditions for sulfate-reducing bacteria, the electron donor was changed to lactate and inorganic nutrients (nitrogen and phosphate sources) were added to the formation water. When nutrient-supplemented formation water with 3.1 mM lactate and 10 mM nitrate was used, the effluent sulfide concentrations of the control core system initially increased to about 3,800 {mu}M, and then decreased to about 1,100 {mu}M after 5 weeks. However, in the test core system inoculated with strain F, the effluent sulfide concentrations were much lower, 160 to 330 {mu}M.

  4. A field demonstration of the microbial treatment of sour produced water

    Energy Technology Data Exchange (ETDEWEB)

    Sublette, K.L. [Univ. of Tulsa, OK (United States); Morse, D.; Raterman, K. [Amoco Production Co., Tulsa, OK (United States)

    1995-12-31

    The potential for detoxification and deodorization of sulfide-laden water (sour water) by microbial treatment was evaluated at a petroleum production site under field conditions. A sulfide-tolerant strain of the chemautotroph and facultative anaerobe, Thiobacillus denitrificans, was introduced into an oil-skimming pit of the Amoco Production Company LACT 10 Unit of the Salt Creek Field, Wyoming. Field-produced water enters this pit from the oil/water separation treatment train at an average flowrate of 5,000 bbl/D (795 m{sup 3}/D) with a potential maximum of 98,000 bbl/D (15,580 m{sup 3}/D). Water conditions at the pit inlet are 4,800 mg/l TDS, 100 mg/l sulfide, pH 7.8, and 107{degrees}F. To this water an aqueous solution of ammonium nitrate and diphosphorous pentoxide was added to provide required nutrients for the bacteria. The first 20% of the pit was aerated to a maximum depth of 5 ft (1.5 m) to facilitate the aerobic oxidation of sulfide. No provisions for pH control or biomass recovery and recycle were made. Pilot operations were initiated in October 1992 with the inoculation of the 19,000 bbl (3,020 m{sup 3}) pit with 40 lb (18.1 kg) of dry weight biomass. After a brief acclimation period, a nearly constant mass flux of 175 lb/D (80 kg/D) sulfide was established to the pit. Bio-oxidation of sulfide to elemental sulfur and sulfate was immediate and complete. Subsequent pilot operations focused upon process optimization and process sensitivity to system upsets. The process appeared most sensitive to large variations in sulfide loading due to maximum water discharge events. However, recoveries from such events could be accomplished within hours. This paper details all pertinent aspects of pilot operation, performance, and economics. Based on this body of evidence, it is suggested that the oxidation of inorganic sulfides by T denitrificans represents a viable concept for the treatment of sour water coproduced with oil and gas.

  5. Modeling the Impact of Biogeochemical Hotspots and Hot Moments on Subsurface Carbon Fluxes from a Flood Plain Site

    Science.gov (United States)

    Arora, B.; Spycher, N.; Steefel, C. I.; King, E.; Conrad, M. E.

    2015-12-01

    Biogeochemical hotspots and hot moments are known to account for a high percentage of carbon and nutrient cycling within flood plain environments. To quantify the impact of these hotspots and hot moments on the carbon cycle, a 2D reactive transport model was developed for the saturated-unsaturated zone of a flood plain site in Rifle, CO. Previous studies have identified naturally reduced zones (NRZs) in the saturated zone of the Rifle site to be hotspots and important regions for subsurface biogeochemical cycling. Wavelet analysis of geochemical concentrations at the site suggested that hydrologic and temperature variations are hot moments and exert an important control on biogeochemical conditions in the Rifle aquifer. Here, we describe the development of a reactive transport model that couples hydrologic and biogeochemical processes to microbial functional distributions inferred from site-specific 'omic' data. The model includes microbial contributions from heterotrophic and chemolithoautotrophic processes. We use Monod based formulations to represent biomass formation and consider energy partitioning between catabolic and anabolic processes. We use this model to explore community emergence at the Rifle site and further constrain the extent and rates of nutrient uptake as well as abiotic and biotic reactions using stable carbon isotopes. Results from 2D model simulations with only abiotic reactions predict lower CO2 partial pressures in the unsaturated zone and severely underpredict (~200%) carbon fluxes to the river compared to simulations with chemolithoautotrophic pathways. δ13C-CO2 profiles also point to biotic sources for the locally observed high CO2 concentrations above NRZs. Results further indicate that groundwater carbon fluxes from the Rifle site to the river are underestimated by almost 180% (to 3.3 g m-2 d-1) when temperature fluctuations are ignored in the simulations. Preliminary results demonstrate the emergence of denitrifiers at specific depths

  6. The Microbiome and Occurrence of Methanotrophy in Carnivorous Sponges

    Science.gov (United States)

    Hestetun, Jon T.; Dahle, Håkon; Jørgensen, Steffen L.; Olsen, Bernt R.; Rapp, Hans T.

    2016-01-01

    As shown by recent studies, filter-feeding sponges are known to host a wide variety of microorganisms. However, the microbial community of the non-filtering carnivorous sponges (Porifera, Cladorhizidae) has been the subject of less scrutiny. Here, we present the results from a comparative study of the methanotrophic carnivorous sponge Cladorhiza methanophila from a mud volcano-rich area at the Barbados Accretionary Prism, and five carnivorous species from the Jan Mayen Vent Field (JMVF) at the Arctic Mid-Ocean Ridge. Results from 16S rRNA microbiome data indicate the presence of a diverse assemblage of associated microorganisms in carnivorous sponges mainly from the Gamma- and Alphaproteobacteria, Flavobacteriaceae, and Thaumarchaeota. While the abundance of particular groups varied throughout the dataset, we found interesting similarities to previous microbiome results from non-carnivorous deep sea sponges, suggesting that the carnivorous sponges share characteristics of a previously hypothesized putative deep-sea sponge microbial community. Chemolithoautotrophic symbiosis was confirmed for C. methanophila through a microbial community with a high abundance of Methylococcales and very light isotopic δ13C and δ15N ratios (-60 to -66‰/3.5 to 5.2‰) compared to the other cladorhizid species (-22 to -24‰/8.5 to 10.5‰). We provide evidence for the presence of putative sulfur-oxidizing Gammaproteobacteria in the arctic cladorhizids; however, δ13C and δ15N signatures did not provide evidence for significant chemoautotrophic symbiosis in this case, and the slightly higher abundance of cladorhizids at the JMVF site compared to the nearby deep sea likely stem from an increased abundance of prey rather than a more direct vent association. The phylogenetic position of C. methanophila in relation to other carnivorous sponges was established using a three-gene phylogenetic analysis, and it was found to be closely related to other non-methanotrophic Cladorhiza species

  7. Community Characterization of Microbial Populations Found at a Cold Water Sulfidic Spring in the Canadian High Arctic

    Science.gov (United States)

    Trivedi, C.; Lau, G. E.; Templeton, A. S.; Grasby, S. E.; Spear, J. R.

    2015-12-01

    The unique environment on Europa makes it an ideal target for astrobiological investigation. One such earth-based analogue to aid in this investigation is the sulfur-dominated glacial spring system found at Borup Fiord Pass (BFP), Ellesmere Island, Nunavut, Canada. In this system, subsurface microbial sulfate reduction produces hydrogen sulfide, which is transported through the glacier along spring channels [1]. As the surface oxidation of H2S occurs, resultant deposition of elemental sulfur (S0) and other minerals becomes visible (attached image). The energy released from these reactions can support potential microbial metabolisms and may be a valuable representation of microbial processes occurring on Europa. The resulting sulfur minerals provide sensitive records of dynamic atmospheric, geological, hydrological, chemical, and biological processes on planetary surfaces. Moreover, we expect that the S0-rich deposits of this glacial spring system will serve as a mineralogical record for biological activity and will provide a valuable tool for recognizing potential sulfur-based life on Europa. During a recent collaborative expedition (2014) to BFP, samples were taken from the toe of the glacier in an area called the 'Blister Crust' (attached image). At this location, glacial channels reach the surface, representing an active interface between subsurface and surface processes. Initial geochemical characterization at the site revealed high amounts of aqueous sulfide (1.8 mM) and hydrogen (29 nM), which likely serve as the electron donation potential in the system. Furthermore, preliminary 16S rRNA gene sequencing has shown a high abundance of the genus Sulfurimonas, which is a known sulfur metabolizer. Our research seeks to further characterize microbial communities found at this interface in order to elucidate information regarding in situ sulfur cycling and the potential to tie this into subsurface/surface processes on Europa. Continued work will provide guidance

  8. Zeta-Proteobacteria dominate the formation of microbial mats in low-temperature hydrothermal vents at Loihi Seamount

    Science.gov (United States)

    Rassa, A. C.; McAllister, S. M.; Safran, S. A.; Moyer, C. L.

    2007-12-01

    Sulfurimonas, which are sulfur- and thiosulfate-oxidizing bacteria.

  9. Identification and characterization of an RTX toxin in the emerging pathogen Kingella kingae.

    Science.gov (United States)

    Kehl-Fie, Thomas E; St Geme, Joseph W

    2007-01-01

    Kingella kingae is an emerging bacterial pathogen that is increasingly recognized as the causative agent of a variety of pediatric diseases, including septic arthritis and osteomyelitis. The pathogenesis of K. kingae disease is believed to begin with colonization of the upper respiratory tract. In the present study, we examined interactions between K. kingae and cultured respiratory epithelial cells and observed potent cytotoxicity, detected by both microscopy and lactic acid dehydrogenase (LDH) release assays. Experiments with synovial and macrophage cell lines revealed cytotoxicity for these cell types as well. Using mariner mutagenesis and a screen for loss of cytotoxicity, a genetic locus encoding an RTX toxin system was identified. Disruption of the K. kingae RTX locus resulted in a loss of cytotoxicity for respiratory epithelial, synovial, and macrophage cell lines. DNA sequence analysis demonstrated that the RTX locus is flanked by insertion elements and has a reduced G+C content compared to that of the whole genome. Two relatively less invasive Kingella species, K. oralis and K. denitrificans, were found to be noncytotoxic and to lack the RTX region, as determined by LDH release assays and Southern blotting. We concluded that K. kingae expresses an RTX toxin that has wide cellular specificity and was likely acquired horizontally. The possible roles for this toxin in the pathogenesis of K. kingae disease include breaching of the epithelial barrier and destruction of target tissues, such as synovium (joint lining).

  10. Analysis of iron- and sulfur-oxidizing bacteria in a treatment plant of acid rock drainage from a Japanese pyrite mine by use of ribulose-1, 5-bisphosphate carboxylase/oxygenase large-subunit gene.

    Science.gov (United States)

    Kamimura, Kazuo; Okabayashi, Ai; Kikumoto, Mei; Manchur, Mohammed Abul; Wakai, Satoshi; Kanao, Tadayoshi

    2010-03-01

    Iron- and sulfur-oxidizing bacteria in a treatment plant of acid rock drainage (ARD) from a pyrite mine in Yanahara, Okayama prefecture, Japan, were analyzed using the gene (cbbL) encoding the large subunit of ribulose-1, 5-bisphosphate carboxylase/oxygenase (RubisCO). Analyses of partial sequences of cbbL genes from Acidithiobacillus ferrooxidans, Acidithiobacillus thiooxidans and Acidithiobacillus caldus strains revealed the diversity in their cbbL gene sequences. In contrast to the presence of two copies of form I cbbL genes (cbbL1 and cbbL2) in A. ferrooxidans genome, A. thiooxidans and A. caldus had a single copy of form I cbbL gene in their genomes. A phylogenetic analysis based on deduced amino acid sequences from cbbL genes detected in the ARD treatment plant and their close relatives revealed that 89% of the total clones were affiliated with A. ferrooxidans. Clones loosely affiliated with the cbbL from A. thiooxidans NB1-3 or Thiobacillus denitrificans was also detected in the treatment plant. cbbL gene sequences of iron- or sulfur-oxidizing bacteria isolated from the ARD and the ARD treatment plant were not detected in the cbbL libraries from the treatment plant, suggesting the low frequencies of isolates in the samples.

  11. Biomimetic Membranes for Multi-Redox Center Proteins

    Directory of Open Access Journals (Sweden)

    Renate L. C. Naumann

    2016-03-01

    Full Text Available His-tag technology was applied for biosensing purposes involving multi-redox center proteins (MRPs. An overview is presented on various surfaces ranging from flat to spherical and modified with linker molecules with nitrile-tri-acetic acid (NTA terminal groups to bind his-tagged proteins in a strict orientation. The bound proteins are submitted to in situ dialysis in the presence of lipid micelles to form a so-called protein-tethered bilayer lipid membrane (ptBLM. MRPs, such as the cytochrome c oxidase (CcO from R. sphaeroides and P. denitrificans, as well as photosynthetic reactions centers (RCs from R. sphaeroides, were thus investigated. Electrochemical and surface-sensitive optical techniques, such as surface plasmon resonance, surface plasmon-enhanced fluorescence, surface-enhanced infrared absorption spectroscopy (SEIRAS and surface-enhanced resonance Raman spectroscopy (SERRS, were employed in the case of the ptBLM structure on flat surfaces. Spherical particles ranging from µm size agarose gel beads to nm size nanoparticles modified in a similar fashion were called proteo-lipobeads (PLBs. The particles were investigated by laser-scanning confocal fluorescence microscopy (LSM and UV/Vis spectroscopy. Electron and proton transfer through the proteins were demonstrated to take place, which was strongly affected by the membrane potential. MRPs can thus be used for biosensing purposes under quasi-physiological conditions.

  12. [Microflora of active ooze participating in the decomposition of sulfanilic acid].

    Science.gov (United States)

    Orshanskaia, F B; Arkad'eva, A Z; Kozlova, E I

    1975-01-01

    Microflora of domestic water can be a source of active ooze adapted to sulphanilic acid. Adaptation of the microflora to sulphanilic acid at a concentration of 170-200 mg/l takes 6 to 8 days. The microflora of active ooze, immediately after adaptation, consists mainly of Pseudomonas species, Ps. denitrificans, Ps. fluorescens, Ps. striata, Ps. putida, etc., and also of Achromobacter stutzeri, Achromobacter flavum, Mycobacterium phlei, Mycobacterium mucosum, Bacillus mesentericus, Bac. cereus, saccharomyces cerevisiae, Schizosaccharomyces pombe, and Rhodotorula glutinus. The number of the species decreased as a result of long cultivation of active ooze on a minimal medium with sulphanilic acid as a sole source of carbon and nitrogen; the following strains prevailed: Ps. putida, Ps. eisenbergii, strains of Mycobacterium phlei and Flavobacterium solare. The isolated strains of Ps. putida and Ps. eisenbergii decomposed sulphanilic acid by 60.0--79.5 percent, and together with Mycobacterium phlei by 100 percent during 4 to 7 days. The ability to oxidize sulphanilic acid decreased after storage. Addition to the medium of other sources of carbon, nitrogen and vitamins did not restore the lost ability of the microorganisms to decompose sulphanilic acid.

  13. Analysis of protein film voltammograms as Michaelis-Menten saturation curves yield the electron cooperativity number for deconvolution.

    Science.gov (United States)

    Heering, Hendrik A

    2012-10-01

    Deconvolution of protein film voltammetric data by fitting multiple components (sigmoids, derivative peaks) often is ambiguous when features are partially overlapping, due to exchangeability between the width and the number of components. Here, a new method is presented to obtain the width of the components. This is based on the equivalence between the sigmoidal catalytic response as function of electrode potential, and the classical saturation curve obtained for the enzyme activity as function of the soluble substrate concentration, which is also sigmoidal when plotted versus log[S]. Thus, analysis of the catalytic voltammogram with Lineweaver-Burk, Eadie-Hofstee, and Hanes-Woolf plots is feasible. This provides a very sensitive measure of the cooperativity number (Hill coefficient), which for electrons equals the apparent (fractional) number of electrons that determine the width, and thereby the number of components (kinetic phases). This analysis is applied to the electrocatalytic oxygen reduction by Paracoccus denitrificans cytochrome aa(3) (cytochrome c oxidase). Four partially overlapping kinetic phases are observed that (stepwise) increase the catalytic efficiency with increasingly reductive potential. Translated to cell biology, the activity of the terminal oxidase stepwise adapts to metabolic demand for oxidative phosphorylation.

  14. Exploring the molecular basis for selective binding of homoserine dehydrogenase from Mycobacterium leprae TN toward inhibitors: a virtual screening study.

    Science.gov (United States)

    Zhan, Dongling; Wang, Dongmei; Min, Weihong; Han, Weiwei

    2014-01-24

    Homoserine dehydrogenase (HSD) from Mycobacterium leprae TN is an antifungal target for antifungal properties including efficacy against the human pathogen. The 3D structure of HSD has been firmly established by homology modeling methods. Using the template, homoserine dehydrogenase from Thiobacillus denitrificans (PDB Id 3MTJ), a sequence identity of 40% was found and molecular dynamics simulation was used to optimize a reliable structure. The substrate and co-factor-binding regions in HSD were identified. In order to determine the important residues of the substrate (L-aspartate semialdehyde (L-ASA)) binding, the ASA was docked to the protein; Thr163, Asp198, and Glu192 may be important because they form a hydrogen bond with HSD through AutoDock 4.2 software. neuraminidaseAfter use of a virtual screening technique of HSD, the four top-scoring docking hits all seemed to cation-π ion pair with the key recognition residue Lys107, and Lys207. These ligands therefore seemed to be new chemotypes for HSD. Our results may be helpful for further experimental investigations.

  15. Exploring the Molecular Basis for Selective Binding of Homoserine Dehydrogenase from Mycobacterium leprae TN toward Inhibitors: A Virtual Screening Study

    Directory of Open Access Journals (Sweden)

    Dongling Zhan

    2014-01-01

    Full Text Available Homoserine dehydrogenase (HSD from Mycobacterium leprae TN is an antifungal target for antifungal properties including efficacy against the human pathogen. The 3D structure of HSD has been firmly established by homology modeling methods. Using the template, homoserine dehydrogenase from Thiobacillus denitrificans (PDB Id 3MTJ, a sequence identity of 40% was found and molecular dynamics simulation was used to optimize a reliable structure. The substrate and co-factor-binding regions in HSD were identified. In order to determine the important residues of the substrate (l-aspartate semialdehyde (l-ASA binding, the ASA was docked to the protein; Thr163, Asp198, and Glu192 may be important because they form a hydrogen bond with HSD through AutoDock 4.2 software. neuraminidaseAfter use of a virtual screening technique of HSD, the four top-scoring docking hits all seemed to cation–π ion pair with the key recognition residue Lys107, and Lys207. These ligands therefore seemed to be new chemotypes for HSD. Our results may be helpful for further experimental investigations.

  16. Technical Progresses of Novel Simultaneous Biological Sulfide and Nitrate Removal%新型生物脱氮除硫工艺研究进展

    Institute of Scientific and Technical Information of China (English)

    蔡靖; 郑平

    2011-01-01

    Some bacteria such as Thiobacillus denitrificans can use sulfide as electron donor and nitrate as electron acceptor. Based on the biological reaction, the simultaneous biological sulfide and nitrate removal process has been developed recently, which can accomplish desulfurization and denitrification at the same time. This paper discussed the development, mechanism and key influencing factors of the simultaneous biological sulfide and nitrate removal process, in order to promote its application and extention.%同步生物脱氮除硫过程是以硫化物为电子供体、硝酸盐为电子受体的生物反应,以此为基础研发的新型生物脱氮除硫工艺,可实现对氮硫两种污染物的同时去除,在环境工程上具有重大应用价值.本文综述了同步生物脱氮除硫工艺的发展概况、原理及其主要影响因素,并展望了该工艺的应用前景.

  17. Enzyme diversity and mosaic gene organization in denitrification.

    Science.gov (United States)

    Zumft, W G; Körner, H

    1997-02-01

    Denitrification is a main branch of the global nitrogen cycle. In the past ten years unravelling the underlying biochemistry and genetics has proceeded at an increasing pace. Fungal denitrification has become a new field. The biochemical investigation of denitrification has culminated in the description of the crystal structures of the two types of nitrite reductases. The N2O reductase shares with cytochrome c oxidase the CuA center as a structurally novel metal site. The cytochrome b subunit of NO reductase has a striking conservation of heme-binding transmembrane segments versus the subunit I of cytochrome c oxidase. Another putative denitrification gene product shows structural relation to the subunit III of the oxidase. N2O reductase and NO reductase may be ancestors of energy-conserving enzymes of the heme-copper oxidase superfamily. More than 30 genes for denitrification are located in a > 30-kb cluster in Pseudomonas stutzeri, and comparable gene clusters have been identified in Pseudomonas aeruginosa and Paracoccus denitrificans. Genes necessary for nitrite reduction and NO reduction have a mosaic arrangement with very few conserved locations within these clusters and relative to each other.

  18. Redox properties of an engineered purple Cu(A) azurin.

    Science.gov (United States)

    Sun, Dapeng; Wang, Xiaotang; Davidson, Victor L

    2002-08-01

    Purple Cu(A) centers are a class of binuclear, mixed-valence copper complexes found in cytochrome c oxidase and nitrous oxide reductase. An engineered Cu(A) protein was formed by replacing a portion of the amino acid sequence that contains three of the ligands to the native type I copper center of Pseudomonas aeruginosa azurin with the corresponding portion of sequence from the Cu(A) center of cytochrome c oxidase from Paracoccus denitrificans [Proc. Natl. Acad. Sci. USA 93 (1996) 461]. Oxidation-reduction midpoint potential (E(m)) values of the Cu(A) azurin of +399+/-10 and +380+/-2mV, respectively, were determined by cyclic voltammetry and spectrochemical titration. An n value of one was obtained, indicating that the redox reaction is cycling between the mixed valence and the fully reduced states. Whereas the E(m) value of native azurin is pH dependent, the E(m) value of Cu(A) azurin is not, as expected for the Cu(A) center. Similarities and differences in the redox properties are discussed in terms of the known crystal structures of Cu(A) centers in cytochrome c oxidase and Cu(A) azurin.

  19. Genetic tools for the investigation of Roseobacter clade bacteria

    Directory of Open Access Journals (Sweden)

    Tielen Petra

    2009-12-01

    Full Text Available Abstract Background The Roseobacter clade represents one of the most abundant, metabolically versatile and ecologically important bacterial groups found in marine habitats. A detailed molecular investigation of the regulatory and metabolic networks of these organisms is currently limited for many strains by missing suitable genetic tools. Results Conjugation and electroporation methods for the efficient and stable genetic transformation of selected Roseobacter clade bacteria including Dinoroseobacter shibae, Oceanibulbus indolifex, Phaeobacter gallaeciensis, Phaeobacter inhibens, Roseobacter denitrificans and Roseobacter litoralis were tested. For this purpose an antibiotic resistance screening was performed and suitable genetic markers were selected. Based on these transformation protocols stably maintained plasmids were identified. A plasmid encoded oxygen-independent fluorescent system was established using the flavin mononucleotide-based fluorescent protein FbFP. Finally, a chromosomal gene knockout strategy was successfully employed for the inactivation of the anaerobic metabolism regulatory gene dnr from D. shibae DFL12T. Conclusion A genetic toolbox for members of the Roseobacter clade was established. This provides a solid methodical basis for the detailed elucidation of gene regulatory and metabolic networks underlying the ecological success of this group of marine bacteria.

  20. Perchlorate and halophilic prokaryotes: implications for possible halophilic life on Mars.

    Science.gov (United States)

    Oren, Aharon; Elevi Bardavid, Rahel; Mana, Lily

    2014-01-01

    In view of the finding of perchlorate among the salts detected by the Phoenix Lander on Mars, we investigated the relationships of halophilic heterotrophic microorganisms (archaea of the family Halobacteriaceae and the bacterium Halomonas elongata) toward perchlorate. All strains tested grew well in NaCl-based media containing 0.4 M perchlorate, but at the highest perchlorate concentrations, tested cells were swollen or distorted. Some species (Haloferax mediterranei, Haloferax denitrificans, Haloferax gibbonsii, Haloarcula marismortui, Haloarcula vallismortis) could use perchlorate as an electron acceptor for anaerobic growth. Although perchlorate is highly oxidizing, its presence at a concentration of 0.2 M for up to 2 weeks did not negatively affect the ability of a yeast extract-based medium to support growth of the archaeon Halobacterium salinarum. These findings show that presence of perchlorate among the salts on Mars does not preclude the possibility of halophilic life. If indeed the liquid brines that may exist on Mars are inhabited by salt-requiring or salt-tolerant microorganisms similar to the halophiles on Earth, presence of perchlorate may even be stimulatory when it can serve as an electron acceptor for respiratory activity in the anaerobic Martian environment.

  1. Electron paramagnetic resonance studies of the soluble CuA protein from the cytochrome ba3 of Thermus thermophilus.

    Science.gov (United States)

    Karpefors, M; Slutter, C E; Fee, J A; Aasa, R; Källebring, B; Larsson, S; Vänngård, T

    1996-11-01

    The electron paramagnetic resonance (EPR) spectrum of the binuclear CuA center in the water-soluble subunit II fragment from cytochrome ba3 of Thermus thermophilus was recorded at 3.93, 9.45, and 34.03 GHz, and the EPR parameters were determined by computer simulations. The frequency and M1 dependence of the linewidth was discussed in terms of g strain superimposed on a correlation between the A and g values. The g values were found to be gx = 1.996, gy = 2.011, gz = 2.187, and the two Cu ions contribute nearly equally to the hyperfine structure, with magnitude of Ax magnitude of approximately 15 G, magnitude of Ay magnitude = 29 G, and magnitude of Az magnitude of = 28.5 G (65Cu). Theoretical CNDO/S calculations, based on the x-ray structure of the Paracoccus denitrificans enzyme, yield a singly occupied antibonding orbital in which each Cu is pi*-bonded to one S and sigma*-bonded to the other. In contrast to the equal spin distribution suggested by the EPR simulations, the calculated contributions from the Cu ions differ by a factor of 2. However, only small changes in the ligand geometry are needed to reproduce the experimental results.

  2. Start-up and microbial communities of a simultaneous nitrogen removal system for high salinity and high nitrogen organic wastewater via heterotrophic nitrification.

    Science.gov (United States)

    Chen, Jiahao; Han, Yi; Wang, Yingmu; Gong, Benzhou; Zhou, Jian; Qing, Xiaoxia

    2016-09-01

    In this study, a simultaneous nitrogen removal system for high salinity and high nitrogen organic wastewater was developed in a pressurized biofilm reactor. The result showed that under the air supply rate of 200Lh(-1), salinity of 3.0±0.2%, organic load of 10kgCODm(-3)d(-1) and nitrogen loading of 0.185kgm(-3)d(-1), the reactor started up rapidly and performed stably after 30days operation. Meanwhile, a simultaneous COD and nitrogen removal was achieved in the single-stage reactor, with COD, NH4(+)-N and TN removal efficiency of 97%, 99% and 98%, respectively. Denaturing gradient gel electrophoresis profile demonstrated that simultaneous nitrogen removal could be achieved through heterotrophic nitrification-aerobic denitrification, and the pivotal microorganisms were Flavobacterium phragmitis and Paracoccus denitrificans. The microbial community of salt-tolerant halophilic microorganisms was developed successfully. This study can provide a more efficient and feasible solution to treat high salinity organic wastewater.

  3. Microbial Diversity of Chromium-Contaminated Soils and Characterization of Six Chromium-Removing Bacteria

    Science.gov (United States)

    He, Zhiguo; Hu, Yuting; Yin, Zhen; Hu, Yuehua; Zhong, Hui

    2016-06-01

    Three soil samples obtained from different sites adjacent to a chromium slag heap in a steel alloy factory were taken to examine the effect of chromium contamination on soil bacterial diversity as determined by construction of 16S rDNA clone libraries and sequencing of selected clones based on restriction fragment length polymorphism (RFLP) analysis. Results revealed that Betaproteobacteria, Gammaproteobacteria, Firmicutes, and Alphaproteobacteria occurred in all three soil samples, although the three samples differed in their total diversity. Sample 1 had the highest microbial diversity covering 12 different classes, while Sample 3 had the lowest microbial diversity. Strains of six different species were successfully isolated, one of which was identified as Zobellella denitrificans. To our knowledge, this is the first report of a strain belonging to the genus Zobellella able to resist and reduce chromium. Among all isolates studied, Bacillus odysseyi YH2 exhibited the highest Cr(VI)-reducing capability, with a total removal of 23.5 % of an initial Cr(VI) concentration of 350 mg L-1.

  4. Clinical evaluation of the Vitek Neisseria-Haemophilus Identification card.

    Science.gov (United States)

    Janda, W M; Malloy, P J; Schreckenberger, P C

    1987-01-01

    A clinical evaluation of the Vitek Neisseria-Haemophilus Identification (NHI) card (Vitek Systems, Inc., Hazelwood, Mo.) was performed with 480 clinical isolates and stock strains of Neisseria spp., Haemophilus spp., and other fastidious microorganisms included in the data base of the system. Identifications obtained with the NHI card were compared with those determined by conventional methods. The card identified 83.2% of 244 Neisseria spp. and Branhamella catarrhalis, 54.9% of 164 Haemophilus spp., and 84.7% of 72 fastidious gram-negative species with no further testing required. Some isolates produced good confidence-marginal separation identifications, in which the correct identification was listed with one or two other possible identifications and extra tests were required and suggested. When isolates producing good confidence-marginal separation identifications were included, correct identifications of these organism groups increased to 97.1, 92.7, and 94.4%, respectively. Among the commonly isolated microorganisms, the NHI card identified 99.1% of 110 N. gonorrhoeae, 98.5% of 68 N. meningitidis, 93.9% of 98 H. influenzae, and 95.6% of 46 H. parainfluenzae strains. All of these organisms produced excellent to very good confidence level identifications except for H. influenzae biotypes II, III, and VII, for which hemolytic reactions were required for differentiation from H. haemolyticus. The NHI card reliably identified other fastidious gram-negative species, including H. aphrophilus, Eikenella corrodens, Gardnerella vaginalis, and Kingella denitrificans.

  5. Bacterial Communities Associated with Different Anthurium andraeanum L. Plant Tissues

    Science.gov (United States)

    Sarria-Guzmán, Yohanna; Chávez-Romero, Yosef; Gómez-Acata, Selene; Montes-Molina, Joaquín Adolfo; Morales-Salazar, Eleacin; Dendooven, Luc; Navarro-Noya, Yendi E.

    2016-01-01

    Plant-associated microbes have specific beneficial functions and are considered key drivers for plant health. The bacterial community structure of healthy Anthurium andraeanum L. plants was studied by 16S rRNA gene pyrosequencing associated with different plant parts and the rhizosphere. A limited number of bacterial taxa, i.e., Sinorhizobium, Fimbriimonadales, and Gammaproteobacteria HTCC2089 were enriched in the A. andraeanum rhizosphere. Endophytes were more diverse in the roots than in the shoots, whereas all shoot endophytes were found in the roots. Streptomyces, Flavobacterium succinicans, and Asteroleplasma were only found in the roots, Variovorax paradoxus only in the stem, and Fimbriimonas 97%-OTUs only in the spathe, i.e., considered specialists, while Brevibacillus, Lachnospiraceae, Pseudomonas, and Pseudomonas pseudoalcaligenes were generalist and colonized all plant parts. The anaerobic diazotrophic bacteria Lachnospiraceae, Clostridium sp., and Clostridium bifermentans colonized the shoot system. Phylotypes belonging to Pseudomonas were detected in the rhizosphere and in the substrate (an equiproportional mixture of soil, cow manure, and peat), and dominated the endosphere. Pseudomonas included nine 97%-OTUs with different patterns of distribution and phylogenetic affiliations with different species. P. pseudoalcaligenes and P. putida dominated the shoots, but were also found in the roots and rhizosphere. P. fluorescens was present in all plant parts, while P. resinovorans, P. denitrificans, P. aeruginosa, and P. stutzeri were only detected in the substrate and rhizosphere. The composition of plant-associated bacterial communities is generally considered to be suitable as an indicator of plant health. PMID:27524305

  6. Food preservative potential of gassericin A-containing concentrate prepared from a cheese whey culture supernatant from Lactobacillus gasseri LA39.

    Science.gov (United States)

    Nakamura, Kiyoshi; Arakawa, Kensuke; Kawai, Yasushi; Yasuta, Narimi; Chujo, Takahiro; Watanabe, Masamichi; Iioka, Hiroyuki; Tanioka, Masashi; Nishimura, Junko; Kitazawa, Haruki; Tsurumi, Koichi; Saito, Tadao

    2013-02-01

    Gassericin A (GA) is a circular bacteriocin produced by Lactobacillus gasseri LA39. In this study, GA-containing concentrate was prepared using a cross-flow membrane filtration device (30 kDa cut-off) from the culture supernatant of Lb. gasseri LA39 cultivated in a cheese whey-based food-grade medium. The bacteriocin activity titer in the concentrate was 16 times as high as that of the culture supernatant and was completely maintained through each incubation at 4°C for 3 months, 37°C for 2 months, 60°C for 5 h, and 100°C for 30 min. The GA-containing concentrate was used with glycine powder to make custard creams, and then four representative strains of custard cream spoilage bacteria (Bacillus cereus, Lactococcus lactis subsp. lactis, Achromobacter denitrificans and Pseudomonas fluorescens) were individually inoculated at c. 10(3) colony forming units/g in the custard creams. Throughout 30 days of incubation at 30°C, all of the inoculated bacteria were completely inhibited by the combination of 5% (w/w) of the GA-containing concentrate and 0.5% (w/w) glycine. This is the first highly practical application of GA to foods as a biopreservative, and the concentration method and the bacteriocin concentrate would contribute to biopreservation of several foods.

  7. Carboxyl-modified single-walled carbon nanotubes negatively affect bacterial growth and denitrification activity

    Science.gov (United States)

    Zheng, Xiong; Su, Yinglong; Chen, Yinguang; Wan, Rui; Li, Mu; Wei, Yuanyuan; Huang, Haining

    2014-07-01

    Single-walled carbon nanotubes (SWNTs) have been used in a wide range of fields, and the surface modification via carboxyl functionalization can further improve their physicochemical properties. However, whether carboxyl-modified SWNT poses potential risks to microbial denitrification after its release into the environment remains unknown. Here we present the possible effects of carboxyl-modified SWNT on the growth and denitrification activity of Paracoccus denitrificans (a model denitrifying bacterium). It was found that carboxyl-modified SWNT were present both outside and inside the bacteria, and thus induced bacterial growth inhibition at the concentrations of 10 and 50 mg/L. After 24 h of exposure, the final nitrate concentration in the presence of 50 mg/L carboxyl-modified SWNT was 21-fold higher than that in its absence, indicating that nitrate reduction was substantially suppressed by carboxyl-modified SWNT. The transcriptional profiling revealed that carboxyl-modified SWNT led to the transcriptional activation of the genes encoding ribonucleotide reductase in response to DNA damage and also decreased the gene expressions involved in glucose metabolism and energy production, which was an important reason for bacterial growth inhibition. Moreover, carboxyl-modified SWNT caused the significant down-regulation and lower activity of nitrate reductase, which was consistent with the decreased efficiency of nitrate reduction.

  8. STRUCTURE AND FUNCTION OF SUBSURFACE MICROBIAL COMMUNITIES AFFECTING RADIONUCLIDE TRANSPORT AND BIOIMMOBILIZATION

    Energy Technology Data Exchange (ETDEWEB)

    Joel E. Kostka; Lee Kerkhof; Kuk-Jeong Chin; Martin Keller; Joseph W. Stucki

    2011-06-15

    are new to science all show high sequence identity to sequences retrieved from ORFRC subsurface. (2) Based on physiological and phylogenetic characterization, two new species of subsurface bacteria were described: the metal-reducer Geobacter daltonii, and the denitrifier Rhodanobacter denitrificans. (3) Strains isolated from the ORFRC show that Rhodanobacter species are well adapted to the contaminated subsurface. Strains 2APBS1 and 116-2 grow at high salt (3% NaCl), low pH (3.5) and tolerate high concentrations of nitrate (400mM) and nitrite (100mM). Strain 2APBS1 was demonstrated to grow at in situ acidic pHs down to 2.5. (4) R. denitrificans strain 2APBS1 is the first described Rhodanobacter species shown to denitrify. Nitrate is almost entirely converted to N2O, which may account for the large accumulation of N2O in the ORFRC subsurface. (5) G. daltonii, isolated from uranium- and hydrocarbon-contaminated subsurface sediments of the ORFRC, is the first organism from the subsurface clade of the genus Geobacter that is capable of growth on aromatic hydrocarbons. (6) High quality draft genome sequences and a complete eco-physiological description are completed for R. denitrificans strain 2APBS1 and G. daltonii strain FRC-32. (7) Given their demonstrated relevance to DOE remediation efforts and the availability of detailed genotypic/phenotypic characterization, Rhodanobacter denitrificans strain 2APBS1 and Geobacter daltonii strain FRC-32 represent ideal model organisms to provide a predictive understanding of subsurface microbial activity through metabolic modeling. Tasks II and III-Diversity and distribution of active anaerobes and Mechanisms linking electron transport and the fate of radionuclides: (1) Our study showed that members of genus Rhodanobacter and Geobacter are abundant and active in the uranium and nitrate contaminated subsurface. In the contaminant source zone of the Oak Ridge site, Rhodanobacter spp. are the predominant, active organisms detected

  9. Proton transfer in ba(3) cytochrome c oxidase from Thermus thermophilus.

    Science.gov (United States)

    von Ballmoos, Christoph; Adelroth, Pia; Gennis, Robert B; Brzezinski, Peter

    2012-04-01

    The respiratory heme-copper oxidases catalyze reduction of O(2) to H(2)O, linking this process to transmembrane proton pumping. These oxidases have been classified according to the architecture, location and number of proton pathways. Most structural and functional studies to date have been performed on the A-class oxidases, which includes those that are found in the inner mitochondrial membrane and bacteria such as Rhodobacter sphaeroides and Paracoccus denitrificans (aa(3)-type oxidases in these bacteria). These oxidases pump protons with a stoichiometry of one proton per electron transferred to the catalytic site. The bacterial A-class oxidases use two proton pathways (denoted by letters D and K, respectively), for the transfer of protons to the catalytic site, and protons that are pumped across the membrane. The B-type oxidases such as, for example, the ba(3) oxidase from Thermus thermophilus, pump protons with a lower stoichiometry of 0.5 H(+)/electron and use only one proton pathway for the transfer of all protons. This pathway overlaps in space with the K pathway in the A class oxidases without showing any sequence homology though. Here, we review the functional properties of the A- and the B-class ba(3) oxidases with a focus on mechanisms of proton transfer and pumping.

  10. Diversity and abundance of bacteria in an underground oil-storage cavity

    Directory of Open Access Journals (Sweden)

    Kodama Yumiko

    2002-08-01

    Full Text Available Abstract Background Microorganisms inhabiting subterranean oil fields have recently attracted much attention. Since intact groundwater can easily be obtained from the bottom of underground oil-storage cavities without contamination by surface water, studies on such oil-storage cavities are expected to provide valuable information to understand microbial ecology of subterranean oil fields. Results DNA was extracted from the groundwater obtained from an oil-storage cavity situated at Kuji in Iwate, Japan, and 16S rRNA gene (16S rDNA fragments were amplified by PCR using combinations of universal and Bacteria-specific primers. The sequence analysis of 154 clones produced 31 different bacterial sequence types (a unique clone or group of clones with sequence similarity of > 98. Major sequence types were related to Desulfotomaculum, Acetobacterium, Desulfovibrio, Desulfobacula, Zoogloea and Thiomicrospira denitrificans. The abundance in the groundwater of bacterial populations represented by these major sequence types was assessed by quantitative competitive PCR using specific primers, showing that five rDNA types except for that related to Desulfobacula shared significant proportions (more than 1% of the total bacterial rDNA. Conclusions Bacteria inhabiting the oil-storage cavity were unexpectedly diverse. A phylogenetic affiliation of cloned 16S rDNA sequences suggests that bacteria exhibiting different types of energy metabolism coexist in the cavity.

  11. Microbial degradation of chloro- and methylphenol mixtures under aerobic and denitrifying conditions. Regulation and interaction in a mixed culture; Mikrobieller Abbau von Chlor- und Methylphenolen im Gemisch unter aeroben und denitrifizierenden Bedingungen. Regulation und Interaktion in einer Mischkultur

    Energy Technology Data Exchange (ETDEWEB)

    Hollender, J.

    1994-12-31

    The mechanism and the regulation of the microbial aerobic degradation of chloro- and methylphenol mixtures were investigated by comparing the mixed culture enriched of these phenolic compounds with the isolated pure cultures. The degradation assays and the characterization of the central degrading enzymes involved show that the degradation potential of the mixed culture was essentially determined by the additive abilities of the three pure cultures Alcaligenes xylosoxidans subspecies denitrificans JH1, Pseudomonas stutzeri JH3 and Comamonas testosteroni JH5. Degradation of some substrate mixtures and avoidance of enrichment of metabolites can only be explained by the interaction of the pure cultures or the participation of other organisms in the mixed culture which were not further investigated. (orig.) [Deutsch] Bei den Untersuchungen zum Mechanismus und der Regulation des aeroben Abbaus von Chlor- und Methylphenolen im Gemisch stand der Vergleich zwischen der auf diesen Phenolen angereicherten Mischkultur und den daraus isolierten Reinkulturen im Vordergrund. Die Abbauversuche und die Charakterisierung der am Metabolismus beteiligten zentralen Enzyme zeigen, dass fuer das Abbaupotential der Mischkultur im wesentlichen die additiven Abbauleistungen der drei Reinkulturen Alcaligenes xylosoxidans subspecies dentrificans JH1, Pseudomonas stutzeri JH3 und Comamonas testosteroni JH5 verantwortlich sind. Der Abbau einiger Substratmischungen und die Vermeidung einer Anreicherung von Metaboliten in der Mischkultur sind jedoch nur durch eine Interaktion der Reinkulturen oder durch eine Beteiligung anderer nicht naeher untersuchter Organismen in der Mischkultur zu erklaeren. (orig.)

  12. Development of PCR primer systems for amplification of nitrite reductase genes (nirK and nirS) to detect denitrifying bacteria in environmental samples

    Energy Technology Data Exchange (ETDEWEB)

    Braker, G.; Witzel, K.P. [Max-Planck-Inst. fuer Limnologie, Ploen (Germany); Fesefeldt, A. [Univ. Kiel (Germany). Inst. fuer Allgemeine Mikrobiologie

    1998-10-01

    A system was developed for the detection of denitrifying bacteria by the application of specific nitrite reductase gene fragments with PCR. Primer sequences were found for the amplification of fragments from both nitrite reductase genes (nirK and nirS) after comparative sequence analysis. Whenever amplification was tried with these primers, the known nir type of denitrifying laboratory cultures could be confirmed. Likewise, the method allowed a determination of the nir type of five laboratory strains. The nirK gene could be amplified from Blastobacter denitrificans, Alcaligenes xylosoxidans, and Alcaligenes sp. (DSM 30128); the nirS gene was amplified from Alcaligenes eutrophus DSM 530 and from the denitrifying isolate IFAM 3698. For each of the two genes, at least one primer combination amplified successfully for all of the test strains. Specific amplification products were not obtained wit h nondenitrifying bacteria or with strains of the other nir type. The specificity of the amplified products was confirmed by subsequent sequencing. These results suggest the suitability of the method for the qualitative detection of denitrifying bacteria in environmental samples. This was shown by applying the generally amplifying primer combination for each nir gene developed in this study to total DNA preparations from aquatic habitats.

  13. 一株降解氨氮菌株的筛选%Isolation of Microorganisms Degrading Ammonia-nitrogen of Wastewater

    Institute of Scientific and Technical Information of China (English)

    刘韶娜; 杨国明; 李卫芬

    2014-01-01

    A high effective microorganism A4 degrading ammonia-nitrogen has been isolated from a fish pond by the selected culture medium with (NH4)2SO4 as exclusive nitrogen source.It can bear the poison of heavy metal ion. In the wastewater,the degrading rate of NH4-N by A4 is over 44%within 48 h.Identified by biochemical methods, gram-stain reaction,PCR and electron microscope,this bacterim is Achromobacter xylosoxidans denitrificans.%以硫酸铵为唯一氮源的选择性培养基,通过富集培养,从天然养鱼池塘中筛选出一株能够降解废水中高浓度氨氮的菌株A4,并能耐受废水中的重金属离子,在造纸废水中48 h的降解速率为44%,通过革兰氏染色、菌落形态和分子生物学鉴定为木糖氧化无色杆菌反硝化亚种。

  14. [Analysis on Diversity of Denitrifying Microorganisms in Sequential Batch Bioreactor Landfill].

    Science.gov (United States)

    Li, Wei-Hua; Sun, Ying-Jie; Liu, Zi-Liang; Ma, Qiang; Yang, Qiang

    2016-01-15

    A denitrification functional microorganism gene clone library (amoA, nosZ) and the PCR-RFLP technology was constructed to investigate the microbial diversity of denitrifying microorganisms in the late period of stabilization of sequential batch bioreactor landfill. The results indicated that: the bacterial diversity of ammonia oxidizing bacteria in the aged refuse reactor was very high, and most of them were unknown groups, also, all bacteria were unculturable or had not been isolated. The phylogenetic analysis suggested that the dominant ammonia oxidizing bacteria were presumably Nitrosomonas of 6-Proteobacteria. The diversity of denitrifying bacteria in fresh refuse reactor was abundant, which mainly included Thauera and Thiobacillus of 6-Proteobacteria. As Thauera sp. has the denitrification characteristics under the condition of aerobic while Thiobacillus denitrificans has the autotrophic denitrification characteristics, it was speculated that aerobic denitrification and autotrophic denitrification might be the main pathways for nitrogen removal in the fresh refuse reactor at the late period of stabilization. Additionally, another group in the gene clone library of denitrifying bacteria may be classified as Bradyrhizobiaceae of alpha-Proteobacteria.

  15. STRUCTURE AND FUNCTION OF SUBSURFACE MICROBIAL COMMUNITIES AFFECTING RADIONUCLIDE TRANSPORT AND BIOIMMOBILIZATION

    Energy Technology Data Exchange (ETDEWEB)

    Joel E. Kostka; Lee Kerkhof; Kuk-Jeong Chin; Martin Keller; Joseph W. Stucki

    2011-06-15

    are new to science all show high sequence identity to sequences retrieved from ORFRC subsurface. (2) Based on physiological and phylogenetic characterization, two new species of subsurface bacteria were described: the metal-reducer Geobacter daltonii, and the denitrifier Rhodanobacter denitrificans. (3) Strains isolated from the ORFRC show that Rhodanobacter species are well adapted to the contaminated subsurface. Strains 2APBS1 and 116-2 grow at high salt (3% NaCl), low pH (3.5) and tolerate high concentrations of nitrate (400mM) and nitrite (100mM). Strain 2APBS1 was demonstrated to grow at in situ acidic pHs down to 2.5. (4) R. denitrificans strain 2APBS1 is the first described Rhodanobacter species shown to denitrify. Nitrate is almost entirely converted to N2O, which may account for the large accumulation of N2O in the ORFRC subsurface. (5) G. daltonii, isolated from uranium- and hydrocarbon-contaminated subsurface sediments of the ORFRC, is the first organism from the subsurface clade of the genus Geobacter that is capable of growth on aromatic hydrocarbons. (6) High quality draft genome sequences and a complete eco-physiological description are completed for R. denitrificans strain 2APBS1 and G. daltonii strain FRC-32. (7) Given their demonstrated relevance to DOE remediation efforts and the availability of detailed genotypic/phenotypic characterization, Rhodanobacter denitrificans strain 2APBS1 and Geobacter daltonii strain FRC-32 represent ideal model organisms to provide a predictive understanding of subsurface microbial activity through metabolic modeling. Tasks II and III-Diversity and distribution of active anaerobes and Mechanisms linking electron transport and the fate of radionuclides: (1) Our study showed that members of genus Rhodanobacter and Geobacter are abundant and active in the uranium and nitrate contaminated subsurface. In the contaminant source zone of the Oak Ridge site, Rhodanobacter spp. are the predominant, active organisms detected

  16. Comparative genome analysis of Pseudomonas knackmussii B13, the first bacterium known to degrade chloroaromatic compounds.

    Science.gov (United States)

    Miyazaki, Ryo; Bertelli, Claire; Benaglio, Paola; Canton, Jonas; De Coi, Nicoló; Gharib, Walid H; Gjoksi, Bebeka; Goesmann, Alexander; Greub, Gilbert; Harshman, Keith; Linke, Burkhard; Mikulic, Josip; Mueller, Linda; Nicolas, Damien; Robinson-Rechavi, Marc; Rivolta, Carlo; Roggo, Clémence; Roy, Shantanu; Sentchilo, Vladimir; Siebenthal, Alexandra Von; Falquet, Laurent; van der Meer, Jan Roelof

    2015-01-01

    Pseudomonas knackmussii B13 was the first strain to be isolated in 1974 that could degrade chlorinated aromatic hydrocarbons. This discovery was the prologue for subsequent characterization of numerous bacterial metabolic pathways, for genetic and biochemical studies, and which spurred ideas for pollutant bioremediation. In this study, we determined the complete genome sequence of B13 using next generation sequencing technologies and optical mapping. Genome annotation indicated that B13 has a variety of metabolic pathways for degrading monoaromatic hydrocarbons including chlorobenzoate, aminophenol, anthranilate and hydroxyquinol, but not polyaromatic compounds. Comparative genome analysis revealed that B13 is closest to Pseudomonas denitrificans and Pseudomonas aeruginosa. The B13 genome contains at least eight genomic islands [prophages and integrative conjugative elements (ICEs)], which were absent in closely related pseudomonads. We confirm that two ICEs are identical copies of the 103 kb self-transmissible element ICEclc that carries the genes for chlorocatechol metabolism. Comparison of ICEclc showed that it is composed of a variable and a 'core' region, which is very conserved among proteobacterial genomes, suggesting a widely distributed family of so far uncharacterized ICE. Resequencing of two spontaneous B13 mutants revealed a number of single nucleotide substitutions, as well as excision of a large 220 kb region and a prophage that drastically change the host metabolic capacity and survivability.

  17. Anodic biofilms in microbial fuel cells harbor low numbers of higher-power-producing bacteria than abundant genera

    KAUST Repository

    Kiely, Patrick D.

    2010-07-15

    Microbial fuel cell (MFC) anode communities often reveal just a few genera, but it is not known to what extent less abundant bacteria could be important for improving performance. We examined the microbial community in an MFC fed with formic acid for more than 1 year and determined using 16S rRNA gene cloning and fluorescent in situ hybridization that members of the Paracoccus genus comprised most (~30%) of the anode community. A Paracoccus isolate obtained from this biofilm (Paracoccus denitrificans strain PS-1) produced only 5.6 mW/m 2, whereas the original mixed culture produced up to 10 mW/m 2. Despite the absence of any Shewanella species in the clone library, we isolated a strain of Shewanella putrefaciens (strain PS-2) from the same biofilm capable of producing a higher-power density (17.4 mW/m2) than the mixed culture, although voltage generation was variable. Our results suggest that the numerical abundance of microorganisms in biofilms cannot be assumed a priori to correlate to capacities of these predominant species for high-power production. Detailed screening of bacterial biofilms may therefore be needed to identify important strains capable of high-power generation for specific substrates. © 2010 Springer-Verlag.

  18. A joint x-ray and neutron study on amicyanin reveals the role of protein dynamics in electron transfer.

    Science.gov (United States)

    Sukumar, N; Mathews, F S; Langan, P; Davidson, V L

    2010-04-13

    The joint x-ray/neutron diffraction model of the Type I copper protein, amicyanin from Paracoccus denitrificans was determined at 1.8 A resolution. The protein was crystallized using reagents prepared in D(2)O. About 86% of the amide hydrogen atoms are either partially or fully exchanged, which correlates well with the atomic depth of the amide nitrogen atom and the secondary structure type, but with notable exceptions. Each of the four residues that provide copper ligands is partially deuterated. The model reveals the dynamic nature of the protein, especially around the copper-binding site. A detailed analysis of the presence of deuterated water molecules near the exchange sites indicates that amide hydrogen exchange is primarily due to the flexibility of the protein. Analysis of the electron transfer path through the protein shows that residues in that region are highly dynamic, as judged by hydrogen/deuterium exchange. This could increase the rate of electron transfer by transiently shortening through-space jumps in pathways or by increasing the atomic packing density. Analysis of C-HX bonding reveals previously undefined roles of these relatively weak H bonds, which, when present in sufficient number can collectively influence the structure, redox, and electron transfer properties of amicyanin.

  19. Metagenomic analysis of nitrogen metabolism genes in the surface of marine sediments

    Science.gov (United States)

    Reyes, Carolina; Schneider, Dominik; Thürmer, Andrea; Dellwig, Olaf; Lipka, Marko; Daniel, Rolf; Böttcher, Michael E.; Friedrich, Michael W.

    2016-04-01

    In this study, we analysed metagenomes along with biogeochemical profiles from Skagerrak (North Sea) and Bothnian Bay (Baltic Sea) sediments, to trace the prevailing nitrogen pathways. NO3- was present in the top 5 cm below the sediment-water interface at both sites. NH4+ increased with depth below 5 cm where it overlapped with the NO3- zone. Steady state modelling of NO3- and NH4+ porewater profiles indicates zones of net nitrogen species transformations. Protease, peptidase, urease and deaminase ammonification genes were detected in metagenomes. Genes involved in ammonia oxidation (amo, hao), nitrite oxidation (nxr), denitrification (nar, nir, nor) and dissimilatory NO3- reduction to NH4+ (nap, nfr and otr) were also present. 16S rRNA gene analysis showed that the nitrifying group Nitrosopumilales and other groups involved in nitrification and denitrification (Nitrobacter, Nitrosomonas, Nitrospira, Nitrosococcus, and Nitrosonomas) appeared less abundant in Skagerrak sediments compared to Bothnian Bay sediments. Beggiatoa and Thiothrix 16S rRNA genes were also present suggesting chemolithoautotrophic NO3- reduction to NO2- or NH4+ as a possible pathway. Although anammox planctomycetes 16S rRNA genes were present in metagenomes, anammox protein-coding genes were not detected. Our results show the metabolic potential for ammonification, nitrification, NO3- reduction, and denitrification activities in Skagerrak and Bothnian Bay sediments.

  20. Evidence of atmospheric sulphur in the martian regolith from sulphur isotopes in meteorites.

    Science.gov (United States)

    Farquhar, J; Savarino, J; Jackson, T L; Thiemens, M H

    2000-03-02

    Sulphur is abundant at the martian surface, yet its origin and evolution over time remain poorly constrained. This sulphur is likely to have originated in atmospheric chemical reactions, and so should provide records of the evolution of the martian atmosphere, the cycling of sulphur between the atmosphere and crust, and the mobility of sulphur in the martian regolith. Moreover, the atmospheric deposition of oxidized sulphur species could establish chemical potential gradients in the martian near-surface environment, and so provide a potential energy source for chemolithoautotrophic organisms. Here we present measurements of sulphur isotopes in oxidized and reduced phases from the SNC meteorites--the group of related achondrite meteorites believed to have originated on Mars--together with the results of laboratory photolysis studies of two important martian atmospheric sulphur species (SO2 and H2S). The photolysis experiments can account for the observed sulphur-isotope compositions in the SNC meteorites, and so identify a mechanism for producing large abiogenic 34S fractionations in the surface sulphur reservoirs. We conclude that the sulphur data from the SNC meteorites reflects deposition of oxidized sulphur species produced by atmospheric chemical reactions, followed by incorporation, reaction and mobilization of the sulphur within the regolith.

  1. Insights into the genome of large sulfur bacteria revealed by analysis of single filaments.

    Directory of Open Access Journals (Sweden)

    Marc Mussmann

    2007-09-01

    Full Text Available Marine sediments are frequently covered by mats of the filamentous Beggiatoa and other large nitrate-storing bacteria that oxidize hydrogen sulfide using either oxygen or nitrate, which they store in intracellular vacuoles. Despite their conspicuous metabolic properties and their biogeochemical importance, little is known about their genetic repertoire because of the lack of pure cultures. Here, we present a unique approach to access the genome of single filaments of Beggiatoa by combining whole genome amplification, pyrosequencing, and optical genome mapping. Sequence assemblies were incomplete and yielded average contig sizes of approximately 1 kb. Pathways for sulfur oxidation, nitrate and oxygen respiration, and CO2 fixation confirm the chemolithoautotrophic physiology of Beggiatoa. In addition, Beggiatoa potentially utilize inorganic sulfur compounds and dimethyl sulfoxide as electron acceptors. We propose a mechanism of vacuolar nitrate accumulation that is linked to proton translocation by vacuolar-type ATPases. Comparative genomics indicates substantial horizontal gene transfer of storage, metabolic, and gliding capabilities between Beggiatoa and cyanobacteria. These capabilities enable Beggiatoa to overcome non-overlapping availabilities of electron donors and acceptors while gliding between oxic and sulfidic zones. The first look into the genome of these filamentous sulfur-oxidizing bacteria substantially deepens the understanding of their evolution and their contribution to sulfur and nitrogen cycling in marine sediments.

  2. Carbon dioxide reduction by mixed and pure cultures in microbial electrosynthesis using an assembly of graphite felt and stainless steel as a cathode.

    Science.gov (United States)

    Bajracharya, Suman; ter Heijne, Annemiek; Dominguez Benetton, Xochitl; Vanbroekhoven, Karolien; Buisman, Cees J N; Strik, David P B T B; Pant, Deepak

    2015-11-01

    Carbon dioxide (CO2) reduction to multi-carbon compounds at the cathode using chemolithoautotrophs is an emerging application of microbial electrosynthesis (MES). In this study, CO2 reduction in MES was investigated at hydrogen evolving potentials, separately by a mixed culture and Clostridium ljungdahlii, using a graphite felt and stainless steel assembly as cathode. The mixed culture reactor produced acetate at the maximum rate of 1.3 mM d(-1), along with methane and hydrogen at -1.1 V/Ag/AgCl. Over 160 days of run-time in four fed-batches, 26% of bicarbonate was converted to acetate between day 28 and 41, whereas in the late batches, methane production prevailed. Out of 45 days of run-time in the C. ljungdahlii reactor, 2.4 mM d(-1) acetate production was achieved at -0.9 V/Ag/AgCl in Batch 1. Simultaneous product degradation occurred when the mixed culture was not selectively enriched. Hydrogen evolution is potentially the rapid way of transferring electrons to the biocatalysts for higher bioproduction rates.

  3. Enrichment and genome sequence of the group I.1a ammonia-oxidizing Archaeon "Ca. Nitrosotenuis uzonensis" representing a clade globally distributed in thermal habitats.

    Directory of Open Access Journals (Sweden)

    Elena V Lebedeva

    Full Text Available The discovery of ammonia-oxidizing archaea (AOA of the phylum Thaumarchaeota and the high abundance of archaeal ammonia monooxygenase subunit A encoding gene sequences in many environments have extended our perception of nitrifying microbial communities. Moreover, AOA are the only aerobic ammonia oxidizers known to be active in geothermal environments. Molecular data indicate that in many globally distributed terrestrial high-temperature habits a thaumarchaeotal lineage within the Nitrosopumilus cluster (also called "marine" group I.1a thrives, but these microbes have neither been isolated from these systems nor functionally characterized in situ yet. In this study, we report on the enrichment and genomic characterization of a representative of this lineage from a thermal spring in Kamchatka. This thaumarchaeote, provisionally classified as "Candidatus Nitrosotenuis uzonensis", is a moderately thermophilic, non-halophilic, chemolithoautotrophic ammonia oxidizer. The nearly complete genome sequence (assembled into a single scaffold of this AOA confirmed the presence of the typical thaumarchaeotal pathways for ammonia oxidation and carbon fixation, and indicated its ability to produce coenzyme F420 and to chemotactically react to its environment. Interestingly, like members of the genus Nitrosoarchaeum, "Candidatus N. uzonensis" also possesses a putative artubulin-encoding gene. Genome comparisons to related AOA with available genome sequences confirmed that the newly cultured AOA has an average nucleotide identity far below the species threshold and revealed a substantial degree of genomic plasticity with unique genomic regions in "Ca. N. uzonensis", which potentially include genetic determinants of ecological niche differentiation.

  4. Arsenic speciation in food chains from mid-Atlantic hydrothermal vents

    Science.gov (United States)

    Taylor, Vivien F.; Jackson, Brian P.; Siegfried, Matthew R.; Navratilova, Jana; Francesconi, Kevin A.; Kirshtein, Julie; Voytek, Mary

    2012-01-01

    Arsenic concentration and speciation were determined in benthic fauna collected from the Mid-Atlantic Ridge hydrothermal vents. The shrimp species, Rimicaris exoculata, the vent chimney-dwelling mussel, Bathymodiolus azoricus, Branchipolynoe seepensis, a commensal worm of B. azoricus and the gastropod Peltospira smaragdina showed variations in As concentration and in stable isotope (δ13C and δ15N) signature between species, suggesting different sources of As uptake. Arsenic speciation showed arsenobetaine to be the dominant species in R. exoculata, whereas in B. azoricus and B. seepensis arsenosugars were most abundant, although arsenobetaine, dimethylarsinate and inorganic arsenic were also observed, along with several unidentified species. Scrape samples from outside the vent chimneys covered with microbial mat, which is a presumed food source for many vent organisms, contained high levels of total As, but organic species were not detectable. The formation of arsenosugars in pelagic environments is typically attributed to marine algae, and the pathway to arsenobetaine is still unknown. The occurrence of arsenosugars and arsenobetaine in these deep sea organisms, where primary production is chemolithoautotrophic and stable isotope analyses indicate food sources are of vent origin, suggests that organic arsenicals can occur in a foodweb without algae or other photosynthetic life.

  5. Arsenite Oxidation and Arsenite Resistance by Bacillus sp. PNKP-S2

    Directory of Open Access Journals (Sweden)

    Pranee Pattanapipitpaisal

    2015-01-01

    Full Text Available Arsenic causes human health problems after accumulate in the body for 10-15 years and arsenite [As(III] is generally regarded as being more mobile and toxic than other oxidation states. In this study, two-hundred and three bacterial strains were isolated from groundwater and soil samples collecting in Ubon Ratchathani Province, Thailand. All strains were screened for arsenic tolerant efficiency at 1-10 mM of sodium arsenite. Eighteen selected strains which had the highest resistance to 10 mM of As(III were further studied for their As(III-oxidizing activity and growth in enrichment and growth medium (EG medium supplemented with 0.58 mM of As(III. It was found that strain PNKP-S2 was able to grow in the medium with As(III as a sole energy source and had 89.11% As(III removal within 48 h. The PCR-based 16S rDNA sequencing analysis revealed that the strain PNKP-S2 was closed relative to Bacillus sp. This is the first report on Bacillus sp. chemolithoautotrophic As(III-oxidizer and this strain could be a potential candidate for application in arsenic remediation of contaminated water.

  6. Molecular and Kinetic Characterization of Planktonic Nitrospira spp. Selectively Enriched from Activated Sludge.

    Science.gov (United States)

    Park, Mee-Rye; Park, Hongkeun; Chandran, Kartik

    2017-03-07

    Nitrospira spp. are chemolithoautotrophic nitrite-oxidizing bacteria (NOB), which are ubiquitous in natural and engineered environments. However, there exist few independent biokinetic studies on Nitrospira spp., likely because their isolation and selective enrichment from environmental consortia such as activated sludge can be challenging. Herein, planktonic Nitrospira spp. cultures closely related to Candidatus Nitrospira defluvii (Nitrospira lineage I) were successfully enriched from activated sludge in a sequencing batch reactor by maintaining sustained limiting extant nitrite and dissolved oxygen concentrations. Morphologically, the enrichment consisted largely of planktonic cells with an average characteristic diameter of 1.3 ± 0.6 μm. On the basis of respirometric assays, estimated maximum specific growth rate (μmax), nitrite half saturation coefficient (KS), oxygen half saturation coefficient (KO), and biomass yield coefficient (Y) of the enriched cultures were 0.69 ± 0.10 d(-1), 0.52 ± 0.14 mg-N/L, 0.33 ± 0.14 mg-O2/L, and 0.14 ± 0.02 mg-COD/mg-N, respectively. These parameters collectively reflect not just higher affinities of this enrichment for nitrite and oxygen, respectively, but also a higher biomass yield and energy transfer efficiency relative to Nitrobacter spp. Used in combination, these kinetic and thermodynamic parameters can help toward the development and application of energy-efficient biological nutrient removal processes through effective Nitrospira out-selection.

  7. Integration of Metagenomic and Stable Carbon Isotope Evidence Reveals the Extent and Mechanisms of Carbon Dioxide Fixation in High-Temperature Microbial Communities.

    Science.gov (United States)

    Jennings, Ryan de Montmollin; Moran, James J; Jay, Zackary J; Beam, Jacob P; Whitmore, Laura M; Kozubal, Mark A; Kreuzer, Helen W; Inskeep, William P

    2017-01-01

    Although the biological fixation of CO2 by chemolithoautotrophs provides a diverse suite of organic compounds utilized by chemoorganoheterotrophs as a carbon and energy source, the relative amounts of autotrophic C in chemotrophic microbial communities are not well-established. The extent and mechanisms of CO2 fixation were evaluated across a comprehensive set of high-temperature, chemotrophic microbial communities in Yellowstone National Park by combining metagenomic and stable (13)C isotope analyses. Fifteen geothermal sites representing three distinct habitat types (iron-oxide mats, anoxic sulfur sediments, and filamentous "streamer" communities) were investigated. Genes of the 3-hydroxypropionate/4-hydroxybutyrate, dicarboxylate/4-hydroxybutyrate, and reverse tricarboxylic acid CO2 fixation pathways were identified in assembled genome sequence corresponding to the predominant Crenarchaeota and Aquificales observed across this habitat range. Stable (13)C analyses of dissolved inorganic and organic C (DIC, DOC), and possible landscape C sources were used to interpret the (13)C content of microbial community samples. Isotope mixing models showed that the minimum fractions of autotrophic C in microbial biomass were >50% in the majority of communities analyzed. The significance of CO2 as a C source in these communities provides a foundation for understanding community assembly and succession, and metabolic linkages among early-branching thermophilic autotrophs and heterotrophs.

  8. Development of reactor configurations for an electrofuels platform utilizing genetically modified iron oxidizing bacteria for the reduction of CO2 to biochemicals.

    Science.gov (United States)

    Guan, Jingyang; Berlinger, Sarah A; Li, Xiaozheng; Chao, Zhongmou; Sousa E Silva, Victor; Banta, Scott; West, Alan C

    2017-03-10

    Electrofuels processes are potentially promising platforms for biochemical production from CO2 using renewable energy. When coupled to solar panels, this approach could avoid the inefficiencies of photosynthesis and there is no competition with food agriculture. In addition, these systems could potentially be used to store intermittent or stranded electricity generated from other renewable sources. Here we develop reactor configurations for continuous electrofuels processes to convert electricity and CO2 to isobutyric acid (IBA) using genetically modified (GM) chemolithoautotrophic Acidithiobacillus ferrooxidans. These cells oxidize ferrous iron which can be electrochemically reduced. During two weeks of cultivation on ferrous iron, stable cell growth and continuous IBA production from CO2 were achieved in a process where media was circulated between electrochemical and biochemical rectors. An alternative process with an additional electrochemical cell for accelerated ferrous production was developed, and this system achieved an almost three-fold increase in steady state cell densities, and an almost 4-fold increase in the ferrous iron oxidation rate. Combined, this led to an almost 8-fold increase in the steady state volumetric productivity of IBA up to 0.063±0.012mg/L/h, without a decline in energy efficiency from previous work. Continued development of reactor configurations which can increase the delivery of energy to the genetically modified cells will be required to increase product titers and volumetric productivities.

  9. Nitrous oxide production by lithotrophic ammonia-oxidizing bacteria and implications for engineered nitrogen-removal systems.

    Science.gov (United States)

    Chandran, Kartik; Stein, Lisa Y; Klotz, Martin G; van Loosdrecht, Mark C M

    2011-12-01

    Chemolithoautotrophic AOB (ammonia-oxidizing bacteria) form a crucial component in microbial nitrogen cycling in both natural and engineered systems. Under specific conditions, including transitions from anoxic to oxic conditions and/or excessive ammonia loading, and the presence of high nitrite (NO₂⁻) concentrations, these bacteria are also documented to produce nitric oxide (NO) and nitrous oxide (N₂O) gases. Essentially, ammonia oxidation in the presence of non-limiting substrate concentrations (ammonia and O₂) is associated with N₂O production. An exceptional scenario that leads to such conditions is the periodical switch between anoxic and oxic conditions, which is rather common in engineered nitrogen-removal systems. In particular, the recovery from, rather than imposition of, anoxic conditions has been demonstrated to result in N₂O production. However, applied engineering perspectives, so far, have largely ignored the contribution of nitrification to N₂O emissions in greenhouse gas inventories from wastewater-treatment plants. Recent field-scale measurements have revealed that nitrification-related N₂O emissions are generally far higher than emissions assigned to heterotrophic denitrification. In the present paper, the metabolic pathways, which could potentially contribute to NO and N₂O production by AOB have been conceptually reconstructed under conditions especially relevant to engineered nitrogen-removal systems. Taken together, the reconstructed pathways, field- and laboratory-scale results suggest that engineering designs that achieve low effluent aqueous nitrogen concentrations also minimize gaseous nitrogen emissions.

  10. Snowmelt induced hydrologic perturbations drive dynamic microbiological and geochemical behaviors across a shallow riparian aquifer

    Science.gov (United States)

    Danczak, Robert; Yabusaki, Steven; Williams, Kenneth; Fang, Yilin; Hobson, Chad; Wilkins, Michael

    2016-05-01

    Shallow riparian aquifers represent hotspots of biogeochemical activity in the arid western US. While these environments provide extensive ecosystem services, little is known of how natural environmental perturbations influence subsurface microbial communities and associated biogeochemical processes. Over a six-month period we tracked the annual snowmelt-driven incursion of groundwater into the vadose zone of an aquifer adjacent to the Colorado River, leading to increased dissolved oxygen (DO) concentrations in the normally suboxic saturated zone. Strong biogeochemical heterogeneity was measured across the site, with abiotic reactions between DO and sulfide minerals driving rapid DO consumption and mobilization of redox active species in reduced aquifer regions. Conversely, extensive DO increases were detected in less reduced sediments. 16S rRNA gene surveys tracked microbial community composition within the aquifer, revealing strong correlations between increases in putative oxygen-utilizing chemolithoautotrophs and heterotrophs and rising DO concentrations. The gradual return to suboxic aquifer conditions favored increasing abundances of 16S rRNA sequences matching members of the Microgenomates (OP11) and Parcubacteria (OD1) that have been strongly implicated in fermentative processes. Microbial community stability measurements indicated that deeper aquifer locations were relatively less affected by geochemical perturbations, while communities in shallower locations exhibited the greatest change. Reactive transport modeling of the geochemical and microbiological results supported field observations, suggesting that a predictive framework can be applied to develop a greater understanding of such environments.

  11. Expression and activity of the Calvin-Benson-Bassham cycle transcriptional regulator CbbR from Acidithiobacillus ferrooxidans in Ralstonia eutropha.

    Science.gov (United States)

    Esparza, Mario; Jedlicki, Eugenia; Dopson, Mark; Holmes, David S

    2015-08-01

    Autotrophic fixation of carbon dioxide into cellular carbon occurs via several pathways but quantitatively, the Calvin-Benson-Bassham cycle is the most important. CbbR regulates the expression of the cbb genes involved in CO2 fixation via the Calvin-Benson-Bassham cycle in a number of autotrophic bacteria. A gene potentially encoding CbbR (cbbR(AF)) has been predicted in the genome of the chemolithoautotrophic, extreme acidophile Acidithiobacillus ferrooxidans. However, this microorganism is recalcitrant to genetic manipulation impeding the experimental validation of bioinformatic predictions. Two novel functional assays were devised to advance our understanding of cbbR(AF) function using the mutated facultative autotroph Ralstonia eutropha H14 ΔcbbR as a surrogate host to test gene function: (i) cbbR(AF) was expressed in R. eutropha and was able to complement ΔcbbR; and (ii) CbbR(AF) was able to regulate the in vivo activity of four A. ferrooxidans cbb operon promoters in R. eutropha. These results open up the use of R. eutropha as a surrogate host to explore cbbR(AF) activity.

  12. New Insight into the Role of the Calvin Cycle: Reutilization of CO2 Emitted through Sugar Degradation.

    Science.gov (United States)

    Shimizu, Rie; Dempo, Yudai; Nakayama, Yasumune; Nakamura, Satoshi; Bamba, Takeshi; Fukusaki, Eiichiro; Fukui, Toshiaki

    2015-07-01

    Ralstonia eutropha is a facultative chemolithoautotrophic bacterium that uses the Calvin-Benson-Bassham (CBB) cycle for CO2 fixation. This study showed that R. eutropha strain H16G incorporated (13)CO2, emitted by the oxidative decarboxylation of [1-(13)C1]-glucose, into key metabolites of the CBB cycle and finally into poly(3-hydroxybutyrate) [P(3HB)] with up to 5.6% (13)C abundance. The carbon yield of P(3HB) produced from glucose by the strain H16G was 1.2 times higher than that by the CBB cycle-inactivated mutants, in agreement with the possible fixation of CO2 estimated from the balance of energy and reducing equivalents through sugar degradation integrated with the CBB cycle. The results proved that the 'gratuitously' functional CBB cycle in R. eutropha under aerobic heterotrophic conditions participated in the reutilization of CO2 emitted during sugar degradation, leading to an advantage expressed as increased carbon yield of the storage compound. This is a new insight into the role of the CBB cycle, and may be applicable for more efficient utilization of biomass resources.

  13. Enhanced Cr bioleaching efficiency from tannery sludge with coinoculation of Acidithiobacillus thiooxidans TS6 and Brettanomyces B65 in an air-lift reactor.

    Science.gov (United States)

    Fang, Di; Zhou, Li-Xiang

    2007-09-01

    Bioleaching process has been demonstrated to be an effective technology in removing Cr from tannery sludge, but a large quantity of dissolved organic matter (DOM) present in tannery sludge often exhibits a marked toxicity to chemolithoautotrophic bioleaching bacteria such as Acidithiobacillus thiooxidans. The purpose of the present study was therefore to enhance Cr bioleaching efficiencies through introducing sludge DOM-degrading heterotrophic microorganism into the sulfur-based sludge bioleaching system. An acid-tolerant DOM-degrading yeast strain Brettanomyces B65 was successfully isolated from a local Haining tannery sludge and it could metabolize sludge DOM as a source of energy and carbon for growth. A combined bioleaching experiment (coupling Brettanomyces B65 and A. thiooxidans TS6) performed in an air-lift reactor indicated that the rates of sludge pH reduction and ORP increase were greatly improved, resulting in enhanced Cr solubilization. Compared with the 5 days required for maximum solubilization of Cr for the control (single bioleaching process without inoculation of Brettanomyces B65), the bioleaching period was significantly shorten to 3 days for the combined bioleaching system. Moreover, little nitrogen and phosphorous were lost and the content of Cr was below the permitted levels for land application after 3 days of bioleaching treatment.

  14. Stoichiometric modeling of oxidation of reduced inorganic sulfur compounds (Riscs) in Acidithiobacillus thiooxidans.

    Science.gov (United States)

    Bobadilla Fazzini, Roberto A; Cortés, Maria Paz; Padilla, Leandro; Maturana, Daniel; Budinich, Marko; Maass, Alejandro; Parada, Pilar

    2013-08-01

    The prokaryotic oxidation of reduced inorganic sulfur compounds (RISCs) is a topic of utmost importance from a biogeochemical and industrial perspective. Despite sulfur oxidizing bacterial activity is largely known, no quantitative approaches to biological RISCs oxidation have been made, gathering all the complex abiotic and enzymatic stoichiometry involved. Even though in the case of neutrophilic bacteria such as Paracoccus and Beggiatoa species the RISCs oxidation systems are well described, there is a lack of knowledge for acidophilic microorganisms. Here, we present the first experimentally validated stoichiometric model able to assess RISCs oxidation quantitatively in Acidithiobacillus thiooxidans (strain DSM 17318), the archetype of the sulfur oxidizing acidophilic chemolithoautotrophs. This model was built based on literature and genomic analysis, considering a widespread mix of formerly proposed RISCs oxidation models combined and evaluated experimentally. Thiosulfate partial oxidation by the Sox system (SoxABXYZ) was placed as central step of sulfur oxidation model, along with abiotic reactions. This model was coupled with a detailed stoichiometry of biomass production, providing accurate bacterial growth predictions. In silico deletion/inactivation highlights the role of sulfur dioxygenase as the main catalyzer and a moderate function of tetrathionate hydrolase in elemental sulfur catabolism, demonstrating that this model constitutes an advanced instrument for the optimization of At. thiooxidans biomass production with potential use in biohydrometallurgical and environmental applications.

  15. Metagenomic investigation of the geologically unique Hellenic Volcanic Arc reveals a distinctive ecosystem with unexpected physiology.

    Science.gov (United States)

    Oulas, Anastasis; Polymenakou, Paraskevi N; Seshadri, Rekha; Tripp, H James; Mandalakis, Manolis; Paez-Espino, A David; Pati, Amrita; Chain, Patrick; Nomikou, Paraskevi; Carey, Steven; Kilias, Stephanos; Christakis, Christos; Kotoulas, Georgios; Magoulas, Antonios; Ivanova, Natalia N; Kyrpides, Nikos C

    2016-04-01

    Hydrothermal vents represent a deep, hot, aphotic biosphere where chemosynthetic primary producers, fuelled by chemicals from Earth's subsurface, form the basis of life. In this study, we examined microbial mats from two distinct volcanic sites within the Hellenic Volcanic Arc (HVA). The HVA is geologically and ecologically unique, with reported emissions of CO2 -saturated fluids at temperatures up to 220°C and a notable absence of macrofauna. Metagenomic data reveals highly complex prokaryotic communities composed of chemolithoautotrophs, some methanotrophs, and to our surprise, heterotrophs capable of anaerobic degradation of aromatic hydrocarbons. Our data suggest that aromatic hydrocarbons may indeed be a significant source of carbon in these sites, and instigate additional research into the nature and origin of these compounds in the HVA. Novel physiology was assigned to several uncultured prokaryotic lineages; most notably, a SAR406 representative is attributed with a role in anaerobic hydrocarbon degradation. This dataset, the largest to date from submarine volcanic ecosystems, constitutes a significant resource of novel genes and pathways with potential biotechnological applications.

  16. Complete genome sequence of Pyrolobus fumarii type strain (1AT)

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, Iain [U.S. Department of Energy, Joint Genome Institute; Goker, Markus [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Nolan, Matt [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; Huntemann, Marcel [U.S. Department of Energy, Joint Genome Institute; Liolios, Konstantinos [U.S. Department of Energy, Joint Genome Institute; Ivanova, N [U.S. Department of Energy, Joint Genome Institute; Pagani, Ioanna [U.S. Department of Energy, Joint Genome Institute; Mavromatis, K [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; Brambilla, Evelyne-Marie [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Huber, Harald [Universitat Regensburg, Regensburg, Germany; Yasawong, Montri [HZI - Helmholtz Centre for Infection Research, 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; Abt, Birte [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Sikorski, Johannes [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Wirth, Reinhard [Universitat Regensburg, Regensburg, Germany; Detter, J. Chris [U.S. Department of Energy, Joint Genome Institute; 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; Lapidus, Alla L. [U.S. Department of Energy, Joint Genome Institute

    2011-01-01

    Pyrolobus fumarii Bl chl et al. 1997 is the type species of the genus Pyrolobus, which be- longs to the crenarchaeal family Pyrodictiaceae. The species is a facultatively microaerophilic non-motile crenarchaeon. It is of interest because of its isolated phylogenetic location in the tree of life and because it is a hyperthermophilic chemolithoautotroph known as the primary producer of organic matter at deep-sea hydrothermal vents. P. fumarii exhibits currently the highest optimal growth temperature of all life forms on earth (106 C). This is the first com- pleted genome sequence of a member of the genus Pyrolobus to be published and only the second genome sequence from a member of the family Pyrodictiaceae. Although Diversa Corporation announced the completion of sequencing of the P. fumarii genome on Septem- ber 25, 2001, this sequence was never released to the public. The 1,843,267 bp long genome with its 1,986 protein-coding and 52 RNA genes is a part of the Genomic Encyclopedia of Bacteria and Archaea project.

  17. Sulfolobus hakonensis sp. nov., a novel species of acidothermophilic archaeon.

    Science.gov (United States)

    Takayanagi, S; Kawasaki, H; Sugimori, K; Yamada, T; Sugai, A; Ito, T; Yamasato, K; Shioda, M

    1996-04-01

    We characterized a microbial strain that was isolated from a hot spring at a geothermal area in Hakone, Japan. This isolate, whose lobed-shaped cells were about 1.0 micron in diameter, was a facultative chemolitho-autotroph that required aerobic conditions for growth. The optimum pH was 3.0 (pH range, 1.0 to 4.0), and the optimum temperature was 70 degrees C (temperature range, 50 to 80 degrees C). Lithotrophically, this strain grew on elemental sulfur and reduced sulfur compounds. The G+C content of the genomic DNA was 38.4 mol%. This organism contained calditoglycerocaldarchaeol, which is characteristic of members of the Sulfolobaceae. The levels of 16S rRNA sequence similarity between the new isolate and Sulfolobus acidocaldarius, Sulfolobus solfataricus, and Sulfolobus shibatae were less than 89.8%. Unlike S. acidocaldarius, S. solfataricus, and S. shibatae, the new isolate utilized sugars and amino acids poorly as sole carbon sources, and the levels of DNA-DNA hybridization between the new isolate and these Sulfolobus species were very low. Phenotypically, the new isolate was also distinct from the obligately lithotrophic organism Sulfolobus metallicus. We concluded that the new organism belongs to a new Sulfolobus species, for which we propose the name Sulfolobus hakonensis.

  18. Comparison of Optimal Thermodynamic Models of the Tricarboxylic Acid Cycle from Heterotrophs, Cyanobacteria, and Green Sulfur Bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Thomas, Dennis G.; Jaramillo Riveri, Sebastian I.; Baxter, Douglas J.; Cannon, William R.

    2014-12-15

    We have applied a new stochastic simulation approach to predict the metabolite levels, energy flow, and material flux in the different oxidative TCA cycles found in E. coli and Synechococcus sp. PCC 7002, and in the reductive TCA cycle typical of chemolithoautotrophs and phototrophic green sulfur bacteria such as Chlorobaculum tepidum. The simulation approach is based on equations of state and employs an assumption similar to that used in transition state theory. The ability to evaluate the thermodynamics of metabolic pathways allows one to understand the relationship between coupling of energy and material gradients in the environment and the selforganization of stable biological systems, and it is shown that each cycle operates in the direction expected due to its environmental niche. The simulations predict changes in metabolite levels and flux in response to changes in cofactor concentrations that would be hard to predict without an elaborate model based on the law of mass action. In fact, we show that a thermodynamically unfavorable reaction can still have flux in the forward direction when it is part of a reaction network. The ability to predict metabolite levels, energy flow and material flux should be significant for understanding the dynamics of natural systems and for understanding principles for engineering organisms for production of specialty chemicals, such as biofuels.

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

    Science.gov (United States)

    Hansen, Moritz; Perner, Mirjam

    2016-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Moritz eHansen

    2016-02-01

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

  1. Metagenomic comparison of two Thiomicrospira lineages inhabiting contrasting deep-sea hydrothermal environments.

    Directory of Open Access Journals (Sweden)

    William J Brazelton

    Full Text Available BACKGROUND: The most widespread bacteria in oxic zones of carbonate chimneys at the serpentinite-hosted Lost City hydrothermal field, Mid-Atlantic Ridge, belong to the Thiomicrospira group of sulfur-oxidizing chemolithoautotrophs. It is unclear why Thiomicrospira-like organisms thrive in these chimneys considering that Lost City hydrothermal fluids are notably lacking in hydrogen sulfide and carbon dioxide. METHODOLOGY/PRINCIPAL FINDINGS: Here we describe metagenomic sequences obtained from a Lost City carbonate chimney that are highly similar to the genome of Thiomicrospira crunogena XCL-2, an isolate from a basalt-hosted hydrothermal vent in the Pacific Ocean. Even though T. crunogena and Lost City Thiomicrospira inhabit different types of hydrothermal systems in different oceans, their genomic contents are highly similar. For example, sequences encoding the sulfur oxidation and carbon fixation pathways (including a carbon concentration mechanism of T. crunogena are also present in the Lost City metagenome. Comparative genomic analyses also revealed substantial genomic changes that must have occurred since the divergence of the two lineages, including large genomic rearrangements, gene fusion events, a prophage insertion, and transposase activity. CONCLUSIONS/SIGNIFICANCE: Our results show significant genomic similarity between Thiomicrospira organisms inhabiting different kinds of hydrothermal systems in different oceans, suggesting that these organisms are widespread and highly adaptable. These data also indicate genomic processes potentially associated with the adaptation of these lineages into strikingly different habitats.

  2. Characterization of Chemosynthetic Microbial Mats Associated with Intertidal Hydrothermal Sulfur Vents in White Point, San Pedro, CA, USA

    Science.gov (United States)

    Miranda, Priscilla J.; McLain, Nathan K.; Hatzenpichler, Roland; Orphan, Victoria J.; Dillon, Jesse G.

    2016-01-01

    The shallow-sea hydrothermal vents at White Point (WP) in Palos Verdes on the southern California coast support microbial mats and provide easily accessed settings in which to study chemolithoautotrophic sulfur cycling. Previous studies have cultured sulfur-oxidizing bacteria from the WP mats; however, almost nothing is known about the in situ diversity and activity of the microorganisms in these habitats. We studied the diversity, micron-scale spatial associations and metabolic activity of the mat community via sequence analysis of 16S rRNA and aprA genes, fluorescence in situ hybridization (FISH) microscopy and sulfate reduction rate (SRR) measurements. Sequence analysis revealed a diverse group of bacteria, dominated by sulfur cycling gamma-, epsilon-, and deltaproteobacterial lineages such as Marithrix, Sulfurovum, and Desulfuromusa. FISH microscopy suggests a close physical association between sulfur-oxidizing and sulfur-reducing genotypes, while radiotracer studies showed low, but detectable, SRR. Comparative 16S rRNA gene sequence analyses indicate the WP sulfur vent microbial mat community is similar, but distinct from other hydrothermal vent communities representing a range of biotopes and lithologic settings. These findings suggest a complete biological sulfur cycle is operating in the WP mat ecosystem mediated by diverse bacterial lineages, with some similarity with deep-sea hydrothermal vent communities. PMID:27512390

  3. Biomass production from electricity using ammonia as an electron carrier in a reverse microbial fuel cell.

    Directory of Open Access Journals (Sweden)

    Wendell O Khunjar

    Full Text Available The storage of renewable electrical energy within chemical bonds of biofuels and other chemicals is a route to decreasing petroleum usage. A critical challenge is the efficient transfer of electrons into a biological host that can covert this energy into high energy organic compounds. In this paper, we describe an approach whereby biomass is grown using energy obtained from a soluble mediator that is regenerated electrochemically. The net result is a separate-stage reverse microbial fuel cell (rMFC that fixes CO₂ into biomass using electrical energy. We selected ammonia as a low cost, abundant, safe, and soluble redox mediator that facilitated energy transfer to biomass. Nitrosomonas europaea, a chemolithoautotroph, was used as the biocatalyst due to its inherent capability to utilize ammonia as its sole energy source for growth. An electrochemical reactor was designed for the regeneration of ammonia from nitrite, and current efficiencies of 100% were achieved. Calculations indicated that overall bioproduction efficiency could approach 2.7±0.2% under optimal electrolysis conditions. The application of chemolithoautotrophy for industrial bioproduction has been largely unexplored, and results suggest that this and related rMFC platforms may enable biofuel and related biochemical production.

  4. Isolation, cultivation and genomic analysis of magnetosome biomineralization genes of a new genus of South-seeking magnetotactic cocci within the Alphaproteobacteria

    Energy Technology Data Exchange (ETDEWEB)

    Morillo, Viviana [Universidade Federal do Rio de Janeiro; Abreu, Fernanda [Universidade Federal do Rio de Janeiro; Araujo, Ana C [Universidade Federal do Rio de Janeiro; de Almeida, Luiz G [Laboratorio Nacional de Computacao Cientifica; Enrich-Prast, Alex [Universidade Federal do Rio de Janeiro; Farina, Marcos [Universidade Federal do Rio de Janeiro; de Vasconcelos, Ana T [Laboratorio Nacional de Computacao Cientifica; Bazylinski, Dennis A [Ames Laboratory; Lins, Ulysses [Universidade Federal do Rio de Janeiro

    2014-01-01

    Although magnetotactic bacteria (MTB) are ubiquitous in aquatic habitats, they are still considered fastidious microorganisms with regard to growth and cultivation with only a relatively low number of axenic cultures available to date. Here, we report the first axenic culture of an MTB isolated in the Southern Hemisphere (Itaipu Lagoon in Rio de Janeiro, Brazil). Cells of this new isolate are coccoid to ovoid in morphology and grow microaerophilically in semi-solid medium containing an oxygen concentration ([O2]) gradient either under chemoorganoheterotrophic or chemolithoautotrophic conditions. Each cell contains a single chain of approximately 10 elongated cuboctahedral magnetite (Fe3O4) magnetosomes. Phylogenetic analysis based on the 16S rRNA gene sequence shows that the coccoid MTB isolated in this study represents a new genus in the Alphaproteobacteria; the name Magnetofaba australis strain IT-1 is proposed. Preliminary genomic data obtained by pyrosequencing shows that M. australis strain IT-1 contains a genomic region with genes involved in biomineralization similar to those found in the most closely related magnetotactic cocci Magnetococcus marinus strain MC-1. However, organization of the magnetosome genes differs from M. marinus.

  5. Geochemical Energy for Catabolism and Anabolism in Hydrothermal Systems

    Science.gov (United States)

    Amend, J. P.; McCollom, T. M.; Bach, W.

    2008-12-01

    Chemically reduced deep-sea vent fluids mixed with oxidized seawater can generate redox disequilibria that serve as energy sources for chemolithoautotrophic (catabolism) and biomass synthesis (anabolism) reactions. Numerical models can be used to evaluate Gibbs energies of such processes on the early Earth and in present-day systems. Here, geochemical data from compositionally diverse vent fluids (Lost City, Rainbow, Logatchev, TAG, 21 °N EPR) are combined with several seawater chemistries to yield a wide range of mixed hydrothermal solutions; this is the starting point for our thermodynamic calculations. In ultramafic-hosted hydrothermal systems, such as Rainbow or Lost City, aerobic chemolithotrophic catabolisms (oxidation of H2, FeII, CH4) are the most energy-yielding at low temperatures (catabolic reaction energetics can then be used to put constraints on the amount of primary biomass production. Under putative early Earth conditions, for example, the net chemoautotrophic synthesis of cellular building blocks is thermodynamically most favorable at moderate temperatures (~50°C), where the energy contributions from HCO3- and H+ in cool seawater coupled to the reducing power in hot vent fluid are optimized. At these conditions, and counter to conventional wisdom, the synthesis of amino acids may even yield small amounts of energy.

  6. High nitrogen removal rate using ANAMMOX process at short hydraulic retention time.

    Science.gov (United States)

    Casagrande, C G; Kunz, A; De Prá, M C; Bressan, C R; Soares, H M

    2013-01-01

    The anaerobic ammonium oxidation (ANAMMOX) is a chemolithoautotrophic process, which converts NH(4)(+) to N(2) using nitrite (NO(2)(-)) as the electron acceptor. This process has very high nitrogen removal rates (NRRs) and is an alternative to classical nitrification/denitrification wastewater treatment. In the present work, a strategy for nitrogen removal using ANAMMOX process was tested evaluating their performance when submitted to high loading rates and very short hydraulic retention times (HRTs). An up-flow ANAMMOX column reactor was inoculated with 30% biomass (v v(-1)) fed from 100 to 200 mg L(-1) of total N (NO(2)(-)-N + NH(4)(+)-N) at 35 °C. After start-up and process stability the maximum NRR in the up-flow anaerobic sludge blanket (UASB) reactor was 18.3 g-N L(-1) d(-1) operated at 0.2 h of HRT. FISH (fluorescence in situ hybridization) analysis and process stoichiometry confirmed that ANAMMOX was the prevalent process for nitrogen removal during the experiments. The results point out that high NRRs can be obtained at very short HRTs using up-flow ANAMMOX column reactor configuration.

  7. Rock weathering creates oases of life in a high Arctic desert.

    Science.gov (United States)

    Borin, Sara; Ventura, Stefano; Tambone, Fulvia; Mapelli, Francesca; Schubotz, Florence; Brusetti, Lorenzo; Scaglia, Barbara; D'Acqui, Luigi P; Solheim, Bjørn; Turicchia, Silvia; Marasco, Ramona; Hinrichs, Kai-Uwe; Baldi, Franco; Adani, Fabrizio; Daffonchio, Daniele

    2010-02-01

    During primary colonization of rock substrates by plants, mineral weathering is strongly accelerated under plant roots, but little is known on how it affects soil ecosystem development before plant establishment. Here we show that rock mineral weathering mediated by chemolithoautotrophic bacteria is associated to plant community formation in sites recently released by permanent glacier ice cover in the Midtre Lovénbreen glacier moraine (78 degrees 53'N), Svalbard. Increased soil fertility fosters growth of prokaryotes and plants at the boundary between sites of intense bacterial mediated chemolithotrophic iron-sulfur oxidation and pH decrease, and the common moraine substrate where carbon and nitrogen are fixed by cyanobacteria. Microbial iron oxidizing activity determines acidity and corresponding fertility gradients, where water retention, cation exchange capacity and nutrient availability are increased. This fertilization is enabled by abundant mineral nutrients and reduced forms of iron and sulfur in pyrite minerals within a conglomerate type of moraine rock. Such an interaction between microorganisms and moraine minerals determines a peculiar, not yet described model for soil genesis and plant ecosystem formation with potential past and present analogues in other harsh environments with similar geochemical settings.

  8. Degradation of carbonyl sulfide by Actinomycetes and detection of clade D of β-class carbonic anhydrase.

    Science.gov (United States)

    Ogawa, Takahiro; Kato, Hiromi; Higashide, Mitsuru; Nishimiya, Mami; Katayama, Yoko

    2016-09-25

    Carbonyl sulfide (COS) is an atmospheric trace gas and one of the sources of stratospheric aerosol contributing to climate change. Although one of the major sinks of COS is soil, the distribution of COS degradation ability among bacteria remains unclear. Seventeen out of 20 named bacteria belonging to Actinomycetales had COS degradation activity at mole fractions of 30 parts per million by volume (ppmv) COS. Dietzia maris NBRC 15801(T) and Mycobacterium sp. THI405 had the activity comparable to a chemolithoautotroph Thiobacillus thioparus THI115 that degrade COS by COS hydrolase for energy production. Among 12 bacteria manifesting rapid degradation at 30 ppmv COS, Dietzia maris NBRC 15801(T) and Streptomyces ambofaciens NBRC 12836(T) degraded ambient COS (∼500 parts per trillion by volume). Geodermatophilus obscurus NBRC 13315(T) and Amycolatopsis orientalis NBRC 12806(T) increased COS concentrations. Moreover, six of eight COS degrading bacteria isolated from soils had partial nucleotide sequences similar to that of the gene encoding clade D of β-class carbonic anhydrase, which included COS hydrolase. These results indicate the potential importance of Actinomycetes in the role of soils as sinks of atmospheric COS.

  9. Potential contribution of planktonic components to ammonium cycling in the coastal area off central-southern Chile during non-upwelling conditions

    Science.gov (United States)

    Molina, Veronica; Morales, Carmen E.; Farías, Laura; Cornejo, Marcela; Graco, Michelle; Eissler, Yoanna; Cuevas, Luis A.

    2012-01-01

    The potential contributions of different microbial components (consumption and production, and carbon assimilation associated with photolithotrophic and chemolithoautotrophic (nitrification) metabolisms in the water column were performed. Despite low water column concentrations of ammonium in wintertime, intense ammonium transformations were registered. Prokaryotes (or bacterioplankton) contributed most to ammonium generation rates over the entire water column; these rates increased with depth (0.4-3.1 μM d -1). In surface waters (10 m depth), aerobic ammonium oxidation (potentially by Bacteria and Archaea) was the dominant consumption process (average 0.7 μM d -1) whereas in the subsurface layer (20 and 50 m depth), unexpectedly, eukaryotes accounted for most of its consumption (average 2.1 μM d -1). Nitrification oxidized an important proportion of the ammonium in both layers (from 25% to 100%) and provided regenerated nitrate. The integrated water column rates of chemosynthesis (0.005 g C m -2 d -1) represented a large proportion (51%) of the total dark carbon fixation during the non-upwelling season when integrated rates of photosynthesis are relatively low (0.42 g C m -2 d -1) and microbial food webs dominate the transfer of carbon within this coastal system.

  10. Chemically Specific Cellular Imaging of Biofilm Formation

    Energy Technology Data Exchange (ETDEWEB)

    Herberg, J L; Schaldach, C; Horn, J; Gjersing, E; Maxwell, R

    2006-02-09

    This document and the accompanying manuscripts summarize the technical accomplishments for our one-year LDRD-ER effort. Biofilm forming microbes have existed on this planet for billions of years and make up 60% of the biological mass on earth. Such microbes exhibit unique biochemical pathways during biofilm formation and play important roles in human health and the environment. Microbial biofilms have been directly implicated in, for example, product contamination, energy losses, and medical infection that cost the loss of human lives and billions of dollars. In no small part due to the lack of detailed understanding, biofilms unfortunately are resistant to control, inhibition, and destruction, either through treatment with antimicrobials or immunological defense mechanisms of the body. Current biofilm research has concentrated on the study of biofilms in the bulk. This is primarily due to the lack of analytical and physical tools to study biofilms non-destructively, in three dimensions, and on the micron or sub-micron scale. This has hindered the development of a clear understanding of either the early stage mechanisms of biofilm growth or the interactions of biofilms with their environment. Enzymatic studies have deduced a biochemical reaction that results in the oxidation of reduced sulfur species with the concomitant reduction of nitrate, a common groundwater pollutant, to dinitrogen gas by the bacterium, Thiobacillus denitrificans (TD). Because of its unique involvement in biologically relevant environmental pathways, TD is scheduled for genome sequencing in the near future by the DOE's Joint Genome Institute and is of interest to DOE's Genomes to Life Program. As our ecosystem is exposed to more and more nitrate contamination large scale livestock and agricultural practices, a further understanding of biofilm formation by organisms that could alleviate these problems is necessary in order to protect out biosphere. However, in order to study this

  11. Screening and Identification of an Achromobacter Strain for Both Arsenic Oxidizing and Denitrifying Abilities%一株具砷氧化和反硝化功能的无色杆菌的筛选和鉴定

    Institute of Scientific and Technical Information of China (English)

    曾琳; 朱琼芳; 卢贯能; 陈来琳; 匡庐峰; 柯林

    2011-01-01

    利用含As(3+)肉汤培养基,从广西河池砷污染地区水样和沉积物样中通过多次分离、纯化获取砷耐受菌.进一步从砷耐受菌中筛选出在好氧条件下可以同时进行砷氧化和反硝化的多功能菌株cll-35.对该菌株进行形态观察,并利用16SrDNA序列分析方法进行鉴定,发现该菌株为革兰氏阴性菌,与Achromobacter denitrificans strain 22426和Achromobacter xylosoxidans strain C8B的同源性均达99%;该菌株在NO(3)-和As(3+)同时存在的条件下好氧反硝化能力和砷氧化速率均得到提高;在只含NO(3)-的条件下,NO(3)-的去除率为53.65%,而在As(3+)和NO(3)-同时存在的条件下,NO(3)-的去除率为75.27%.在不含NO(3)-和含NO(3)-的条件下,As(3+)的转化率都在99%以上,而在含NONO(3)-的条件下,As(3+)的氧化速率更快.这种相互促进可能与反硝化过程中的电子传递和砷氧化过程中的动态平衡有关.%Water and sediment samples were taken from an arsenic-contaminated region in Hechi, Cuangxi Province. Arsenite-resisting strains were isolated and purified several times from broth medium containing As3+. Cll-35, the arsenite-oxidizer and aerobic denitrifier was further screened from arsenite-resisting strains. Morphological studies and 16S rDNA sequencing revealed that the isolate was Gram negative, and had 99% homogeneity to Achromobacter denitrificans strain 22426 and Achromobacter xylosoxidans strain C8B. The aerobic denitrifying and arsenite-oxidizing ability of this strain was enhanced when arsenite and nitrate were present together. The removal rate of nitrate was 53. 65% when only nitrate was present, while the removal rate increased to 75. 27% when both arsenite and nitrate were present. The conversion rate of As3+ was above 99% with and without nitrate, and the oxidation rate was enhanced by the presence of nitrate. This phenomenon may be related to the electron transmission process of aerobic denitrification and the dynamic balance

  12. Determination of substrate specificity of polyamine transporters in roseobacter species

    Science.gov (United States)

    Madhuri, S.; Mou, X.

    2012-12-01

    Polyamines, such as cadaverine, putrescine, spermidine, spermine and norspermine are a class of dissolved organic nitrogen (DON) that is ubiquitously found in marine environments. Intracellular polyamines are important in a variety of biological reactions, such as nucleic acid synthesis and protein synthesis. Free polyamines in seawater can be transported into bacterial cells by ABC transporter systems, each of which consists of four components including one substrate binding protein, one ATPase and two permeases. In silico analysis of marine bacterial genomes has revealed that roseobacter, a numerically and ecologically important taxa of marine bacteria, have at least two sets of polyamine transporter genes. This study was to examine the potential preference of roseobacter to different polyamine compounds and the substrate specificity of different polyamine transporters. Eleven roseobacter species, which genomes have been sequenced, were grown in defined media supplied with single polyamine compound as the sole carbon and nitrogen source. Growth assay showed a small number of roseobacter isolates to be generalist showing no preference among the tested polyamines (Ruegeria pomeroyi DSS-3, Roseovarius sp. TM1035, Roseovarius nubinhibens ISM, Jannaschia sp. CCS1 and Sagittula stellata E-37), whereas other isolates were specilists and were specific on polyamine compounds (Roseobacter sp. CCS2 and Roseobacter denitrificans OCh 114). Primers that probe poly-1 and pot-D genes, the two genes that encode common polyamine-binding genes of polyamine transporter systems were designed using net primer and primer design program. The specificity of the primers was validated by PCR followed by amplicon sequencing. Single step reverse transcription quantitative polymerase chain reactions (RT-qPCR) was performed to investigate substrate specificity of poly-1 and pot-D genes. Key-words Roseobacter, polyamine, polyamine transporter, dissolved organic nitrogen

  13. Sequence and structure-based comparative analysis to assess, identify and improve the thermostability of penicillin G acylases.

    Science.gov (United States)

    Panigrahi, Priyabrata; Chand, Deepak; Mukherji, Ruchira; Ramasamy, Sureshkumar; Suresh, C G

    2015-11-01

    Penicillin acylases are enzymes employed by the pharmaceutical industry for the manufacture of semi-synthetic penicillins. There is a continuous demand for thermostable and alkalophilic enzymes in such applications. We have carried out a computational analysis of known penicillin G acylases (PGAs) in terms of their thermostable nature using various protein-stabilizing factors. While the presence of disulfide bridges was considered initially to screen putative thermostable PGAs from the database, various other factors such as high arginine to lysine ratio, less content of thermolabile amino acids, presence of proline in β-turns, more number of ion-pair and other non-bonded interactions were also considered for comparison. A modified consensus approach designed could further identify stabilizing residue positions by site-specific comparison between mesostable and thermostable PGAs. A most likely thermostable enzyme identified from the analysis was PGA from Paracoccus denitrificans (PdPGA). This was cloned, expressed and tested for its thermostable nature using biochemical and biophysical experiments. The consensus site-specific sequence-based approach predicted PdPGA to be more thermostable than Escherichia coli PGA, but not as thermostable as the PGA from Achromobacter xylosoxidans. Experimental data showed that PdPGA was comparatively less thermostable than Achromobacter xylosoxidans PGA, although thermostability factors favored a much higher stability. Despite being mesostable, PdPGA being active and stable at alkaline pH is an advantage. Finally, several residue positions could be identified in PdPGA, which upon mutation selectively could improve the thermostability of the enzyme.

  14. Identification and Metabolic Mechanism of Non-fermentative Short-cut Denitrifying Phosphorus-removing Bacteria

    Institute of Scientific and Technical Information of China (English)

    LIU Hui; SUN Yanfu; JIA Xiaoshan; LI Jun; ZHOU Kangqun; QU Xiangdong; TAO Xueqin

    2013-01-01

    To investigate the characteristics and metabolic mechanism of short-cut denitrifying phosphorus-removing bacteria (SDPB) that are capable of enhanced biological phosphorus removal (EBPR) using nitrite as an electron acceptor,an aerobic/anoxic sequencing batch reactor was operated under three phases.An SDPB-strain YC was screened after the sludge enrichment and was identified by morphological,physiological,biochemical properties and 16S rDNA gene sequence analysis.Denitrifying phosphorus-removing experiments were conducted to study anaerobic and anoxic metabolic mechanisms by analyzing the changes of chemical oxygen demand (COD),phosphate,nitrite,poly-fβ-hydroxybutyrate (PHB),and glycogen.The results show that strain YC is a non-fermentative SDPB similar to Paracoccus denitrificans.As a kind of non-fermentative bacteria,the energy of strain YC was mainly generated from phosphorus release (96.2%) under anaerobic conditions with 0.32 mg P per mg synthesized PHB.Under anoxic conditions,strain YC accumulated 0.45 mg P per mg degraded PHB,which produced most of energy for phosphate accumulation (91.3%) and a little for glycogen synthesis (8.7%).This metabolic mechanism of strain YC is different from that of traditional phosphorus-accumulating organisms (PAOs).It is also found that PHB,a kind of intracellular polymer,plays a very important role in denitrifying and accumulating phosphorus by supplying sufficient energy for phosphorous accumulation and carbon sources for denitrification.Therefore,monitoring △P/△PHB and △NO2--N/△PHB is more necessary than monitoring △P/△COD,△NO2--N/△COD,or △P/△NO2--N.

  15. Dialects of the DNA uptake sequence in Neisseriaceae.

    Directory of Open Access Journals (Sweden)

    Stephan A Frye

    2013-04-01

    Full Text Available In all sexual organisms, adaptations exist that secure the safe reassortment of homologous alleles and prevent the intrusion of potentially hazardous alien DNA. Some bacteria engage in a simple form of sex known as transformation. In the human pathogen Neisseria meningitidis and in related bacterial species, transformation by exogenous DNA is regulated by the presence of a specific DNA Uptake Sequence (DUS, which is present in thousands of copies in the respective genomes. DUS affects transformation by limiting DNA uptake and recombination in favour of homologous DNA. The specific mechanisms of DUS-dependent genetic transformation have remained elusive. Bioinformatic analyses of family Neisseriaceae genomes reveal eight distinct variants of DUS. These variants are here termed DUS dialects, and their effect on interspecies commutation is demonstrated. Each of the DUS dialects is remarkably conserved within each species and is distributed consistent with a robust Neisseriaceae phylogeny based on core genome sequences. The impact of individual single nucleotide transversions in DUS on meningococcal transformation and on DNA binding and uptake is analysed. The results show that a DUS core 5'-CTG-3' is required for transformation and that transversions in this core reduce DNA uptake more than two orders of magnitude although the level of DNA binding remains less affected. Distinct DUS dialects are efficient barriers to interspecies recombination in N. meningitidis, N. elongata, Kingella denitrificans, and Eikenella corrodens, despite the presence of the core sequence. The degree of similarity between the DUS dialect of the recipient species and the donor DNA directly correlates with the level of transformation and DNA binding and uptake. Finally, DUS-dependent transformation is documented in the genera Eikenella and Kingella for the first time. The results presented here advance our understanding of the function and evolution of DUS and genetic

  16. Bacterial Sugar 3,4-Ketoisomerases: Structural Insight into Product Stereochemistry.

    Science.gov (United States)

    Thoden, James B; Vinogradov, Evgeny; Gilbert, Michel; Salinger, Ari J; Holden, Hazel M

    2015-07-28

    3-Acetamido-3,6-dideoxy-d-galactose (Fuc3NAc) and 3-acetamido-3,6-dideoxy-d-glucose (Qui3NAc) are unusual sugars found on the lipopolysaccharides of Gram-negative bacteria and on the S-layers of Gram-positive bacteria. The 3,4-ketoisomerases, referred to as FdtA and QdtA, catalyze the third steps in the respective biosynthetic pathways for these sugars. Whereas both enzymes utilize the same substrate, the stereochemistries of their products are different. Specifically, the hydroxyl groups at the hexose C-4' positions assume the "galactose" and "glucose" configurations in the FdtA and QdtA products, respectively. In 2007 we reported the structure of the apoform of FdtA from Aneurinibacillus thermoaerophilus, which was followed in 2014 by the X-ray analysis of QdtA from Thermoanaerobacterium thermosaccharolyticum as a binary complex. Both of these enzymes belong to the cupin superfamily. Here we report a combined structural and enzymological study to explore the manner in which these enzymes control the stereochemistry of their products. Various site-directed mutant proteins of each enzyme were constructed, and their dTDP-sugar products were analyzed by NMR spectroscopy. In addition, the kinetic parameters for these protein variants were measured, and the structure of one, namely, the QdtA Y17R/R97H double mutant form, was determined to 2.3-Å resolution. Finally, in an attempt to obtain a model of FdtA with a bound dTDP-linked sugar, the 3,4-ketoisomerase domain of a bifunctional enzyme from Shewanella denitrificans was cloned, purified, and crystallized in the presence of a dTDP-linked sugar analogue. Taken together, the results from this investigation demonstrate that it is possible to convert a "galacto" enzyme into a "gluco" enzyme and vice versa.

  17. Enhanced rate of intramolecular electron transfer in an engineered purple CuA azurin.

    Science.gov (United States)

    Farver, O; Lu, Y; Ang, M C; Pecht, I

    1999-02-02

    The recent expression of an azurin mutant where the blue type 1 copper site is replaced by the purple CuA site of Paracoccus denitrificans cytochrome c oxidase has yielded an optimal system for examining the unique electron mediation properties of the binuclear CuA center, because both type 1 and CuA centers are placed in the same location in the protein while all other structural elements remain the same. Long-range electron transfer is induced between the disulfide radical anion, produced pulse radiolytically, and the oxidized binuclear CuA center in the purple azurin mutant. The rate constant of this intramolecular process, kET = 650 +/- 60 s-1 at 298 K and pH 5.1, is almost 3-fold faster than for the same process in the wild-type single blue copper azurin from Pseudomonas aeruginosa (250 +/- 20 s-1), in spite of a smaller driving force (0.69 eV for purple CuA azurin vs. 0.76 eV for blue copper azurin). The reorganization energy of the CuA center is calculated to be 0.4 eV, which is only 50% of that found for the wild-type azurin. These results represent a direct comparison of electron transfer properties of the blue and purple CuA sites in the same protein framework and provide support for the notion that the binuclear purple CuA center is a more efficient electron transfer agent than the blue single copper center because reactivity of the former involves a lower reorganization energy.

  18. Microbiology and potential applications of aerobic methane oxidation coupled to denitrification (AME-D) process: A review.

    Science.gov (United States)

    Zhu, Jing; Wang, Qian; Yuan, Mengdong; Tan, Giin-Yu Amy; Sun, Faqian; Wang, Cheng; Wu, Weixiang; Lee, Po-Heng

    2016-03-01

    Aerobic methane oxidation coupled to denitrification (AME-D) is an important link between the global methane and nitrogen cycles. This mini-review updates discoveries regarding aerobic methanotrophs and denitrifiers, as a prelude to spotlight the microbial mechanism and the potential applications of AME-D. Until recently, AME-D was thought to be accomplished by a microbial consortium where denitrifying bacteria utilize carbon intermediates, which are excreted by aerobic methanotrophs, as energy and carbon sources. Potential carbon intermediates include methanol, citrate and acetate. This mini-review presents microbial thermodynamic estimations and postulates that methanol is the ideal electron donor for denitrification, and may serve as a trophic link between methanotrophic bacteria and denitrifiers. More excitingly, new discoveries have revealed that AME-D is not only confined to the conventional synergism between methanotrophic bacteria and denitrifiers. Specifically, an obligate aerobic methanotrophic bacterium, Methylomonas denitrificans FJG1, has been demonstrated to couple partial denitrification with methane oxidation, under hypoxia conditions, releasing nitrous oxide as a terminal product. This finding not only substantially advances the understanding of AME-D mechanism, but also implies an important but unknown role of aerobic methanotrophs in global climate change through their influence on both the methane and nitrogen cycles in ecosystems. Hence, further investigation on AME-D microbiology and mechanism is essential to better understand global climate issues and to develop niche biotechnological solutions. This mini-review also presents traditional microbial techniques, such as pure cultivation and stable isotope probing, and powerful microbial techniques, such as (meta-) genomics and (meta-) transcriptomics, for deciphering linked methane oxidation and denitrification. Although AME-D has immense potential for nitrogen removal from wastewater, drinking

  19. Engineering of alanine dehydrogenase from Bacillus subtilis for novel cofactor specificity.

    Science.gov (United States)

    Lerchner, Alexandra; Jarasch, Alexander; Skerra, Arne

    2016-09-01

    The l-alanine dehydrogenase of Bacillus subtilis (BasAlaDH), which is strictly dependent on NADH as redox cofactor, efficiently catalyzes the reductive amination of pyruvate to l-alanine using ammonia as amino group donor. To enable application of BasAlaDH as regenerating enzyme in coupled reactions with NADPH-dependent alcohol dehydrogenases, we alterated its cofactor specificity from NADH to NADPH via protein engineering. By introducing two amino acid exchanges, D196A and L197R, high catalytic efficiency for NADPH was achieved, with kcat /KM  = 54.1 µM(-1)  Min(-1) (KM  = 32 ± 3 µM; kcat  = 1,730 ± 39 Min(-1) ), almost the same as the wild-type enzyme for NADH (kcat /KM  = 59.9 µM(-1)  Min(-1) ; KM  = 14 ± 2 µM; kcat  = 838 ± 21 Min(-1) ). Conversely, recognition of NADH was much diminished in the mutated enzyme (kcat /KM  = 3 µM(-1)  Min(-1) ). BasAlaDH(D196A/L197R) was applied in a coupled oxidation/transamination reaction of the chiral dicyclic dialcohol isosorbide to its diamines, catalyzed by Ralstonia sp. alcohol dehydrogenase and Paracoccus denitrificans ω-aminotransferase, thus allowing recycling of the two cosubstrates NADP(+) and l-Ala. An excellent cofactor regeneration with recycling factors of 33 for NADP(+) and 13 for l-Ala was observed with the engineered BasAlaDH in a small-scale biocatalysis experiment. This opens a biocatalytic route to novel building blocks for industrial high-performance polymers.

  20. Formation of palladium(0) nanoparticles at microbial surfaces.

    Science.gov (United States)

    Bunge, Michael; Søbjerg, Lina S; Rotaru, Amelia-Elena; Gauthier, Delphine; Lindhardt, Anders T; Hause, Gerd; Finster, Kai; Kingshott, Peter; Skrydstrup, Troels; Meyer, Rikke L

    2010-10-01

    The increasing demand and limited natural resources for industrially important platinum-group metal (PGM) catalysts render the recovery from secondary sources such as industrial waste economically interesting. In the process of palladium (Pd) recovery, microorganisms have revealed a strong potential. Hitherto, bacteria with the property of dissimilatory metal reduction have been in focus, although the biochemical reactions linking enzymatic Pd(II) reduction and Pd(0) deposition have not yet been identified. In this study we investigated Pd(II) reduction with formate as the electron donor in the presence of Gram-negative bacteria with no documented capacity for reducing metals for energy production: Cupriavidus necator, Pseudomonas putida, and Paracoccus denitrificans. Only large and close-packed Pd(0) aggregates were formed in cell-free buffer solutions. Pd(II) reduction in the presence of bacteria resulted in smaller, well-suspended Pd(0) particles that were associated with the cells (called "bioPd(0)" in the following). Nanosize Pd(0) particles (3-30 nm) were only observed in the presence of bacteria, and particles in this size range were located in the periplasmic space. Pd(0) nanoparticles were still deposited on autoclaved cells of C. necator that had no hydrogenase activity, suggesting a hydrogenase-independent formation mechanism. The catalytic properties of Pd(0) and bioPd(0) were determined by the amount of hydrogen released in a reaction with hypophosphite. Generally, bioPd(0) demonstrated a lower level of activity than the Pd(0) control, possibly due to the inaccessibility of the Pd(0) fraction embedded in the cell envelope. Our results demonstrate the suitability of bacterial cells for the recovery of Pd(0), and formation and immobilization of Pd(0) nanoparticles inside the cell envelope. However, procedures to make periplasmic Pd(0) catalytically accessible need to be developed for future nanobiotechnological applications.

  1. Comparison of the MBBR denitrification carriers for advanced nitrogen removal of wastewater treatment plant effluent.

    Science.gov (United States)

    Yuan, Quan; Wang, Haiyan; Hang, Qianyu; Deng, Yangfan; Liu, Kai; Li, Chunmei; Zheng, Shengzhi

    2015-09-01

    The moving bed biofilm reactors (MBBRs) were used to remove the residual NO3(-)-N of wastewater treatment plant (WWTP) effluent, and the MBBR carriers for denitrification were compared. The results showed that high denitrification efficiency can be achieved with polyethylene, polypropylene, polyurethane foam, and haydite carriers under following conditions: 7.2 to 8.0 pH, 24 to 26 °C temperature, 12 h hydraulic retention time (HRT), and 25.5 mg L(-1) external methanol dosage, while the WWTP effluent total nitrogen (TN) was between 2.6 and 15.4 mg L(-1) and NO3(-)-N was between 0.2 and 12.6 mg L(-1). The MBBR filled with polyethylene carriers had higher TN and NO3(-)-N removal rate (44.9 ± 19.1 and 83.4 ± 13.0%, respectively) than those with other carriers. The minimum effluent TN and NO3(-)-N of polyethylene MBBR were 1.6 and 0.1 mg L(-1), respectively, and the maximum denitrification rate reached 23.0 g m(-2) day(-1). When chemical oxygen demand (COD)/TN ratio dropped from 6 to 4, the NO3(-)- N and TN removal efficiency decreased significantly in all reactors except for that filled with polyethylene, which indicated that the polyethylene MBBR can resist influent fluctuation much better. The three-dimensional excitation-emission matrix analysis showed that all the influent and effluent of MBBRs contain soluble microbial products (SMPs)-like organics and biochemical oxygen demand (BOD), which can be removed better by MBBRs filled with haydite and polyethylene carriers. The nitrous oxide reductase (nosZ)-based terminal restriction fragment length polymorphism (T-RFLP) analysis suggested that the dominant bacteria in polyethylene MBBR are the key denitrificans.

  2. Structure of azurin from Achromobacter xylosoxidans NCIB11015 at 2.5 A resolution.

    Science.gov (United States)

    Inoue, T; Shibata, N; Nakanishi, H; Koyama, S; Ishii, H; Kai, Y; Harada, S; Kasai, N; Ohshiro, Y; Suzuki, S

    1994-12-01

    The crystal structure of azurin from a denitrifying bacterium, Achromobacter xylosoxidans NCIB11015, has been refined at 2.5 A resolution using diffraction data obtained by means of synchrotron radiation at KEK. Crystals suitable for X-ray experiment were obtained by the macro-seeding method and an intensity data were obtained on imaging plates mounted on a Weissenberg camera (Rmerge = 0.09). The initial model was obtained by the molecular replacement method using the structure of azurin from Alcaligenes denitrificans NCTC8582 as a starting model. The structure was refined by molecular dynamics optimization and the restrained least-squares method to a crystallographic R-value of 0.205. However, the current model gave an electron-density of the side-chain regions of several residues close to the N-terminus quite different from those expected from the amino acid sequences reported. Very recently, two kinds of azurins (Az-I and Az-II) were isolated from this bacterium by a slightly modified purification method and have been characterized and found to have different CD spectra. On analysis of amino acid sequences around the N-terminus, the second azurin (Az-II) was proved to be a new type of azurin in this bacterium. It was consequently revealed that the current model corresponds to a new type of azurin because of the complete agreement between the electron-density and the amino acid sequence of the newly determined 20 residues from the N-terminus. Determination of the whole amino acid sequence of this azurin and further refinement are in progress.

  3. Anoxic growth of Ensifer meliloti 1021 by N2O-reduction, a potential mitigation strategy

    Directory of Open Access Journals (Sweden)

    Emilio eBueno

    2015-05-01

    Full Text Available Denitrification in agricultural soils is a major source of N2O. Legume crops enhance N2O emission by providing N-rich residues, thereby stimulating denitrification, both by free-living denitrifying bacteria and by the symbiont (rhizobium within the nodules. However, there are limited data concerning N2O production and consumption by endosymbiotic bacteria associated with legume crops. It has been reported that the alfalfa endosymbiont Ensifer meliloti strain 1021, despite possessing and expressing the complete set of denitrification enzymes, is unable to grow via nitrate respiration under anoxic conditions. In the present study, we have demonstrated by using a robotized incubation system that this bacterium is able to grow through anaerobic respiration of N2O to N2. N2O reductase (N2OR activity was not dependent on the presence of nitrogen oxyanions or NO, thus the expression could be induced by oxygen depletion alone. When incubated at pH 6, E. meliloti was unable to reduce N2O, corroborating previous observations found in both, extracted soil bacteria and Paracoccus denitrificans pure cultures, where expression of functional N2O reductase is difficult at low pH. Furthermore, the presence in the medium of highly reduced C-substrates, such as butyrate, negatively affected N2OR activity. The emission of N2O from soils can be lowered if legumes plants are inoculated with rhizobial strains overexpressing N2O reductase. This study demonstrates that strains like E. meliloti 1021, which do not produce N2O but are able to reduce the N2O emitted by other organisms, could act as even better N2O sinks.

  4. Theoretical studies of the functional role of the cross-linked histidine-tyrosine copper-B ligand of cytochrome c oxidase

    Science.gov (United States)

    McDonald, William J.

    In the present work density functional theory calculations were performed to explore the vibrational and electronic spectral changes associated with proton and electron transfer from the CuB center at the active site of cytochrome c oxidase that lead to the formation of either the PR or PM intermediates in the catalytic reduction of dioxygen to water. Furthermore, the thermodynamics of proton and electron transfer from the cross-linked histidine-tyrosine Cu B ligand were explored to assess the possible role of this ligand as a proton and/or electron donor during enzymatic turnover. Characteristic calculated cross-linked imidazole-phenolate and imidazole-phenoxyl radical vibrational frequencies and isotope shifts are in good agreement with the vibrational spectra of the PR and PM intermediates of the bo3 quinol oxidase from E. coli and R. sphaeroides and P. denitrificans suggesting that the Y244 (bovine numbering) is deprotonated in the PR intermediate, and is a tyrosyl radical in P M. Furthermore, using isodesmic reactions, the cross-linked phenol is found to be a significantly stronger acid than an unmodified phenol in the gas-phase, and likely in the inhomogeneous low-dielectric environment of the membrane bound enzyme, supporting the conclusion that the cross-linked tyrosine is a proton donor during enzymatic turnover. Time-dependent density functional theory calculations qualitatively reproduce the red-shift in the UV/visible absorption spectrum of a cross-linked imidazole-phenolate anion compared to the imidazole-phenol. Furthermore, the unique ˜500 nm absorption of a cross-linked imidazole-phenoxyl radical is correctly predicted using TDDFT and may be assigned as an imidazole-phenoxyl radical pi-to-pi* transition. Furthermore, this absorption is predicted in the spectrum of a Cu2+-imidazole-phenoxyl biradical model.

  5. Characterizing the proton loading site in cytochrome c oxidase.

    Science.gov (United States)

    Lu, Jianxun; Gunner, M R

    2014-08-26

    Cytochrome c oxidase (CcO) uses the energy released by reduction of O2 to H2O to drive eight charges from the high pH to low pH side of the membrane, increasing the electrochemical gradient. Four electrons and protons are used for chemistry, while four more protons are pumped. Proton pumping requires that residues on a pathway change proton affinity through the reaction cycle to load and then release protons. The protonation states of all residues in CcO are determined in MultiConformational Continuum Electrostatics simulations with the protonation and redox states of heme a, a3, Cu(B), Y288, and E286 used to define the catalytic cycle. One proton is found to be loaded and released from residues identified as the proton loading site (PLS) on the P-side of the protein in each of the four CcO redox states. Thus, the same proton pumping mechanism can be used each time CcO is reduced. Calculations with structures of Rhodobacter sphaeroides, Paracoccus denitrificans, and bovine CcO derived by crystallography and molecular dynamics show the PLS functions similarly in different CcO species. The PLS is a cluster rather than a single residue, as different structures show 1-4 residues load and release protons. However, the proton affinity of the heme a3 propionic acids primarily determines the number of protons loaded into the PLS; if their proton affinity is too low, less than one proton is loaded.

  6. Survey of Ability of Activated Sludge Isolated Bacteria in Removal of RB-B Dyestuff from Aqueous Medium

    Directory of Open Access Journals (Sweden)

    Z Javadi

    2011-01-01

    Full Text Available "n "n "nBackgrounds and Objectives: Reactive dyestuff has potential of toxicity, carcinogenesis and mutagenesis for mammals and aquatic organisms. The current physical and chemical methods such as adsorption, coagulation, precipitation, filtration and ... can been used for removing of dyestuff. Biological treatment which is effective and economic for decontamination of dyestuff wastewaters was preferred because of limitation and difficulty of physicochemical methods. In order to investigate the trend of pollution reduction of color compounds, ability of Remazol Black-B dyestuff removal from aqueous medium by bacterial consortium under anoxic conditions was studied."nMaterials and Methods: The mix culture of bacteria from textile industries activated sludge was enriched in luria broth medium containing RB-B dyestuff as a carbon source. Then biodegradation was assessed in 4 batch reactors. Microbial population of bacterial and decolorization quantities of samples were detected by MPN and UV-Vis spectrophotometer."nResults: Decolorization efficiency by the bacterial consortium was obtained more than 99% for 50 and 250 mg/L concentrations in 72 and 144 h (3 and 6 days respectively, while for the initial concentration of 500 mg/L was 98.1in 240 h (10 days of biodegradation period. Dyestuff reduction rate after completed removal was about 0.69, 1.74,2 mg/L/h for initial concentration of 50, 250, 500 mg/L respectively."nConclusion: Results showed that Alcaligenes denitrificans and Alcaligenes xylosoxidans bacteria"nwhich were isolated from activated sludge have good potential of RB-B dyestuff removal and this removal is depending on primary concentration of dye. Removal efficiency increased as primary concentration went up.

  7. MALDI-TOF Mass Spectrometry: A Powerful Tool for Clinical Microbiology at Hopital Principal de Dakar, Senegal (West Africa.

    Directory of Open Access Journals (Sweden)

    Cheikh I Lo

    Full Text Available Our team in Europe has developed the routine clinical laboratory identification of microorganisms by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF mass spectrometry (MS. To evaluate the utility of MALDI-TOF MS in tropical Africa in collaboration with local teams, we installed an apparatus in the Hôpital Principal de Dakar (Senegal, performed routine identification of isolates, and confirmed or completed their identification in France. In the case of discordance or a lack of identification, molecular biology was performed. Overall, 153/191 (80.1% and 174/191 (91.1% isolates yielded an accurate and concordant identification for the species and genus, respectively, with the 2 different MALDI-TOF MSs in Dakar and Marseille. The 10 most common bacteria, representing 94.2% of all bacteria routinely identified in the laboratory in Dakar (Escherichia coli, Klebsiella pneumoniae, Streptococcus agalactiae, Acinetobacter baumannii, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus haemolyticus, Enterobacter cloacae, Enterococcus faecalis, and Staphylococcus epidermidis were accurately identified with the MALDI-TOF MS in Dakar. The most frequent misidentification in Dakar was at the species level for Achromobacter xylosoxidans, which was inaccurately identified as Achromobacter denitrificans, and the bacteria absent from the database, such as Exiguobacterium aurientacum or Kytococcus schroeteri, could not be identified. A few difficulties were observed with MALDI-TOF MS for Bacillus sp. or oral streptococci. 16S rRNA sequencing identified a novel bacterium, "Necropsobacter massiliensis." The robust identification of microorganisms by MALDI-TOF MS in Dakar and Marseille demonstrates that MALDI-TOF MS can be used as a first-line tool in clinical microbiology laboratories in tropical countries.

  8. Characterization and expression of genes from the RubisCO gene cluster of the chemoautotrophic symbiont of Solemya velum: cbbLSQO.

    Science.gov (United States)

    Schwedock, Julie; Harmer, Tara L; Scott, Kathleen M; Hektor, Harm J; Seitz, Angelica P; Fontana, Matthew C; Distel, Daniel L; Cavanaugh, Colleen M

    2004-09-01

    Chemoautotrophic endosymbionts residing in Solemya velum gills provide this shallow water clam with most of its nutritional requirements. The cbb gene cluster of the S. velum symbiont, including cbbL and cbbS, which encode the large and small subunits of the carbon-fixing enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO), was cloned and expressed in Escherichia coli. The recombinant RubisCO had a high specific activity, approximately 3 micromol min(-1) mg protein (-1), and a KCO2 of 40.3 microM. Based on sequence identity and phylogenetic analyses, these genes encode a form IA RubisCO, both subunits of which are closely related to those of the symbiont of the deep-sea hydrothermal vent gastropod Alviniconcha hessleri and the photosynthetic bacterium Allochromatium vinosum. In the cbb gene cluster of the S. velum symbiont, the cbbLS genes were followed by cbbQ and cbbO, which are found in some but not all cbb gene clusters and whose products are implicated in enhancing RubisCO activity post-translationally. cbbQ shares sequence similarity with nirQ and norQ, found in denitrification clusters of Pseudomonas stutzeri and Paracoccus denitrificans. The 3' region of cbbO from the S. velum symbiont, like that of the three other known cbbO genes, shares similarity to the 3' region of norD in the denitrification cluster. This is the first study to explore the cbb gene structure for a chemoautotrophic endosymbiont, which is critical both as an initial step in evaluating cbb operon structure in chemoautotrophic endosymbionts and in understanding the patterns and forces governing RubisCO evolution and physiology.

  9. The microbial sulfur cycle at extremely haloalkaline conditions of soda lakes.

    Science.gov (United States)

    Sorokin, Dimitry Y; Kuenen, J Gijs; Muyzer, Gerard

    2011-01-01

    Soda lakes represent a unique ecosystem with extremely high pH (up to 11) and salinity (up to saturation) due to the presence of high concentrations of sodium carbonate in brines. Despite these double extreme conditions, most of the lakes are highly productive and contain a fully functional microbial system. The microbial sulfur cycle is among the most active in soda lakes. One of the explanations for that is high-energy efficiency of dissimilatory conversions of inorganic sulfur compounds, both oxidative and reductive, sufficient to cope with costly life at double extreme conditions. The oxidative part of the sulfur cycle is driven by chemolithoautotrophic haloalkaliphilic sulfur-oxidizing bacteria (SOB), which are unique for soda lakes. The haloalkaliphilic SOB are present in the surface sediment layer of various soda lakes at high numbers of up to 10(6) viable cells/cm(3). The culturable forms are so far represented by four novel genera within the Gammaproteobacteria, including the genera Thioalkalivibrio, Thioalkalimicrobium, Thioalkalispira, and Thioalkalibacter. The latter two were only found occasionally and each includes a single species, while the former two are widely distributed in various soda lakes over the world. The genus Thioalkalivibrio is the most physiologically diverse and covers the whole spectrum of salt/pH conditions present in soda lakes. Most importantly, the dominant subgroup of this genus is able to grow in saturated soda brines containing 4 M total Na(+) - a so far unique property for any known aerobic chemolithoautotroph. Furthermore, some species can use thiocyanate as a sole energy source and three out of nine species can grow anaerobically with nitrogen oxides as electron acceptor. The reductive part of the sulfur cycle is active in the anoxic layers of the sediments of soda lakes. The in situ measurements of sulfate reduction rates and laboratory experiments with sediment slurries using sulfate, thiosulfate, or elemental sulfur as

  10. Microbial Response to a Deep-Sea Volcanic Eruption at 9N on the East Pacific Rise

    Science.gov (United States)

    Sievert, S. M.; Gulmann, L. K.; Hugler, M.; Taylor, C. D.; Molyneaux, S. J.; Sylva, S. P.; Beaulieu, S. E.; Shank, T. M.; Summons, R. E.; Wirsen, C. O.

    2006-12-01

    sulfidicus, a chemolithoautotrophic sulfur oxidizing bacterium that forms filamentous sulfur, dominating. Arcobacter related sequences were not observed on the basalt samples, indicating that these organisms might be early colonizers, which subsequently get replaced by other bacteria. This supports our hypothesis that autotrophic epsilon- proteobacteria, such as Arcobacter, will be the first colonizers, followed by increasing numbers of heterotrophic microorganisms over time. These autotrophic microbes appear to predominantly live in the subsurface at vents, allowing the rapid colonization of newly exposed surfaces. To confirm this hypothesis we also filtered large volumes of water in situ to access the composition of the microbial communities associated with the expelled fluids. Shipboard incubations with 13C-labeled bicarbonate were further carried out to identify chemolithoautotrophic microbes. The data obtained in this study will further be compared with recent colonization experiments that were carried out at 9 ° N EPR prior to the eruption. Overall, our studies show that microorganisms rapidly colonize newly exposed surfaces at sites where warm water emanates from the sub-seafloor. Furthermore there appears to be a succession of microbes, with epsilon-proteobacteria and in particular Arcobacter species as primary colonizers, possibly triggering the settlement of larvae. Finally, the data obtained during this cruise will serve as a benchmark for future cruises and analyses to document the changes in the microbial communities occurring over time.

  11. Microbial Community Structure of Subglacial Lake Whillans, West Antarctica

    Science.gov (United States)

    Achberger, Amanda M.; Christner, Brent C.; Michaud, Alexander B.; Priscu, John C.; Skidmore, Mark L.; Vick-Majors, Trista J.; Adkins, W.

    2016-01-01

    Subglacial Lake Whillans (SLW) is located beneath ∼800 m of ice on the Whillans Ice Stream in West Antarctica and was sampled in January of 2013, providing the first opportunity to directly examine water and sediments from an Antarctic subglacial lake. To minimize the introduction of surface contaminants to SLW during its exploration, an access borehole was created using a microbiologically clean hot water drill designed to reduce the number and viability of microorganisms in the drilling water. Analysis of 16S rRNA genes (rDNA) amplified from samples of the drilling and borehole water allowed an evaluation of the efficacy of this approach and enabled a confident assessment of the SLW ecosystem inhabitants. Based on an analysis of 16S rDNA and rRNA (i.e., reverse-transcribed rRNA molecules) data, the SLW community was found to be bacterially dominated and compositionally distinct from the assemblages identified in the drill system. The abundance of bacteria (e.g., Candidatus Nitrotoga, Sideroxydans, Thiobacillus, and Albidiferax) and archaea (Candidatus Nitrosoarchaeum) related to chemolithoautotrophs was consistent with the oxidation of reduced iron, sulfur, and nitrogen compounds having important roles as pathways for primary production in this permanently dark ecosystem. Further, the prevalence of Methylobacter in surficial lake sediments combined with the detection of methanogenic taxa in the deepest sediment horizons analyzed (34–36 cm) supported the hypothesis that methane cycling occurs beneath the West Antarctic Ice Sheet. Large ratios of rRNA to rDNA were observed for several operational taxonomic units abundant in the water column and sediments (e.g., Albidiferax, Methylobacter, Candidatus Nitrotoga, Sideroxydans, and Smithella), suggesting a potentially active role for these taxa in the SLW ecosystem. Our findings are consistent with chemosynthetic microorganisms serving as the ecological foundation in this dark subsurface environment, providing new

  12. Sulfur metabolizing microbes dominate microbial communities in Andesite-hosted shallow-sea hydrothermal systems.

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    Yao Zhang

    Full Text Available To determine microbial community composition, community spatial structure and possible key microbial processes in the shallow-sea hydrothermal vent systems off NE Taiwan's coast, we examined the bacterial and archaeal communities of four samples collected from the water column extending over a redoxocline gradient of a yellow and four from a white hydrothermal vent. Ribosomal tag pyrosequencing based on DNA and RNA showed statistically significant differences between the bacterial and archaeal communities of the different hydrothermal plumes. The bacterial and archaeal communities from the white hydrothermal plume were dominated by sulfur-reducing Nautilia and Thermococcus, whereas the yellow hydrothermal plume and the surface water were dominated by sulfide-oxidizing Thiomicrospira and Euryarchaeota Marine Group II, respectively. Canonical correspondence analyses indicate that methane (CH(4 concentration was the only statistically significant variable that explains all community cluster patterns. However, the results of pyrosequencing showed an essential absence of methanogens and methanotrophs at the two vent fields, suggesting that CH(4 was less tied to microbial processes in this shallow-sea hydrothermal system. We speculated that mixing between hydrothermal fluids and the sea or meteoric water leads to distinctly different CH(4 concentrations and redox niches between the yellow and white vents, consequently influencing the distribution patterns of the free-living Bacteria and Archaea. We concluded that sulfur-reducing and sulfide-oxidizing chemolithoautotrophs accounted for most of the primary biomass synthesis and that microbial sulfur metabolism fueled microbial energy flow and element cycling in the shallow hydrothermal systems off the coast of NE Taiwan.

  13. The genome of Nitrospina gracilis illuminates the metabolism and evolution of the major marine nitrite oxidizer

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    Sebastian eLuecker

    2013-02-01

    Full Text Available In marine systems, nitrate is the major reservoir of inorganic fixed nitrogen. The only known biological nitrate-forming reaction is nitrite oxidation, but despite its importance, our knowledge of the organisms catalyzing this key process in the marine N-cycle is very limited. The most frequently encountered marine NOB are related to Nitrospina gracilis, an aerobic chemolithoautotrophic bacterium isolated from ocean surface waters. To date, limited physiological and genomic data for this organism were available and its phylogenetic affiliation was uncertain. In this study, the draft genome sequence of Nitrospina gracilis strain 3/211 was obtained. Unexpectedly for an aerobic organism, N. gracilis lacks classical reactive oxygen defense mechanisms and uses the reductive tricarboxylic acid cycle for carbon fixation. These features indicate microaerophilic ancestry and are consistent with the presence of Nitrospina in marine oxygen minimum zones. Fixed carbon is stored intracellularly as glycogen, but genes for utilizing external organic carbon sources were not identified. N. gracilis also contains a full gene set for oxidative phosphorylation with oxygen as terminal electron acceptor and for reverse electron transport from nitrite to NADH. A novel variation of complex I may catalyze the required reverse electron flow to low-potential ferredoxin. Interestingly, comparative genomics indicated a strong evolutionary link between Nitrospina, the nitrite-oxidizing genus Nitrospira, and anaerobic ammonium oxidizers, apparently including the horizontal transfer of a periplasmically oriented nitrite oxidoreductase and other key genes for nitrite oxidation at an early evolutionary stage. Further, detailed phylogenetic analyses using concatenated marker genes provided evidence that Nitrospina forms a novel bacterial phylum, for which we propose the name Nitrospinae.

  14. The Genome of Nitrospina gracilis Illuminates the Metabolism and Evolution of the Major Marine Nitrite Oxidizer.

    Science.gov (United States)

    Lücker, Sebastian; Nowka, Boris; Rattei, Thomas; Spieck, Eva; Daims, Holger

    2013-01-01

    In marine systems, nitrate is the major reservoir of inorganic fixed nitrogen. The only known biological nitrate-forming reaction is nitrite oxidation, but despite its importance, our knowledge of the organisms catalyzing this key process in the marine N-cycle is very limited. The most frequently encountered marine NOB are related to Nitrospina gracilis, an aerobic chemolithoautotrophic bacterium isolated from ocean surface waters. To date, limited physiological and genomic data for this organism were available and its phylogenetic affiliation was uncertain. In this study, the draft genome sequence of N. gracilis strain 3/211 was obtained. Unexpectedly for an aerobic organism, N. gracilis lacks classical reactive oxygen defense mechanisms and uses the reductive tricarboxylic acid cycle for carbon fixation. These features indicate microaerophilic ancestry and are consistent with the presence of Nitrospina in marine oxygen minimum zones. Fixed carbon is stored intracellularly as glycogen, but genes for utilizing external organic carbon sources were not identified. N. gracilis also contains a full gene set for oxidative phosphorylation with oxygen as terminal electron acceptor and for reverse electron transport from nitrite to NADH. A novel variation of complex I may catalyze the required reverse electron flow to low-potential ferredoxin. Interestingly, comparative genomics indicated a strong evolutionary link between Nitrospina, the nitrite-oxidizing genus Nitrospira, and anaerobic ammonium oxidizers, apparently including the horizontal transfer of a periplasmically oriented nitrite oxidoreductase and other key genes for nitrite oxidation at an early evolutionary stage. Further, detailed phylogenetic analyses using concatenated marker genes provided evidence that Nitrospina forms a novel bacterial phylum, for which we propose the name Nitrospinae.

  15. Insights into the Quorum Sensing Regulon of the Acidophilic Acidithiobacillus ferrooxidans Revealed by Transcriptomic in the Presence of an Acyl Homoserine Lactone Superagonist Analog.

    Science.gov (United States)

    Mamani, Sigde; Moinier, Danielle; Denis, Yann; Soulère, Laurent; Queneau, Yves; Talla, Emmanuel; Bonnefoy, Violaine; Guiliani, Nicolas

    2016-01-01

    While a functional quorum sensing system has been identified in the acidophilic chemolithoautotrophic Acidithiobacillus ferrooxidans ATCC 23270(T) and shown to modulate cell adhesion to solid substrates, nothing is known about the genes it regulates. To address the question of how quorum sensing controls biofilm formation in A. ferrooxidans (T), the transcriptome of this organism in conditions in which quorum sensing response is stimulated by a synthetic superagonist AHL (N-acyl homoserine lactones) analog has been studied. First, the effect on biofilm formation of a synthetic AHL tetrazolic analog, tetrazole 9c, known for its agonistic QS activity, was assessed by fluorescence and electron microscopy. A fast adherence of A. ferrooxidans (T) cells on sulfur coupons was observed. Then, tetrazole 9c was used in DNA microarray experiments that allowed the identification of genes regulated by quorum sensing signaling, and more particularly, those involved in early biofilm formation. Interestingly, afeI gene, encoding the AHL synthase, but not the A. ferrooxidans quorum sensing transcriptional regulator AfeR encoding gene, was shown to be regulated by quorum sensing. Data indicated that quorum sensing network represents at least 4.5% (141 genes) of the ATCC 23270(T) genome of which 42.5% (60 genes) are related to biofilm formation. Finally, AfeR was shown to bind specifically to the regulatory region of the afeI gene at the level of the palindromic sequence predicted to be the AfeR binding site. Our results give new insights on the response of A. ferrooxidans to quorum sensing and on biofilm biogenesis.

  16. Dissolved inorganic carbon uptake in Thiomicrospira crunogena XCL-2 is Δp- and ATP-sensitive and enhances RubisCO-mediated carbon fixation.

    Science.gov (United States)

    Menning, Kristy J; Menon, Balaraj B; Fox, Gordon; Scott, Kathleen M

    2016-03-01

    The gammaproteobacterium Thiomicrospira crunogena XCL-2 is an aerobic sulfur-oxidizing hydrothermal vent chemolithoautotroph that has a CO2 concentrating mechanism (CCM), which generates intracellular dissolved inorganic carbon (DIC) concentrations much higher than extracellular, thereby providing substrate for carbon fixation at sufficient rate. This CCM presumably requires at least one active DIC transporter to generate the elevated intracellular concentrations of DIC measured in this organism. In this study, the half-saturation constant (K CO2) for purified carboxysomal RubisCO was measured (276 ± 18 µM) which was much greater than the K CO2 of whole cells (1.03 µM), highlighting the degree to which the CCM facilitates CO2 fixation under low CO2 conditions. To clarify the bioenergetics powering active DIC uptake, cells were incubated in the presence of inhibitors targeting ATP synthesis (DCCD) or proton potential (CCCP). Incubations with each of these inhibitors resulted in diminished intracellular ATP, DIC, and fixed carbon, despite an absence of an inhibitory effect on proton potential in the DCCD-incubated cells. Electron transport complexes NADH dehydrogenase and the bc 1 complex were found to be insensitive to DCCD, suggesting that ATP synthase was the primary target of DCCD. Given the correlation of DIC uptake to the intracellular ATP concentration, the ABC transporter genes were targeted by qRT-PCR, but were not upregulated under low-DIC conditions. As the T. crunogena genome does not include orthologs of any genes encoding known DIC uptake systems, these data suggest that a novel, yet to be identified, ATP- and proton potential-dependent DIC transporter is active in this bacterium. This transporter serves to facilitate growth by T. crunogena and other Thiomicrospiras in the many habitats where they are found.

  17. The complete genome sequence of Thermoproteus tenax: a physiologically versatile member of the Crenarchaeota.

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    Bettina Siebers

    Full Text Available Here, we report on the complete genome sequence of the hyperthermophilic Crenarchaeum Thermoproteus tenax (strain Kra1, DSM 2078(T a type strain of the crenarchaeotal order Thermoproteales. Its circular 1.84-megabase genome harbors no extrachromosomal elements and 2,051 open reading frames are identified, covering 90.6% of the complete sequence, which represents a high coding density. Derived from the gene content, T. tenax is a representative member of the Crenarchaeota. The organism is strictly anaerobic and sulfur-dependent with optimal growth at 86°C and pH 5.6. One particular feature is the great metabolic versatility, which is not accompanied by a distinct increase of genome size or information density as compared to other Crenarchaeota. T. tenax is able to grow chemolithoautotrophically (CO₂/H₂ as well as chemoorganoheterotrophically in presence of various organic substrates. All pathways for synthesizing the 20 proteinogenic amino acids are present. In addition, two presumably complete gene sets for NADH:quinone oxidoreductase (complex I were identified in the genome and there is evidence that either NADH or reduced ferredoxin might serve as electron donor. Beside the typical archaeal A₀A₁-ATP synthase, a membrane-bound pyrophosphatase is found, which might contribute to energy conservation. Surprisingly, all genes required for dissimilatory sulfate reduction are present, which is confirmed by growth experiments. Mentionable is furthermore, the presence of two proteins (ParA family ATPase, actin-like protein that might be involved in cell division in Thermoproteales, where the ESCRT system is absent, and of genes involved in genetic competence (DprA, ComF that is so far unique within Archaea.

  18. Aerobic bacterial pyrite oxidation and acid rock drainage during the Great Oxidation Event.

    Science.gov (United States)

    Konhauser, Kurt O; Lalonde, Stefan V; Planavsky, Noah J; Pecoits, Ernesto; Lyons, Timothy W; Mojzsis, Stephen J; Rouxel, Olivier J; Barley, Mark E; Rosìere, Carlos; Fralick, Phillip W; Kump, Lee R; Bekker, Andrey

    2011-10-19

    The enrichment of redox-sensitive trace metals in ancient marine sedimentary rocks has been used to determine the timing of the oxidation of the Earth's land surface. Chromium (Cr) is among the emerging proxies for tracking the effects of atmospheric oxygenation on continental weathering; this is because its supply to the oceans is dominated by terrestrial processes that can be recorded in the Cr isotope composition of Precambrian iron formations. However, the factors controlling past and present seawater Cr isotope composition are poorly understood. Here we provide an independent and complementary record of marine Cr supply, in the form of Cr concentrations and authigenic enrichment in iron-rich sedimentary rocks. Our data suggest that Cr was largely immobile on land until around 2.48 Gyr ago, but within the 160 Myr that followed--and synchronous with independent evidence for oxygenation associated with the Great Oxidation Event (see, for example, refs 4-6)--marked excursions in Cr content and Cr/Ti ratios indicate that Cr was solubilized at a scale unrivalled in history. As Cr isotope fractionations at that time were muted, Cr must have been mobilized predominantly in reduced, Cr(III), form. We demonstrate that only the oxidation of an abundant and previously stable crustal pyrite reservoir by aerobic-respiring, chemolithoautotrophic bacteria could have generated the degree of acidity required to solubilize Cr(III) from ultramafic source rocks and residual soils. This profound shift in weathering regimes beginning at 2.48 Gyr ago constitutes the earliest known geochemical evidence for acidophilic aerobes and the resulting acid rock drainage, and accounts for independent evidence of an increased supply of dissolved sulphate and sulphide-hosted trace elements to the oceans around that time. Our model adds to amassing evidence that the Archaean-Palaeoproterozoic boundary was marked by a substantial shift in terrestrial geochemistry and biology.

  19. Genome Analysis of Thermosulfurimonas dismutans, the First Thermophilic Sulfur-Disproportionating Bacterium of the Phylum Thermodesulfobacteria

    Science.gov (United States)

    Mardanov, Andrey V.; Beletsky, Alexey V.; Kadnikov, Vitaly V.; Slobodkin, Alexander I.; Ravin, Nikolai V.

    2016-01-01

    Thermosulfurimonas dismutans S95T, isolated from a deep-sea hydrothermal vent is the first bacterium of the phylum Thermodesulfobacteria reported to grow by the disproportionation of elemental sulfur, sulfite, or thiosulfate with carbon dioxide as the sole carbon source. In contrast to its phylogenetically close relatives, which are dissimilatory sulfate-reducers, T. dismutans is unable to grow by sulfate respiration. The features of this organism and its 2,1 Mb draft genome sequence are described in this report. Genome analysis revealed that the T. dismutans genome contains the set of genes for dissimilatory sulfate reduction including ATP sulfurylase, the AprA and B subunits of adenosine-5′-phosphosulfate reductase, and dissimilatory sulfite reductase. The oxidation of elemental sulfur to sulfite could be enabled by APS reductase-associated electron transfer complex QmoABC and heterodisulfide reductase. The genome also contains several membrane-linked molybdopterin oxidoreductases that are thought to be involved in sulfur metabolism as subunits of thiosulfate, polysulfide, or tetrathionate reductases. Nitrate could be used as an electron acceptor and reduced to ammonium, as indicated by the presence of periplasmic nitrate and nitrite reductases. Autotrophic carbon fixation is enabled by the Wood–Ljungdahl pathway, and the complete set of genes that is required for nitrogen fixation is also present in T. dismutans. Overall, our results provide genomic insights into energy and carbon metabolism of chemolithoautotrophic sulfur-disproportionating bacterium that could be important primary producer in microbial communities of deep-sea hydrothermal vents. PMID:27379079

  20. Chemolithotrophy in the continental deep subsurface: Sanford Underground Research Facility (SURF, USA

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    Magdalena Rose Osburn

    2014-11-01

    Full Text Available The deep subsurface is an enormous repository of microbial life. However, the metabolic capabilities of these microorganisms and the degree to which they are dependent on surface processes are largely unknown. Due to the logistical difficulty of sampling and inherent heterogeneity, the microbial populations of the terrestrial subsurface are poorly characterized. In an effort to better understand the biogeochemistry of deep terrestrial habitats, we evaluate the energetic yield of chemolithotrophic metabolisms and microbial diversity in the Sanford Underground Research Facility (SURF in the former Homestake Gold Mine, SD, USA. Geochemical data, energetic modeling, and DNA sequencing were combined with principle component analysis to describe this deep (down to 8100 ft below surface, terrestrial environment. SURF provides access into an iron-rich Paleoproterozoic metasedimentary deposit that contains deeply circulating groundwater. Geochemical analyses of subsurface fluids reveal enormous geochemical diversity ranging widely in salinity, oxidation state (ORP 330 to -328 mV, and concentrations of redox sensitive species (e.g., Fe2+ from near 0 to 6.2 mg/L and ΣS2- from 7 to 2778 μg/L. As a direct result of this compositional buffet, Gibbs energy calculations reveal an abundance of energy for microorganisms from the oxidation of sulfur, iron, nitrogen, methane, and manganese. Pyrotag DNA sequencing reveals diverse communities of chemolithoautotrophs, thermophiles, aerobic and anaerobic heterotrophs, and numerous uncultivated clades. Extrapolated across the mine footprint, these data suggest a complex spatial mosaic of subsurface primary productivity that is in good agreement with predicted energy yields. Notably, we report Gibbs energy normalized both per mole of reaction and per kg fluid (energy density and find the later to be more consistent with observed physiologies and environmental conditions. Further application of this approach will

  1. Biogenic Carbon on Mars: A Subsurface Chauvinistic Viewpoint

    Science.gov (United States)

    Onstott, T. C.; Lau, C. Y. M.; Magnabosco, C.; Harris, R.; Chen, Y.; Slater, G.; Sherwood Lollar, B.; Kieft, T. L.; van Heerden, E.; Borgonie, G.; Dong, H.

    2015-12-01

    A review of 150 publications on the subsurface microbiology of the continental subsurface provides ~1,400 measurements of cellular abundances down to 4,800 meter depth. These data suggest that the continental subsurface biomass is comprised of ~1016-17 grams of carbon, which is higher than the most recent estimates of ~1015 grams of carbon (1 Gt) for the marine deep biosphere. If life developed early in Martian history and Mars sustained an active hydrological cycle during its first 500 million years, then is it possible that Mars could have developed a subsurface biomass of comparable size to that of Earth? Such a biomass would comprise a much larger fraction of the total known Martian carbon budget than does the subsurface biomass on Earth. More importantly could a remnant of this subsurface biosphere survive to the present day? To determine how sustainable subsurface life could be in isolation from the surface we have been studying subsurface fracture fluids from the Precambrian Shields in South Africa and Canada. In these environments the energetically efficient and deeply rooted acetyl-CoA pathway for carbon fixation plays a central role for chemolithoautotrophic primary producers that form the base of the biomass pyramid. These primary producers appear to be sustained indefinitely by H2 generated through serpentinization and radiolytic reactions. Carbon isotope data suggest that in some subsurface locations a much larger population of secondary consumers are sustained by the primary production of biogenic CH4 from a much smaller population of methanogens. These inverted biomass and energy pyramids sustained by the cycling of CH4 could have been and could still be active on Mars. The C and H isotopic signatures of Martian CH4 remain key tools in identifying potential signatures of an extant Martian biosphere. Based upon our results to date cavity ring-down spectroscopic technologies provide an option for making these measurements on future rover missions.

  2. Activity and abundance of denitrifying bacteria in the subsurface biosphere of diffuse hydrothermal vents of the Juan de Fuca Ridge

    Science.gov (United States)

    Bourbonnais, A.; Juniper, S. K.; Butterfield, D. A.; Devol, A. H.; Kuypers, M. M. M.; Lavik, G.; Hallam, S. J.; Wenk, C. B.; Chang, B. X.; Murdock, S. A.; Lehmann, M. F.

    2012-11-01

    Little is known about fixed nitrogen (N) transformation and elimination at diffuse hydrothermal vents where anoxic fluids are mixed with oxygenated crustal seawater prior to discharge. Oceanic N sinks that remove bio-available N ultimately affect chemosynthetic primary productivity in these ecosystems. Using 15N paired isotope techniques, we determined potential rates of fixed N loss pathways (denitrification, anammox) and dissimilatory nitrate reduction to ammonium (DNRA) in sulfidic hydrothermal vent fluids discharging from the subsurface at several sites at Axial Volcano and the Endeavour Segment on the Juan de Fuca Ridge. We also measured physico-chemical parameters (i.e., temperature, pH, nutrients, H2S and N2O concentrations) as well as the biodiversity and abundance of chemolithoautotrophic nitrate-reducing, sulfur-oxidizing γ-proteobacteria (SUP05 cluster) using sequence analysis of amplified small subunit ribosomal RNA (16S rRNA) genes in combination with taxon-specific quantitative polymerase chain reaction (qPCR) assays. Denitrification was the dominant N loss pathway in the subsurface biosphere of the Juan de Fuca Ridge, with rates of up to ~1000 nmol N l-1 day-1. In comparison, anammox rates were always waters. Taxon-specific qPCR revealed that γ-proteobacteria of the SUP05 cluster sometimes dominated the microbial community (SUP05/total bacteria up to 38%). Significant correlations were found between fixed N loss (i.e., denitrification, anammox) rates and in situ nitrate and dissolved inorganic nitrogen (DIN) deficits in the fluids, indicating that DIN availability may ultimately regulate N loss in the subsurface. Based on our rate measurements, and on published data on hydrothermal fluid fluxes and residence times, we estimated that up to ~10 Tg N yr-1 could globally be removed in the subsurface biosphere of hydrothermal vents systems, thus, representing a small fraction of the total marine N loss (~275 to > 400 Tg N yr-1).

  3. Bacteria, fungi and biokarst in Lechuguilla Cave, Carlsbad Caverns National Park, New Mexico

    Science.gov (United States)

    Cunningham, K. I.; Northup, D. E.; Pollastro, R. M.; Wright, W. G.; Larock, E. J.

    1995-02-01

    Lechuguilla Cave is a deep, extensive, gypsumand sulfur-bearing hypogenic cave in Carlsbad Caverns National Park, New Mexico, most of which (>90%) lies more than 300 m beneath the entrance. Located in the arid Guadalupe Mountains, Lechuguilla's remarkable state of preservation is partially due to the locally continuous Yates Formation siltstone that has effectively diverted most vadose water away from the cave. Allocthonous organic input to the cave is therefore very limited, but bacterial and fungal colonization is relatively extensive: (1) Aspergillus sp. fungi and unidentified bacteria are associated with iron-, manganese-, and sulfur-rich encrustations on calcitic folia near the suspected water table 466 m below the entrance; (2) 92 species of fungi in 19 genera have been identified throughout the cave in oligotrophic (nutrient-poor) “soils” and pools; (3) cave-air condensate contains unidentified microbes; (4) indigenous chemoheterotrophic Seliberius and Caulobacter bacteria are known from remote pool sites; and (5) at least four genera of heterotrophic bacteria with population densities near 5×105 colony-forming units (CFU) per gram are present in ceiling-bound deposits of supposedly abiogenic condensation-corrosion residues. Various lines of evidence suggest that autotrophic bacteria are present in the ceiling-bound residues and could act as primary producers in a unique subterranean microbial food chain. The suspected autotrophic bacteria are probably chemolithoautotrophic (CLA), utilizing trace iron, manganese, or sulfur in the limestone and dolomitic bedrock to mechanically (and possibly biochemically) erode the substrate to produce residual floor deposits. Because other major sources of organic matter have not been detected, we suggest that these CLA bacteria are providing requisite organic matter to the known heterotrophic bacteria and fungi in the residues. The cavewide bacterial and fungal distribution, the large volumes of corrosion residues

  4. Influence of hydrological, biogeochemical and temperature transients on subsurface carbon fluxes in a flood plain environment

    Energy Technology Data Exchange (ETDEWEB)

    Arora, Bhavna; Spycher, Nicolas F.; Steefel, Carl I.; Molins, Sergi; Bill, Markus; Conrad, Mark E.; Dong, Wenming; Faybishenko, Boris; Tokunaga, Tetsu K.; Wan, Jiamin; Williams, Kenneth H.; Yabusaki, Steven B.

    2016-02-01

    Flood plains play a potentially important role in the global carbon cycle. The accumulation of organic matter in flood plains often induces the formation of chemically reduced groundwater and sediments along riverbanks. In this study, our objective is to evaluate the cumulative impact of such reduced zones, water table fluctuations, and temperature gradients on subsurface carbon fluxes in a flood plain at Rifle, Colorado located along the Colorado River. 2-D coupled variably-saturated, non-isothermal flow and biogeochemical reactive transport modeling was applied to improve our understanding of the abiotic and microbially mediated reactions controlling carbon dynamics at the Rifle site. Model simulations considering only abiotic reactions (thus ignoring microbial reactions) underestimated CO2 partial pressures observed in the unsaturated zone and severely underestimated inorganic (and overestimated organic) carbon fluxes to the river compared to simulations with biotic pathways. Both model simulations and field observations highlighted the need to include microbial contributions from chemolithoautotrophic processes (e.g., Fe?2 and S-2 oxidation) to match locally-observed high CO2 concentrations above reduced zones. Observed seasonal variations in CO2 concentrations in the unsaturated zone could not be reproduced without incorporating temperature gradients in the simulations. Incorporating temperature fluctuations resulted in an increase in the annual groundwater carbon fluxes to the river by 170 % to 3.3 g m-2 d-1, while including water table variations resulted in an overall decrease in the simulated fluxes. We conclude that spatial microbial and redox zonation as well as temporal fluctuations of temperature and water table depth contribute significantly to subsurface carbon fluxes in flood plains and need to be represented appropriately in model simulations.

  5. Chemolithotrophic acetogenic H2/CO2 utilization in Italian rice field soil.

    Science.gov (United States)

    Liu, Fanghua; Conrad, Ralf

    2011-09-01

    Acetate oxidation in Italian rice field at 50 °C is achieved by uncultured syntrophic acetate oxidizers. As these bacteria are closely related to acetogens, they may potentially also be able to synthesize acetate chemolithoautotrophically. Labeling studies using exogenous H(2) (80%) and (13)CO(2) (20%), indeed demonstrated production of acetate as almost exclusive primary product not only at 50 °C but also at 15 °C. Small amounts of formate, propionate and butyrate were also produced from (13)CO(2). At 50 °C, acetate was first produced but later on consumed with formation of CH(4). Acetate was also produced in the absence of exogenous H(2) albeit to lower concentrations. The acetogenic bacteria and methanogenic archaea were targeted by stable isotope probing of ribosomal RNA (rRNA). Using quantitative PCR, (13)C-labeled bacterial rRNA was detected after 20 days of incubation with (13)CO(2). In the heavy fractions at 15 °C, terminal restriction fragment length polymorphism, cloning and sequencing of 16S rRNA showed that Clostridium cluster I and uncultured Peptococcaceae assimilated (13)CO(2) in the presence and absence of exogenous H(2), respectively. A similar experiment showed that Thermoanaerobacteriaceae and Acidobacteriaceae were dominant in the (13)C treatment at 50 °C. Assimilation of (13)CO(2) into archaeal rRNA was detected at 15 °C and 50 °C, mostly into Methanocellales, Methanobacteriales and rice cluster III. Acetoclastic methanogenic archaea were not detected. The above results showed the potential for acetogenesis in the presence and absence of exogenous H(2) at both 15 °C and 50 °C. However, syntrophic acetate oxidizers seemed to be only active at 50 °C, while other bacterial groups were active at 15 °C.

  6. Assembly and Succession of Iron Oxide Microbial Mat Communities in Acidic Geothermal Springs

    Energy Technology Data Exchange (ETDEWEB)

    Beam, Jake; Bernstein, Hans C.; Jay, Z.; Kozubal, Mark; Jennings, Ryan; Tringe, Susannah G.; Inskeep, William P.

    2016-02-15

    Iron oxide microbial mats are ubiquitous geobiological features on Earth and occur in extant acidic hot springs of Yellowstone National Park (YNP), WY, USA, and form as a result of microbial processes. The relative contribution of different organisms to the development of these mat ecosystems is of specific interest. We hypothesized that chemolithoautotrophic organisms contribute to the early development and production of Fe(III)-oxide mats, which could support later-colonizing heterotrophic microorganisms. Sterile glass slides were incubated in the outflow channels of two acidic geothermal springs in YNP, and spatiotemporal changes in Fe(III)-oxide accretion and abundance of relevant community members were measured. Lithoautotrophic Hydrogenobaculum spp. were first colonizers and the most abundant taxa identified during early successional stages (7 – 40 days). Populations of M. yellowstonensis colonized after ~ 7 days, corresponding to visible Fe(III)-oxide accretion. Heterotrophic archaea colonized after 30 days, and emerge as the dominant functional guild in mature iron oxide mats (1 – 2 cm thick) that form after 70 – 120 days. First-order rate constants of iron oxide accretion ranged from 0.05 – 0.046 day-1, and reflected the absolute amount of iron accreted. Micro- and macroscale microterracettes were identified during iron oxide mat development, and suggest that the mass transfer of oxygen limits microbial growth. This was also demonstrated using microelectrode measurements of oxygen as a function of mat depth, which showed steep gradients in oxygen from the aqueous mat interface to ~ 1 mm. The formation and succession of amorphous Fe(III)-oxide mat communities follows a predictable pattern of distinct stages and growth. The successional stages and microbial signatures observed in these extant Fe(III)-oxide mat communities may be relevant to other past or present Fe(III)-oxide mineralizing systems.

  7. Carbon dioxide fixation by Metallosphaera yellowstonensis and acidothermophilic iron-oxidizing microbial communities from Yellowstone National Park

    Energy Technology Data Exchange (ETDEWEB)

    Jennings, Ryan; Whitmore, Laura M.; Moran, James J.; Kreuzer, Helen W.; Inskeep, William P.

    2014-05-01

    The fixation of inorganic carbon (as carbon dioxide) has been documented in all three domains of life and results in the biosynthesis of a diverse suite of organic compounds that support the growth of heterotrophic organisms. The primary aim of this study was to assess the importance of carbon dioxide fixation in high-temperature Fe(III)-oxide mat communities and in pure cultures of one of the dominant Fe(II)-oxidizing organisms (Metallosphaera yellowstonensis strain MK1) present in situ. Protein-encoding genes of the complete 3-hydroxypropionate/4-hydroxybutyrate (3-HP/4-HB) carbon fixation pathway were identified in pure-cultures of M. yellowstonensis strain MK1. Metagenome sequencing from the same environments also revealed genes for the 3-HP/4-HB pathway belonging to M. yellowstonensis populations, as well as genes for a complete reductive TCA cycle from Hydrogenobaculum spp. (Aquificales). Stable isotope (13CO2) labeling was used to measure the fixation of CO2 by M. yellowstonensis strain MK1, and in ex situ assays containing live Fe(III)-oxide microbial mats. Results showed that M. yellowstonensis strain MK1 fixes CO2 via the 3-HP/4-HB pathway with a fractionation factor of ~ 2.5 ‰. Direct analysis of the 13C composition of dissolved inorganic C (DIC), dissolved organic C (DOC), landscape C and microbial mat C showed that mat C is comprised of both DIC and non-DIC sources. The estimated contribution of DIC carbon to biomass C (> ~ 35%) is reasonably consistent with the relative abundance of known chemolithoautotrophs and corresponding CO2 fixation pathways detected in metagenome sequence. The significance of DIC as a major source of carbon for Fe-oxide mat communities provides a foundation for examining microbial interactions in these systems that are dependent on the activity of autotrophic organisms such as Hydrogenobaculum and Metallosphaera spp.

  8. Complete genome sequence of Nitrobacter hamburgensis X14 and comparative genomic analysis of species within the genus Nitrobacter.

    Energy Technology Data Exchange (ETDEWEB)

    Starkenburg, Shawn R [Oregon State University; Larimer, Frank W [ORNL; Stein, Lisa Y [University of California, Riverside; Klotz, Martin G [University of Louisville, Louisville; Chain, Patrick S. G. [Lawrence Livermore National Laboratory (LLNL); Sayavedra-Soto, LA [Oregon State University; Poret-Peterson, Amisha T. [University of Louisville, Louisville; Gentry, ME [University of Louisville, Louisville; Arp, D J [Oregon State University; Ward, Bess B. [Princeton University; Bottomley, Peter J [Oregon State University

    2008-05-01

    The alphaproteobacterium Nitrobacter hamburgensis X14 is a gram-negative facultative chemolithoautotroph that conserves energy from the oxidation of nitrite to nitrate. Sequencing and analysis of the Nitrobacter hamburgensis X14 genome revealed four replicons comprised of one chromosome (4.4 Mbp) and three plasmids (294, 188, and 121 kbp). Over 20% of the genome is composed of pseudogenes and paralogs. Whole-genome comparisons were conducted between N. hamburgensis and the finished and draft genome sequences of Nitrobacter winogradskyi and Nitrobacter sp. strain Nb-311A, respectively. Most of the plasmid-borne genes were unique to N. hamburgensis and encode a variety of functions (central metabolism, energy conservation, conjugation, and heavy metal resistance), yet approximately 21 kb of a approximately 28-kb "autotrophic" island on the largest plasmid was conserved in the chromosomes of Nitrobacter winogradskyi Nb-255 and Nitrobacter sp. strain Nb-311A. The N. hamburgensis chromosome also harbors many unique genes, including those for heme-copper oxidases, cytochrome b(561), and putative pathways for the catabolism of aromatic, organic, and one-carbon compounds, which help verify and extend its mixotrophic potential. A Nitrobacter "subcore" genome was also constructed by removing homologs found in strains of the closest evolutionary relatives, Bradyrhizobium japonicum and Rhodopseudomonas palustris. Among the Nitrobacter subcore inventory (116 genes), copies of genes or gene clusters for nitrite oxidoreductase (NXR), cytochromes associated with a dissimilatory nitrite reductase (NirK), PII-like regulators, and polysaccharide formation were identified. Many of the subcore genes have diverged significantly from, or have origins outside, the alphaproteobacterial lineage and may indicate some of the unique genetic requirements for nitrite oxidation in Nitrobacter.

  9. Genome-Enabled Modeling of Biogeochemical Processes Predicts Metabolic Dependencies that Connect the Relative Fitness of Microbial Functional Guilds

    Science.gov (United States)

    Brodie, E.; King, E.; Molins, S.; Karaoz, U.; Steefel, C. I.; Banfield, J. F.; Beller, H. R.; Anantharaman, K.; Ligocki, T. J.; Trebotich, D.

    2015-12-01

    Pore-scale processes mediated by microorganisms underlie a range of critical ecosystem services, regulating carbon stability, nutrient flux, and the purification of water. Advances in cultivation-independent approaches now provide us with the ability to reconstruct thousands of genomes from microbial populations from which functional roles may be assigned. With this capability to reveal microbial metabolic potential, the next step is to put these microbes back where they belong to interact with their natural environment, i.e. the pore scale. At this scale, microorganisms communicate, cooperate and compete across their fitness landscapes with communities emerging that feedback on the physical and chemical properties of their environment, ultimately altering the fitness landscape and selecting for new microbial communities with new properties and so on. We have developed a trait-based model of microbial activity that simulates coupled functional guilds that are parameterized with unique combinations of traits that govern fitness under dynamic conditions. Using a reactive transport framework, we simulate the thermodynamics of coupled electron donor-acceptor reactions to predict energy available for cellular maintenance, respiration, biomass development, and enzyme production. From metagenomics, we directly estimate some trait values related to growth and identify the linkage of key traits associated with respiration and fermentation, macromolecule depolymerizing enzymes, and other key functions such as nitrogen fixation. Our simulations were carried out to explore abiotic controls on community emergence such as seasonally fluctuating water table regimes across floodplain organic matter hotspots. Simulations and metagenomic/metatranscriptomic observations highlighted the many dependencies connecting the relative fitness of functional guilds and the importance of chemolithoautotrophic lifestyles. Using an X-Ray microCT-derived soil microaggregate physical model combined

  10. Microbial diversity in the deep-subsurface hydrothermal aquifer feeding the giant gypsum crystal-bearing Naica Mine, Mexico.

    Science.gov (United States)

    Ragon, Marie; Van Driessche, Alexander E S; García-Ruíz, Juan M; Moreira, David; López-García, Purificación

    2013-01-01

    The Naica Mine in northern Mexico is famous for its giant gypsum crystals, which may reach up to 11 m long and contain fluid inclusions that might have captured microorganisms during their formation. These crystals formed under particularly stable geochemical conditions in cavities filled by low salinity hydrothermal water at 54-58°C. We have explored the microbial diversity associated to these deep, saline hydrothermal waters collected in the deepest (ca. 700-760 m) mineshafts by amplifying, cloning and sequencing small-subunit ribosomal RNA genes using primers specific for archaea, bacteria, and eukaryotes. Eukaryotes were not detectable in the samples and the prokaryotic diversity identified was very low. Two archaeal operational taxonomic units (OTUs) were detected in one sample. They clustered with, respectively, basal Thaumarchaeota lineages and with a large clade of environmental sequences branching at the base of the Thermoplasmatales within the Euryarchaeota. Bacterial sequences belonged to the Candidate Division OP3, Firmicutes and the Alpha- and Beta-proteobacteria. Most of the lineages detected appear autochthonous to the Naica system, since they had as closest representatives environmental sequences retrieved from deep sediments or the deep subsurface. In addition, the high GC content of 16S rRNA gene sequences belonging to the archaea and to some OP3 OTUs suggests that at least these lineages are thermophilic. Attempts to amplify diagnostic functional genes for methanogenesis (mcrA) and sulfate reduction (dsrAB) were unsuccessful, suggesting that those activities, if present, are not important in the aquifer. By contrast, genes encoding archaeal ammonium monooxygenase (AamoA) were amplified, suggesting that Naica Thaumarchaeota are involved in nitrification. These organisms are likely thermophilic chemolithoautotrophs adapted to thrive in an extremely energy-limited environment.

  11. Microbial diversity in the deep-subsurface hydrothermal aquifer feeding the giant gypsum crystal-bearing Naica mine, Mexico

    Directory of Open Access Journals (Sweden)

    Marie eRagon

    2013-03-01

    Full Text Available The Naica mine in Northern Mexico is famous for its giant gypsum crystals, which may reach up to 11 m long and contain fluid inclusions that might have captured microorganisms during their formation. These crystals formed under particularly stable geochemical conditions in cavities filled by low salinity hydrothermal water at 54-58°C. We have explored the microbial diversity associated to these deep, saline hydrothermal waters collected in the deepest (ca. 700-760 m mineshafts by amplifying, cloning and sequencing small-subunit ribosomal RNA genes using primers specific for archaea, bacteria and eukaryotes. Eukaryotes were not detectable in the samples and the prokaryotic diversity identified was very low. Two archaeal operational taxonomic units (OTUs were detected in one sample. They clustered with, respectively, basal Thaumarchaeota lineages and with a large clade of environmental sequences branching at the base of the Thermoplasmatales within the Euryarchaeota. Bacterial sequences belonged to the Candidate Division OP3, Firmicutes and the Alpha- and Beta-Proteobacteria. Most of the lineages detected appear autochthonous to the Naica system, since they had as closest representatives environmental sequences retrieved from deep sediments or the deep subsurface. In addition, the high GC content of 16S rRNA gene sequences belonging to the archaea and to some OP3 OTUs suggests that at least these lineages are thermophilic. Attempts to amplify diagnostic functional genes for methanogenesis (mcrA and sulfate reduction (dsrAB were unsuccessful, suggesting that those activities, if present, are not important in the aquifer. By contrast, genes encoding archaeal ammonium monooxygenase (AamoA were amplified, suggesting that Naica Thaumarchaeota are involved in nitrification. These organisms are likely thermophilic chemolithoautotrophs adapted to thrive in an extremely energy-limited environment.

  12. Deferrisoma paleochoriense sp. nov., a thermophilic, iron(III)-reducing bacterium from a shallow-water hydrothermal vent in the Mediterranean Sea

    Science.gov (United States)

    Perez-Rodriguez, Ileana M.; Rawls, Matthew; Coykendall, Dolly K.; Foustoukos, Dionysis I.

    2016-01-01

    A novel thermophilic, anaerobic, mixotrophic bacterium, designated strain MAG-PB1T, was isolated from a shallow-water hydrothermal vent system in Palaeochori Bay off the coast of the island of Milos, Greece. The cells were Gram-negative, rugose, short rods, approximately 1.0 μm long and 0.5 μm wide. Strain MAG-PB1T grew at 30–70 °C (optimum 60 °C), 0–50 g NaCl l− 1 (optimum 15–20 g l− 1) and pH 5.5–8.0 (optimum pH 6.0). Generation time under optimal conditions was 2.5 h. Optimal growth occurred under chemolithoautotrophic conditions with H2 as the energy source and CO2 as the carbon source. Fe(III), Mn(IV), arsenate and selenate were used as electron acceptors. Peptone, tryptone, Casamino acids, sucrose, yeast extract, d-fructose, α-d-glucose and ( − )-d-arabinose also served as electron donors. No growth occurred in the presence of lactate or formate. The G+C content of the genomic DNA was 66.7 mol%. Phylogenetic analysis of the 16S rRNA gene sequence indicated that this organism is closely related to Deferrisoma camini, the first species of a recently described genus in the Deltaproteobacteria. Based on the 16S rRNA gene phylogenetic analysis and on physiological, biochemical and structural characteristics, the strain was found to represent a novel species, for which the name Deferrisoma palaeochoriense sp. nov. is proposed. The type strain is MAG-PB1T ( = JCM 30394T = DSM 29363T). 

  13. Snowmelt Induced Hydrologic Perturbations Drive Dynamic Microbiological and Geochemical Behaviors across a Shallow Riparian Aquifer

    Energy Technology Data Exchange (ETDEWEB)

    Danczak, Robert E.; Yabusaki, Steven B.; Williams, Kenneth H.; Fang, Yilin; Hobson, Chad; Wilkins, Michael J.

    2016-05-11

    Shallow riparian aquifers represent hotspots of biogeochemical activity in the arid western US. While these environments provide extensive ecosystem services, little is known of how natural environmental perturbations influence subsurface microbial communities and associated biogeochemical processes. Over a six-month period we tracked the annual snowmelt-driven incursion of groundwater into the vadose zone of an aquifer adjacent to the Colorado River, leading to increased dissolved oxygen (DO) concentrations in the normally suboxic saturated zone. Strong biogeochemical heterogeneity was measured across the site, with abiotic reactions between DO and sulfide minerals driving rapid DO consumption and mobilization of redox active species in reduced aquifer regions. Conversely, extensive DO increases were detected in less reduced sediments. 16S rRNA gene surveys tracked microbial community composition within the aquifer, revealing strong correlations between increases in putative oxygen-utilizing chemolithoautotrophs and heterotrophs and rising DO concentrations. The gradual return to suboxic aquifer conditions favored increasing abundances of 16S rRNA sequences matching members of the Microgenomates (OP11) and Parcubacteria (OD1) that have been strongly implicated in fermentative processes. Microbial community stability measurements indicated that deeper aquifer locations were relatively less affected by geochemical perturbations, while communities in shallower locations exhibited the greatest change. Reactive transport modeling of the geochemical and microbiological results supported field observations, suggesting that a predictive framework can be applied to develop a greater understanding of such environments. Frontiers in Earth Science Journal Impact & Description - ResearchGate - Impact Rankings ( 2015 and 2016 ). Available from: https://www.researchgate.net/journal/2296-6463_Frontiers_in_Earth_Science [accessed Jul 25, 2016].

  14. Addition of citrate to Acidithiobacillus ferrooxidans cultures enables precipitate-free growth at elevated pH and reduces ferric inhibition.

    Science.gov (United States)

    Li, Xiaozheng; Mercado, Roel; Kernan, Timothy; West, Alan C; Banta, Scott

    2014-10-01

    Acidithiobacillus ferrooxidans is an acidophilic chemolithoautotroph that is important in biomining and other biotechnological operations. The cells are able to oxidize inorganic iron, but the insolubility and product inhibition by Fe(3+) complicates characterization of these cultures. Here we explore the growth kinetics of A. ferrooxidans in iron-based medium in a pH range from 1.6 to 2.2. It was found that as the pH was increased from 1.6 to 2.0, the maintenance coefficient decreased while both the growth kinetics and maximum cell yield increased in the precipitate-free, low Fe(2+) concentration medium. In higher iron media a similar trend was observed at low pH, but the formation of precipitates at higher pH (2.0) hampered cell growth and lowered the specific growth rate and maximum cell yield. In order to eliminate ferric precipitates, chelating agents were introduced into the medium. Citric acid was found to be relatively non-toxic and did not appear to interfere with iron oxidation at a maximum concentration of 70 mM. Inclusion of citric acid prevented precipitation and A. ferrooxidans growth parameters resumed their trends as a function of pH. The addition of citrate also decreased the apparent substrate saturation constant (KS ) indicating a reduction in the competitive inhibition of growth by ferric ions. These results indicate that continuous cultures of A. ferrooxidans in the presence of citrate at elevated pH will enable enhanced cell yields and productivities. This will be critical as these cells are used in the development of new biotechnological applications such as electrofuel production.

  15. The small heat shock proteins from Acidithiobacillus ferrooxidans: gene expression, phylogenetic analysis, and structural modeling

    Directory of Open Access Journals (Sweden)

    Ribeiro Daniela A

    2011-12-01

    Full Text Available Abstract Background Acidithiobacillus ferrooxidans is an acidophilic, chemolithoautotrophic bacterium that has been successfully used in metal bioleaching. In this study, an analysis of the A. ferrooxidans ATCC 23270 genome revealed the presence of three sHSP genes, Afe_1009, Afe_1437 and Afe_2172, that encode proteins from the HSP20 family, a class of intracellular multimers that is especially important in extremophile microorganisms. Results The expression of the sHSP genes was investigated in A. ferrooxidans cells submitted to a heat shock at 40°C for 15, 30 and 60 minutes. After 60 minutes, the gene on locus Afe_1437 was about 20-fold more highly expressed than the gene on locus Afe_2172. Bioinformatic and phylogenetic analyses showed that the sHSPs from A. ferrooxidans are possible non-paralogous proteins, and are regulated by the σ32 factor, a common transcription factor of heat shock proteins. Structural studies using homology molecular modeling indicated that the proteins encoded by Afe_1009 and Afe_1437 have a conserved α-crystallin domain and share similar structural features with the sHSP from Methanococcus jannaschii, suggesting that their biological assembly involves 24 molecules and resembles a hollow spherical shell. Conclusion We conclude that the sHSPs encoded by the Afe_1437 and Afe_1009 genes are more likely to act as molecular chaperones in the A. ferrooxidans heat shock response. In addition, the three sHSPs from A. ferrooxidans are not recent paralogs, and the Afe_1437 and Afe_1009 genes could be inherited horizontally by A. ferrooxidans.

  16. Sulfur metabolism in the extreme acidophile Acidithiobacillus caldus

    Directory of Open Access Journals (Sweden)

    Stefanie eMangold

    2011-02-01

    Full Text Available Given the challenges to life at low pH, an analysis of inorganic sulfur compound oxidation was initiated in the chemolithoautotrophic extremophile Acidithiobacillus caldus. A. caldus is able to metabolize elemental sulfur and a broad range of inorganic sulfur compounds. It has been implicated in the production of environmentally damaging acidic solutions as well as participating in industrial bioleaching operations where it forms part of microbial consortia used for the recovery of metal ions. Based upon the recently published A. caldus type strain genome sequence, a bioinformatic reconstruction of elemental sulfur and inorganic sulfur compound metabolism predicted genes included: sulfide quinone reductase (sqr, tetrathionate hydrolase (tth, two sox gene clusters potentially involved in thiosulfate oxidation (soxABXYZ, sulfur oxygenase reductase (sor, and various electron transport components. RNA transcript profiles by semi-quantitative reverse transcription PCR suggested up-regulation of sox genes in the presence of tetrathionate. Extensive gel based proteomic comparisons of total soluble and membrane enriched protein fractions during growth on elemental sulfur and tetrathionate identified differential protein levels from the two Sox clusters as well as several chaperone and stress proteins up-regulated in the presence of elemental sulfur. Proteomics results also suggested the involvement of heterodisulfide reductase (HdrABC in A. caldus inorganic sulfur compound metabolism. A putative new function of Hdr in acidophiles is discussed. Additional proteomic analysis evaluated protein expression differences between cells grown attached to solid, elemental sulfur versus planktonic cells. This study has provided insights into sulfur metabolism of this acidophilic chemolithotroph and gene expression during attachment to solid elemental sulfur.

  17. Palagonitization of Basalt Glass in the Flanks of Mid-Ocean Ridges: Implications for the Bioenergetics of Oceanic Intracrustal Ecosystems.

    Science.gov (United States)

    Türke, Andreas; Nakamura, Kentaro; Bach, Wolfgang

    2015-10-01

    When basalt is exposed to oxygenated aqueous solutions, rims of palagonite form along fractures at the expense of glass. We employed electron microprobe and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) analyses of fresh glass and adjacent palagonite crusts to determine the geochemical changes involved in palagonite formation. Samples were retrieved from drill cores taken in the North Pond Area, located on the western flank of the Mid-Atlantic Ridge at 22°45'N and 46°05'W. We also analyzed whole rock powders to determine the overall crust-seawater exchange in a young ridge flank. Radioactive elements are enriched in palagonite relative to fresh glass, reaching concentrations where radiolytic production of molecular hydrogen (H2) may be a significant energy source. Based on these results, we hypothesize that microbial ecosystems in ridge flank habitats undergo a transition in the principal energy carrier, fueling carbon fixation from Fe oxidation in very young crust to H2 consumption in older crust. Unless the H2 is swept away by rapid fluid flow (i.e., in young flanks), it may easily accumulate to levels high enough to support chemolithoautotrophic life. In older flanks, crustal sealing and sediment accumulation have slowed down seawater circulation, and the significance of radiolytically produced H2 for catalytic energy supply is expected to increase greatly. Similar habitats on other planetary surfaces are theoretically possible, as accumulation of radiolytically produced hydrogen merely requires the presence of H2O molecules and a porous medium, from which the hydrogen is not lost.

  18. Diffuse flow environments within basalt- and sediment-based hydrothermal vent ecosystems harbor specialized microbial communities.

    Science.gov (United States)

    Campbell, Barbara J; Polson, Shawn W; Zeigler Allen, Lisa; Williamson, Shannon J; Lee, Charles K; Wommack, K Eric; Cary, S Craig

    2013-01-01

    Hydrothermal vents differ both in surface input and subsurface geochemistry. The effects of these differences on their microbial communities are not clear. Here, we investigated both alpha and beta diversity of diffuse flow-associated microbial communities emanating from vents at a basalt-based hydrothermal system along the East Pacific Rise (EPR) and a sediment-based hydrothermal system, Guaymas Basin. Both Bacteria and Archaea were targeted using high throughput 16S rRNA gene pyrosequencing analyses. A unique aspect of this study was the use of a universal set of 16S rRNA gene primers to characterize total and diffuse flow-specific microbial communities from varied deep-sea hydrothermal environments. Both surrounding seawater and diffuse flow water samples contained large numbers of Marine Group I (MGI) Thaumarchaea and Gammaproteobacteria taxa previously observed in deep-sea systems. However, these taxa were geographically distinct and segregated according to type of spreading center. Diffuse flow microbial community profiles were highly differentiated. In particular, EPR dominant diffuse flow taxa were most closely associated with chemolithoautotrophs, and off axis water was dominated by heterotrophic-related taxa, whereas the opposite was true for Guaymas Basin. The diversity and richness of diffuse flow-specific microbial communities were strongly correlated to the relative abundance of Epsilonproteobacteria, proximity to macrofauna, and hydrothermal system type. Archaeal diversity was higher than or equivalent to bacterial diversity in about one third of the samples. Most diffuse flow-specific communities were dominated by OTUs associated with Epsilonproteobacteria, but many of the Guaymas Basin diffuse flow samples were dominated by either OTUs within the Planctomycetes or hyperthermophilic Archaea. This study emphasizes the unique microbial communities associated with geochemically and geographically distinct hydrothermal diffuse flow environments.

  19. Metagenomic analysis of microbial consortium from natural crude oil that seeps into the marine ecosystem offshore Southern California.

    Science.gov (United States)

    Hawley, Erik R; Piao, Hailan; Scott, Nicole M; Malfatti, Stephanie; Pagani, Ioanna; Huntemann, Marcel; Chen, Amy; Glavina Del Rio, Tijana; Foster, Brian; Copeland, Alex; Jansson, Janet; Pati, Amrita; Tringe, Susannah; Gilbert, Jack A; Lorenson, Thomas D; Hess, Matthias

    2014-06-15

    Crude oils can be major contaminants of the marine ecosystem and microorganisms play a significant role in the degradation of its main constituents. To increase our understanding of the microbial hydrocarbon degradation process in the marine ecosystem, we collected crude oil from an active seep area located in the Santa Barbara Channel (SBC) and generated a total of about 52 Gb of raw metagenomic sequence data. The assembled data comprised ~500 Mb, representing ~1.1 million genes derived primarily from chemolithoautotrophic bacteria. Members of Oceanospirillales, a bacterial order belonging to the Deltaproteobacteria, recruited less than 2% of the assembled genes within the SBC metagenome. In contrast, the microbial community associated with the oil plume that developed in the aftermath of the Deepwater Horizon (DWH) blowout in 2010, was dominated by Oceanospirillales, which comprised more than 60% of the metagenomic data generated from the DWH oil plume. This suggests that Oceanospirillales might play a less significant role in the microbially mediated hydrocarbon conversion within the SBC seep oil compared to the DWH plume oil. We hypothesize that this difference results from the SBC oil seep being mostly anaerobic, while the DWH oil plume is aerobic. Within the Archaea, the phylum Euryarchaeota, recruited more than 95% of the assembled archaeal sequences from the SBC oil seep metagenome, with more than 50% of the sequences assigned to members of the orders Methanomicrobiales and Methanosarcinales. These orders contain organisms capable of anaerobic methanogenesis and methane oxidation (AOM) and we hypothesize that these orders - and their metabolic capabilities - may be fundamental to the ecology of the SBC oil seep.

  20. Microbiological oxidation of antimony(III) with oxygen or nitrate by bacteria isolated from contaminated mine sediments

    Science.gov (United States)

    Terry, Lee R.; Kulp, Thomas R.; Wiatrowski, Heather A.; Miller, Laurence G.; Oremland, Ronald S.

    2015-01-01

    Bacterial oxidation of arsenite [As(III)] is a well-studied and important biogeochemical pathway that directly influences the mobility and toxicity of arsenic in the environment. In contrast, little is known about microbiological oxidation of the chemically similar anion antimonite [Sb(III)]. In this study, two bacterial strains, designated IDSBO-1 and IDSBO-4, which grow on tartrate compounds and oxidize Sb(III) using either oxygen or nitrate, respectively, as a terminal electron acceptor, were isolated from contaminated mine sediments. Both isolates belonged to the Comamonadaceae family and were 99% similar to previously described species. We identify these novel strains as Hydrogenophagataeniospiralis strain IDSBO-1 and Variovorax paradoxus strain IDSBO-4. Both strains possess a gene with homology to the aioA gene, which encodes an As(III)-oxidase, and both oxidize As(III) aerobically, but only IDSBO-4 oxidized Sb(III) in the presence of air, while strain IDSBO-1 could achieve this via nitrate respiration. Our results suggest that expression of aioA is not induced by Sb(III) but may be involved in Sb(III) oxidation along with an Sb(III)-specific pathway. Phylogenetic analysis of proteins encoded by the aioA genes revealed a close sequence similarity (90%) among the two isolates and other known As(III)-oxidizing bacteria, particularly Acidovorax sp. strain NO1. Both isolates were capable of chemolithoautotrophic growth using As(III) as a primary electron donor, and strain IDSBO-4 exhibited incorporation of radiolabeled [14C]bicarbonate while oxidizing Sb(III) from Sb(III)-tartrate, suggesting possible Sb(III)-dependent autotrophy. Enrichment cultures produced the Sb(V) oxide mineral mopungite and lesser amounts of Sb(III)-bearing senarmontite as precipitates.

  1. Microbiological oxidation of antimony(III) with oxygen or nitrate by bacteria isolated from contaminated mine sediments.

    Science.gov (United States)

    Terry, Lee R; Kulp, Thomas R; Wiatrowski, Heather; Miller, Laurence G; Oremland, Ronald S

    2015-12-01

    Bacterial oxidation of arsenite [As(III)] is a well-studied and important biogeochemical pathway that directly influences the mobility and toxicity of arsenic in the environment. In contrast, little is known about microbiological oxidation of the chemically similar anion antimonite [Sb(III)]. In this study, two bacterial strains, designated IDSBO-1 and IDSBO-4, which grow on tartrate compounds and oxidize Sb(III) using either oxygen or nitrate, respectively, as a terminal electron acceptor, were isolated from contaminated mine sediments. Both isolates belonged to the Comamonadaceae family and were 99% similar to previously described species. We identify these novel strains as Hydrogenophaga taeniospiralis strain IDSBO-1 and Variovorax paradoxus strain IDSBO-4. Both strains possess a gene with homology to the aioA gene, which encodes an As(III)-oxidase, and both oxidize As(III) aerobically, but only IDSBO-4 oxidized Sb(III) in the presence of air, while strain IDSBO-1 could achieve this via nitrate respiration. Our results suggest that expression of aioA is not induced by Sb(III) but may be involved in Sb(III) oxidation along with an Sb(III)-specific pathway. Phylogenetic analysis of proteins encoded by the aioA genes revealed a close sequence similarity (90%) among the two isolates and other known As(III)-oxidizing bacteria, particularly Acidovorax sp. strain NO1. Both isolates were capable of chemolithoautotrophic growth using As(III) as a primary electron donor, and strain IDSBO-4 exhibited incorporation of radiolabeled [(14)C]bicarbonate while oxidizing Sb(III) from Sb(III)-tartrate, suggesting possible Sb(III)-dependent autotrophy. Enrichment cultures produced the Sb(V) oxide mineral mopungite and lesser amounts of Sb(III)-bearing senarmontite as precipitates.

  2. Diversity and phylogenetic analyses of bacteria from a shallow-water hydrothermal vent in Milos island (Greece).

    Science.gov (United States)

    Giovannelli, Donato; d'Errico, Giuseppe; Manini, Elena; Yakimov, Michail; Vetriani, Costantino

    2013-01-01

    Studies of shallow-water hydrothermal vents have been lagging behind their deep-sea counterparts. Hence, the importance of these systems and their contribution to the local and regional diversity and biogeochemistry is unclear. This study analyzes the bacterial community along a transect at the shallow-water hydrothermal vent system of Milos island, Greece. The abundance and biomass of the prokaryotic community is comparable to areas not affected by hydrothermal activity and was, on average, 1.34 × 10(8) cells g(-1). The abundance, biomass and diversity of the prokaryotic community increased with the distance from the center of the vent and appeared to be controlled by the temperature gradient rather than the trophic conditions. The retrieved 16S rRNA gene fragments matched sequences from a variety of geothermal environments, although the average similarity was low (94%), revealing previously undiscovered taxa. Epsilonproteobacteria constituted the majority of the population along the transect, with an average contribution to the total diversity of 60%. The larger cluster of 16S rRNA gene sequences was related to chemolithoautotrophic Sulfurovum spp., an Epsilonproteobacterium so far detected only at deep-sea hydrothermal vents. The presence of previously unknown lineages of Epsilonproteobacteria could be related to the abundance of organic matter in these systems, which may support alternative metabolic strategies to chemolithoautotrophy. The relative contribution of Gammaproteobacteria to the Milos microbial community increased along the transect as the distance from the center of the vent increased. Further attempts to isolate key species from these ecosystems will be critical to shed light on their evolution and ecology.

  3. Diversity and phylogenetic analyses of bacteria from a shallow-waterhydrothermal vent in Milos island (Greece

    Directory of Open Access Journals (Sweden)

    Donato eGiovannelli

    2013-07-01

    Full Text Available Studies of shallow-water hydrothermal vents have been lagging behind their deep-sea counterparts. Hence, the importance of these systems and their contribution to the local and regional diversity and biogeochemistry is unclear. This study analyzes the bacterial community along a transect at the shallow-water hydrothermal vent system of Milos island, Greece. The abundance and biomass of the prokaryotic community is comparable to areas not affected by hydrothermal activity and was, on average, 1.34×108 cells g-1. The abundance, biomass and diversity of the prokaryotic community increased with the distance from the center of the vent and appeared to be controlled by the temperature gradient rather than the trophic conditions. The retrieved 16S rRNA gene fragments matched sequences from a variety of geothermal environments, although the average similarity was low (94 %, revealing previously undiscovered taxa. Epsilonproteobacteria constituted the majority of the population along the transect, with an average contribution to the total diversity of 60%. The larger cluster of 16S rRNA gene sequences was related to chemolithoautotrophic Sulfurovum spp., an Epsilonproteobacterium so far detected only at deep-sea hydrothermal vents. The presence of previously unknown lineages of Epsilonproteobacteria could be related to the abundance of organic matter in these systems, which may support alternative metabolic strategies to chemolithoautotrophy. The relative contribution of Gammaproteobacteria to the Milos microbial community increased along the transect as the distance from the center of the vent increased. Further attempts to isolate key species from these ecosystems will be critical to shed light on their evolution and ecology.

  4. Genomic diversity within the haloalkaliphilic genus Thioalkalivibrio

    Science.gov (United States)

    Ahn, Anne-Catherine; Meier-Kolthoff, Jan P.; Overmars, Lex; Richter, Michael; Woyke, Tanja; Sorokin, Dimitry Y.

    2017-01-01

    Thioalkalivibrio is a genus of obligate chemolithoautotrophic haloalkaliphilic sulfur-oxidizing bacteria. Their habitat are soda lakes which are dual extreme environments with a pH range from 9.5 to 11 and salt concentrations up to saturation. More than 100 strains of this genus have been isolated from various soda lakes all over the world, but only ten species have been effectively described yet. Therefore, the assignment of the remaining strains to either existing or novel species is important and will further elucidate their genomic diversity as well as give a better general understanding of this genus. Recently, the genomes of 76 Thioalkalivibrio strains were sequenced. On these, we applied different methods including (i) 16S rRNA gene sequence analysis, (ii) Multilocus Sequence Analysis (MLSA) based on eight housekeeping genes, (iii) Average Nucleotide Identity based on BLAST (ANIb) and MUMmer (ANIm), (iv) Tetranucleotide frequency correlation coefficients (TETRA), (v) digital DNA:DNA hybridization (dDDH) as well as (vi) nucleotide- and amino acid-based Genome BLAST Distance Phylogeny (GBDP) analyses. We detected a high genomic diversity by revealing 15 new “genomic” species and 16 new “genomic” subspecies in addition to the ten already described species. Phylogenetic and phylogenomic analyses showed that the genus is not monophyletic, because four strains were clearly separated from the other Thioalkalivibrio by type strains from other genera. Therefore, it is recommended to classify the latter group as a novel genus. The biogeographic distribution of Thioalkalivibrio suggested that the different “genomic” species can be classified as candidate disjunct or candidate endemic species. This study is a detailed genome-based classification and identification of members within the genus Thioalkalivibrio. However, future phenotypical and chemotaxonomical studies will be needed for a full species description of this genus. PMID:28282461

  5. Transition of microbiological and sedimentological features associated with the geochemical gradient in a travertine mound in northern Sumatra, Indonesia

    Science.gov (United States)

    Sugihara, Chiya; Yanagawa, Katsunori; Okumura, Tomoyo; Takashima, Chizuru; Harijoko, Agung; Kano, Akihiro

    2016-08-01

    Modern travertines, carbonate deposits in Ca-rich hydrothermal water with high pCO2, often display a changing environment along the water path, with corresponding variability in the microbial communities. We investigated a travertine-bearing hot spring at the Blue Pool in northern Sumatra, Indonesia. The thermal water of 62 °C with high H2S (200 μM) and pCO2 ( 1 atm) developed a travertine mound 70 m wide. The concentrations of the gas components H2S and CO2, decrease immediately after the water is discharged, while the dissolved oxygen, pH, and aragonite saturation increase in the downstream direction. Responding to the geochemical gradient in the water, the surface biofilms change color from white to pink, light-green, dark-green, and brown as the water flows from the vent; this corresponds to microbial communities characterized by chemolithoautotrophs (Halothiobacillaceae), purple sulfur bacteria (Chromatiaceae), Anaerolineaceae, and co-occurrence of green non-sulfur bacteria (Chloroflexales)-Cyanobacteria, and green sulfur bacteria (Chlorobiales), respectively. In an environment with a certain level of H2S (> 1 μM), sulfur digestion and anoxygenic photosynthesis can be more profitable than oxygenic photosynthesis by Cyanobacteria. The precipitated carbonate mineral consists of aragonite and calcite, with the proportion of aragonite increasing downstream due to the larger Mg2 +/Ca2 + ratio in the water or the development of thicker biofilm. Where the biofilm is well developed, the aragonite travertines often exhibit laminated structures that were likely associated with the daily metabolism of these bacteria. The microbiological and sedimentological features at the Blue Pool may be the modern analogs of geomicrobiological products in the early Earth. Biofilm of anoxygenic photosynthetic bacteria had the potential to form ancient stromatolites that existed before the appearance of cyanobacteria.

  6. Metagenomic analysis of microbial consortium from natural crude oil that seeps into the marine ecosystem offshore Southern California

    Energy Technology Data Exchange (ETDEWEB)

    Hawley, Erik R.; Piao, Hailan; Scott, Nicole M.; Malfatti, Stephanie; Pagani, Ioanna; Huntemann, Marcel; Chen, Amy; del Rio, Tijana G.; Foster, Brian; Copeland, A.; Jansson, Janet K.; Pati, Amrita; Gilbert, Jack A.; Tringe, Susannah G.; Lorenson, Thomas D.; Hess, Matthias

    2014-01-02

    Crude oils can be major contaminants of the marine ecosystem and microorganisms play a significant role in the degradation of the main constituents of crude oil. To increase our understanding of the microbial hydrocarbon degradation process in the marine ecosystem, we collected crude oil from an active seep area located in the Santa Barbara Channel (SBC) and generated a total of about 52 Gb of raw metagenomic sequence data. The assembled data comprised ~500 Mb, representing ~1.1 million genes derived primarily from chemolithoautotrophic bacteria. Members of Oceanospirillales, a bacterial order belonging to the Deltaproteobacteria, recruited less than 2% of the assembled genes within the SBC metagenome. In contrast, the microbial community associated with the oil plume that developed in the aftermath of the Deepwater Horizon (DWH) blowout in 2010, was dominated by Oceanospirillales, which comprised more than 60% of the metagenomic data generated from the DWH oil plume. This suggests that Oceanospirillales might play a less significant role in the microbially mediated hydrocarbon conversion within the SBC seep oil compared to the DWH plume oil. We hypothesize that this difference results from the SBC oil seep being mostly anaerobic, while the DWH oil plume is aerobic. Within the Archaea, the phylum Euryarchaeota, recruited more than 95% of the assembled archaeal sequences from the SBC oil seep metagenome, with more than 50% of the sequences assigned to members of the orders Methanomicrobiales and Methanosarcinales. These orders contain organisms capable of anaerobic methanogenesis and methane oxidation (AOM) and we hypothesize that these orders and their metabolic capabilities may be fundamental to the ecology of the SBC oil seep.

  7. Novel sulfur-oxidizing streamers thriving in perennial cold saline springs of the Canadian high Arctic.

    Science.gov (United States)

    Niederberger, Thomas D; Perreault, Nancy N; Lawrence, John R; Nadeau, Jay L; Mielke, Randall E; Greer, Charles W; Andersen, Dale T; Whyte, Lyle G

    2009-03-01

    The perennial springs at Gypsum Hill (GH) and Colour Peak (CP), situated at nearly 80 degrees N on Axel Heiberg Island in the Canadian high Arctic, are one of the few known examples of cold springs in thick permafrost on Earth. The springs emanate from deep saline aquifers and discharge cold anoxic brines rich in both sulfide and sulfate. Grey-coloured microbial streamers form during the winter months in snow-covered regions of the GH spring run-off channels (-1.3 degrees C to 6.9 degrees C, approximately 7.5% NaCl, 0-20 p.p.m. dissolved sulfide, 1 p.p.m. dissolved oxygen) but disappear during the Arctic summer. Culture- and molecular-based analyses of the 16S rRNA gene (FISH, DGGE and clone libraries) indicated that the streamers were uniquely dominated by chemolithoautotrophic sulfur-oxidizing Thiomicrospira species. The streamers oxidized both sulfide and thiosulfate and fixed CO(2) under in situ conditions and a Thiomicrospira strain isolated from the streamers also actively oxidized sulfide and thiosulfate and fixed CO(2) under cold, saline conditions. Overall, the snow-covered spring channels appear to represent a unique polar saline microhabitat that protects and allows Thiomicrospira streamers to form and flourish via chemolithoautrophic, phototrophic-independent metabolism in a high Arctic winter environment characterized by air temperatures commonly below -40 degrees C and with an annual average air temperature of -15 degrees C. These results broaden our knowledge of the physical and chemical boundaries that define life on Earth and have astrobiological implications for the possibility of life existing under similar Martian conditions.

  8. Assessing microbial processes in deep-sea hydrothermal systems by incubation at in situ temperature and pressure

    Science.gov (United States)

    McNichol, Jesse; Sylva, Sean P.; Thomas, François; Taylor, Craig D.; Sievert, Stefan M.; Seewald, Jeffrey S.

    2016-09-01

    At deep-sea hydrothermal vents, a large source of potential chemical energy is created when reducing vent fluid and oxidizing seawater mix. In this environment, chemolithoautotrophic microbes catalyze exergonic redox reactions which in turn provide the energy needed to fuel their growth and the fixation of CO2 into biomass. In addition to producing new organic matter, this process also consumes compounds contained both in vent fluid and entrained seawater (e.g. H2, NO3-). Despite their biogeochemical importance, such reactions have remained difficult to quantify due to methodological limitations. To address this knowledge gap, this study reports a novel application of isobaric gas-tight fluid samplers for conducting incubations of hydrothermal vent fluids at in situ temperature and pressure. Eighteen 24 h incubations were carried out, representing seven distinct conditions that examine amendments consisting of different electron donors and acceptors. Microbial activity was observed in all treatments, and time series chemical measurements showed that activity was limited by electron acceptor supply, confirming predictions based on geochemical data. Also consistent with these predictions, the presence of nitrate increased rates of hydrogen consumption and yielded ammonium as a product of nitrate respiration. The stoichiometry of predicted redox reactions was also determined, revealing that the sulfur and nitrogen cycles are incompletely understood at deep-sea vents, and likely involve unknown intermediate redox species. Finally, the measured rates of redox processes were either equal to or far greater than what has been reported in previous studies where in situ conditions were not maintained. In addition to providing insights into deep-sea hydrothermal vent biogeochemistry, the methods described herein also offer a practical approach for the incubation of any deep-sea pelagic sample under in situ conditions.

  9. Modeling Microbial Biogeochemistry from Terrestrial to Aquatic Ecosystems Using Trait-Based Approaches

    Science.gov (United States)

    King, E.; Molins, S.; Karaoz, U.; Johnson, J. N.; Bouskill, N.; Hug, L. A.; Thomas, B. C.; Castelle, C. J.; Beller, H. R.; Banfield, J. F.; Steefel, C. I.; Brodie, E.

    2014-12-01

    Currently, there is uncertainty in how climate or land-use-induced changes in hydrology and vegetation will affect subsurface carbon flux, the spatial and temporal distribution of flow and transport, biogeochemical cycling, and microbial metabolic activity. Here we focus on the initial development of a Genome-Enabled Watershed Simulation Capability (GEWaSC), which provides a predictive framework for understanding how genomic information stored in a subsurface microbiome affects biogeochemical watershed functioning, how watershed-scale processes affect microbial function, and how these interactions co-evolve. This multiscale framework builds on a hierarchical approach to multiscale modeling, which considers coupling between defined microscale and macroscale components of a system (e.g., a catchment being defined as macroscale and biogeofacies as microscale). Here, we report our progress in the development of a trait-based modeling approach within a reactive transport framework that simulates coupled guilds of microbes. Guild selection is driven by traits extracted from, and physiological properties inferred from, large-scale assembly of metagenome data. Meta-genomic, -transcriptomic and -proteomic information are also used to complement our existing biogeochemical reaction networks and contributes key reactions where biogeochemical analyses are unequivocal. Our approach models the rate of nutrient uptake and the thermodynamics of coupled electron donors and acceptors for a range of microbial metabolisms including heterotrophs and chemolitho(auto)trophs. Metabolism of exogenous substrates fuels catabolic and anabolic processes, with the proportion of energy used for each based upon dynamic intracellular and environmental conditions. In addition to biomass development, anabolism includes the production of key enzymes, such as nitrogenase for nitrogen fixation or exo-enzymes for the hydrolysis of extracellular polymers. This internal resource partitioning represents a

  10. Tumebacillus permanentifrigoris gen. nov., sp. nov., an aerobic, spore-forming bacterium isolated from Canadian high Arctic permafrost.

    Science.gov (United States)

    Steven, Blaire; Chen, Min Qun; Greer, Charles W; Whyte, Lyle G; Niederberger, Thomas D

    2008-06-01

    A Gram-positive, aerobic, rod-shaped bacterium (strain Eur1 9.5(T)) was isolated from a 9-m-deep permafrost sample from the Canadian high Arctic. Strain Eur1 9.5(T) could not be cultivated in liquid medium and grew over the temperature range 5-37 degrees C; no growth was observed at 42 degrees C and only slow growth was observed at 5 degrees C following 1 month of incubation. Eur1 9.5(T) grew over the pH range 5.5-8.9 and tolerated NaCl concentrations of 0-0.5 % (w/v). Eur1 9.5(T) grew heterotrophically on complex carbon substrates and chemolithoautotrophically on inorganic sulfur compounds, as demonstrated by growth on sodium thiosulfate and sulfite as sole electron donors. Eur1 9.5(T) contained iso-C(15 : 0) as the major cellular fatty acid and menaquinone 7 (MK-7) as the major respiratory quinone. The cell-wall peptidoglycan was of type A1gamma. The DNA G+C content was 53.1 mol%. The 16S rRNA gene sequence of strain Eur1 9.5(T) was only distantly related (

  11. Subseafloor fluid mixing and fossilized microbial life in a Cretaceous 'Lost City'-type hydrothermal system at the Iberian Margin

    Science.gov (United States)

    Klein, F.; Humphris, S. E.; Guo, W.; Schubotz, F.; Schwarzenbach, E. M.; Orsi, W.

    2015-12-01

    Subseafloor mixing of reduced hydrothermal fluids with seawater is believed to provide the energy and substrates needed to support autotrophic microorganisms in the hydrated oceanic mantle (serpentinite). Despite the potentially significant implications for the distribution of microbial life on Earth and other water-bearing planetary bodies, our understanding of such environments remains elusive. In the present study we examined fossilized microbial communities and fluid mixing processes in the subseafloor of a Cretaceous 'Lost City'-type hydrothermal system at the passive Iberia Margin (ODP Leg 149, Hole 897D). Brucite and calcite co-precipitated from mixed fluids ca. 65m below the Cretaceous palaeo-seafloor at temperatures of 32±4°C within steep chemical gradients (fO2, pH, CH4, SO4, ΣCO2, etc) between weathered, carbonate-rich serpentinite breccia and serpentinite. Mixing of oxidized seawater and strongly reducing hydrothermal fluid at moderate temperatures created conditions capable of supporting microbial activity within the oceanic basement. Dense microbial colonies are fossilized in brucite-calcite veins that are strongly enriched in organic carbon but depleted in 13C. We detected a combination of bacterial diether lipid biomarkers, archaeol and archaeal tetraethers analogous to those found in brucite-carbonate chimneys at the active Lost City hydrothermal field. The exposure of mantle rocks to seawater during the breakup of Pangaea fueled chemolithoautotrophic microbial communities at the Iberia Margin during the Cretaceous, possibly before the onset of seafloor spreading in the Atlantic. 'Lost City'-type serpentinization systems have been discovered at mid-ocean ridges, in forearc settings of subduction zones and at continental margins. It appears that, wherever they occur, they can support microbial life, even in deep subseafloor environments as demonstrated in the present study. Because equivalent systems have likely existed throughout most of Earth

  12. Metagenomic Reconstruction of a Microbial Community from a CO2-driven Geyser

    Science.gov (United States)

    Emerson, J. B.; Banfield, J. F.; Thomas, B. C.

    2012-12-01

    Given that only ~1% of microorganisms are cultivable, and because microbes naturally exist in interactive consortia, it is important to include culture-independent analyses of microbial communities as we evaluate the role that microbes may play in geologic carbon sequestration. Through metagenomics, we report near-complete genomes from a microbial community dominated by iron-oxidizing Zetaproteobacteria from a CO2-driven geyser. Our study site, Crystal Geyser (Green River, Utah), is a cold (17 °C), iron-rich geyser that erupts due to pressure from soluble and free-phase CO2 accumulation in an aquifer ~500 m below the surface, and it is an established natural analog for geologic carbon sequestration. We collected 65 L of geyser water as it precipitated during an eruption in November 2009, and we sequentially filtered the water through 3.0 and 0.2 μm. DNA was extracted from the 0.2 μm filter, from which we generated 13 million 150 bp paired-end Illumina sequencing reads. We assembled near-complete genomes of neutrophilic, iron-oxidizing Mariprofundus ferrooxydans and sulfur-oxidizing Thiomicrospira crunogena. Significant genomic reconstruction was also achieved for other chemolithoautotrophic bacteria, for representatives of Candidate Division TM7 (an as yet uncultivable bacterial phylum), and for a small number of low-abundance archaea. We see evidence for a variety of metabolisms, including iron, sulfur, and complex carbon oxidation, carbon and nitrogen fixation, hydrogen metabolism, aerobic and anaerobic respiration (e.g., of sulfate and nitrate), and methanogenesis. Although the geyser community contains only ~20 populations at 0.5% or higher abundance, our results demonstrate that a CO2-saturated solution can be conducive to a thriving microbial community of diverse phylogeny and broad metabolic potential.

  13. Microbial Community Structure of Subglacial Lake Whillans, West Antarctica

    Directory of Open Access Journals (Sweden)

    Amanda M Achberger

    2016-09-01

    Full Text Available Subglacial Lake Whillans (SLW, located beneath ~800 m of ice on the Whillans Ice Stream in West Antarctica was sampled in January of 2013, providing the first opportunity to directly examine water and sediments from an Antarctic subglacial lake. To minimize the introduction of surface contaminants to SLW during its exploration, an access borehole was created using a microbiologically clean hot water drill designed to reduce the number and viability of microorganisms in the drilling water. Analysis of 16S rRNA genes (rDNA amplified from samples of the drilling and borehole water allowed an evaluation of the efficacy of this approach and enabled a confident assessment of the SLW ecosystem inhabitants. Based on an analysis of 16S rDNA and rRNA (i.e., reverse-transcribed rRNA molecules data, the SLW community was found to be bacterially dominated and compositionally distinct from the assemblages identified in the drill system. The abundance of bacteria (e.g., Candidatus Nitrotoga, Sideroxydans, Thiobacillus, and Albidiferax and archaea (Candidatus Nitrosoarcheaum related to chemolithoautotrophs was consistent with the oxidation of reduced iron, sulfur, and nitrogen compounds having important roles as pathways for primary production in this permanently dark ecosystem. Further, the prevalence of Methylobacter in surficial lake sediments combined with the detection of methanogenic taxa in the deepest sediment horizons analyzed (34-36 cm provided evidence for methane cycling beneath the West Antarctic Ice Sheet. Large ratios of rRNA to rDNA were observed for several OTUs abundant in the water column and sediments (e.g., Albidiferax, Methylobacter, Candidatus Nitrotoga, Sideroxydans, and Smithella, suggesting a potentially active role for these taxa in the SLW ecosystem. Our findings are consistent with chemosynthetic microorganisms serving as the ecological foundation in this dark subsurface environment, providing new organic matter that sustains a

  14. Enhanced metabolic versatility of planktonic sulfur-oxidizing γ-proteobacteria in an oxygen-deficient coastal ecosystem

    Directory of Open Access Journals (Sweden)

    Alejandro A. Murillo

    2014-07-01

    Full Text Available Sulfur-oxidizing Gamma-proteobacteria are abundant in marine oxygen-deficient waters, and appear to play a key role in a previously unrecognized cryptic sulfur cycle. Metagenomic analyses of members of the uncultured SUP05 lineage in the Canadian seasonally anoxic fjord Saanich Inlet (SI, hydrothermal plumes in the Guaymas Basin (GB and single cell genomics analysis of two ARCTIC96BD-19 representatives from the South Atlantic Sub-Tropical Gyre (SASG have shown them to be metabolically versatile. However, SI and GB SUP05 bacteria seem to be obligate chemolithoautotrophs, whereas ARCTIC96BD-19 has the genetic potential for aerobic respiration. Here, we present results of a metagenomic analysis of sulfur-oxidizing Gamma-proteobacteria (GSO, closely related to the SUP05/ARCTIC96BD-19 clade, from a coastal ecosystem in the eastern South Pacific (ESP. This ecosystem experiences seasonal anoxia and accumulation of nitrite and ammonium at depth, with a corresponding increase in the abundance of GSO representatives. The ESP-GSOs appear to have a significantly different gene complement than those from Saanich Inlet, Guaymas Basin and SASG. Genomic analyses of de novo assembled contigs indicate the presence of a complete aerobic respiratory complex based on the cytochrome bc1 oxidase. Furthermore, they appear to encode a complete TCA cycle and several transporters for dissolved organic carbon species, suggesting a mixotrophic lifestyle. Thus, the success of sulfur-oxidizing Gamma-proteobacteria in oxygen-deficient marine ecosystems appears due not only to their previously recognized anaerobic metabolic versatility, but also to their capacity to function under aerobic conditions using different carbon sources. Finally, members of ESP-GSO cluster also have the genetic potential for reducing nitrate to ammonium based on the nirBD genes, and may therefore facilitate a tighter coupling of the nitrogen and sulfur cycles in oxygen-deficient waters.

  15. ENVIROMENTALLY BENIGN MITIGATION OF MICROBIOLOGICALLY INFLUENCED CORROSION (MIC)

    Energy Technology Data Exchange (ETDEWEB)

    J. Robert Paterek; Gemma Husmillo; Amrutha Daram; Vesna Trbovic

    2003-10-31

    The overall program objective is to develop and evaluate environmentally benign agents or products that are effective in the prevention, inhibition, and mitigation of microbially influenced corrosion (MIC) in the internal surfaces of metallic natural gas pipelines. The goal is to develop one or more environmentally benign (a.k.a. ''green'') products that can be applied to maintain the structure and dependability of the natural gas infrastructure. The technical approach for this quarter includes the application of the method of fractionation of the extracts by high performance liquid chromatography (HPLC); determination of antimicrobial activities of the new extracts and fractions using a growth inhibition assay, and evaluation of the extracts' ability to inhibit biofilm formation. We initiated the delivery system for these new biocides in the test cell and in mixtures of foam components and biocides/anti-biofilms. A total of 51 fractions collected by HPLC from crude extracts that were obtained from three varieties of Capsicum sp. (Serrano, Habanero, Chile de Arbol) were subjected to growth inhibition tests against two SRB strains, D. vulgaris and D. desulfuricans. Five fractions showed growth inhibition against both strains while seven inhibited D. desulfuricans only. The crude extracts did not show growth inhibition on both strains but were proven to be potent in preventing the formation of biofilm. Growth inhibition tests of the same set of crude extracts against Comamonas denitrificans did not show positive results. The fractions will be subjected to biofilm inhibition and dissociation assay as well. The delivery system to be evaluated first was foam. The ''foam pig'' components of surfactants and water were tested with the biocide addition. The first chemical and physical parameters to be tested were pH and surfactants. Tests using the fractionated pepper extracts are progressing rapidly. Gas chromatographic analysis

  16. Rates of N2 production and diversity and abundance of functional genes associated with denitrification and anaerobic ammonium oxidation in the sediment of the Amundsen Sea Polynya, Antarctica

    Science.gov (United States)

    Choi, Ayeon; Cho, Hyeyoun; Kim, Sung-Han; Thamdrup, Bo; Lee, SangHoon; Hyun, Jung-Ho

    2016-01-01

    A combination of molecular microbiological analyses and metabolic rate measurements was conducted to elucidate the diversity and abundance of denitrifying and anaerobic ammonium oxidation (anammox) bacteria and the nitrogen gas (N2) production rates in sediment underlying the highly productive polynya (Stns. 10 and 17) and the sea-ice zone on the outer shelf (Stn. 83) of the Amundsen Sea, Antarctica. Despite the high water column productivity, the N2 production rates by denitrification (0.04-0.31 nmol N cm-3sed. h-1) and anammox (0.13-0.26 nmol N cm-3 sed. h-1) were lower than those measured in other polar regions. In contrast, gene copy number (106-107 copies cm-3 of nirS and nosZ genes targeting denitirifiers and 105-107 copies cm-3 of 16S rRNA genes related to anammox bacteria) of the two bacterial groups at Stn. 17 was similar compared to those of other organic-rich environments. The majority of the nirS sequences were affiliated with Gammaproteobacteria (54% and 61% of the total nirS gene at Stns. 17 and 83, respectively), which were closely related to Pseudomonas aeruginosa. Most nosZ sequences (92% and 72% of the total nosZ genes at Stns. 17 and 83, respectively) were related to the Alphaproteobacteria, which were closely related to Ruegeria pomeroyi and Roseobacter denitrificans. Most (98%) of the sequences related to anammox bacteria were affiliated with Candidatus Scalindua at Stn. 17. Consequently, despite the low metabolic activity, the abundance and composition of most denitrifying and anammox bacteria detected from the ASP were similar to those reported from a variety of marine environments. Our results further imply that increased labile organic matter production resulting from a shift of the phytoplankton community from Phaeocystis to diatoms in response to rapid melting of sea ice stimulates metabolic activities of the denitrifying and anammox bacteria, thereby enhancing the N removal process in the ASP.

  17. Microbial control of hydrogen sulfide production in a porous medium

    Energy Technology Data Exchange (ETDEWEB)

    McInerney, M.J.; Wofford, N.Q. [Univ. of Oklahoma, Norman, OK (United States); Sublette, K.L. [Univ. of Tulsa, OK (United States)

    1996-12-31

    The ability of a sulfide- and glutaraldehyde-tolerant strain of Thiobacillus denitrificans (strain F) to control sulfide production in an experimental system of cores and formation water from the Redfield, Iowa natural gas storage facility was investigated. A stable, sulfide-producing biofilm was established in two separate core systems, one of which was inoculated with strain F, and the other core system (control) was treated in an identical manner, but was not inoculated with strain F. When formation water with 10 mM acetate and 5mM nitrate was injected into both core systems, the effluent sulfide concentrations in the control core system ranged from 200-460 {mu}M. In the test core system inoculated with strain F, the effluent sulfide concentrations were lower, ranging from 70-110 {mu}M. In order to determine whether strain F could control sulfide production under optimal conditions for sulfate-reducing bacteria, the electron donor was changed to lactate, and inorganic nutrients (nitrogen and phosphate sources) were added to the formation water. When nutrient-supplemented formation water with 3.1 mM lactate and 10 mM nitrate was used, the effluent sulfide concentrations of the control core system initially increased to about 3800 pM, and then decreased to about 1100 {mu}M after 5 wk. However, in the test core system inoculated with strain F, the effluent sulfide concentrations were much lower, 160-330 {mu}M. Nitrate consumption (5 mM) and high concentrations (101-1011 cells/mL) of strain F were detected in the test core system. An accumulation of biomass occurred in the influent lines during 2 mo of continuous operation, but only a small increase in injection pressure was observed. These studies showed that inoculation with strain F was needed for effective control of sulfide production, and that significant plugging or loss of injectivity owing to microbial inoculation did not occur. 7 refs., 3 figs., 1 tab.

  18. Open source approaches to establishing Roseobacter clade bacteria as synthetic biology chassis for biogeoengineering.

    Science.gov (United States)

    Borg, Yanika; Grigonyte, Aurelija Marija; Boeing, Philipp; Wolfenden, Bethan; Smith, Patrick; Beaufoy, William; Rose, Simon; Ratisai, Tonderai; Zaikin, Alexey; Nesbeth, Darren N

    2016-01-01

    Aim. The nascent field of bio-geoengineering stands to benefit from synthetic biologists' efforts to standardise, and in so doing democratise, biomolecular research methods. Roseobacter clade bacteria comprise 15-20% of oceanic bacterio-plankton communities, making them a prime candidate for establishment of synthetic biology chassis for bio-geoengineering activities such as bioremediation of oceanic waste plastic. Developments such as the increasing affordability of DNA synthesis and laboratory automation continue to foster the establishment of a global 'do-it-yourself' research community alongside the more traditional arenas of academe and industry. As a collaborative group of citizen, student and professional scientists we sought to test the following hypotheses: (i) that an incubator capable of cultivating bacterial cells can be constructed entirely from non-laboratory items, (ii) that marine bacteria from the Roseobacter clade can be established as a genetically tractable synthetic biology chassis using plasmids conforming to the BioBrick(TM) standard and finally, (iii) that identifying and subcloning genes from a Roseobacter clade species can readily by achieved by citizen scientists using open source cloning and bioinformatic tools. Method. We cultivated three Roseobacter species, Roseobacter denitrificans, Oceanobulbus indolifexand Dinoroseobacter shibae. For each species we measured chloramphenicol sensitivity, viability over 11 weeks of glycerol-based cryopreservation and tested the effectiveness of a series of electroporation and heat shock protocols for transformation using a variety of plasmid types. We also attempted construction of an incubator-shaker device using only publicly available components. Finally, a subgroup comprising citizen scientists designed and attempted a procedure for isolating the cold resistance anf1 gene from Oceanobulbus indolifexcells and subcloning it into a BioBrick(TM) formatted plasmid. Results. All species were stable

  19. Open source approaches to establishing Roseobacter clade bacteria as synthetic biology chassis for biogeoengineering

    Directory of Open Access Journals (Sweden)

    Yanika Borg

    2016-07-01

    Full Text Available Aim. The nascent field of bio-geoengineering stands to benefit from synthetic biologists’ efforts to standardise, and in so doing democratise, biomolecular research methods. Roseobacter clade bacteria comprise 15–20% of oceanic bacterio-plankton communities, making them a prime candidate for establishment of synthetic biology chassis for bio-geoengineering activities such as bioremediation of oceanic waste plastic. Developments such as the increasing affordability of DNA synthesis and laboratory automation continue to foster the establishment of a global ‘do-it-yourself’ research community alongside the more traditional arenas of academe and industry. As a collaborative group of citizen, student and professional scientists we sought to test the following hypotheses: (i that an incubator capable of cultivating bacterial cells can be constructed entirely from non-laboratory items, (ii that marine bacteria from the Roseobacter clade can be established as a genetically tractable synthetic biology chassis using plasmids conforming to the BioBrickTM standard and finally, (iii that identifying and subcloning genes from a Roseobacter clade species can readily by achieved by citizen scientists using open source cloning and bioinformatic tools. Method. We cultivated three Roseobacter species, Roseobacter denitrificans, Oceanobulbus indolifexand Dinoroseobacter shibae. For each species we measured chloramphenicol sensitivity, viability over 11 weeks of glycerol-based cryopreservation and tested the effectiveness of a series of electroporation and heat shock protocols for transformation using a variety of plasmid types. We also attempted construction of an incubator-shaker device using only publicly available components. Finally, a subgroup comprising citizen scientists designed and attempted a procedure for isolating the cold resistance anf1 gene from Oceanobulbus indolifexcells and subcloning it into a BioBrickTM formatted plasmid. Results. All

  20. Far from superficial: microbial diversity associated with the skin and mucus of fish

    Science.gov (United States)

    Cipriano, Rocco C.; Dove, Alistair; Cipriano, R.C.; Bruckner, A.W.; Shchelkunov, I.S.

    2011-01-01

    During horizontal or water-borne infection involving an obligate pathogen (e.g. – Aeromonas salmonicida, cause of furunculosis), the pathogen interacted with and influenced the microbial diversity of the dermal mucus of fish. Prior to infection, the prevalent bacterial flora cultured from juvenile Atlantic salmon (Salmo salar) included Pseudomonas fluorescens, Comomonas terrigenia, Acinetobacter sp., Moraxella sp., Pseudomonas dimunita, Alcaligenes denitrificans, Pseudomonas pseudoalcaligenes, and Pseudomonas alcaligenes, Serratia liquefaciens, Aeromonas hydrophila, other motile Aeromonas spp., and Corynebacterium aquaticum. After A. salmonicida was initially detected in this population as an external mucus infection, Acinetobacter sp., Moraxella sp., C. terrigenia, P. fluorescens, and P. dimunita, Staphylococcus sp., and A. hydrophila, were also present in appreciable numbers. Within several weeks, however, the A. salmonicida infection amplified and composed 78% of the total flora in the mucus. Only P. dimunita (4%). P. fluorescens (2%), and C. terrigenia (1%) were cultured at that time and more than a third of these fish showed evidence of a systemic A. salmonicida infection within their kidneys. Eight weeks after oral oxytetracycline treatments, A. salmonicida was no longer isolated from the mucus or kidneys of any fish and glucose inert or other oxidative microbes (e.g., P. fluorescens, C. terrigenia, Acinetobacter sp., Moraxella sp.) were beginning to repopulate the external surface of the salmon in increasing frequency. Still present and composing fairly large percentages of the total flora were A. hydrophila, as well as Enterobacter sp., and P. putrefaciens. A normal microbial diversity was re-established as the fish recovered. In another investigation, reduced biological diversity was noted in the dermal mucus among smallmouth bass that were sampled from the Jackson River (Covington, VA). In these fish, A. hydrophila and P. putrefaciens were the two

  1. Isolation and Identification of Hydrogen-oxidizing Bacteria in Medicago sativa Rhizosphere%紫花苜蓿根际氢氧化细菌的分离与鉴定

    Institute of Scientific and Technical Information of China (English)

    付博; 王卫卫; 郝莹; 王莉娟; 唐明; 陈兴都

    2009-01-01

    利用气体循环培养体系从陕西乾县HUP-豆科植物紫花苜蓿(Medicago sativa)根际土壤中分离获得37株细菌.菌株氧化氢能力测定结果表明,8株菌氧化氢和自养生长能力较强,初步确定为氢氧化细菌类群;根据其形态特征、培养特征和生理生化特性,鉴定为7个不同属:假单胞菌属(Pseudomonas)、邻单胞菌属(Plesiomonas)、脂肪杆菌属(Pimelobacter)、黄色杆菌属(Xanthobacter)、勒米诺氏菌属(Leminorella)、地杆菌属(Terrabacter)和稀有杆菌属(Rarobacter);其中氧化氢能力最强的优势菌株WMQ-7 16S rDNA序列(GenBank登录号为EU807744)长度为1 451bp,GC含量为53.8%,其核苷酸序列与假单胞菌属同源性高于99%,在系统发育树上位于同一分支,将WMQ-7菌株鉴定为假单胞菌属(Pseudomonas).图2表5参16%Gas-cycle incubation system was used to enrich the Medicago sativa rhizosphere soil collected in Shaanxi Province, China and 37 strains of bacteria were isolated. Among them, 8 stains had stronger ability to oxidize hydrogen and could grow chemolithoautotrophically, so they were preliminarily identified as hydrogen-oxidizing bacteria. According to their cells, colony forms and biochemical characteristics, the 8 strains were identified as Pseudomonas, Plesiomonas, Pimelobacter, Xanthobacter, Leminorella, Terrabacter and Rarobacte. Strain WMQ-7 had the strongest ability to oxidize hydrogen, and its 16S rDNA sequence (GenBank accession number EU807744) was 1 451 bp and GC content was 53.8%. Strain WMQ-7 was clustered together with Pseuclomonas in phylogenic tree, with the sequence identity of 99%, Therefore, strain WMQ-7 was identified as Pseudomonas. Fig 2, Tab 5, Ref 16

  2. Ubiquity and diversity of heterotrophic bacterial nasA genes in diverse marine environments.

    Science.gov (United States)

    Jiang, Xuexia; Dang, Hongyue; Jiao, Nianzhi

    2015-01-01

    Nitrate uptake by heterotrophic bacteria plays an important role in marine N cycling. However, few studies have investigated the diversity of environmental nitrate assimilating bacteria (NAB). In this study, the diversity and biogeographical distribution of NAB in several global oceans and particularly in the western Pacific marginal seas were investigated using both cultivation and culture-independent molecular approaches. Phylogenetic analyses based on 16S rRNA and nasA (encoding the large subunit of the assimilatory nitrate reductase) gene sequences indicated that the cultivable NAB in South China Sea belonged to the α-Proteobacteria, γ-Proteobacteria and CFB (Cytophaga-Flavobacteria-Bacteroides) bacterial groups. In all the environmental samples of the present study, α-Proteobacteria, γ-Proteobacteria and Bacteroidetes were found to be the dominant nasA-harboring bacteria. Almost all of the α-Proteobacteria OTUs were classified into three Roseobacter-like groups (I to III). Clone library analysis revealed previously underestimated nasA diversity; e.g. the nasA gene sequences affiliated with β-Proteobacteria, ε-Proteobacteria and Lentisphaerae were observed in the field investigation for the first time, to the best of our knowledge. The geographical and vertical distributions of seawater nasA-harboring bacteria indicated that NAB were highly diverse and ubiquitously distributed in the studied marginal seas and world oceans. Niche adaptation and separation and/or limited dispersal might mediate the NAB composition and community structure in different water bodies. In the shallow-water Kueishantao hydrothermal vent environment, chemolithoautotrophic sulfur-oxidizing bacteria were the primary NAB, indicating a unique nitrate-assimilating community in this extreme environment. In the coastal water of the East China Sea, the relative abundance of Alteromonas and Roseobacter-like nasA gene sequences responded closely to algal blooms, indicating that NAB may be

  3. Genome implosion elicits host-confinement in Alcaligenaceae: evidence from the comparative genomics of Tetrathiobacter kashmirensis, a pathogen in the making.

    Directory of Open Access Journals (Sweden)

    Wriddhiman Ghosh

    Full Text Available This study elucidates the genomic basis of the evolution of pathogens alongside free-living organisms within the family Alcaligenaceae of Betaproteobacteria. Towards that end, the complete genome sequence of the sulfur-chemolithoautotroph Tetrathiobacter kashmirensis WT001(T was determined and compared with the soil isolate Achromobacter xylosoxidans A8 and the two pathogens Bordetella bronchiseptica RB50 and Taylorella equigenitalis MCE9. All analyses comprehensively indicated that the RB50 and MCE9 genomes were almost the subsets of A8 and WT001(T, respectively. In the immediate evolutionary past Achromobacter and Bordetella shared a common ancestor, which was distinct from the other contemporary stock that gave rise to Tetrathiobacter and Taylorella. The Achromobacter-Bordetella precursor, after diverging from the family ancestor, evolved through extensive genome inflation, subsequent to which the two genera separated via differential gene losses and acquisitions. Tetrathiobacter, meanwhile, retained the core characteristics of the family ancestor, and Taylorella underwent massive genome degeneration to reach an evolutionary dead-end. Interestingly, the WT001(T genome, despite its conserved architecture, had only 85% coding density, besides which 578 out of its 4452 protein-coding sequences were found to be pseudogenized. Translational impairment of several DNA repair-recombination genes in the first place seemed to have ushered the rampant and indiscriminate frame-shift mutations across the WT001(T genome. Presumably, this strain has just come out of a recent evolutionary bottleneck, representing a unique transition state where genome self-degeneration has started comprehensively but selective host-confinement has not yet set in. In the light of this evolutionary link, host-adaptation of Taylorella clearly appears to be the aftereffect of genome implosion in another member of the same bottleneck. Remarkably again, potent virulence factors

  4. Where microorganisms meet rocks in the Earth's Critical Zone

    Directory of Open Access Journals (Sweden)

    D. M. Akob

    2011-03-01

    Full Text Available The Earth's Critical Zone (CZ is the critical, outer shell of the Earth that provides an arena for the interplay of diverse physical, chemical, and biological processes that are fundamental for sustaining life. As microbes are the principle drivers of biogeochemical cycles, it is necessary to understand the biodiversity of the CZ unseen majority and their impact on life-sustaining processes. This review aims to summarize the factors controlling where microbes (prokaryotes and micro-eukaryotes live within the CZ and what is known to date about their diversity and function. Microbes live in all regions of the CZ down to 5 km depth, but due to changing habitat complexity, e.g., variability in pore spaces, water, oxygen, and nutrients, their functional role changes with depth. The abundance of prokaryotes and micro-eukaryotes decreases from a maximum of 1010 or 107 cells g soil−1 up to eight orders of magnitude with depth. Symbiotic mycorrhizal fungi and free-living decomposers are best understood in soil habitats, where they are up to 103 cells g soil−1. However, little is known about their identity and impact on weathering in the deep subsurface. The relatively low abundance of micro-eukaryotes in the deep subsurface suggests that these organisms are either limited in space or nutrients or unable to cope with oxygen limitations. Since deep regions of the CZ are limited in the recent input of photosynthesis-derived carbon, microbes are dependent on deposited organic material or on chemolithoautotrophic metabolism that allows for the establishment of a complete food chain independent from the surface. However, the energy flux available might only allow cell growth over tens to thousands of years. The recent development of "omics" technologies has provided microbial ecologists with methods to link the composition and function of in situ microbial communities. We should expect new metabolic

  5. Metagenomic Assembly of the Dominant Zetaproteobacteria in an Iron-oxidizing Hydrothermal Microbial Mat

    Science.gov (United States)

    Moyer, C. L.; Fullerton, H.

    2013-12-01

    Iron is the fourth most abundant element in the Earth's crust and is potentially one of the most abundant energy sources on the earth as an electron donor for chemolithoautotrophic growth coupled to Fe(II) oxidation. Despite the rapid abiotic oxidation rate of iron, many microbes have adapted to feeding off this fleeting energy source. One such bacterial class is the Zetaproteobacteria. Iron-dominated microbial mat material was collected with a small-scale syringe sampler from Loihi Seamount, Hawaii. From this sample, gDNA was extracted and prepared for paired-end Illumina sequencing. Reconstruction of SSU rDNA genes using EMERGE allowed for comparison to previous SSU rDNA surveys. Clone libraries and qPCR show these microbial mats to be dominated by Zetaproteobacteria. Results from our in silico reconstruction confirm these initial findings. RDP classification of the EMERGE reconstructed sequences resulted in 44% of the community being identified as Zetaproteobacteria. The most abundant SSU rDNA has 99% similarity to Zeta OTU-2, and only a 94% similarity to M. ferrooxidans PV-1. Zeta OTU-2 has been shown to be the most cosmopolitan population in iron-dominated hydrothermal systems from across Pacific Ocean. Metagenomic assembly has resulted in many contigs with high identity to M. ferrooxidans as identified, by BLAST. However, with large differences in SSU rRNA similarity, M. ferrooxidans PV-1 is not an adequate reference. Current work is focusing on reconstruction of the dominant microbial mat member, without the use of a reference genome through an iterative assembly approach. The resulting 'pan-genome' will be compared to other Zetaproteobacteria (at the class level) and the functional ecology of this cosmopolitan microbial mat community member will be extrapolated. Thus far, we have detected multiple housekeeping genes involved in DNA replication, transcription and translation. The most abundant metabolic gene we have found is Aconitase, a key enzyme in the

  6. Comets, Carbonaceous Meteorites, and the Origin of the Biosphere

    Science.gov (United States)

    Hoover, Richard B.

    2005-01-01

    The biosphere comprises the Earth s crust, atmosphere, oceans, and ice caps and the living organisms that survive within this habitat. The discoveries of barophilic chemolithoautotrophic thermophiles living deep within the crust and in deep-sea hydrothermal vents, and psychrophiles in permafrost and deep within the Antarctic Ice Sheet indicate the Earth s biosphere is far more extensive than previously recognized. Molecular biomarkers and Bacterial Paleontology provide evidence that life appeared very early on the primitive Earth and the origin of the biosphere is closely linked with the emergence of life. The role of comets, meteorites, and interstellar dust in the delivery of water, organics and prebiotic chemicals has long been recognized. Deuterium enrichment of seawater and comets indicates that comets delivered oceans to the early Earth. Furthermore, the similarity of the D/H ratios and the chemical compositions of CI carbonaceous meteorites and comets indicate that the CI meteorites may be remnants of cometary nuclei with most volatiles removed. Comets, meteorites, and interstellar dust also contain complex organic chemicals, amino acids, macromolecules, and kerogen-like biopolymers and may have played a crucial role in the delivery of complex organics and prebiotic chemicals during the Hadean (4.5-3.8 Gyr) period of heavy bombardment. The existence of indigenous microfossils of morphotypes of cyanobacteria in the CI and CM carbonaceous meteorites suggests that the paradigm that life originated endogenously in the primitive oceans of early Earth may require re-consideration. Recent data on the hot (300-400 K) black crust on comet P/Halley and Stardust images of P/Wild 2 showing depressions, tall cliffs, and pinnacles, indicate the presence of thick, durable, dark crusts on comets. If cavities within the ice and crust sustain vapor pressures in excess of 10 millibar, then localized pools of liquid water and brines could exist within the comet. Since life

  7. Oxygen cycling in the northern Benguela Upwelling System: Modelling oxygen sources and sinks

    Science.gov (United States)

    Schmidt, Martin; Eggert, Anja

    2016-12-01

    This paper elucidates the oxygen dynamics in the northern Benguela Upwelling System by means of process oriented, numerical modelling. Owing to the complex physical-biological interaction in this system, a coupled hydrodynamic-biogeochemical model is required to grasp the various aspects of the oxygen dynamics. We used high-resolution atmospheric fields derived from observations to force our model, available since 1999. The model results represent a 15 years, consistent data set of realistic hydrographic and ecosystem variables, including oxygen distribution patterns. After a concise description of the main aspects of the model, we use the model data to analyse the components contributing to the oxygen dynamics, namely, the ocean circulation, the exchange between ocean and atmosphere as well as the local biogeochemical oxygen cycling in the system. We thoroughly validate the model with available field observations and remote sensing data. The strengths of coastal upwelling, which controls the nutrient supply to the euphotic zone, as well as the poleward undercurrent that carries oxygen and nutrients to the shelf in the northern Benguela Upwelling System are well reproduced in the model. Among the biological oxygen sinks, mineralisation in the sediment, respiration of zooplankton and nitrification in the water column are important. We also found that vertical migration of zooplankton in response to the oxygen conditions provides a regulating feedback, which may prevent a complete deoxygenation of suboxic waters. As long as oxygen or nitrate are available in the bottom waters, the activities of chemolithoautotrophic sulphur bacteria on the sediment surface keep the redoxcline within the sediment and prevent the release of hydrogen sulphide into the water column. By horizontal integration of the simulated ocean-atmosphere oxygen flux, it can be shown that the Kunene upwelling cell between 16 ° S and 18 ° S is a boundary between the equatorial ocean, characterise by

  8. Metabolism of hyperthermophiles.

    Science.gov (United States)

    Schönheit, P; Schäfer, T

    1995-01-01

    Hyperthermophiles are characterized by a temperature optimum for growth between 80 and 110°C. They are considered to represent the most ancient phenotype of living organisms and thus their metabolic design might reflect the situation at an early stage of evolution. Their modes of metabolism are diverse and include chemolithoautotrophic and chemoorganoheterotrophic. No extant phototrophic hyperthermophiles are known. Lithotrophic energy metabolism is mostly anaerobic or microaerophilic and based on the oxidation of H2 or S coupled to the reduction of S, SO inf4 (sup2-) , CO2 and NO inf3 (sup-) but rarely to O2. the substrates are derived from volcanic activities in hyperthermophilic habitats. The lithotrophic energy metabolism of hyperthermophiles appears to be similar to that of mesophiles. Autotrophic CO2 fixation proceeds via the reductive citric acid cycle, considered to be one of the first metabolic cycles, and via the reductive acetyl-CoA/carbon monoxide dehydrogenase pathway. The Calvin cycle has not been found in hyperthermophiles (or any Archaea). Organotrophic metabolism mainly involves peptides and sugars as substrates, which are either oxidized to CO2 by external electron acceptors or fermented to acetate and other products. Sugar catabolism in hyperthermophiles involves non-phosphorylated versions of the Entner-Doudoroff pathway and modified versions of the Embden-Meyerhof pathway. The 'classical' Embden-Meyerhof pathway is present in hyperthermophilic Bacteria (Thermotoga) but not in Archaea. All hyperthermophiles (and Archaea) tested so far utilize pyruvate:ferredoxin oxidoreductase for acetyl-CoA formation from pyruvate. Acetyl-CoA oxidation in anaerobic sulphur-reducing and aerobic hyperthermophiles proceeds via the citric acid cycle; in the hyperthermophilic sulphate-reducer Archaeoglobus an oxidative acetyl-CoA/carbon monoxide dehydrogenase pathway is operative. Acetate formation from acetyl-CoA in Archaea, including hyperthermophiles, is

  9. Isolation of iron-oxidizing bacteria from corroded concretes of sewage treatment plants.

    Science.gov (United States)

    Maeda, T; Negishi, A; Komoto, H; Oshima, Y; Kamimura, K; Sugio, T

    1999-01-01

    Thirty-six strains of iron-oxidizing bacteria were isolated from corroded concrete samples obtained at eight sewage treatment plants in Japan. All of the strains isolated grew autotrophically in ferrous sulfate (3.0%), elemental sulfur (1.0%) and FeS (1.0%) media (pH 1.5). Washed intact cells of the 36 isolates had activities to oxidize both ferrous iron and elemental sulfur. Strain SNA-5, a representative of the isolated strains, was a gram-negative, rod-shaped bacterium (0.5-0.6x0.9-1.5 microm). The mean G+C content of its DNA was 55.9 mol%. The pH and temperature optima for growth were 1.5 and 30 degrees C, and the bacterium had activity to assimilate 14CO2 into the cells when ferrous iron or elemental sulfur was used as a sole source of energy. These results suggest that SNA-5 is Thiobacillus ferrooxidans strain. The pHs and numbers of iron-oxidizing bacteria in corroded concrete samples obtained by boring to depths of 0-1, 1-3, and 3-5 cm below the concrete surface were respectively 1.4, 1.7, and 2.0, and 1.2 x 10(8), 5 x 10(7), and 5 x 10(6) cells/g concrete. The degree of corrosion in the sample obtained nearest to the surface was more severe than in the deeper samples. The findings indicated that the levels of acidification and corrosion of the concrete structure corresponded with the number of iron-oxidizing bacteria in a concrete sample. Sulfuric acid produced by the chemolithoautotrophic sulfur-oxidizing bacterium Thiobacillus thiooxidansis known to induce concrete corrosion. Since not only T. thiooxidans but also T. ferrooxidans can oxidize reduced sulfur compounds and produce sulfuric acid, the results strongly suggest that T. ferrooxidans as well as T. thiooxidans is involved in concrete corrosion.

  10. Accessing the Martian deep subsurface to search for life

    Science.gov (United States)

    Mancinelli, Rocco L.

    2000-09-01

    To date there has been no data indicating that the surface of Mars is inhabited. Research conducted on Earth has revealed that life can exist deep beneath the surface of a planet. Current data from Mars missions suggesting the presence of liquid water early in Mars' history and mathematical modeling of the fate of water on Mars imply that liquid water may exist deep beneath the surface of Mars. The existence of liquid water beneath the Martian surface, combined with life's ability to live chemolithoautotrophically, leads to the hypothesis that life may exist deep beneath the Martian surface. Acquisition and analyses of Martian subsurface samples will shed light on the possibility of extant or extinct life, in permafrost and liquid water, on Mars, the processes leading to the origin of life, and the size of the Solar System's habitable zone. The results of a workshop convened by NASA in 1998 suggest that no fewer than two missions could be considered for accessing the deep subsurface of Mars. This two mission scenario includes a mission penetrating to a depth of ˜300 m and a mission penetrating to a depth of ˜3 km. As in all space missions the power, mass and volume of the penetration system and all associated equipment to be used to gain access to the Martian subsurface must be kept to a minimum. One technique that may be applicable, but in need of further development, is an electrically heated probe (bit) that penetrates and cores by melting through rock. The probe is attached to an umbilical cable to provide power and to allow periodic retrieval of the probe. This approach appears to address planetary protection concerns and may decrease the mass, power and volume requirements of conventional drilling systems by eliminating the need for casing material, drilling fluids, and handling potentially contaminated debris from the bore hole. Deep drilling on Mars presents several planetary protection forward and back contamination issues. These issues include sample

  11. Single-cell Sequencing of Thiomargarita Reveals Genomic Flexibility for Adaptation to Dynamic Redox Conditions

    Science.gov (United States)

    Winkel, Matthias; Salman-Carvalho, Verena; Woyke, Tanja; Richter, Michael; Schulz-Vogt, Heide N.; Flood, Beverly E.; Bailey, Jake V.; Mußmann, Marc

    2016-01-01

    Large, colorless sulfur-oxidizing bacteria (LSB) of the family Beggiatoaceae form thick mats at sulfidic sediment surfaces, where they efficiently detoxify sulfide before it enters the water column. The genus Thiomargarita harbors the largest known free-living bacteria with cell sizes of up to 750 μm in diameter. In addition to their ability to oxidize reduced sulfur compounds, some Thiomargarita spp. are known to store large amounts of nitrate, phosphate and elemental sulfur internally. To date little is known about their energy yielding metabolic pathways, and how these pathways compare to other Beggiatoaceae. Here, we present a draft single-cell genome of a chain-forming “Candidatus Thiomargarita nelsonii Thio36”, and conduct a comparative analysis to five draft and one full genome of other members of the Beggiatoaceae. “Ca. T. nelsonii Thio36” is able to respire nitrate to both ammonium and dinitrogen, which allows them to flexibly respond to environmental changes. Genes for sulfur oxidation and inorganic carbon fixation confirmed that “Ca. T. nelsonii Thio36” can function as a chemolithoautotroph. Carbon can be fixed via the Calvin–Benson–Bassham cycle, which is common among the Beggiatoaceae. In addition we found key genes of the reductive tricarboxylic acid cycle that point toward an alternative CO2 fixation pathway. Surprisingly, “Ca. T. nelsonii Thio36” also encodes key genes of the C2-cycle that convert 2-phosphoglycolate to 3-phosphoglycerate during photorespiration in higher plants and cyanobacteria. Moreover, we identified a novel trait of a flavin-based energy bifurcation pathway coupled to a Na+-translocating membrane complex (Rnf). The coupling of these pathways may be key to surviving long periods of anoxia. As other Beggiatoaceae “Ca. T. nelsonii Thio36” encodes many genes similar to those of (filamentous) cyanobacteria. In summary, the genome of “Ca. T. nelsonii Thio36” provides additional insight into the ecology of

  12. Autotrophic fixation of geogenic CO2 by microorganisms contributes to soil organic matter formation and alters isotope signatures in a wetland mofette

    Directory of Open Access Journals (Sweden)

    M. E. Nowak

    2015-09-01

    Full Text Available To quantify the contribution of autotrophic microorganisms to organic matter formation (OM in soils, we investigated natural CO2 vents (mofettes situated in a wetland in NW Bohemia (Czech Republic. Mofette soils had higher SOM concentrations than reference soils due to restricted decomposition under high CO2 levels. We used radiocarbon (Δ14C and stable carbon isotope ratios (δ13C to characterize SOM and its sources in two moffetes and compared it with respective reference soils, which were not influenced by geogenic CO2. The geogenic CO2 emitted at these sites is free of radiocarbon and enriched in δ13C compared to atmospheric CO2. Together, these isotopic signals allow us to distinguish C fixed by plants from C fixed by autotrophic microorganisms using their differences in δ13C discrimination. We can then estimate that up to 27 % of soil organic matter in the 0–10 cm layer of these soils was derived from microbially assimilated CO2. Isotope values of bulk SOM were shifted towards more positive δ13C and more negative Δ14C values in mofettes compared to reference soils, suggesting that geogenic CO2 emitted from the soil atmosphere is incorporated into SOM. To distinguish whether geogenic CO2 was fixed by plants or by CO2 assimilating microorganisms, we first used the proportional differences in radiocarbon and δ13C values to indicate the magnitude of discrimination of the stable isotopes in living plants. Deviation from this relationship was taken to indicate the presence of microbial CO2 fixation, as microbial discrimination should differ from that of plants. 13CO2-labelling experiments confirmed high activity of CO2 assimilating microbes in the top 10 cm, where δ13C values of SOM were shifted up to 2 ‰ towards more negative values. Uptake rates of microbial CO2 fixation ranged up to 1.59 ± 0.16 μg gdw−1 d−1. We inferred that the negative δ13C shift was caused by the activity of chemo-lithoautotrophic microorganisms, as

  13. Conversion of 4-Hydroxybutyrate to Acetyl Coenzyme A and Its Anapleurosis in the Metallosphaera sedula 3-Hydroxypropionate/4-Hydroxybutyrate Carbon Fixation Pathway

    Energy Technology Data Exchange (ETDEWEB)

    Hawkins, AB; Adams, MWW; Kelly, RM

    2014-03-25

    The extremely thermoacidophilic archaeon Metallosphaera sedula (optimum growth temperature, 73 degrees C, pH 2.0) grows chemolithoautotrophically on metal sulfides or molecular hydrogen by employing the 3-hydroxypropionate/4-hydroxybutyrate (3HP/4HB) carbon fixation cycle. This cycle adds two CO2 molecules to acetyl coenzyme A (acetyl-CoA) to generate 4HB, which is then rearranged and cleaved to form two acetyl-CoA molecules. Previous metabolic flux analysis showed that two-thirds of central carbon precursor molecules are derived from succinyl-CoA, which is oxidized to malate and oxaloacetate. The remaining one-third is apparently derived from acetyl-CoA. As such, the steps beyond succinyl-CoA are essential for completing the carbon fixation cycle and for anapleurosis of acetyl-CoA. Here, the final four enzymes of the 3HP/4HB cycle, 4-hydroxybutyrate-CoA ligase (AMP forming) (Msed_0406), 4-hydroxybutyryl-CoA dehydratase (Msed_1321), crotonyl-CoA hydratase/(S)-3-hydroxybutyryl-CoA dehydrogenase (Msed_0399), and acetoacetyl-CoA beta-ketothiolase (Msed_0656), were produced recombinantly in Escherichia coli, combined in vitro, and shown to convert 4HB to acetyl-CoA. Metabolic pathways connecting CO2 fixation and central metabolism were examined using a gas-intensive bioreactor system in which M. sedula was grown under autotrophic (CO2-limited) and heterotrophic conditions. Transcriptomic analysis revealed the importance of the 3HP/4HB pathway in supplying acetyl-CoA to anabolic pathways generating intermediates in M. sedula metabolism. The results indicated that flux between the succinate and acetyl-CoA branches in the 3HP/4HB pathway is governed by 4-hydroxybutyrate-CoA ligase, possibly regulated posttranslationally by the protein acetyltransferase (Pat)/Sir2-dependent system. Taken together, this work confirms the final four steps of the 3HP/4HB pathway, thereby providing the framework for examining connections between CO2 fixation and central metabolism in M. sedula.

  14. Detecting Organic Compounds Released from Iron Oxidizing Bacteria using Sample Analysis at Mars (SAM) Like Instrument Protocols

    Science.gov (United States)

    Glavin, D. P.; Popa, R.; Martin, M. G.; Freissinet, C.; Fisk, M. R.; Dworkin, J. P.; Mahaffy, P. R.

    2012-01-01

    Mars is a planet of great interest for Astrobiology since its past environmental conditions are thought to have been favourable for the emergence life. At present, the Red Planet is extremely cold and dry and the surface is exposed to intense UV and ionizing radiation, conditions generally considered to be incompatible with life as we know it on Earth. It was proposed that the shallow subsurface of Mars, where temperatures can be above freezing and liquid water can exist on rock surfaces, could harbor chemolithoautotrophic bacteria such as the iron oxidizing microorganism Pseudomonas sp. HerB. The Mars Science Laboratory (MSL) mission will provide the next opportunity to carry out in situ measurements for organic compounds of possible biological origin on Mars. One instrument onboard MSL, called the Sample Analysis at Mars (SAM) instrument suite, will carry out a broad and sensitive search for organic compounds in surface samples using either high temperature pyrolysis or chemical extraction followed by gas chromatography mass spectrometry. We present gas chromatograph mass spectrometer (GC/MS) data on crushed olivine rock powders that have been inoculated with Pseudomonas sp. HerB at different concentrations ranging from approx 10(exp 2) to 10(exp 7) cells per gram. The inoculated olivine samples were heated under helium carrier gas flow at 500 C and the pyrolysis products concentrated using a SAM-like hydrocarbon trap set at -20 C followed by trap heating and analysis by GC/Ms. In addition, the samples were also extracted using a low temperature "one-pot" chemical extraction technique using N-methyl, N-(tert-butyldimethylsilyl) trifluoroacetamide (MTBSTFA) as the silylating agent prior to GC/MS analysis. We identified several aldehydes, thiols, and alkene nitriles after pyrolysis GC/MS analysis of the bacteria that were not found in the olivine control samples that had not been inoculated with bacteria. The distribution of pyrolysis products extracted from the

  15. Is Mars a habitable environment for extremophilic microorganisms from Earth?

    Science.gov (United States)

    Rettberg, Petra; Reitz, Guenther; Flemming, Hans-Curt; Bauermeister, Anja

    A. ferrooxidans in a Mars subsurface simulation experiment could be demonstrated. Thus, from a geochemical perspective, these chemolithoautotrophic bacteria are relevant candidates for a hypothetical underground Martian food chain, despite their limited ability to tolerate the harsh physical conditions of the surface of Mars today.

  16. Organic chemistry of fluids from sediment-buried young basement: discrete sampling from ODP borehole 1301A & 1025C

    Science.gov (United States)

    Lin, H.; Cowen, J. P.; Amend, J. P.; Albert, D. B.; Glazer, B. T.; Rappe, M.; Jungbluth, S.; Matzinger, M.

    2010-12-01

    Hydrothermal systems profoundly influence seawater chemistry. However, the extent to which hydrothermal systems impact the quantity and the quality of the deep ocean dissolved organic carbon (DOC) pool remains unclear. To study the organic chemistry within sedimented ridge flank basement aquifers, discrete low temperature (~65°C & ~40°C) basement fluids were collected from Integrated Ocean Drilling Program boreholes via Circulation Obviation Retrofit Kit (CORK) observatories at 1301A (47°45N, 127°45W) and 1025C (47°53N, 128°39W) on the eastern flank of Juan de Fuca Ridge. The basement age is 1.24 Ma and 3.5 Ma at 1025C and 1301A, respectively. Basement fluids were collected using a new clean pumping system (Mobile Pump Valve Unit, or MPVU) and stored in acid-cleaned 60 L Large Volume Tedlar Bags (LVBS). A stainless steel fluid delivery line extends from the basement to the seafloor at 1301A; whereas fluids rise up the middle of the iron casing at 1025C. Here, concentration and preliminary characterization of the DOC will be presented, including labile organic components such as dissolved free and combined amino acids (DFAA & DCAA) and low molecular weight organic acids (LMW-OA). Our data show that compared to seawater levels, the DOC concentration in the younger basement fluid (1025C) was about one-half and in the older (1031A) was less than one-third. Relative to bottom seawater, the basement fluids are also depleted in SO42-, PO43-, NO3- and NO2-, but enriched in NH4+, H2S, Mn and Fe. Microbially mediated reduction of nitrate, sulfate, iron, and manganese and an array of heterotrophic metabolisms may explain these observations. Phylogenic trees based on 16S rRNA gene sequences from borehole 1301A fluids further support the presence of chemolithoautotrophic and heterotrophic organisms. Our preliminary results show that some dissolved free amino acids were slightly enriched in the 1301A basement fluid relative to bottom seawater. Thermodynamic calculations

  17. Hydrothermal mixing: Fuel for life in the deep-sea

    Science.gov (United States)

    Hentscher, M.; Bach, W.; Amend, J.; McCollom, T.

    2009-04-01

    Deep-sea hydrothermal vent systems show a wide range of fluid compositions and temperatures. They reach from highly alkaline and reducing, like the Lost City hydrothermal field, to acidic and reducing conditions, (e. g., the Logatchev hydrothermal field) to acidic and oxidizing conditions (e. g., island arc hosted systems). These apparently hostile vent systems are generally accompanied by high microbial activity forming the base of a food-web that often includes higher organisms like mussels, snails, or shrimp. The primary production is boosted by mixing of chemically reduced hydrothermal vent fluids with ambient seawater, which generates redox disequilibria that serve as energy source for chemolithoautotrophic microbial life. We used geochemical reaction path models to compute the affinities of catabolic (energy-harvesting) and anabolic (biosynthesis) reactions along trajectories of batch mixing between vent fluids and 2 °C seawater. Geochemical data of endmember hydrothermal fluids from 12 different vent fields (Lost City, Rainbow, Logatchev, TAG, EPR 21 °N, Manus Basin, Mariana Arc, etc.) were included in this reconnaissance study of the variability in metabolic energetics in global submarine vent systems. The results show a distinction between ultramafic-hosted and basalt-hosted hydrothermal systems. The highest energy yield for chemolithotrophic catabolism in ultramafic-hosted hydrothermal systems is reached at low temperature and under slightly aerobic to aerobic conditions. The dominant reactions, for example at Rainbow or Lost City, are the oxidation of H2, Fe2+ and methane. At temperatures >60 °C, anaerobic metabolic reactions, e. g., sulphate reduction and methanogenesis, become more profitable. In contrast, basalt-hosted systems, such as TAG and 21 °N EPR uniformly indicate H2S oxidation to be the catabolically dominant reaction over the entire microbial-relevant temperature range. Affinities were calculated for the formation of individual cellular

  18. Where microorganisms meet rocks in the Earth's Critical Zone

    Directory of Open Access Journals (Sweden)

    D. M. Akob

    2011-12-01

    Full Text Available The Critical Zone (CZ is the Earth's outer shell where all the fundamental physical, chemical, and biological processes critical for sustaining life occur and interact. As microbes in the CZ drive many of these biogeochemical cycles, understanding their impact on life-sustaining processes starts with an understanding of their biodiversity. In this review, we summarize the factors controlling where terrestrial CZ microbes (prokaryotes and micro-eukaryotes live and what is known about their diversity and function. Microbes are found throughout the CZ, down to 5 km below the surface, but their functional roles change with depth due to habitat complexity, e.g. variability in pore spaces, water, oxygen, and nutrients. Abundances of prokaryotes and micro-eukaryotes decrease from 1010 or 107 cells g soil−1 or rock−1, or ml water−1 by up to eight orders of magnitude with depth. Although symbiotic mycorrhizal fungi and free-living decomposers have been studied extensively in soil habitats, where they occur up to 103 cells g soil−1, little is known regarding their identity or impact on weathering in the deep subsurface. The relatively low abundance of micro-eukaryotes in the deep subsurface suggests that they are limited in space, nutrients, are unable to cope with oxygen limitations, or some combination thereof. Since deep regions of the CZ have limited access to recent photosynthesis-derived carbon, microbes there depend on deposited organic material or a chemolithoautotrophic metabolism that allows for a complete food chain, independent from the surface, although limited energy flux means cell growth may take tens to thousands of years. Microbes are found in all regions of the CZ and can mediate important biogeochemical processes, but more work is needed to understand how microbial populations influence the links between different regions of the CZ and weathering

  19. PROSPECTS FOR LIFE IN THE SUBGLACIAL LAKE VOSTOK, EAST ANTARCTICA

    Directory of Open Access Journals (Sweden)

    S. A. Bulat

    2012-01-01

    Full Text Available The objective was to estimate the genuine microbial content of ice samples from refrozen water (accretion ice from the subglacialLakeVostok(Antarctica buried beneath the 4-km thick East Antarctic ice sheet as well as surface snow nearby Vostok station. The lake ice samples were extracted by heavy deep ice drilling from3764 mbelow the surface reaching the depth3769.3 mby February 2011 (lake entering. High pressure, an ultra low carbon and chemical content, isolation, complete darkness and the probable excess of oxygen in water for millions of years characterize this extreme environment. A decontamination protocol was first applied to samples selected for the absence of cracks to remove the outer part contaminated by handling and drilling fluid. Preliminary indications showed the accretion ice samples to be almost gas free with the very low impurity content. Flow cytometry showed the very low unevenly distributed biomass in both accretion (0–19 cells per ml and glacier (0–24 cells per ml ice and surface snow (0–0.02 cells per ml as well while repeated microscopic observations were unsuccessful meaning that the whole Central East Antarctic ice sheet seems to be microbial cell-free.We used strategies of Ancient DNA research that include establishing contaminant databases and criteria to validate the amplification results. To date, positive results that passed the artifacts and contaminant databases have been obtained for a few bacterial phylotypes only in accretion ice samples featured by some bedrock sediments. Amongst them are the chemolithoautotrophic thermophile Hydrogenophilus thermoluteolus of beta-Proteobacteria, the actinobacterium rather related (95% to Ilumatobacter luminis and one unclassified phylotype distantly related (92% to soil-inhabiting uncultured bacteria. Combined with geochemical and geophysical considerations, our results suggest the presence of a deep biosphere, possibly thriving within some active faults of the bedrock

  20. Microbiology and optimization of hydrogen fermentation and bioelectricity production

    Energy Technology Data Exchange (ETDEWEB)

    Makinen, A.

    2013-11-01

    yield. Pentose fermentation was accompanied by production of acetate, butyrate and formate, while in hexose fermentation the main soluble end product was lactate. In CSTR Hisarkoy enrichment culture produced H{sub 2} from xylose with the maximum average H{sub 2} yield and production rate of 1.97 mol/mol xylose and 7.3 mmol/h/L, respectively, at suboptimal temperature of 45 deg C for meso- and thermophiles. At 45 deg C microbial community consisted of only two bacterial strains affiliated to Clostridium acetobutylicum and Cirtobacter freundii. An exoeletrogenic culture was enriched on xylose from compost sample in MFCs. In enrichment phase electricity production in MFCs was accompanied with ethanol production. The main bacterium responsible of xylose degradation was xylanolytic Ruminobacillus xylanolyticum and the main bacteria responsible for electricity production were denitrifiers Paracoccus pantotrophus, Comamonas denitrificans and Alicycliphilus denitrificans. Anode potential had a significant effect on current production in MFCs. Compost enrichment culture was able to produce electricity from xylose at poised anode potential of 0.4 V vs standard hydrogen electrode (SHE) having the maximum current density and Coulombic efficiency (CE) of 1.65 A/m{sup 2} and 37 %, respectively. Lower anode potentials of 0.1 or -0.2 V vs SHE didn't lead to current production. Optimum operational parameters for bioelectricity production from xylose by compost enrichment culture were without mixing, external resistance of 100 ohm, 0.5 g/L xylose and pH 7. High current density and CE of 1.74 A/m{sup 2} and 82 %, respectively, were obtained. This is the highest CE obtained with xylose in two-chamber MFC reported in the literature. This very efficient exoelectrogenic culture was dominated by Geobacter species, including G. sulfurreducens, which were enriched on anode biofilm. Xylose fermenters, including Escherichia coli, Sphaerochaeta sp. TQ1, and Bacteroides sp. were present in

  1. Chemically Specific Cellular Imaging of Biofilm Formation

    Energy Technology Data Exchange (ETDEWEB)

    Herberg, J L; Schaldach, C; Horn, J; Gjersing, E; Maxwell, R

    2006-02-09

    This document and the accompanying manuscripts summarize the technical accomplishments for our one-year LDRD-ER effort. Biofilm forming microbes have existed on this planet for billions of years and make up 60% of the biological mass on earth. Such microbes exhibit unique biochemical pathways during biofilm formation and play important roles in human health and the environment. Microbial biofilms have been directly implicated in, for example, product contamination, energy losses, and medical infection that cost the loss of human lives and billions of dollars. In no small part due to the lack of detailed understanding, biofilms unfortunately are resistant to control, inhibition, and destruction, either through treatment with antimicrobials or immunological defense mechanisms of the body. Current biofilm research has concentrated on the study of biofilms in the bulk. This is primarily due to the lack of analytical and physical tools to study biofilms non-destructively, in three dimensions, and on the micron or sub-micron scale. This has hindered the development of a clear understanding of either the early stage mechanisms of biofilm growth or the interactions of biofilms with their environment. Enzymatic studies have deduced a biochemical reaction that results in the oxidation of reduced sulfur species with the concomitant reduction of nitrate, a common groundwater pollutant, to dinitrogen gas by the bacterium, Thiobacillus denitrificans (TD). Because of its unique involvement in biologically relevant environmental pathways, TD is scheduled for genome sequencing in the near future by the DOE's Joint Genome Institute and is of interest to DOE's Genomes to Life Program. As our ecosystem is exposed to more and more nitrate contamination large scale livestock and agricultural practices, a further understanding of biofilm formation by organisms that could alleviate these problems is necessary in order to protect out biosphere. However, in order to study this

  2. Development of biological platform for the autotrophic production of biofuels

    Science.gov (United States)

    Khan, Nymul

    The research described herein is aimed at developing an advanced biofuel platform that has the potential to surpass the natural rate of solar energy capture and CO2 fixation. The underlying concept is to use the electricity from a renewable source, such as wind or solar, to capture CO 2 via a biological agent, such as a microbe, into liquid fuels that can be used for the transportation sector. In addition to being renewable, the higher rate of energy capture by photovoltaic cells than natural photosynthesis is expected to facilitate higher rate of liquid fuel production than traditional biofuel processes. The envisioned platform is part of ARPA-E's (Advanced Research Projects Agency - Energy) Electrofuels initiative which aims at supplementing the country's petroleum based fuel production with renewable liquid fuels that can integrate easily with the existing refining and distribution infrastructure (http://arpae. energy.gov/ProgramsProjects/Electrofuels.aspx). The Electrofuels initiative aimed to develop liquid biofuels that avoid the issues encountered in the current generation of biofuels: (1) the reliance of biomass-derived technologies on the inefficient process of photosynthesis, (2) the relatively energy- and resource-intensive nature of agronomic processes, and (3) the occupation of large areas of arable land for feedstock production. The process proceeds by the capture of solar energy into electrical energy via photovoltaic cells, using the generated electricity to split water into molecular hydrogen (H2) and oxygen (O2), and feeding these gases, along with carbon dioxide (CO2) emitted from point sources such as a biomass or coal-fired power plant, to a microbial bioprocessing platform. The proposed microbial bioprocessing platform leverages a chemolithoautotrophic microorganism (Rhodobacter capsulatus or Ralstonia eutropha) naturally able to utilize these gases as growth substrates, and genetically modified to produce a triterpene hydrocarbon fuel

  3. Start-up of a completely autotrophic nitrogen removal process in a three- dimensional electrode-biofilm reactor%三维电极生物膜反应器全程自养脱氮的启动研究

    Institute of Scientific and Technical Information of China (English)

    郭劲松; 杨琳; 陈猷鹏; 方芳; 唐金晶

    2012-01-01

    A completely autotrophic nitrogen removal process was started up in a three-dimensional electrode-biofilm reactor for artificial ammonia wastewater treatment. The titanium rod coated with a thin layer of ruthenium was used as anode to generate oxygen. In the aerobic area, NH4^+-N was oxidized to NO3^- -N or NO2^- -N by nitrifying bacteria. The active carbon fiber-felt was used as cathode to generate hydrogen. And in this anaerobic area, the denitrification was completed while hydrogen was acted as the electron donor. A lot of carbon particles were filled in tbe cathode area used as three- dimensional electrode. Nitrification and denitrification process were controlled by adjusting dissolved oxygen and pH values under the condition that the initial concentration of ammonia-nitrogen was 30 mg·L^-1 , the hydraulic retention time was 24h and the temperature was 30℃. After biofilm was formed and stabilized, the removal rate of NH4^+-N and TN achieved 97.8% and 92.4% respectively. It was indicated that the completely autotrophic nitrogen removal was started up successfully. The scanning electron microscopy showed that the bacteria on surface of activated carbon fiber felt were mainly short rod-shaped Pseudomonas, while the bacteria on the surface of the activated carbon particles were Micrococcus denitrificans. They both belong to hydrogen autotrophic denitrifying bacteria. In the reactor, the stable autotrophic nitrogen system was gradually established.%采用人工配制氨氮废水,对三维电极生物膜反应器进行全程自养脱氮的启动研究.反应器中阳极采用钌涂层钛棒,在阳极区电解水产氧供硝化菌进行硝化反应;阴极采用活性炭纤维毡,并在阴极区填充活性炭颗粒构建三维电极,在阴极区电解水产氢供反硝化菌完成反硝化过程.在进水NH4^+-N浓度30mg·L^-1、温度30℃、HRT为24h的试验条件下,通过调节DO和pH实现对硝化和反硝化反应的控制.结果

  4. 城市污水厂活性污泥强化自养反硝化菌研究%Experimental Study of Autotrophic Denitrification Bacteria Through Bioaugmentation of Activated Sludge from Municipal Wastewater Plant

    Institute of Scientific and Technical Information of China (English)

    常玉梅; 杨琦; 郝春博; 尚海涛; 姜体胜

    2011-01-01

    Activated sludge of municipal wastewater treatment plant was domesticated by sulfur as the electron donor under autotrophic.The sludge activity was determined by measuring growth rate of sludge. The removal efficiency of nitrate and sulfate production efficiency were analyzed by continuously measuring the concentration of NO3- -N and S024-. When the removal efficiency of nitrate was more than 90%, 16S rRNA genetic libraries were built up to compare their microbial biodiversity. The growth rate of sludge is 0.177 g/( L· d). The relation between concentration of nitrate and time meets first order reaction kinetics. The bacteria in the sludge affiliated with Beta-Proteobacteria, Deta-Proteobacteria, Gamma-Proteobacteria and Unclassified bacteria. Beta-Proteobacteria is the main phylum in the sludge. Bacteria related to Thiobacillus denitrificans from denitrifying bioreaetor perform 48.65%. In addition, the bacteria of Denitratisoma sp. , Curvibacter sp. , Thermomonas sp. Geobacter sp. are existed in the sludge. The study of autotrophic denitrifying bacteria diversity is conducive to optimization of reaction conditions and efficient removal of nitrate.%采集北京高碑店城市污水厂的反硝化污泥样品,以硫磺作为电子供体进行驯化培养.测定污泥的增长率来确定污泥活性,分别测定NO-N、SO浓度来确定硝酸盐的去除效率和硫酸盐生成速率.当硝酸盐去除率达到90%以上时,提取污泥中微生物总DNA,构建16S rRNA基因片段克隆文库来分析细菌群落结构,结果表明,污泥的增长率为0.177 g/(L·d),污泥中硝酸盐浓度与时间的关系符合一级反应.污泥中细菌类群主要为Beta-Proteobacteria、Deta-Proteobacteria、Gamma-Proteobacteria和Unclassified bacteria,其中Beta-Proteobacteria类细菌占主导地位.在成熟的反硝化污泥中,自养反硝化菌Thiobacillusdenitrificans占所占比例高达48.65%.此外,反应器中还存在Denitratisoma sp.、Curvibacter sp

  5. Effect of Injected Leachate C/N Ratio on the Denitrification of a Bioreactor Filled with Landfilled Refuse%回灌渗滤液C/N对填埋垃圾生物反应器反硝化特性的影响

    Institute of Scientific and Technical Information of China (English)

    黎乾; 吴松维; 吴伟祥; 孙法迁; 刘晶静; 蔡传钰; 陈敏

    2011-01-01

    The denitrification capacity of a landfill bioreactor was investigated under different ratios of injected leachate C/N;the denitrifying bacteria community compositions over four injection times were also studied using molecular approaches and functional gene nirS served as marker.Results showed that COD/NO-3-N ratio of the injected landfill leachate had a significant impact on the denitrification capacity of the bioreactor.Nitrate reduction rate increased from 1.14 mg·(kg·h)-1 to 11.40 mg mg·(kg·h)-1 when the injected leachate COD/NO-3-N ratio raised from 3.11 to 13.08.It suggested that a stable and rapid denitrification could be obtained when bioavailable COD/NO-3-N ratio in the injected leachate was 6.37.In the bioreactor,the main denitrifying bacteria was similar to β-proteobacteria,and others belonged to uncultured bacteria.Thiobacillus denitrificans and Azoarcus tolulyticus known as β-proteobacteria were the dominant species and played an important role in NO-3-N consumption during the leachate injection.%利用模拟填埋垃圾生物反应器(landfill bioreactor)研究了回灌渗滤液C/N对填埋垃圾堆体反硝化性能的影响,并采用PCR扩增、克隆测序等分子生物学技术,以功能基因nirS为分子标记考察了4次回灌过程中反应器内反硝化微生物群落结构变化.结果表明,回灌渗滤液的COD/NO 3--N比例对反应器的反硝化活性具有显著性影响.当COD/NO 3--N从3.11提高到13.08时,反应器内硝酸盐还原速率可从1.14 mg.(kg.h)-1提高到11.40 mg.(kg.h)-1.当回灌渗滤液生物可利用性COD与NO 3--N比值达到6.37时,可以实现填埋垃圾生物反应器反硝化作用的快速、稳定运行.4次回灌,反应器内的反硝化微生物大部分与β-变形菌纲(β-proteobacteria)细菌相似,少数属于非培养微生物(uncultured bacteria),其中Thiobacillus denitrificans和Azoarcus tolulyticus是反硝化过程的主要功能微生物,在回灌渗

  6. SBR亚硝化处理化肥厂氨氮废水影响因素分析%Influence factors of treatment by nitrosation in sequencing batch reactor with ammonia-nitrogen wastewater of fertilizer plant

    Institute of Scientific and Technical Information of China (English)

    庞荣辉; 黄智宁; 曹蔓

    2014-01-01

    A lab-scale batch sequencing reactor (SBR) was used to treat ammonia-nitrogen wastewater of fertilizer plant and factors affecting the short-cut biological nitrogen removal, such as pH, DO, effluent of NH4+-N and temperature etc., were investigated. The optimum operating condition of nitrosation by SBR was confirmed. It reveals that pH value over low restrained generation of nitrite when pH value over high prejudiced the removal of TN through denitrification by denitrificans. Nitrosation rate remained high level and effluent of TN was well controlled as the pH value was approximately maintained 8.0. When DO was 0.2~0.3 mg/L, the reaction rate of nitrosation was slow as the partial nitrification was still going on. The nitrosation in system couldn’t achieved and nitrification became maim reaction while DO was 1.5~2.8 mg/l. Nitrite was abundantly accumulated at high ammonia nitrogen load from inflow while the removal efficiency of NH4+-N was not good. High nitrosation rate can be obtained at relatively high temperature. The optimum operating conditions were controlled as follows: temperature at 35℃, pH of initial inflow at 7.8 to 8.2, DO at 0.5 to 0.6 mg/L and influent concentration of NH4+-N at 100mg/L. The nitrosation rate can be retained above 90%in optimum operating conditions.%利用SBR亚硝化处理化肥厂实际废水,研究其短程生物脱氮过程中pH、溶解氧、进水氨氮负荷和温度等因素影响,并确定亚硝化处理的最佳操作条件。结果表明,pH过低会抑制亚硝酸盐的生成,过高则不利于反硝化菌反硝化过程TN的去除;当pH值控制在8.0左右时,亚硝化率保持较高水平,同时出水TN浓度控制较好。当DO浓度为0.2~0.3 mg/L时,亚硝化反应继续进行,但NH4+-N亚硝化反应速率较慢;当DO浓度为1.5~2.8 mg/l时,无法实现系统中亚硝化的运行,硝化作用成为主要反应。高氨氮负荷进水有利于亚硝酸盐的积累,但对出水氨氮

  7. DDE remediation and degradation.

    Science.gov (United States)

    Thomas, John E; Ou, Li-Tse; All-Agely, Abid

    2008-01-01

    breakdown of DDE by the extracellular lignolytic enzymes produced by white rot fungi. The addition of adjutants such as sodium ion, surfactants, and cellulose increased the rate of DDT aerobic or anaerobic degradation but did little to enhance the rate of DDE disappearance under anaerobic conditions. Only in the past decade has it been demonstrated that DDE can undergo reductive dechlorination under methanogenic and sulfidogenic conditions to form the degradation product DDMU, 1-chloro-2,2'-bis-(4'-chlorophenyl)ethane. The only pure culture reported to degrade DDE under anaerobic conditions was the denitrifier Alcaligens denitrificans. The degradation of DDE by this bacterium was enhanced by glucose, whereas biphenyl fumes had no effect. Abiotic remediation by DDE volatilization was enhanced by flooding and irrigation and deepplowing inhibited the volatilization. The use of zero-valent iron and surfactants in flooded soils enhanced DDT degradation but did not significantly alter the rate of DDE removal. Other catalysts (palladized magnesium, palladium on carbon, and nickel/aluminum alloys) degraded DDT and its metabolites, including DDE. However, these systems are often biphasic or involve explosive gases or both. Safer abiotic alternatives use UV light with titanium oxide or visible light with methylene green to degrade DDT, DDD, and DDE in aqueous or mixed solvent systems. Remediation and degradation of DDE in soil and water by phytoextraction, aerobic and anaerobic microorganisms, or abiotic methods can be accomplished. However, success has been limited, and great care must be taken that the method does not transfer the contaminants to another locale (by volatilization, deep plowing, erosion, or runoff) or to another species (by ingestion of accumulating plants or contaminated water). Although the remediation of DDT-, DDD-, and DDE-contaminated soil and water is beset with myriad problems, there remain many open avenues of research.

  8. Land-use controls on sources and fate of nitrate in shallow groundwater of an agricultural area revealed by multiple environmental tracers

    Science.gov (United States)

    Koh, Dong-Chan; Mayer, Bernhard; Lee, Kwang-Sik; Ko, Kyung-Seok,

    2010-10-01

    Sources and transformation processes of nitrate in groundwater from shallow aquifers were investigated in an agricultural area in the mid-western part of South Korea using a multi-tracer approach including δ 2H and δ 18O values of water, δ 15N and δ 18O values of nitrate, Cl/Br ratios and chlorofluorocarbons (CFCs). The study area was comprised of four land-use types with natural areas at higher altitudes, upland areas with fruit orchards, paddy fields and residential areas at lower elevations. The isotopic composition of water was suitable for distinguishing groundwater that had infiltrated in the higher elevation natural areas with lower δ 2H and δ 18O values from groundwater underneath paddy fields that was characterized by elevated δ 2H and δ 18O values due to evaporation. δ 18O-H 2O values and Cl - concentrations indicated that groundwater and contaminant sources were derived from three land-use types: natural areas, residential areas and paddy fields. Groundwater age determination based on CFCs showed that nitrate contamination of groundwater is primarily controlled by historic nitrogen loadings at least in areas with higher nitrate contamination. Nitrate sources were identified using the stable isotope composition of nitrate and Cl/Br ratios. Higher δ 15N-NO 3- values and Cl/Br ratios of 300 to 800 in residential areas indicated that waste water and septic effluents were major nitrate sources whereas lower δ 15N-NO 3- values and Cl/Br ratios of 100 to 700 in upland areas suggested that synthetic fertilizers constituted a major source of nitrate contamination of aquifers. With only few exceptions in the natural area, contributions of atmospheric nitrate were insignificant due to the resetting of δ 18O-NO 3- values via immobilization and re-mineralization of nitrate in the soil zone. In groundwater underneath paddy fields, 30% of samples had δ 18O-NO 3- values at least 2‰ higher than expected for nitrate formed by chemolithoautotrophic

  9. Low-temperature hydration, oxidation and hydrogen production from Oman peridotite

    Science.gov (United States)

    Miller, H. M.; Mayhew, L.; Templeton, A. S.

    2013-12-01

    Peridotite in the shallow subsurface undergoes hydration and oxidation (serpentinization) during reactions with percolating fluids, generating hydrogen gas and releasing magnesium, iron, and calcium into solution. In the presence of fluids enriched in dissolved carbon dioxide, extensive precipitation of carbonate minerals occurs. This reaction has large-scale implications for mitigating climate change by providing a stable, geological carbon repository. The Samail Ophiolite in Oman contains large quantities of ultramafic rocks that are currently undergoing serpentinization at low temperatures (30°C) and forming carbonate minerals. The production of hydrogen gas provides an electron donor for subsurface chemolithoautotrophic life which can contribute to carbon cycling in the subsurface as microorganisms utilize carbon dioxide as an inorganic carbon source. Serpentinization reactions require the oxidation of Fe (II) to Fe (III) to reduce water to H2, but the mechanisms of hydrogen generation in low-temperature systems is poorly characterized. To address this question, we conducted low temperature (100°C) water-rock reactions with Oman peridotite, measured H2 and characterized the speciation of Fe-bearing minerals before and after water-rock interaction using micro-X-ray Absorption Near Edge Structure (μXANES) spectra obtained from Stanford Synchrotron Radiation Lightsource. The experimental water-rock reactions produce H2 at a pH of 9, which corresponds with observations of ultrabasic springs in the Samail ophiolite and the presence of H2 in these spring waters. Significant hydrogen production occurs for two and a half months of reaction, peaking at 400 nmol/gram of reacted peridotite and then steadily decreases with time. These maximum values of hydrogen production from Oman peridotite are greater than observed by our laboratory and others during aqueous alteration of San Carlos peridotite and isolated pyroxenes and olivines (e.g. Mayhew et al. 2013 [1]). The

  10. Destruction and Sequestration of H2O on Mars

    Science.gov (United States)

    Clark, Benton

    2016-07-01

    The availability of water in biologically useable form on any planet is a quintessential resource, even if the planet is in a zone habitable with temperature regimes required for growth of organisms (above -18 °C). Mars and most other planetary objects in the solar system do not have sufficient liquid water at their surfaces that photosynthesis or chemolithoautotrophic metabolism could occur. Given clear evidence of hydrous mineral alteration and geomorphological constructs requiring abundant supplies of liquid water in the past, the question arises whether this H2O only became trapped physically as ice, or whether there could be other, more or less accessible reservoirs that it has evolved into. Salts containing S or Cl appear to be ubiquitous on Mars, having been measured in soils by all six Mars landed missions, and detected in additional areas by orbital investigations. Volcanoes emit gaseous H2S, S, SO2, HCl and Cl2. A variety of evidence indicates the geochemical fate of these gases is to be transformed into sulfates, chlorides, chlorates and perchlorates. Depending on the gas, the net reaction causes the destruction of between one and up to eight molecules of H2O per atom of S or Cl (although hydrogen atoms are also released, they are lost relatively rapidly to atmospheric escape). Furthermore, the salt minerals formed often incorporate H2O into their crystalline structures, and can result in the sequestration of up to yet another six (sometimes, more) molecules of H2O. In addition, if the salts are microcrystalline or amorphous, they are potent adsorbents for H2O. In certain cases, they are even deliquescent under martian conditions. Finally, the high solubility of the vast majority of these salts (with notable exception of CaSO4) can result in dense brines with low water activity, aH, as well as cations which can be inimical to microbial metabolism, effectively "poisoning the well." The original geologic materials on Mars, igneous rocks, also provide some

  11. Deliberations on Microbial Life in the Subglacial Lake Vostok, East Antarctica

    Science.gov (United States)

    Bulat, S.; Alekhina, I.; Lipenkov, V.; Lukin, V.; Marie, D.; Petit, J.

    2004-12-01

    The objective was to estimate microbial contents of accretion (lake originating) ice from the Lake Vostok buried beneath 4-km thick East Antarctic ice sheet with the ultimate goal to discover microbial life in this extreme icy environment featured by no light, close to freezing point temperature, ultra-low DOC contents, and an excess of oxygen. The PCR based bacterial and archaeal 16S ribosomal RNA gene sequencing constrained by Forensic Biology and Ancient DNA research criteria was used as a main approach. Epifluorescent and confocal microscopies as well as flow cytometry were implemented. DNA study showed that the accretion ice is essentially bacteria- and archaea-free. Up to now, the only accretion ice type 1 featured by mica-clay sediments presence and namely one horizon of four studied (3607m) allowed the recovery a few bacterial phylotypes. This unexpectedly included the chemolithoautotrophic thermophile Hydrogenophilus thermoluteolus and two more unclassified phylotypes all passing numerous contaminant controls. In contrast, the deeper and cleaner accretion ice 2 (three cores) with no sediments presence and near detection limit gas contents gave no reliable signals. The microbes detected in accretion ice 1 are unbelievable to resist an excess of oxygen in the lake water body (700 - 1300 mg O2/l). They are supposed to be thriving in rather warm anoxic sediments in deep faults at the lake bottom and sporadically flushing out along with sediments to the lake veins in a shallow depth bay due to a seismotectonic activity likely operating in the lake environment. A few geophysical and geological evidences support this scenario. In the bay the presence of mica-clay sediments, higher accretion rate due to relief rise and likely oxygen-depleted upper layer of water can provide microbes with a chance to escape the high oxygen tension by the rapid entrapment into accretion ice 1. Sediment-free accretion ice 2, which forms above a deeper part of the lake, shows no

  12. Benthic mineralization and nutrient exchange over the inner continental shelf of western India

    Science.gov (United States)

    Pratihary, A. K.; Naqvi, S. W. A.; Narvenkar, G.; Kurian, S.; Naik, H.; Naik, R.; Manjunatha, B. R.

    2014-05-01

    The western Indian continental shelf is one of the most productive coastal systems of the world ocean. This system experiences extreme changes in its oxygen regime, being normoxic from November to May and suboxic (denitrifying)/anoxic from June to October, owing to the biogeochemical response to cyclical monsoonal influence. In order to understand the impact of the seasonally varying oxygen regime on benthic mineralization, nutrient exchange and, in turn, on the shelf ecosystem, we carried out the first ever intact-core incubations during two contrasting seasons - spring intermonsoon and fall intermonsoon (late southwest monsoon) at a 28 m-deep fixed site on the inner shelf off Goa, dominated by fine-grained cohesive sediments. The results showed that incomplete sediment oxygen consumption (SOC) occurred during April as opposed to the complete SOC and subsequent sulfide flux observed in the fall intermonsoon incubations. The sediments acted as a perennial net source of DIN (dissolved inorganic nitrogen i.e. NO3- + NO2- + NH4+), PO43- and SiO44- to the overlying water column. The efflux of DIN increased from 1.4 to 3.74 mmol m-2 d-1 from April to October, of which NH4+ flux comprised 59-100%. During the oxic regime, ∼75% of diffusing NH4+ appeared to be nitrified (2.55 mmol m-2 d-1), of which ∼77% remained coupled to benthic denitrification. Consequently, 58% of NH4+ flux was lost in active coupled nitrification-denitrification, resulting in substantial N loss (1.98 mmol m-2 d-1) in the sediments. The continental shelf sediments switched over from being a NO3- source during the oxic regime to a NO3- sink during the anoxic regime. During suboxia, benthic denitrification that is fed by NO3- from the overlying water caused N loss at the rate of 1.04 mmol m-2 d-1. Nitrogen loss continued even under sulfidic conditions during October, possibly through the chemolithoautotrophic denitrification, at a potential rate of 3.21 mmol m-2 d-1. Phosphate flux increased more

  13. Detecting Organic Compounds Released from Iron Oxidizing Bacteria using Sample Analysis at Mars (SAM)-like Instrument Protocols

    Science.gov (United States)

    Glavin, D. P.; Popa, R.; Martin, M. G.; Freissinet, C.; Fisk, M. R.; Dworkin, J. P.; Mahaffy, P. R.

    2012-12-01

    Mars is a planet of great interest for Astrobiology since its past environmental conditions are thought to have been favourable for the emergence life. At present, the Red Planet is extremely cold and dry and the surface is exposed to intense UV and ionizing radiation, conditions generally considered to be incompatible with life as we know it on Earth. It was proposed that the shallow subsurface of Mars, where temperatures can be above freezing and liquid water can exist on rock surfaces, could harbor chemolithoautotrophic bacteria such as the iron oxidizing microorganism Pseudomonas sp. HerB [Popa et al. 2012]. The Mars Science Laboratory (MSL) mission will provide the next opportunity to carry out in situ measurements for organic compounds of possible biological origin on Mars. One instrument onboard MSL, called the Sample Analysis at Mars (SAM) instrument suite, will carry out a broad and sensitive search for organic compounds in surface samples using either high temperature pyrolysis or chemical extraction followed by gas chromatography mass spectrometry [Mahaffy et al. 2012]. We present gas chromatograph mass spectrometer (GC/MS) data on crushed olivine rock powders that have been inoculated with Pseudomonas sp. HerB at different concentrations ranging from ~102 to 107 cells per gram. The inoculated olivine samples were heated under helium carrier gas flow at 500°C and the pyrolysis products concentrated using a SAM-like hydrocarbon trap set at -20°C followed by trap heating and analysis by GC/MS. In addition, the samples were also extracted using a low temperature "one-pot" chemical extraction technique using N-methyl, N-(tert-butyldimethylsilyl) trifluoroacetamide (MTBSTFA) as the silylating agent prior to GC/MS analysis [Stalport et al. 2012]. We identified several aldehydes, thiols, and alkene nitriles after pyrolysis GC/MS analysis of the bacteria that were not found in the olivine control samples that had not been inoculated with bacteria. The

  14. Energy and Carbon Flow: Comparing ultramafic- and basalt-hosted vents

    Science.gov (United States)

    Perner, M.; Bach, W.; Seifert, R.; Strauss, H.; Laroche, J.

    2010-12-01

    In deep-sea vent habitats hydrothermal fluids provide the grounds for life by supplying reduced inorganic compounds (e.g. H2, sulfide). Chemolithoautotrophs can oxidize these substrates hereby yielding energy, which can then be used to fuel autotrophic CO2 fixation. Depending on the type of host rocks (and the degree of admixed ambient seawater) the availability of inorganic electron donors can vary considerably. While in ultramafic-hosted vents H2 levels are high and H2-oxidizing metabolisms are thought to dominate, in basalt-hosted vents, H2 is much lower and microbial sulfide oxidation is considered to prevail [1, 2]. We have investigated the effect of H2 and sulfide availability on the microbial community of distinct H2-rich and H2-poor vent sites along the Mid-Atlantic Ridge. Hydrothermally influenced samples were collected from the H2-rich ultramafic-hosted Logatchev field (15°N) and the comparatively H2-poor basalt-hosted vents from 5°S and 9°S. We conducted catabolic energy calculations to estimate the potential of various electron donors to function as microbial energy sources. We performed incubation experiments with hydrothermal fluids amended with H2 or sulfide and radioactively labelled bicarbonate and determined H2 and sulfide consumption and carbon incorporation rates. We constructed metagenomic libraries for sequence-based screening of genes encoding key enzymes for H2 uptake (NiFe uptake hydrogenases, group 1), sulfide oxidation (sulfide quinone oxidoreductase, sqr) and CO2 fixation pathways (RubisCOs of the Calvin cycle [CBB] and beta-subunit of the ATP citrate lyase of the reductive tricarboxylic acid cycle [rTCA]). We evaluated parts of the metagenomes from basalt-hosted sites by pyrosequencing. Based on our incubation experiments - under the conditions applied - we could not confirm that generally H2 consumption rates and biomass syntheses in fluids derived from ultramafic-hosted locations are significantly enhanced over those from basalt

  15. Engineering Ralstonia eutropha for Production of Isobutanol (IBT) Motor Fuel from Carbon Dioxide, Hydrogen, and Oxygen Project Final Report

    Energy Technology Data Exchange (ETDEWEB)

    Sinskey, Anthony J. [MIT; Worden, Robert Mark [Michigan State University MSU; Brigham, Christopher [MIT; Lu, Jingnan [MIT; Quimby, John Westlake [MIT; Gai, Claudia [MIT; Speth, Daan [MIT; Elliott, Sean [Boston University; Fei, John Qiang [MIT; Bernardi, Amanda [MIT; Li, Sophia [MIT; Grunwald, Stephan [MIT; Grousseau, Estelle [MIT; Maiti, Soumen [MSU; Liu, Chole [MSU

    2013-12-16

    This research project is a collaboration between the Sinskey laboratory at MIT and the Worden laboratory at Michigan State University. The goal of the project is to produce Isobutanol (IBT), a branched-chain alcohol that can serve as a drop-in transportation fuel, through the engineered microbial biosynthesis of Carbon Dioxide, Hydrogen, and Oxygen using a novel bioreactor. This final technical report presents the findings of both the biological engineering work at MIT that extended the native branched-chain amino acid pathway of the wild type Ralstonia eutropha H16 to perform this biosynthesis, as well as the unique design, modeling, and construction of a bioreactor for incompatible gasses at Michigan State that enabled the operational testing of the complete system. This 105 page technical report summarizing the three years of research includes 72 figures and 11 tables of findings. Ralstonia eutropha (also known as Cupriavidus necator) is a Gram-negative, facultatively chemolithoautotrophic bacteria. It has been the principle organism used for the study of polyhydroxybutyrate (PHB) polymer biosynthesis. The wild-type Ralstonia eutropha H16 produces PHB as an intracellular carbon storage material while under nutrient stress in the presence of excess carbon. Under this stress, it can accumulate approximately 80 % of its cell dry weight (CDW) as this intracellular polymer. With the restoration of the required nutrients, the cells are then able to catabolize this polymer. If extracted from the cell, this PHB polymer can be processed into biodegradable and biocompatible plastics, however for this research, it is the efficient metabolic pathway channeling the captured carbon that is of interest. R. eutropha is further unique in that it contains two carbon-fixation Calvin–Benson–Bassham cycle operons, two oxygen-tolerant hydrogenases, and several formate dehydrogenases. It has also been much studied for its ability in the presence of oxygen, to fix carbon dioxide

  16. 'Low-acid' sulfide oxidation using nitrate-enriched groundwater

    Science.gov (United States)

    Donn, Michael; Boxall, Naomi; Reid, Nathan; Meakin, Rebecca; Gray, David; Kaksonen, Anna; Robson, Thomas; Shiers, Denis

    2016-04-01

    Acid drainage (AMD/ARD) is undoubtedly one of the largest environmental, legislative and economic challenges facing the mining industry. In Australia alone, at least 60m is spent on AMD related issues annually, and the global cost is estimated to be in the order of tens of billions US. Furthermore, the challenge of safely and economically storing or treating sulfidic wastes will likely intensify because of the trend towards larger mines that process increasingly higher volumes of lower grade ores and the associated sulfidic wastes and lower profit margins. While the challenge of managing potentially acid forming (PAF) wastes will likely intensify, the industrial approaches to preventing acid production or ameliorating the effects has stagnated for decades. Conventionally, PAF waste is segregated and encapsulated in non-PAF tips to limit access to atmospheric oxygen. Two key limitations of the 'cap and cover' approach are: 1) the hazard (PAF) is not actually removed; only the pollutant linkage is severed; and, 2) these engineered structures are susceptible to physical failure in short-to-medium term, potentially re-establishing that pollutant linkage. In an effort to address these concerns, CSIRO is investigating a passive, 'low-acid' oxidation mechanism for sulfide treatment, which can potentially produce one quarter as much acidity compared with pyrite oxidation under atmospheric oxygen. This 'low-acid' mechanism relies on nitrate, rather than oxygen, as the primary electron accepter and the activity of specifically cultured chemolithoautotrophic bacteria and archaea communities. This research was prompted by the observation that, in deeply weathered terrains of Australia, shallow (oxic to sub-oxic) groundwater contacting weathering sulfides are commonly inconsistent with the geochemical conditions produced by ARD. One key characteristic of these aquifers is the natural abundance of nitrate on a regional scale, which becomes depleted around the sulfide bodies, and

  17. Survivability and growth kinetics of methanogenic archaea at various pHs and pressures: Implications for deep subsurface life on Mars

    Science.gov (United States)

    Sinha, Navita; Nepal, Sudip; Kral, Timothy; Kumar, Pradeep

    2017-02-01

    different pHs and pressures on the metabolic activities of M. wolfeii, we also calculated their growth rate by measuring methane concentration in the headspace gas samples at regular intervals. In acidic conditions, the growth rate (γ) of M. wolfeii increased with the increase in pressure. In neutral and alkaline conditions, the growth rate (γ) of M. wolfeii initially increased with pressure, but decreased upon further increase of pressure. To investigate the effect of combined pH, pressure, and temperature on the morphology of M. wolfeii, we took phase contrast images of the cells. We did not find any obvious significant alteration in the morphology of M. wolfeii cells. Methanogens, chemolithoautotrophic anaerobic microorganisms, are considered as ideal model microorganisms for Mars. In light of research presented here, we suggest that at least one methanogen, M. wolfeii, could survive in the deep subsurface environment of Mars.

  18. 洋底热液喷口系统的微生物成矿研究进展%THE RECENT PROGRESS OF SUBMARINE HYDROTHERMAL BIOMINERALIZATION

    Institute of Scientific and Technical Information of China (English)

    孙治雷; 何拥军; 李军; 齐崇阳; 李季伟; 刘维亮

    2011-01-01

    The study of biomineralization in modern hydrothermal vent system is one of the keys to the research and the exploring of the early history of the earth, the evolution of life, the subsurface biosphere and the study of terrestrial planets (such as the Mar). It has in the past decade become one of the focuses of geobiological research with the application of the microelectronic technology and molecular biology technology. Available information indicates that microorganisms play a critical role in the formation of oxy-hydroxides (for instance, Fe, Mn, S or Si oxyhydroxide) and silicates in the hydrothermal systems of the earth. Furthermore, the biomineralization of modern chemolithoautotrophic microorganisms has been iden-tified to be the nexus of the interaction between the geoshpere and the biosphere and one of the forces to push forward the in-depth study of bioscience and geosciences In this paper, we summarized the ongoing research of hydrothermal bionmieralzaiton, including the biogenic minerals, the microbial biodiversity and the interactions between the minerals and microorganisms. In the foreseeable future, the research of hydrothermal biomineralization will inspire both the development of geosciences and biosciences and deepen our understanding of the earth history, life evolution and even astrobiology.%现代洋底热液喷口系统的微生物成矿研究进展是地球自身发展、生命演化、洋底下的生物圈层以及天文微生物探索工作得以有效开展的重要保障.近10余年来,随着微电子和分子生物学技术的发展,热液系统微生物成矿研究得以拓展和深入,逐渐成为地质微生物学研究的一个热点领域.当前在全球的热液喷口系统已经发现微生物在包括Fe、Mn、S、Si的氧化物以及硅酸盐矿物的形成过程中起到了重要甚至关键性的作用,同时热液喷口系统依赖于无机化能代谢活动存在的微生物的矿化成为人们理解生命形式与无机

  19. Biooxidación de concentrados de arsenopirita por Acidithiobacillus ferrooxidans en erlenmeyer agitados

    Directory of Open Access Journals (Sweden)

    Juan David Ospina

    2012-03-01

    los microorganismos, sugiriendo que a menor tamaño del sustrato empleado mayor dificultad se le presenta al microorganismo para oxidar el mineral. Palabras clave: arsénico; lixiviación; quimiolitótrofos; ATP. Abstract Arsenopyrite biooxidation process was evaluated with A. ferrooxidans ATCC 23270. The microorganisms were previously adapted to mineral and two different Tyler mesh sizes, 200 (~75μm and 325 (~45μm. Also, the mineral concentration was made by DRX and MOLPP/LR under ASTM D 2799. The microorganisms were adapted through gradual decreasing of ferrous sulphate in successive state and subsequent arsenopyrite concentration increase. Finally, biooxidation process was carried out without Fe2+. After thirty days of process, Arsenic bioleaching was 7550 mgL-1(18,7% and 2850 mgL-1 (7,1% for the 200 and 325 Tyler meshes, respectively. On the other hand, bacterial growth curve showed, between 6 and 21 days of process that the average bacterial population was 1,70x108 cel.mL‐1 y de 8,00x107 cel.mL‐1for 200 and 325 Tyler mesh respectively. For this reason, the particle size played an important role in the adaption kinetics of microorganism. The results showed that the microorganism oxide the larger particle size of the mineral easier.  Keywords: arsenic; lixiviation; chemolithoautotrophic; ATP.

  20. New Understanding on Metabolism of Anaerobic Ammonium Oxidation Bacteria Based on Metagenomics Technology%基于宏基因组技术获得的对厌氧氨氧化菌代谢的新理解

    Institute of Scientific and Technical Information of China (English)

    丁爽; 郑平; 陆慧锋; 唐崇俭

    2012-01-01

    厌氧氨氧化菌(Anaerobic ammonium oxidation bacteria,AAOB)是化能自养菌,由于其生理代谢的奇异性、细胞结构的特殊性以及对氮素循环的重要性,已成为环境工程、微生物以及海洋生物学等领域的研究热点.然而.AAOB未能实现纯培养的现状已成为AAOB代谢途径研究的巨大障碍近年来兴起的宏基因组技术(Metagenomics)为AAOB代谢途径的研究提供了新手段.采用宏基因组技术,可直接研究微生物群体中某特定微生物基因组的结构与功能,摆脱了传统微生物学研究对纯培养的依赖,使未培养微生物的认识和开发成为可能本文首先简述获取AAOB宏基因组信息的过程,然后通过比较由传统代谢研究方法和宏基因组技术获得的AAOB代谢途径的研究成果,论述基于宏基因组技术获得的对AAOB代谢的新理解,得出以下结果和结论:1)AAOB的碳素固定途径为乙酰辅酶A途径,碳素固定的还原力来自NADH或者QH2;2)AAOB氮素转化的重要中问产物是NO,而非NH2OH,并提出了以NO为核心的AAOB代谢的改进模型;3)AAOB的ATP合成途径为氧化磷酸化,推测的电子传递途径为N2H4-QH2-细胞色素bc1 复合体;细胞色素bc1复合体再将电子用于NO2还原和N2H4合成AAOB的宏基因组技术使AAOB代谢途径的研究更具方向性.随着分子生物学理论和技术的不断发展,宏基因组学的升级技术(如宏转录组学、宏蛋白质组学)将为AAOB代谢途径的研究提供新的方法与平台.%Anaerobic ammonium oxidation bacteria (AAOB) belong to chemolitho-autotrophs. AAOB have become one of the research hotspots in the field of environmental engineering, microbiology and oceanography because of their specificities in metabolism, cell structure and nitrogen cycle. However, AAOB can not been cultivated in pure culture, which has become a great obstacle to study their metabolic pathways in further. Nowadays, fast-developing metagenomics provides

  1. Site Selection for Mars Exopaleontology in 2001

    Science.gov (United States)

    Farmer, Jack

    1998-01-01

    subsequent diagenetic change or metamorphism. In this context, host rocks composed of highly ordered, chemically-stable mineral phases, like silica (forming cherts) or phosphate (forming phosphorites), are especially favored. Such lithologies tend to have very long crustal residence times and (along with carbonates and shales), are the most common host rocks for the Precambrian microfossil record on Earth. If we assume that a subsurface hydrosphere has been present throughout martian history, then life could have originated there at any time, perhaps emerging at the surface periodically when climate changes, induced by external forcing or endogenous processes (e.g. volcanism), allowed liquid water to exist at the surface. The recent discovery of subsurface chemolithoautotrophic organisms which are capable of synthesizing organic substrates from C02 and H2 liberated from the aqueous weathering of basalt, is especially. relevant as a model for martian life. While a subsurface habitable zone may yet exist on Mars, access to such environments will likely require drilling to depths of several kilometers. Given the technological challenge of deep drilling, this is unlikely to occur prior to human missions. So, even if there is extant life on Mars today in subsurface habitats, it may be much easier to find its fossil counterparts in ancient deposits exposed at the surface. In exploring for a fossil record in subsurface environments on Mars there are several geological situations that may provide access to the appropriate materials. These include 1) ejecta from impact craters, 2) talus slopes, debris flows or alluvial fans developed below the walls of deep canyons, and 3) the deposits of outflood channels. Examples of aqueous mineral deposits of formed in subsurface environments that could harbor a microbial fossil record include such things as cements in detrital sedimentary rocks, low temperature diagenetic minerals deposited in veins, or filling vesicles in volcanic rocks, and

  2. جداسازی و شناسایی باکتری‌های تجزیه کننده گازوئیل از خاک‌های آلوده به مواد نفتی در استان خوزستان

    Directory of Open Access Journals (Sweden)

    مهدی حسن شاهیان

    2016-11-01

    cloacae strain L2 (72درصد بود. بیشترین آب‌گریزی سطح سلولی مربوط به باکتری مربوط به سویۀstrain K2 Klebsiella oxytoca (76درصد و بیشترین فعالیت امولسیون‌کنندگی مربوط به سویۀ  strain B2  Achromobacter denitrificans (58درصد بود. ‌ بحث و نتیجه‌گیری: نتایج این پژوهش ثابت کرد که باکتری‌های تجزیه‌کنندۀ گازوئیل در خاک‌های آلوده در ایران از تنوع مناسبی برخوردار هستند و پتانسیل استفاده در حذف آلودگی گازوئیلی از خاک را دارند.

  3. Engineering Ralstonia eutropha for Production of Isobutanol (IBT) Motor Fuel from Carbon Dioxide, Hydrogen, and Oxygen Project Final Report

    Energy Technology Data Exchange (ETDEWEB)

    Sinskey, Anthony J. [MIT; Worden, Robert Mark [Michigan State University MSU; Brigham, Christopher [MIT; Lu, Jingnan [MIT; Quimby, John Westlake [MIT; Gai, Claudia [MIT; Speth, Daan [MIT; Elliott, Sean [Boston University; Fei, John Qiang [MIT; Bernardi, Amanda [MIT; Li, Sophia [MIT; Grunwald, Stephan [MIT; Grousseau, Estelle [MIT; Maiti, Soumen [MSU; Liu, Chole [MSU

    2013-12-16

    This research project is a collaboration between the Sinskey laboratory at MIT and the Worden laboratory at Michigan State University. The goal of the project is to produce Isobutanol (IBT), a branched-chain alcohol that can serve as a drop-in transportation fuel, through the engineered microbial biosynthesis of Carbon Dioxide, Hydrogen, and Oxygen using a novel bioreactor. This final technical report presents the findings of both the biological engineering work at MIT that extended the native branched-chain amino acid pathway of the wild type Ralstonia eutropha H16 to perform this biosynthesis, as well as the unique design, modeling, and construction of a bioreactor for incompatible gasses at Michigan State that enabled the operational testing of the complete system. This 105 page technical report summarizing the three years of research includes 72 figures and 11 tables of findings. Ralstonia eutropha (also known as Cupriavidus necator) is a Gram-negative, facultatively chemolithoautotrophic bacteria. It has been the principle organism used for the study of polyhydroxybutyrate (PHB) polymer biosynthesis. The wild-type Ralstonia eutropha H16 produces PHB as an intracellular carbon storage material while under nutrient stress in the presence of excess carbon. Under this stress, it can accumulate approximately 80 % of its cell dry weight (CDW) as this intracellular polymer. With the restoration of the required nutrients, the cells are then able to catabolize this polymer. If extracted from the cell, this PHB polymer can be processed into biodegradable and biocompatible plastics, however for this research, it is the efficient metabolic pathway channeling the captured carbon that is of interest. R. eutropha is further unique in that it contains two carbon-fixation Calvin–Benson–Bassham cycle operons, two oxygen-tolerant hydrogenases, and several formate dehydrogenases. It has also been much studied for its ability in the presence of oxygen, to fix carbon dioxide