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Sample records for moderately thermophilic chlorobium

  1. Two exopolyphosphatases with distinct molecular architectures and substrate specificities from the thermophilic green-sulfur bacterium Chlorobium tepidum TLS.

    PubMed

    Albi, Tomás; Serrano, Aurelio

    2014-09-01

    The genome of the thermophilic green-sulfur bacterium Chlorobium tepidum TLS possesses two genes encoding putative exopolyphosphatases (PPX; EC 3.6.1.11), namely CT0099 (ppx1, 993 bp) and CT1713 (ppx2, 1557 bp). The predicted polypeptides of 330 and 518 aa residues are Ppx-GppA phosphatases of different domain architectures - the largest one has an extra C-terminal HD domain - which may represent ancient paralogues. Both ppx genes were cloned and overexpressed in Escherichia coli BL21(DE3). While CtPPX1 was validated as a monomeric enzyme, CtPPX2 was found to be a homodimer. Both PPX homologues were functional, K(+)-stimulated phosphohydrolases, with an absolute requirement for divalent metal cations and a marked preference for Mg(2+). Nevertheless, they exhibited remarkably different catalytic specificities with regard to substrate classes and chain lengths. Even though both enzymes were able to hydrolyse the medium-size polyphosphate (polyP) P13-18 (polyP mix with mean chain length of 13-18 phosphate residues), CtPPX1 clearly reached its highest catalytic efficiency with tripolyphosphate and showed substantial nucleoside triphosphatase (NTPase) activity, while CtPPX2 preferred long-chain polyPs (>300 Pi residues) and did not show any detectable NTPase activity. These catalytic features, taken together with the distinct domain architectures and molecular phylogenies, indicate that the two PPX homologues of Chl. tepidum belong to different Ppx-GppA phosphatase subfamilies that should play specific biochemical roles in nucleotide and polyP metabolisms. In addition, these results provide an example of the remarkable functional plasticity of the Ppx-GppA phosphatases, a family of proteins with relatively simple structures that are widely distributed in the microbial world. PMID:24969471

  2. Bioleaching of multiple metals from contaminated sediment by moderate thermophiles.

    PubMed

    Gan, Min; Jie, Shiqi; Li, Mingming; Zhu, Jianyu; Liu, Xinxing

    2015-08-15

    A moderately thermophilic consortium was applied in bioleaching multiple metals from contaminated sediment. The consortium got higher acidification and metals soubilization efficiency than that of the pure strains. The synergistic effect of the thermophilic consortium accelerated substrates utilization. The utilization of substrate started with sulfur in the early stage, and then the pH declined, giving rise to making use of the pyrite. Community dynamic showed that A. caldus was the predominant bacteria during the whole bioleaching process while the abundance of S. thermotolerans increased together with pyrite utilization. Solubilization efficiency of Zn, Cu, Mn and Cd reached 98%, 94%, 95%, and 89% respectively, while As, Hg, Pb was only 45%, 34%, 22%. Logistic model was used to simulate the bioleaching process, whose fitting degree was higher than 90%. Correlation analysis revealed that metal leaching was mainly an acid solubilization process. Fraction analysis revealed that metals decreased in mobility and bioavailability. PMID:26140749

  3. First Insights into the Genome of the Moderately Thermophilic Bacterium Clostridium tepidiprofundi SG 508T.

    PubMed

    Poehlein, Anja; Friedrich, Ines; Krüger, Larissa; Daniel, Rolf

    2016-01-01

    The moderately thermophilic bacterium Clostridium tepidiprofundi is Gram-positive and belongs to clostridial cluster I. It was isolated from a hydrothermal vent chimney. Substrates utilized by C. tepidiprofundi include casein, peptone, tryptone, yeast extract, beef extract, starch, maltose, and glucose. The genome consists of one replicon (3.06 Mb). PMID:27174286

  4. First Insights into the Genome of the Moderately Thermophilic Bacterium Clostridium tepidiprofundi SG 508T

    PubMed Central

    Poehlein, Anja; Friedrich, Ines; Krüger, Larissa

    2016-01-01

    The moderately thermophilic bacterium Clostridium tepidiprofundi is Gram-positive and belongs to clostridial cluster I. It was isolated from a hydrothermal vent chimney. Substrates utilized by C. tepidiprofundi include casein, peptone, tryptone, yeast extract, beef extract, starch, maltose, and glucose. The genome consists of one replicon (3.06 Mb). PMID:27174286

  5. Draft Genome Sequence of the Moderately Thermophilic Bacterium Schleiferia thermophila Strain Yellowstone (Bacteroidetes).

    PubMed

    Thiel, Vera; Hamilton, Trinity L; Tomsho, Lynn P; Burhans, Richard; Gay, Scott E; Ramaley, Robert F; Schuster, Stephan C; Steinke, Laurey; Bryant, Donald A

    2014-01-01

    The draft genome sequence of the moderately thermophilic bacterium Schleiferia thermophila strain Yellowstone (Bacteroidetes), isolated from Octopus Spring (Yellowstone National Park, WY, USA) was sequenced and comprises 2,617,694 bp in 35 contigs. The draft genome is predicted to encode 2,457 protein coding genes and 37 tRNA encoding genes and two rRNA operons. PMID:25169864

  6. Draft Genome Sequence of the Moderately Thermophilic Bacterium Schleiferia thermophila Strain Yellowstone (Bacteroidetes)

    PubMed Central

    Thiel, Vera; Hamilton, Trinity L.; Tomsho, Lynn P.; Burhans, Richard; Gay, Scott E.; Ramaley, Robert F.; Schuster, Stephan C.; Steinke, Laurey

    2014-01-01

    The draft genome sequence of the moderately thermophilic bacterium Schleiferia thermophila strain Yellowstone (Bacteroidetes), isolated from Octopus Spring (Yellowstone National Park, WY, USA) was sequenced and comprises 2,617,694 bp in 35 contigs. The draft genome is predicted to encode 2,457 protein coding genes and 37 tRNA encoding genes and two rRNA operons. PMID:25169864

  7. Bioleaching of metals from spent refinery petroleum catalyst using moderately thermophilic bacteria: effect of particle size.

    PubMed

    Srichandan, Haragobinda; Singh, Sradhanjali; Pathak, Ashish; Kim, Dong-Jin; Lee, Seoung-Won; Heyes, Graeme

    2014-01-01

    The present work investigated the leaching potential of moderately thermophilic bacteria in the recovery of metals from spent petroleum catalyst of varying particle sizes. The batch bioleaching experiments were conducted by employing a mixed consortium of moderate thermophilic bacteria at 45°C and by using five different particle sizes (from 45 to >2000 μm) of acetone-washed spent catalyst. The elemental mapping by FESEM confirmed the presence of Al, Ni, V and Mo along with sulfur in the spent catalyst. During bioleaching, Ni (92-97%) and V (81-91%) were leached in higher concentrations, whereas leaching yields of Al (23-38%) were found to be lowest in all particle sizes investigated. Decreasing the particle size from >2000 μm to 45-106 μm caused an increase in leaching yields of metals during initial hours. However, the final metals leaching yields were almost independent of particle sizes of catalyst. Leaching kinetics was observed to follow the diffusion-controlled model showing the linearity more close than the chemical control. The results of the present study suggested that bioleaching using moderate thermophilic bacteria was highly effective in removing the metals from spent catalyst. Moreover, bioleaching can be conducted using spent catalyst of higher particle size (>2000 μm), thus saving the grinding cost and making process attractive for larger scale application. PMID:24679088

  8. Bioleaching of electronic scrap by mixed culture of moderately thermophilic microorganisms

    NASA Astrophysics Data System (ADS)

    Ivǎnuş, D.; ǎnuş, R. C., IV; Cǎlmuc, F.

    2010-06-01

    A process for the metal recovery from electronic scrap using bacterial leaching was investigated. A mixed culture of moderately thermophilic microorganisms was enriched from acid mine drainages (AMDs) samples collected from several sulphide mines in Romania, and the bioleaching of electronic scrap was conducted both in shake flask and bioreactor. The results show that in the shake flask, the mixture can tolerate 50 g/L scrap after being acclimated to gradually increased concentrations of scrap. The copper extraction increases obviously in bioleaching of scrap with moderately thermophilic microorganisms supplemented with 0.4 g/L yeast extract at 180 r/min, 74% copper can be extracted in the pulp of 50 g/L scrap after 20 d. Compared with copper extractions of mesophilic culture, unacclimated culture and acclimated culture without addition of yeast extract, that of accliniated culture with addition of yeast extract is increased by 53%, 44% and 16%, respectively. In a completely stirred tank reactor, the mass fraction of copper and total iron extraction reach up to 81% and 56%, respectively. The results also indicate that it is necessary to add a large amount of acid to the pulp to extract copper from electronic scrap effectively.

  9. Single-step bioconversion of lignocellulose to hydrogen using novel moderately thermophilic bacteria

    PubMed Central

    2014-01-01

    Background Consolidated bioprocessing (CBP) of lignocellulosic biomass to hydrogen offers great potential for lower cost and higher efficiency compared to processes featuring dedicated cellulase production. Current studies on CBP-based hydrogen production mainly focus on using the thermophilic cellulolytic bacterium Clostridium thermocellum and the extremely thermophilic cellulolytic bacterium Caldicellulosiruptor saccharolyticus. However, no studies have demonstrated that the strains in the genus Thermoanaerobacterium could be used as the sole microorganism to accomplish both cellulose degradation and H2 generation. Results We have specifically screened for moderately thermophilic cellulolytic bacteria enabling to produce hydrogen directly from conversion of lignocellulosic materials. Three new strains of thermophilic cellulolytic bacteria in the genus Thermoanaerobacterium growing at a temperature of 60°C were isolated. All of them grew well on various plant polymers including microcrystalline cellulose, filter paper, xylan, glucose, and xylose. In particular, the isolated bacterium, designated as Thermoanaerobacterium thermosaccharolyticum M18, showed high cellulolytic activity and a high yield of H2. When it was grown in 0.5% microcrystalline cellulose, approximately 82% cellulose was consumed, and the H2 yield and maximum production rate reached 10.86 mmol/g Avicel and 2.05 mmol/L/h, respectively. Natural lignocellulosic materials without any physicochemical or biological pretreatment also supported appreciable growth of strain M18, which resulted in 56.07% to 62.71% of insoluble cellulose and hemicellulose polymer degradation in corn cob, corn stalk, and wheat straw with a yield of 3.23 to 3.48 mmol H2/g substrate and an average production rate of 0.10 to 0.13 mmol H2/L/h. Conclusions The newly isolated strain T. thermosaccharolyticum M18 displayed effective degradation of lignocellulose and produced large amounts of hydrogen. This is the first report

  10. Reduction of Soluble Iron and Reductive Dissolution of Ferric Iron-Containing Minerals by Moderately Thermophilic Iron-Oxidizing Bacteria

    PubMed Central

    Bridge, Toni A. M.; Johnson, D. Barrie

    1998-01-01

    Five moderately thermophilic iron-oxidizing bacteria, including representative strains of the three classified species (Sulfobacillus thermosulfidooxidans, Sulfobacillus acidophilus, and Acidimicrobium ferrooxidans), were shown to be capable of reducing ferric iron to ferrous iron when they were grown under oxygen limitation conditions. Iron reduction was most readily observed when the isolates were grown as mixotrophs or heterotrophs with glycerol as an electron donor; in addition, some strains were able to couple the oxidation of tetrathionate to the reduction of ferric iron. Cycling of iron between the ferrous and ferric states was observed during batch culture growth in unshaken flasks incubated under aerobic conditions, although the patterns of oxidoreduction of iron varied in different species of iron-oxidizing moderate thermophiles and in strains of a single species (S. acidophilus). All three bacterial species were able to grow anaerobically with ferric iron as a sole electron acceptor; the growth yields correlated with the amount of ferric iron reduced when the isolates were grown in the absence of oxygen. One of the moderate thermophiles (identified as a strain of S. acidophilus) was able to bring about the reductive dissolution of three ferric iron-containing minerals (ferric hydroxide, jarosite, and goethite) when it was grown under restricted aeration conditions with glycerol as a carbon and energy source. The significance of iron reduction by moderately thermophilic iron oxidizers in both environmental and applied contexts is discussed. PMID:9603832

  11. Bioleaching of chalcopyrite and bornite by moderately thermophilic bacteria: an emphasis on their interactions

    NASA Astrophysics Data System (ADS)

    Zhao, Hong-bo; Wang, Jun; Gan, Xiao-wen; Qin, Wen-qing; Hu, Ming-hao; Qiu, Guan-zhou

    2015-08-01

    Interactions between chalcopyrite and bornite during bioleaching by moderately thermophilic bacteria were investigated mainly by X-ray diffraction, scanning electron microscopy, and electrochemical measurements performed in conjunction with bioleaching experiments. The results showed that a synergistic effect existed between chalcopyrite and bornite during bioleaching by both Acidithiobacillus caldus and Leptospirillum ferriphilum and that extremely high copper extraction could be achieved when chalcopyrite and bornite coexisted in a bioleaching system. Bornite dissolved preferentially because of its lower corrosion potential, and its dissolution was accelerated by the galvanic current during the initial stage of bioleaching. The galvanic current and optimum redox potential of 390-480 mV vs. Ag/AgCl promoted the reduction of chalcopyrite to chalcocite (Cu2S), thus accelerating its dissolution.

  12. Multiple Serotypes of the Moderate Thermophile Thiobacillus caldus, a Limitation of Immunological Assays for Biomining Microorganisms.

    PubMed

    Hallberg, K B; Lindstrom, E B

    1996-11-01

    Phylogenetic and phenotypic analysis indicates that a moderately thermophilic isolate, C-SH12, from Australia belongs to the species Thiobacillus caldus. Antiserum generated against whole cells of T. caldus KU recognized protein antigens common to cell lysates of the three T. caldus strains KU, BC13, and C-SH12 but did not recognize whole cells of isolate C-SH12. Differences in the lipopolysaccharide (LPS) of strain C-SH12 and those of the other two T. caldus strains were found, and the anti-KU antiserum did not recognize the LPS from strain C-SH12. These data indicate that this T. caldus isolate belongs to a serotype different from that of strains KU and BC13. PMID:16535449

  13. Escherichia coli inactivation kinetics in anaerobic digestion of dairy manure under moderate, mesophilic and thermophilic temperatures.

    PubMed

    Pandey, Pramod K; Soupir, Michelle L

    2011-01-01

    Batch anaerobic digestion experiments using dairy manure as feedstocks were performed at moderate (25°C), mesophilic (37°C), and thermophilic (52.5°C) temperatures to understand E. coli, an indicator organism for pathogens, inactivation in dairy manure. Incubation periods at 25, 37, and 52.5°C, were 61, 41, and 28 days respectively. Results were used to develop models for predicting E. coli inactivation and survival in anaerobic digestion. For modeling we used the decay of E. coli at each temperature to calculate the first-order inactivation rate coefficients, and these rates were used to formulate the time - temperature - E. coli survival relationships. We found the inactivation rate coefficient at 52.5°C was 17 and 15 times larger than the inactivation rate coefficients at 25 and 37°C, respectively. Decimal reduction times (D10; time to achieve one log removal) at 25, 37, and 52.5°C, were 9 -10, 7 - 8 days, and < 1 day, respectively. The Arrhenius correlation between inactivation rate coefficients and temperatures over the range 25 -52.5°C was developed to understand the impacts of temperature on E. coli inactivation rate. Using this correlation, the time - temperature - E. coli survival relationships were derived. Besides E. coli inactivation, impacts of temperature on biogas production, methane content, pH change, ORP, and solid reduction were also studied. At higher temperatures, biogas production and methane content was greater than that at low temperatures. While at thermophilic temperature pH was increased, at mesophilic and moderate temperatures pH were reduced over the incubation period. These results can be used to understand pathogen inactivation during anaerobic digestion of dairy manure, and impacts of temperatures on performance of anaerobic digesters treating dairy manure. PMID:21906374

  14. Ruegeria intermedia sp. nov., a moderately thermophilic bacterium isolated from a coastal hot spring.

    PubMed

    Kämpfer, P; Arun, A B; Rekha, P D; Busse, H-J; Young, C-C; Glaeser, S P

    2013-07-01

    A cream-coloured, Gram-negative, aerobic, non-motile moderately thermophilic, rod-to-irregular-shaped bacterium, CC-GIMAT-2(T), was isolated from a coastal hot spring of Green Island (Lutao), located off Taituang, Taiwan, on marine agar 2216. The 16S rRNA gene sequence analysis and subsequent comparisons showed that it was placed into the genus Ruegeria with 97.4 % similarity to Ruegeria lacuscaerulensis ITI-1157(T), and a lower sequence similarity to all other species of the genus Ruegeria. Reconstruction of phylogenetic trees indicated that strain CC-GIMAT-2(T) clustered within the genus Ruegeria. Robust tree topology for the genus Ruegeria including the new strain was only obtained by including all Rhodobacteraceae type strains but not if the analysis was limited to few selected taxa. The quinone system contained exclusively ubiquinone Q-10 and the fatty acid profile consisted mainly of C18 : 1ω7c, 11-methyl C18 : 1ω7c and C12 : 0 3-OH. The predominant polar lipids were diphosphatidylglycerol, phosphatidylglycerol and an unidentified aminolipid. Other lipids were detected in moderate to minor amounts. The characteristic feature of the polyamine pattern was the predominant triamine spermidine. On the basis of the 16S rRNA gene sequence analysis and of the chemotaxonomic and physiological data, strain CC-GIMAT-2(T) represents a novel species of the genus Ruegeria, for which the name Ruegeria intermedia sp. nov. is proposed. The type strain is CC-GIMAT-2(T) ( = CCUG 59209(T) = LMG 25539(T) = CCM 7758(T)). PMID:23243093

  15. Differential expression of extracellular thiol groups of moderately thermophilic Sulfobacillus thermosulfidooxidans and extremely thermophilic Acidianus manzaensis grown on S(0) and Fe (2.).

    PubMed

    Liu, Hong-Chang; Xia, Jin-Lan; Nie, Zhen-Yuan; Zhen, Xiang-Jun; Zhang, Li-Juan

    2015-08-01

    Bio-oxidation of elemental sulfur (S(0)) is very important in bioleaching and sulfur cycle. S(0) was proposed to be first activated by reacting with reactive thiol groups (-SH) of outer membrane proteins, forming -S n H (n ≥ 2) complexes. The differential expression of -SH of moderately thermophilic Sulfobacillus thermosulfidooxidans and extremely thermophilic Acidianus manzaensis grown on Fe(2+) and S(0) was investigated by synchrotron radiation-based scanning transmission X-ray microscopy (STXM) imaging and micro-beam X-ray fluorescence (μ-XRF) mapping. The STXM imaging and μ-XRF mapping of extracellular -SH were based on the analysis of Ca(2+) bound on the cell. By comparing Ca(2+) of the cells with and without labeling by Ca(2+), the distribution and content of thiol groups were obtained. The results showed that, for both S. thermosulfidooxidans and A. manzaensis, the expression of extracellular -SH of S(0)-grown cells was higher than that of Fe(2+)-grown cells. Statistical analysis indicated that the expression of extracellular -SH for S. thermosulfidooxidans and A. manzaensis grown on S(0) was 2.37 times and 2.14 times, respectively, to that on Fe(2+). These results evidently demonstrate that the extracellular thiol groups are most probably involved in elemental sulfur activation and oxidation of the acidophilic sulfur-oxidizing microorganisms. PMID:25983134

  16. Copper extraction from coarsely ground printed circuit boards using moderate thermophilic bacteria in a rotating-drum reactor

    SciTech Connect

    Rodrigues, Michael L.M.; Leão, Versiane A.; Gomes, Otavio; Lambert, Fanny; Bastin, David; Gaydardzhiev, Stoyan

    2015-07-15

    Highlights: • Copper bioleaching from PCB (20 mm) by moderate thermophiles was demonstrated. • Larger PCB sheets enable a cost reduction due to the elimination of fine grinding. • Crushing generated cracks in PCB increasing the copper extraction. • A pre-treatment step was necessary to remove the lacquer coating. • High copper extractions (85%) were possible with pulp density of up to 25.0 g/L. - Abstract: The current work reports on a new approach for copper bioleaching from Printed Circuit Board (PCB) by moderate thermophiles in a rotating-drum reactor. Initially leaching of PCB was carried out in shake flasks to assess the effects of particle size (−208 μm + 147 μm), ferrous iron concentration (1.25–10.0 g/L) and pH (1.5–2.5) on copper leaching using mesophile and moderate thermophile microorganisms. Only at a relatively low solid content (10.0 g/L) complete copper extraction was achieved from the particle size investigated. Conversely, high copper extractions were possible from coarse-ground PCB (20 mm-long) working with increased solids concentration (up to 25.0 g/L). Because there was as the faster leaching kinetics at 50 °C Sulfobacillus thermosulfidooxidans was selected for experiments in a rotating-drum reactor with the coarser-sized PCB sheets. Under optimal conditions, copper extraction reached 85%, in 8 days and microscopic observations by SEM–EDS of the on non-leached and leached material suggested that metal dissolution from the internal layers was restricted by the fact that metal surface was not entirely available and accessible for the solution in the case of the 20 mm-size sheets.

  17. Anoxybacillus suryakundensis sp. nov, a Moderately Thermophilic, Alkalitolerant Bacterium Isolated from Hot Spring at Jharkhand, India

    PubMed Central

    Deep, Kamal; Poddar, Abhijit; Das, Subrata K.

    2013-01-01

    Four closely related facultative anaerobe, moderately thermophilic, Gram positive rods (JS1T, JS5, JS11, and JS15) were isolated from sediment samples from a hot spring at Suryakund, Jharkhand, India. Colonies were pale yellow, rough surface with uneven edges on TSA after 72 h incubation. Heterotrophic growth was observed at 40-60°C and pH 5.5-11.5; optimum growth occurred at 55°C and pH 7.5. 16S rRNA gene sequence analysis revealed the strains belong to genus Anoxybacillus. DNA-DNA homology values among strains were above 70% and showed distinct ERIC and REP PCR profile. On the basis of morphology and biochemical characteristics, strain JS1T was studied further. Strain JS1T showed 99.30% sequence similarity with A. flavithermus subsp. yunnanensis, 99.23% with A. mongoliensis, 99.16% with A. eryuanensis, 98.74% with A. flavithermus subsp. flavithermus, 98.54% with A. tengchongensis, 98.51% with A. pushchinoensis, 97.91% with A. thermarum, 97.82% with A. kaynarcensis, 97.77% with A. ayderensis and A. kamchatkensis, 97.63% with A. salavatliensis, 97.55% with A. kestanbolensis, 97.48% with A. contaminans, 97.27% with A. gonensis and 97.17% with A. voinovskiensis. In 16S rRNA secondary structure based phylogenetic comparison, strain JS1T was clustered with Anoxybacillus eryuanensis, A. mongoliensis, and A. flavithermus subsp. yunnanensis and showed 15 species specific base substitutions with maximum variability in helix 6. Moreover, DNA-DNA relatedness between JS1T and the closely related type strains were well below 70%. The DNA G+C content was 42.1 mol%. The major fatty acids were C15:0 iso, C16:0 iso and C17:0iso. The polar lipids were a phosphatidylgylycerol, a diphosphatidylglycerol, a phosphatidylethnolamine, a phosphatidylcholine, a phosphatidyl monomethylethanolamine and four unknown lipids. Based on polyphasic approach, strain JS1T represent a novel species of the genus Anoxybacillus for which Anoxybacillus suryakundensis sp. nov. is proposed. The type

  18. Caloramator quimbayensis sp. nov., an anaerobic, moderately thermophilic bacterium isolated from a terrestrial hot spring.

    PubMed

    Rubiano-Labrador, Carolina; Baena, Sandra; Díaz-Cárdenas, Carolina; Patel, Bharat K C

    2013-04-01

    An anaerobic, moderately thermophilic, terminal-spore-forming bacterium, designated strain USBA A(T), was isolated from a terrestrial hot spring located at an altitude of 2683 m in the Andean region of Colombia (04° 50' 14.0″ N 75° 32' 53.4″ W). Cells of strain USBA A(T) were Gram-stain-positive, straight to slightly curved rods (0.9×2.5 µm), that were arranged singly or in pairs, and were motile by means of flagella. Growth occurred at 37-55 °C and pH 6.0-8.0, with a doubling time of 2 h under the optimal conditions (50 °C and pH 7.0). Glucose fermentation in strain USBA A(T) required yeast extract or peptone (each at 0.2 %, w/v). The novel strain fermented sugars, amino acids, Casamino acids, propanol, propionate, starch and dextrin, but no growth was observed on galactose, lactose, xylose, histidine, serine, threonine, benzoate, butyrate, lactate, pyruvate, succinate, methanol, ethanol, glycerol, casein, gelatin or xylan. The end products of glucose fermentation were formate, acetate, ethanol and lactate. Strain USBA A(T) did not grow autotrophically (with CO2 as carbon source and H2 as electron donor) and did not reduce thiosulfate, sulfate, elemental sulfur, sulfite, vanadium (V) or Fe (III) citrate. Growth of strain USBA A(T) was inhibited by ampicillin, chloramphenicol, kanamycin, penicillin and streptomycin (each at 10 µg ml(-1)). The predominant fatty acids were iso-C15 : 0, C16 : 0 and iso-C17 : 0 and the genomic DNA G+C content was 32.6 mol%. 16S rRNA gene sequence analysis indicated that strain USBA A(T) belonged in the phylum Firmicutes and that its closest relative was Caloramator viterbiensis JW/MS-VS5(T) (95.0 % sequence similarity). A DNA-DNA relatedness value of only 30 % was recorded in hybridization experiments between strain USBA A(T) and Caloramator viterbiensis DSM 13723(T). Based on the phenotypic, chemotaxonomic and phylogenetic evidence and the results of the DNA-DNA hybridization experiments, strain USBA A

  19. Fermentation of Corn Fiber Hydrolysate to Lactic Acid by the Moderate Thermophile Bacillus coagulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Composted manure from a dairy farm in Texas was examined for thermophilic microorganisms by enrichment in xylose broth medium. Forty randomly picked isolates were identified as strains of Bacillus coagulans by sequence analysis of rRNA genes. One strain, designated as MXL-9, could convert mixed su...

  20. Acid-Tolerant Moderately Thermophilic Methanotrophs of the Class Gammaproteobacteria Isolated From Tropical Topsoil with Methane Seeps

    PubMed Central

    Islam, Tajul; Torsvik, Vigdis; Larsen, Øivind; Bodrossy, Levente; Øvreås, Lise; Birkeland, Nils-Kåre

    2016-01-01

    Terrestrial tropical methane seep habitats are important ecosystems in the methane cycle. Methane oxidizing bacteria play a key role in these ecosystems as they reduce methane emissions to the atmosphere. Here, we describe the isolation and initial characterization of two novel moderately thermophilic and acid-tolerant obligate methanotrophs, assigned BFH1 and BFH2 recovered from a tropical methane seep topsoil habitat. The new isolates were strictly aerobic, non-motile, coccus-shaped and utilized methane and methanol as sole carbon and energy source. Isolates grew at pH range 4.2–7.5 (optimal 5.5–6.0) and at a temperature range of 30–60°C (optimal 51–55°C). 16S rRNA gene phylogeny placed them in a well-separated branch forming a cluster together with the genus Methylocaldum as the closest relatives (93.1–94.1% sequence similarity). The genes pmoA, mxaF, and cbbL were detected, but mmoX was absent. Strains BFH1 and BFH2 are, to our knowledge, the first isolated acid-tolerant moderately thermophilic methane oxidizers of the class Gammaproteobacteria. Each strain probably denotes a novel species and they most likely represent a novel genus within the family Methylococcaceae of type I methanotrophs. Furthermore, the isolates increase our knowledge of acid-tolerant aerobic methanotrophs and signify a previously unrecognized biological methane sink in tropical ecosystems. PMID:27379029

  1. Copper extraction from coarsely ground printed circuit boards using moderate thermophilic bacteria in a rotating-drum reactor.

    PubMed

    Rodrigues, Michael L M; Leão, Versiane A; Gomes, Otavio; Lambert, Fanny; Bastin, David; Gaydardzhiev, Stoyan

    2015-07-01

    The current work reports on a new approach for copper bioleaching from Printed Circuit Board (PCB) by moderate thermophiles in a rotating-drum reactor. Initially leaching of PCB was carried out in shake flasks to assess the effects of particle size (-208μm+147μm), ferrous iron concentration (1.25-10.0g/L) and pH (1.5-2.5) on copper leaching using mesophile and moderate thermophile microorganisms. Only at a relatively low solid content (10.0g/L) complete copper extraction was achieved from the particle size investigated. Conversely, high copper extractions were possible from coarse-ground PCB (20mm-long) working with increased solids concentration (up to 25.0g/L). Because there was as the faster leaching kinetics at 50°C Sulfobacillus thermosulfidooxidans was selected for experiments in a rotating-drum reactor with the coarser-sized PCB sheets. Under optimal conditions, copper extraction reached 85%, in 8days and microscopic observations by SEM-EDS of the on non-leached and leached material suggested that metal dissolution from the internal layers was restricted by the fact that metal surface was not entirely available and accessible for the solution in the case of the 20mm-size sheets. PMID:25899037

  2. Acid-Tolerant Moderately Thermophilic Methanotrophs of the Class Gammaproteobacteria Isolated From Tropical Topsoil with Methane Seeps.

    PubMed

    Islam, Tajul; Torsvik, Vigdis; Larsen, Øivind; Bodrossy, Levente; Øvreås, Lise; Birkeland, Nils-Kåre

    2016-01-01

    Terrestrial tropical methane seep habitats are important ecosystems in the methane cycle. Methane oxidizing bacteria play a key role in these ecosystems as they reduce methane emissions to the atmosphere. Here, we describe the isolation and initial characterization of two novel moderately thermophilic and acid-tolerant obligate methanotrophs, assigned BFH1 and BFH2 recovered from a tropical methane seep topsoil habitat. The new isolates were strictly aerobic, non-motile, coccus-shaped and utilized methane and methanol as sole carbon and energy source. Isolates grew at pH range 4.2-7.5 (optimal 5.5-6.0) and at a temperature range of 30-60°C (optimal 51-55°C). 16S rRNA gene phylogeny placed them in a well-separated branch forming a cluster together with the genus Methylocaldum as the closest relatives (93.1-94.1% sequence similarity). The genes pmoA, mxaF, and cbbL were detected, but mmoX was absent. Strains BFH1 and BFH2 are, to our knowledge, the first isolated acid-tolerant moderately thermophilic methane oxidizers of the class Gammaproteobacteria. Each strain probably denotes a novel species and they most likely represent a novel genus within the family Methylococcaceae of type I methanotrophs. Furthermore, the isolates increase our knowledge of acid-tolerant aerobic methanotrophs and signify a previously unrecognized biological methane sink in tropical ecosystems. PMID:27379029

  3. Naphthalene metabolism in Nocardia otitidiscaviarum strain TSH1, a moderately thermophilic microorganism.

    PubMed

    Zeinali, Majid; Vossoughi, Manouchehr; Ardestani, Sussan K

    2008-06-01

    The thermophilic bacterium Nocardia otitidiscaviarum strain TSH1, originally isolated in our laboratory from a petroindustrial wastewater contaminated soil in Iran, grows at 50 degrees C on a broad range of hydrocarbons. Transformation of naphthalene by strain TSH1 which is able to use this two ring-polycyclic aromatic hydrocarbon (PAH) as a sole source of carbon and energy was investigated. The metabolic pathway was elucidated by identifying metabolites, biotransformation studies and monitoring enzyme activities in cell-free extracts. The identification of metabolites suggests that strain TSH1 initiates its attack on naphthalene by dioxygenation at its C-1 and C-2 positions to give 1,2-dihydro-1,2-dihydroxynaphthalene. The intermediate 2-hydroxycinnamic acid, characteristic of the meta-cleavage of the resulting diol was identified in the acidic extract. Apart from typical metabolites of naphthalene degradation known from mesophiles, benzoic acid was identified as an intermediate for the naphthalene pathway of this Nocardia strain. Neither phthalic acid nor salicylic acid metabolites were detected in culture extracts. Enzymatic experiments with cell extract showed the catechol 1,2-dioxygenase activity while transformation of phthalic acid and protocatechuic acid was not observed. The results of enzyme activity assays and identification of benzoic acid in culture extract provide strong indications that further degradation goes through benzoate and beta-ketoadipate pathway. Our results indicate that naphthalene degradation by thermophilic N. otitidiscaviarum strain TSH1 differs from the known pathways found for the thermophilic Bacillus thermoleovorans Hamburg 2 and mesophilic bacteria. PMID:18471862

  4. Draft Genome Sequence of Tepidibacillus decaturensis Strain Z9, an Anaerobic, Moderately Thermophilic, and Heterotrophic Bacterium from the Deep Subsurface of the Illinois Basin, USA.

    PubMed

    Dong, Yiran; Chang, Yun-Juan; Sanford, Robert A; Fouke, Bruce W

    2016-01-01

    The genome of the moderately thermophilic and halotolerant bacteriumTepidibacillus decaturensisstrain Z9 was sequenced. The draft genome comprises three scaffolds, for a total of 2.95 Mb. As the first sequenced genome within the genusTepidibacillus, 2,895 protein-coding genes, 52 tRNA genes, and 3 rRNA operons were predicted. PMID:27056217

  5. Draft Genome Sequence of Tepidibacillus decaturensis Strain Z9, an Anaerobic, Moderately Thermophilic, and Heterotrophic Bacterium from the Deep Subsurface of the Illinois Basin, USA

    PubMed Central

    Chang, Yun-Juan; Sanford, Robert A.; Fouke, Bruce W.

    2016-01-01

    The genome of the moderately thermophilic and halotolerant bacterium Tepidibacillus decaturensis strain Z9 was sequenced. The draft genome comprises three scaffolds, for a total of 2.95 Mb. As the first sequenced genome within the genus Tepidibacillus, 2,895 protein-coding genes, 52 tRNA genes, and 3 rRNA operons were predicted. PMID:27056217

  6. Moderately thermophilic, hydrocarbonoclastic bacterial communities in Kuwaiti desert soil: enhanced activity via Ca(2+) and dipicolinic acid amendment.

    PubMed

    Al-Mailem, D M; Kansour, M K; Radwan, S S

    2015-05-01

    Pristine and oil-contaminated desert soil samples from Kuwait harbored between 10 and 100 cells g(-1) of hydrocarbonoclastic bacteria capable of growth at 50 °C. Enrichment by incubation of moistened soils for 6 months at 50 °C raised those numbers to the magnitude of 10(3) cells g(-1). Most of these organisms were moderately thermophilic and belonged to the genus Bacillus; they grew at 40-50 °C better than at 30 °C. Species belonging to the genera Amycolatopsis, Chelativorans, Isoptericola, Nocardia, Aeribacillus, Aneurinibacillus, Brevibacillus, Geobacillus, Kocuria, Marinobacter and Paenibacillus were also found. This microbial diversity indicates a good potential for hydrocarbon removal in soil at high temperature. Analysis of the same desert soil samples by a culture-independent method (combined, DGGE and 16S rDNA sequencing) revealed dramatically different lists of microorganisms, many of which had been recorded as hydrocarbonoclastic. Many species were more frequent in the oil contaminated than in the pristine soil samples, which may reflect their hydrocarbonoclastic activity in situ. The growth and hydrocarbon consumption potential of all tested isolates were dramatically enhanced by amendment of the cultures with Ca(2+) (up to 2.5 M CaSO4). This enhanced effect was even amplified when in addition 8 % w/v dipicolinic acid was amended. These novel findings are useful in suggesting biotechnologies for waste hydrocarbon remediation at moderately high temperature. PMID:25716145

  7. Construction of conjugative gene transfer system between E. coli and moderately thermophilic, extremely acidophilic Acidithiobacillus caldus MTH-04.

    PubMed

    Liu, Xiangmei; Lin, Jianqun; Zhang, Zheng; Bian, Jiang; Zhao, Qing; Liu, Ying; Lin, Jianqiang; Yan, Wangming

    2007-01-01

    A genetic transfer system for introducing foreign genes to biomining microorganisms is urgently needed. Thus, a conjugative gene transfer system was investigated for a moderately thermophilic, extremely acidophilic biomining bacterium, Acidithiobacillus caldus MTH-04. The broad-host-range IncP plasmids RP4 and R68.45 were transferred directly into A. caldus MTH-04 from Escherichia coli by conjugation at relatively high frequencies. Additionally the broad-host-range IncQ plasmids pJRD215, pVLT33, and pVLT35 were also transferred into A. caldus MTH-04 with the help of plasmid RP4 or strains with plasmid RP4 integrated into their chromosome, such as E. coli SM10. The Km(r) and Sm(r) selectable markers from these plasmids were successfully expressed in A. caldus MTH-04. Futhermore, the IncP and IncQ plasmids were transferred back into E. coli cells from A. caldus MTH-04, thereby confirming the initial transfer of these plasmids from E. coli to A. caldus MTH-04. All the IncP and IncQ plasmids studied were stable in A. caldus MTH-04. Consequently, this development of a conjugational system for A. caldus MTH-04 will greatly facilitate its genetic study. PMID:18051368

  8. Oxidative Dissolution of Arsenopyrite by Mesophilic and Moderately Thermophilic Acidophiles †

    PubMed Central

    Tuovinen, Olli H.; Bhatti, Tariq M.; Bigham, Jerry M.; Hallberg, Kevin B.; Garcia, Oswaldo; Lindström, E. Börje

    1994-01-01

    The purpose of this work was to determine solution- and solid-phase changes associated with the oxidative leaching of arsenopyrite (FeAsS) by Thiobacillus ferrooxidans and a moderately thermoacidophilic mixed culture. Jarosite [KFe3(SO4)2(OH)6], elemental sulfur (S0), and amorphous ferric arsenate were detected by X-ray diffraction as solid-phase products. The oxidation was not a strongly acid-producing reaction and was accompanied by a relatively low redox level. The X-ray diffraction lines of jarosite increased considerably when ferrous sulfate was used as an additional substrate for T. ferroxidans. A moderately thermoacidophilic mixed culture oxidized arsenopyrite faster at 45°C than did T. ferroxidans at 22°C, and the oxidation was accompanied by a nearly stoichiometric release of Fe and As. The redox potential was initially low but subsequently increased during arsenopyrite oxidation by the thermoacidophiles. Jarosite, S0, and amorphous ferric arsenate were also formed under these conditions. PMID:16349379

  9. Expression of Critical Sulfur- and Iron-Oxidation Genes and the Community Dynamics During Bioleaching of Chalcopyrite Concentrate by Moderate Thermophiles.

    PubMed

    Zhou, Dan; Peng, Tangjian; Zhou, Hongbo; Liu, Xueduan; Gu, Guohua; Chen, Miao; Qiu, Guanzhou; Zeng, Weimin

    2015-07-01

    Sulfate adenylyltransferase gene and 4Fe-4S ferredoxin gene are the key genes related to sulfur and iron oxidations during bioleaching system, respectively. In order to better understand the bioleaching and microorganism synergistic mechanism in chalcopyrite bioleaching by mixed culture of moderate thermophiles, expressions of the two energy metabolism genes and community dynamics of free and attached microorganisms were investigated. Specific primers were designed for real-time quantitative PCR to study the expression of these genes. Real-time PCR results showed that sulfate adenylyltransferase gene was more highly expressed in Sulfobacillus thermosulfidooxidans than that in Acidithiobacillus caldus, and expression of 4Fe-4S ferredoxin gene was higher in Ferroplasma thermophilum than that in S. thermosulfidooxidans and Leptospirillum ferriphilum. The results indicated that in the bioleaching system of chalcopyrite concentrate, sulfur and iron oxidations were mainly performed by S. thermosulfidooxidans and F. thermophilum, respectively. The community dynamics results revealed that S. thermosulfidooxidans took up the largest proportion during the whole period, followed by F. thermophilum, A. caldus, and L. ferriphilum. The CCA analysis showed that 4Fe-4S ferredoxin gene expression was mainly affected (positively correlated) by high pH and elevated concentration of ferrous ion, while no factor was observed to prominently influence the expression of sulfate adenylyltransferase gene. PMID:25941022

  10. Genome sequence of the moderately thermophilic sulfur-reducing bacterium Thermanaerovibrio velox type strain (Z-9701(T)) and emended description of the genus Thermanaerovibrio.

    PubMed

    Palaniappan, Krishna; Meier-Kolthoff, Jan P; Teshima, Hazuki; Nolan, Matt; Lapidus, Alla; Tice, Hope; Del Rio, Tijana Glavina; Cheng, Jan-Fang; Han, Cliff; Tapia, Roxanne; Goodwin, Lynne A; Pitluck, Sam; Liolios, Konstantinos; Mavromatis, Konstantinos; Pagani, Ioanna; Ivanova, Natalia; Mikhailova, Natalia; Pati, Amrita; Chen, Amy; Rohde, Manfred; Mayilraj, Shanmugam; Spring, Stefan; Detter, John C; Göker, Markus; Bristow, James; Eisen, Jonathan A; Markowitz, Victor; Hugenholtz, Philip; Kyrpides, Nikos C; Klenk, Hans-Peter; Woyke, Tanja

    2013-10-16

    Thermanaerovibrio velox Zavarzina et al. 2000 is a member of the Synergistaceae, a family in the phylum Synergistetes that is already well-characterized at the genome level. Members of this phylum were described as Gram-negative staining anaerobic bacteria with a rod/vibrioid cell shape and possessing an atypical outer cell envelope. They inhabit a large variety of anaerobic environments including soil, oil wells, wastewater treatment plants and animal gastrointestinal tracts. They are also found to be linked to sites of human diseases such as cysts, abscesses, and areas of periodontal disease. The moderately thermophilic and organotrophic T. velox shares most of its morphologic and physiologic features with the closely related species, T. acidaminovorans. In addition to Su883(T), the type strain of T. acidaminovorans, stain Z-9701(T) is the second type strain in the genus Thermanaerovibrio to have its genome sequence published. Here we describe the features of this organism, together with the non-contiguous genome sequence and annotation. The 1,880,838 bp long chromosome (non-contiguous finished sequence) with its 1,751 protein-coding and 59 RNA genes is a part of the G enomic E ncyclopedia of Bacteria and Archaea project. PMID:24501645

  11. Complete Genome Sequences of Two Interactive Moderate Thermophiles, Paenibacillus napthalenovorans 32O-Y and Paenibacillus sp. 32O-W

    PubMed Central

    Wang, Jia; Stark, Benjamin C.

    2016-01-01

    Microorganisms with the capability to desulfurize petroleum are in high demand with escalating restrictions currently placed on fuel purity. Thermophilic desulfurizers are particularly valuable in high-temperature industrial applications. We report the whole-genome sequences of Paenibacillus napthalenovorans 32O-Y and Paenibacillus sp. 32O-W, which can and cannot, respectively, metabolize dibenzothiophene. PMID:26868401

  12. Biodiversity of acidophilic moderate thermophiles isolated from two sites in Yellowstone National Park and their roles in the dissimilatory oxido-reduction of iron

    SciTech Connect

    D. B. Johnson; D.A. Body; T. A. M. Bridge; D. F. Bruhn; F. F. Roberto

    2001-07-01

    Some of the thermal sites within Yellowstone National Park are extremely acidic and are therefore potential sites for isolating novel strains of acidophilic thermophiles, including those that are involved in the biogeochemical cycling of iron. This paper describes the isolation and characterization of thermotolerant, acidophilic “iron bacteria” isolated from two such sites in Yellowstone National Park, and reports the biodiversity of isolates in terms of their physiological traits and their phylogenetic make-up.

  13. Anaerobic Thermophiles

    PubMed Central

    Canganella, Francesco; Wiegel, Juergen

    2014-01-01

    The term “extremophile” was introduced to describe any organism capable of living and growing under extreme conditions. With the further development of studies on microbial ecology and taxonomy, a variety of “extreme” environments have been found and an increasing number of extremophiles are being described. Extremophiles have also been investigated as far as regarding the search for life on other planets and even evaluating the hypothesis that life on Earth originally came from space. The first extreme environments to be largely investigated were those characterized by elevated temperatures. The naturally “hot environments” on Earth range from solar heated surface soils and water with temperatures up to 65 °C, subterranean sites such as oil reserves and terrestrial geothermal with temperatures ranging from slightly above ambient to above 100 °C, to submarine hydrothermal systems with temperatures exceeding 300 °C. There are also human-made environments with elevated temperatures such as compost piles, slag heaps, industrial processes and water heaters. Thermophilic anaerobic microorganisms have been known for a long time, but scientists have often resisted the belief that some organisms do not only survive at high temperatures, but actually thrive under those hot conditions. They are perhaps one of the most interesting varieties of extremophilic organisms. These microorganisms can thrive at temperatures over 50 °C and, based on their optimal temperature, anaerobic thermophiles can be subdivided into three main groups: thermophiles with an optimal temperature between 50 °C and 64 °C and a maximum at 70 °C, extreme thermophiles with an optimal temperature between 65 °C and 80 °C, and finally hyperthermophiles with an optimal temperature above 80 °C and a maximum above 90 °C. The finding of novel extremely thermophilic and hyperthermophilic anaerobic bacteria in recent years, and the fact that a large fraction of them belong to the Archaea has

  14. Anaerobic thermophiles.

    PubMed

    Canganella, Francesco; Wiegel, Juergen

    2014-01-01

    The term "extremophile" was introduced to describe any organism capable of living and growing under extreme conditions. With the further development of studies on microbial ecology and taxonomy, a variety of "extreme" environments have been found and an increasing number of extremophiles are being described. Extremophiles have also been investigated as far as regarding the search for life on other planets and even evaluating the hypothesis that life on Earth originally came from space. The first extreme environments to be largely investigated were those characterized by elevated temperatures. The naturally "hot environments" on Earth range from solar heated surface soils and water with temperatures up to 65 °C, subterranean sites such as oil reserves and terrestrial geothermal with temperatures ranging from slightly above ambient to above 100 °C, to submarine hydrothermal systems with temperatures exceeding 300 °C. There are also human-made environments with elevated temperatures such as compost piles, slag heaps, industrial processes and water heaters. Thermophilic anaerobic microorganisms have been known for a long time, but scientists have often resisted the belief that some organisms do not only survive at high temperatures, but actually thrive under those hot conditions. They are perhaps one of the most interesting varieties of extremophilic organisms. These microorganisms can thrive at temperatures over 50 °C and, based on their optimal temperature, anaerobic thermophiles can be subdivided into three main groups: thermophiles with an optimal temperature between 50 °C and 64 °C and a maximum at 70 °C, extreme thermophiles with an optimal temperature between 65 °C and 80 °C, and finally hyperthermophiles with an optimal temperature above 80 °C and a maximum above 90 °C. The finding of novel extremely thermophilic and hyperthermophilic anaerobic bacteria in recent years, and the fact that a large fraction of them belong to the Archaea has definitely

  15. Unusually Stable Spinae from a Freshwater Chlorobium sp

    PubMed Central

    Brooke, J. S.; Koval, S. F.; Beveridge, T. J.

    1995-01-01

    A green Chlorobium sp. with spinae, strain JSB1, was isolated from an enrichment culture previously obtained from Fayetteville Green Lake, N.Y. (J. S. Brooke, J. B. Thompson, T. J. Beveridge, and S. F. Koval, Arch. Microbiol. 157:319-322, 1992). Cells were gram-negative, nonmotile rods which contained bacteriochlorophyll c and chlorosomes. Spinae were best seen by transmission electron microscopy in thin sections of cells fixed in the presence of tannic acid. High-resolution scanning electron microscopy showed the spinae randomly distributed at the cell surface and at the junctions between cells. Spinae were physically sheared from cells and isolated from the culture supernatant by ultrafiltration. As observed by electron microscopy, spinae demonstrated unusual structural stability when exposed for 1 h at 37 deg C to chemical treatments such as hydrogen bond-breaking agents, detergents, metal-chelating agents, proteases, and organic solvents. They were stable for 1 h at 37 deg C over the pH range 2.3 to 9.9 and in 1 M HCl and 1 M NaOH. The structural integrity of the spinae was also maintained when spinae were subjected to harsher treatments of autoclaving in 2% (wt/vol) sodium dodecyl sulfate and exposure to dithiothreitol at pH 9 for 1 h at 100 deg C. Partially dissociated spinae were obtained after 5 h at 100 deg C in 1 M HCl and 1 M NaOH. In acid, the tubular spinae became amorphous structures, with no helical striations visible. In alkali, the spinae had dissociated into irregular aggregates of disks. Since both high temperature and extremes of pH were required to achieve partial dissociation of the spinae, the strength of the structure presumably comes from covalent bonding. PMID:16534897

  16. Draft Genome Sequence of Lampenflora Chlorobium limicola Strain Frasassi in a Sulfidic Cave System.

    PubMed

    Mansor, Muammar; Macalady, Jennifer L

    2016-01-01

    The draft genome sequence of Chlorobium limicola strain Frasassi was assembled from metagenomic sequencing of a green mat in an artificially lighted aquarium inside the Frasassi caves in Italy. The genome is 2.08 Mbp in size and contains the necessary genes for anoxygenic photosynthesis and CO2 fixation. PMID:27174272

  17. Draft Genome Sequence of Lampenflora Chlorobium limicola Strain Frasassi in a Sulfidic Cave System

    PubMed Central

    2016-01-01

    The draft genome sequence of Chlorobium limicola strain Frasassi was assembled from metagenomic sequencing of a green mat in an artificially lighted aquarium inside the Frasassi caves in Italy. The genome is 2.08 Mbp in size and contains the necessary genes for anoxygenic photosynthesis and CO2 fixation. PMID:27174272

  18. Mechanism of biosynthesis of 2-oxo-3-methylvalerate in Chlorobium vibrioforme.

    PubMed

    Nesbakken, T; Kolsaker, P; Ormerod, J

    1988-07-01

    The biosynthesis of 2-oxo-3-methylvalerate in Chlorobium vibrioforme was investigated by 13C nuclear magnetic resonance spectroscopy of the oxoacid formed from 13C-labeled acetate by washed suspensions. The threonine pathway could be excluded, and the results are in accord with a mechanism for the formation of 2-oxobutyrate from acetyl coenzyme A and pyruvate via citramalate. PMID:3384813

  19. An immunological assay for detection and enumeration of thermophilic biomining microorganisms.

    PubMed

    Amaro, A M; Hallberg, K B; Lindström, E B; Jerez, C A

    1994-09-01

    A specific, fast, and sensitive nonradioactive immunobinding assay for the detection and enumeration of the moderate thermophile Thiobacillus caldus and the thermophilic archaeon Sulfolobus acidocaldarius was developed. It employs enhanced chemiluminescence or peroxidase-conjugated immunoglobulins in a dot or slot blotting system and is very convenient for monitoring thermophilic bioleaching microorganisms in effluents from industrial bioleaching processes. PMID:16349398

  20. Structural and functional studies of the glycoside hydrolase family 3 β-glucosidase Cel3A from the moderately thermophilic fungus Rasamsonia emersonii.

    PubMed

    Gudmundsson, Mikael; Hansson, Henrik; Karkehabadi, Saeid; Larsson, Anna; Stals, Ingeborg; Kim, Steve; Sunux, Sergio; Fujdala, Meredith; Larenas, Edmund; Kaper, Thijs; Sandgren, Mats

    2016-07-01

    The filamentous fungus Hypocrea jecorina produces a number of cellulases and hemicellulases that act in a concerted fashion on biomass and degrade it into monomeric or oligomeric sugars. β-Glucosidases are involved in the last step of the degradation of cellulosic biomass and hydrolyse the β-glycosidic linkage between two adjacent molecules in dimers and oligomers of glucose. In this study, it is shown that substituting the β-glucosidase from H. jecorina (HjCel3A) with the β-glucosidase Cel3A from the thermophilic fungus Rasamsonia emersonii (ReCel3A) in enzyme mixtures results in increased efficiency in the saccharification of lignocellulosic materials. Biochemical characterization of ReCel3A, heterologously produced in H. jecorina, reveals a preference for disaccharide substrates over longer gluco-oligosaccharides. Crystallographic studies of ReCel3A revealed a highly N-glycosylated three-domain dimeric protein, as has been observed previously for glycoside hydrolase family 3 β-glucosidases. The increased thermal stability and saccharification yield and the superior biochemical characteristics of ReCel3A compared with HjCel3A and mixtures containing HjCel3A make ReCel3A an excellent candidate for addition to enzyme mixtures designed to operate at higher temperatures. PMID:27377383

  1. Structural and functional studies of the glycoside hydrolase family 3 β-glucosidase Cel3A from the moderately thermophilic fungus Rasamsonia emersonii

    PubMed Central

    Gudmundsson, Mikael; Hansson, Henrik; Karkehabadi, Saeid; Larsson, Anna; Stals, Ingeborg; Kim, Steve; Sunux, Sergio; Fujdala, Meredith; Larenas, Edmund; Kaper, Thijs; Sandgren, Mats

    2016-01-01

    The filamentous fungus Hypocrea jecorina produces a number of cellulases and hemicellulases that act in a concerted fashion on biomass and degrade it into monomeric or oligomeric sugars. β-Glucosidases are involved in the last step of the degradation of cellulosic biomass and hydrolyse the β-glycosidic linkage between two adjacent molecules in dimers and oligomers of glucose. In this study, it is shown that substituting the β-glucosidase from H. jecorina (HjCel3A) with the β-glucosidase Cel3A from the thermophilic fungus Rasamsonia emersonii (ReCel3A) in enzyme mixtures results in increased efficiency in the saccharification of lignocellulosic materials. Biochemical characterization of ReCel3A, heterologously produced in H. jecorina, reveals a preference for disaccharide substrates over longer gluco-oligosaccharides. Crystallographic studies of ReCel3A revealed a highly N-glycosylated three-domain dimeric protein, as has been observed previously for glycoside hydrolase family 3 β-glucosidases. The increased thermal stability and saccharification yield and the superior biochemical characteristics of ReCel3A compared with HjCel3A and mixtures containing HjCel3A make ReCel3A an excellent candidate for addition to enzyme mixtures designed to operate at higher temperatures. PMID:27377383

  2. Thermophilic Fungi: Their Physiology and Enzymes†

    PubMed Central

    Maheshwari, Ramesh; Bharadwaj, Girish; Bhat, Mahalingeshwara K.

    2000-01-01

    Thermophilic fungi are a small assemblage in mycota that have a minimum temperature of growth at or above 20°C and a maximum temperature of growth extending up to 60 to 62°C. As the only representatives of eukaryotic organisms that can grow at temperatures above 45°C, the thermophilic fungi are valuable experimental systems for investigations of mechanisms that allow growth at moderately high temperature yet limit their growth beyond 60 to 62°C. Although widespread in terrestrial habitats, they have remained underexplored compared to thermophilic species of eubacteria and archaea. However, thermophilic fungi are potential sources of enzymes with scientific and commercial interests. This review, for the first time, compiles information on the physiology and enzymes of thermophilic fungi. Thermophilic fungi can be grown in minimal media with metabolic rates and growth yields comparable to those of mesophilic fungi. Studies of their growth kinetics, respiration, mixed-substrate utilization, nutrient uptake, and protein breakdown rate have provided some basic information not only on thermophilic fungi but also on filamentous fungi in general. Some species have the ability to grow at ambient temperatures if cultures are initiated with germinated spores or mycelial inoculum or if a nutritionally rich medium is used. Thermophilic fungi have a powerful ability to degrade polysaccharide constituents of biomass. The properties of their enzymes show differences not only among species but also among strains of the same species. Their extracellular enzymes display temperature optima for activity that are close to or above the optimum temperature for the growth of organism and, in general, are more heat stable than those of the mesophilic fungi. Some extracellular enzymes from thermophilic fungi are being produced commercially, and a few others have commercial prospects. Genes of thermophilic fungi encoding lipase, protease, xylanase, and cellulase have been cloned and

  3. Anaerobic thermophilic culture

    DOEpatents

    Ljungdahl, Lars G.; Wiegel, Jurgen K. W.

    1981-01-01

    A newly discovered thermophilic anaerobe is described that was isolated in a biologically pure culture and designated Thermoanaerobacter ethanolicus ATCC 3/550. T. Ethanolicus is cultured in aqueous nutrient medium under anaerobic, thermophilic conditions and is used in a novel process for producing ethanol by subjecting carbohydrates, particularly the saccharides, to fermentation action of the new microorganism in a biologically pure culture.

  4. Uncultivated thermophiles: current status and spotlight on 'Aigarchaeota'.

    PubMed

    Hedlund, Brian P; Murugapiran, Senthil K; Alba, Timothy W; Levy, Asaf; Dodsworth, Jeremy A; Goertz, Gisele B; Ivanova, Natalia; Woyke, Tanja

    2015-06-01

    Meta-analysis of cultivation-independent sequence data shows that geothermal systems host an abundance of novel organisms, representing a vast unexplored phylogenetic and functional diversity among yet-uncultivated thermophiles. A number of thermophiles have recently been interrogated using metagenomic and/or single-cell genomic approaches, including members of taxonomic groups that inhabit both thermal and non-thermal environments, such as 'Acetothermia' (OP1) and 'Atribacteria' (OP9/JS1), as well as the exclusively thermophilic lineages 'Korarchaeota', 'Calescamantes' (EM19), 'Fervidibacteria' (OctSpA1-106), and 'Aigarchaeota' (HWCG-I). The 'Aigarchaeota', a sister lineage to the Thaumarchaeota, likely includes both hyperthermophiles and moderate thermophiles. They inhabit terrestrial, marine, and subsurface thermal environments and comprise at least nine genus-level lineages, several of which are globally distributed. PMID:26113243

  5. Anaerobic thermophilic culture system

    DOEpatents

    Ljungdahl, Lars G.; Wiegel, Jurgen K. W.

    1981-01-01

    A mixed culture system of the newly discovered microorganism Thermoanaerobacter ethanolicus ATCC31550 and the microorganism Clostridium thermocellum ATCC31549 is described. In a mixed nutrient culture medium that contains cellulose, these microorganisms have been coupled and cultivated to efficiently ferment cellulose to produce recoverable quantities of ethanol under anaerobic, thermophilic conditions.

  6. Thermophilic Beta-Glycosidase

    NASA Technical Reports Server (NTRS)

    Grogan, Dennis W.

    1992-01-01

    Report describes identification of thermophilic Beta-glycosidase enzyme from isolate of Sulfolobus solfataricus, sulfur-metabolizing archaebacteria growing aerobically and heterotrophically to relatively high cell yields. Enzyme useful in enzymatic conversion of cellulose to D-glucose and important in recycling of biomass. Used for removal of lactose from milk products. Offers promise as model substance for elucidation of basic principles of structural stabilization of proteins.

  7. Comparative genomic analysis of the thermophilic biomass-degrading fungi Myceliophthora thermophila and Thielavia terrestris

    SciTech Connect

    Berka, Randy M.; Grigoriev, Igor V.; Otillar, Robert; Salamov, Asaf; Grimwood, Jane; Reid, Ian; Ishmael, Nadeeza; John, Tricia; Darmond, Corinne; Moisan, Marie-Claude; Henrissat, Bernard; Coutinho, Pedro M.; Lombard, Vincent; Natvig, Donald O.; Lindquist, Erika; Schmutz, Jeremy; Lucas, Susan; Harris, Paul; Powlowski, Justin; Bellemare, Annie; Taylor, David; Butler, Gregory; de Vries, Ronald P.; Allijn, Iris E.; van den Brink, Joost; Ushinsky, Sophia; Storms, Reginald; Powell, Amy J.; Paulsen, Ian T.; Elbourne, Liam D. H.; Baker, Scott. E.; Magnuson, Jon; LaBoissiere, Sylvie; Clutterbuck, A. John; Martinez, Diego; Wogulis, Mark; Lopez de Leon, Alfredo; Rey, Michael W.; Tsang, Adrian

    2011-05-16

    Thermostable enzymes and thermophilic cell factories may afford economic advantages in the production of many chemicals and biomass-based fuels. Here we describe and compare the genomes of two thermophilic fungi, Myceliophthora thermophila and Thielavia terrestris. To our knowledge, these genomes are the first described for thermophilic eukaryotes and the first complete telomere-to-telomere genomes for filamentous fungi. Genome analyses and experimental data suggest that both thermophiles are capable of hydrolyzing all major polysaccharides found in biomass. Examination of transcriptome data and secreted proteins suggests that the two fungi use shared approaches in the hydrolysis of cellulose and xylan but distinct mechanisms in pectin degradation. Characterization of the biomass-hydrolyzing activity of recombinant enzymes suggests that these organisms are highly efficient in biomass decomposition at both moderate and high temperatures. Furthermore, we present evidence suggesting that aside from representing a potential reservoir of thermostable enzymes, thermophilic fungi are amenable to manipulation using classical and molecular genetics.

  8. Comparative genomic analysis of the thermophilic biomass-degrading fungi Myceliophthora thermophila and Thielavia terrestris.

    PubMed

    Berka, Randy M; Grigoriev, Igor V; Otillar, Robert; Salamov, Asaf; Grimwood, Jane; Reid, Ian; Ishmael, Nadeeza; John, Tricia; Darmond, Corinne; Moisan, Marie-Claude; Henrissat, Bernard; Coutinho, Pedro M; Lombard, Vincent; Natvig, Donald O; Lindquist, Erika; Schmutz, Jeremy; Lucas, Susan; Harris, Paul; Powlowski, Justin; Bellemare, Annie; Taylor, David; Butler, Gregory; de Vries, Ronald P; Allijn, Iris E; van den Brink, Joost; Ushinsky, Sophia; Storms, Reginald; Powell, Amy J; Paulsen, Ian T; Elbourne, Liam D H; Baker, Scott E; Magnuson, Jon; Laboissiere, Sylvie; Clutterbuck, A John; Martinez, Diego; Wogulis, Mark; de Leon, Alfredo Lopez; Rey, Michael W; Tsang, Adrian

    2011-10-01

    Thermostable enzymes and thermophilic cell factories may afford economic advantages in the production of many chemicals and biomass-based fuels. Here we describe and compare the genomes of two thermophilic fungi, Myceliophthora thermophila and Thielavia terrestris. To our knowledge, these genomes are the first described for thermophilic eukaryotes and the first complete telomere-to-telomere genomes for filamentous fungi. Genome analyses and experimental data suggest that both thermophiles are capable of hydrolyzing all major polysaccharides found in biomass. Examination of transcriptome data and secreted proteins suggests that the two fungi use shared approaches in the hydrolysis of cellulose and xylan but distinct mechanisms in pectin degradation. Characterization of the biomass-hydrolyzing activity of recombinant enzymes suggests that these organisms are highly efficient in biomass decomposition at both moderate and high temperatures. Furthermore, we present evidence suggesting that aside from representing a potential reservoir of thermostable enzymes, thermophilic fungi are amenable to manipulation using classical and molecular genetics. PMID:21964414

  9. Comparative genomic analysis of the thermophilic biomass-degrading fungi Myceliophthora thermophila and thielavia terrestris

    SciTech Connect

    Berka, Randy; Grigoriev, Igor V.; Otillar, Robert P.; Salamov, Asaf; Grimwood, Jane; Reid, Ian; Ishmael, Nadeeza; john, tricia; Darmond, Corinne; Moisan, Marie-Claude; Henrissat, Bernard; Coutinho, Pedro M.; Lombard, Vincent; Natvig, Donald O.; Lindquist, Erika; Schmutz, Jeremy; Lucas, Susan; Harris, Paul; Powlowski, Justin; Bellemare, Annie; Taylor, David; Butler, Gregory; de Vries, Ronald P.; Allijn, Iris E.; van den Brink, Joost; Ushinsky, Sophia; Storms, Reginald; Powell, Amy J.; Paulsen, Ian T.; Elbourne, Liam D. H.; Baker, Scott E.; Magnuson, Jon K.; LaBoissiere, Sylvie; Martinez, Diego; Wogulis, Mark; Lopez de Leon, Alfredo; Rey, Michael; Tsang, Adrian

    2011-10-02

    Thermostable enzymes and thermophilic cell factories may afford economic advantages in the production of many chemicals and biomass-based fuels. Here we describe and compare the genomes of two thermophilic fungi, Myceliophthora thermophila and Thielavia terrestris. To our knowledge, these genomes are the first described for thermophilic eukaryotes and the first complete telomere-to-telomere genomes for filamentous fungi. Genome analyses and experimental data suggest that both thermophiles are capable of hydrolyzing all major polysaccharides found in biomass. Examination of transcriptome data and secreted proteins suggests that the two fungi use shared approaches in the hydrolysis of cellulose and xylan but distinct mechanisms in pectin degradation. Characterization of the biomass-hydrolyzing activity of recombinant enzymes suggests that these organisms are highly efficient in biomass decomposition at both moderate and high temperatures. Furthermore, we present evidence suggesting that aside from representing a potential reservoir of thermostable enzymes, thermophilic fungi are amenable to manipulation using classical and molecular genetics.

  10. Crystallization and preliminary X-ray studies of ferredoxin-NAD(P)+ reductase from Chlorobium tepidum

    PubMed Central

    Muraki, Norifumi; Seo, Daisuke; Shiba, Tomoo; Sakurai, Takeshi; Kurisu, Genji

    2008-01-01

    Ferredoxin-NAD(P)+ reductase (FNR) is a key enzyme that catalyzes the photoreduction of NAD(P)+ to generate NAD(P)H during the final step of the photosynthetic electron-transport chain. FNR from the green sulfur bacterium Chlorobium tepidum is a homodimeric enzyme with a molecular weight of 90 kDa; it shares a high level of amino-acid sequence identity to thioredoxin reductase rather than to conventional plant-type FNRs. In order to understand the structural basis of the ferredoxin-dependency of this unique photosynthetic FNR, C. tepidum FNR has been heterologously expressed, purified and crystallized in two forms. Form I crystals belong to space group C2221 and contain one dimer in the asymmetric unit, while form II crystals belong to space group P4122 or P4322. Diffraction data were collected from a form I crystal to 2.4 Å resolution on the synchrotron-radiation beamline NW12 at the Photon Factory. PMID:18323604

  11. MECHANISMS OF THERMOPHILIC SURVIVAL, DEGRADATION BY THERMOPHILICS AND OPTIMIZATION OF THERMOPHILIC BACTERIA FOR BIODEGRADATION

    EPA Science Inventory

    We have developed a consortium of thermophilic methanotrophic bacteria from Yellowstone National Park that degrades TCE by the use of methyl monooxygenase. We are going to isolate the thermophiles in patent 5,858,763,determine their mechanisms of surviving extreme temperatures, d...

  12. Thermophilic degradation of cellulosic biomass

    NASA Astrophysics Data System (ADS)

    Ng, T.; Zeikus, J. G.

    1982-12-01

    The conversion of cellulosic biomass to chemical feedstocks and fuel by microbial fermentation is an important objective of developing biotechnology. Direct fermentation of cellulosic derivatives to ethanol by thermophilic bacteria offers a promising approach to this goal. Fermentations at elevated temperatures lowers the energy demand for cooling and also facilitates the recovery of volatile products. In addition, thermophilic microorganisms possess enzymes with greater stability than those from mesophilic microorganisms. Three anaerobic thermophilic cocultures that ferment cellulosic substrate mainly to ethanol have been described: Clostridium thermocellum/Clostriidium thermohydrosulfuricum, C. thermocellum/Clostridium thermosaccharolyticum, and C. thermocellum/Thermoanaerobacter ethanolicus sp. nov. The growth characteristics and metabolic features of these cocultures are reviewed.

  13. Thermophilic molds: Biology and applications.

    PubMed

    Singh, Bijender; Poças-Fonseca, Marcio J; Johri, B N; Satyanarayana, Tulasi

    2016-11-01

    Thermophilic molds thrive in a variety of natural habitats including soils, composts, wood chip piles, nesting materials of birds and other animals, municipal refuse and others, and ubiquitous in their distribution. These molds grow in simple media containing carbon and nitrogen sources and mineral salts. Polyamines are synthesized in these molds and the composition of lipids varies considerably, predominantly containing palmitic, oleic and linoleic acids with low levels of lauric, palmiotoleic and stearic acids. Thermophilic molds are capable of efficiently degrading organic materials by secreting thermostable enzymes, which are useful in the bioremediation of industrial wastes and effluents that are rich in oil, heavy metals, anti-nutritional factors such as phytic acid and polysaccharides. Thermophilic molds synthesize several antimicrobial substances and biotechnologically useful miscellaneous enzymes. The analysis of genomes of thermophilic molds reveals high G:C contents, shorter introns and intergenic regions with lesser repetitive sequences, and further confirms their ability to degrade agro-residues efficiently. Genetic engineering has aided in ameliorating the characteristics of the enzymes of thermophilic molds. This review is aimed at focusing on the biology of thermophilic molds with emphasis on recent developments in the analysis of genomes, genetic engineering and potential applications. PMID:26777293

  14. Final Report - "CO2 Sequestration in Cell Biomass of Chlorobium Thiosulfatophilum"

    SciTech Connect

    James L. Gaddy, PhD; Ching-Whan Ko, PhD

    2009-05-04

    World carbon dioxide emissions from the combustion of fossil fuels have increased at a rate of about 3 percent per year during the last 40 years to over 24 billion tons today. While a number of methods have been proposed and are under study for dealing with the carbon dioxide problem, all have advantages as well as disadvantages which limit their application. The anaerobic bacterium Chlorobium thiosulfatophilum uses hydrogen sulfide and carbon dioxide to produce elemental sulfur and cell biomass. The overall objective of this project is to develop a commercial process for the biological sequestration of carbon dioxide and simultaneous conversion of hydrogen sulfide to elemental sulfur. The Phase I study successfully demonstrated the technical feasibility of utilizing this bacterium for carbon dioxide sequestration and hydrogen sulfide conversion to elemental sulfur by utilizing the bacterium in continuous reactor studies. Phase II studies involved an advanced research and development to develop the engineering and scale-up parameters for commercialization of the technology. Tasks include culture isolation and optimization studies, further continuous reactor studies, light delivery systems, high pressure studies, process scale-up, a market analysis and economic projections. A number of anaerobic and aerobic microorgansims, both non-photosynthetic and photosynthetic, were examined to find those with the fastest rates for detailed study to continuous culture experiments. C. thiosulfatophilum was selected for study to anaerobically produce sulfur and Thiomicrospira crunogena waws selected for study to produce sulfate non-photosynthetically. Optimal conditions for growth, H2S and CO2 comparison, supplying light and separating sulfur were defined. The design and economic projections show that light supply for photosynthetic reactions is far too expensive, even when solar systems are considered. However, the aerobic non-photosynthetic reaction to produce sulfate with T

  15. Were the original eubacteria thermophiles?

    NASA Technical Reports Server (NTRS)

    Achenbach-Richter, L.; Gupta, R.; Stetter, K. O.; Woese, C. R.; Johnson, P. C. (Principal Investigator)

    1987-01-01

    Thermotoga maritima is one of the more unusual eubacteria: It is highly thermophilic, growing at temperatures higher than any other eubacterium; its cell wall appears to have a unique structure and its lipids a unique composition; and the organism is surrounded by a loose-fitting sheath of unknown function. Its phenotypic uniqueness is matched by its phylogenetic position; Thermotoga maritima represents the deepest known branching in the eubacterial line of descent, as measured by ribosomal RNA sequence comparisons. T. maritima also represents the most slowly evolving of eubacterial lineages. The fact that the two deepest branchings in the eubacterial line of descent (the other, the green non-sulfur bacteria and relatives, i.e. Chloroflexus, Thermomicrobium, etc.) are both basically thermophilic and slowly evolving, strongly suggests that all eubacteria have ultimately arisen from a thermophilic ancestor.

  16. Internal Structure of Chlorosomes from Brown-Colored Chlorobium Species and the Role of Carotenoids in Their Assembly

    PubMed Central

    Pšenčík, Jakub; Arellano, Juan B.; Ikonen, Teemu P.; Borrego, Carles M.; Laurinmäki, Pasi A.; Butcher, Sarah J.; Serimaa, Ritva E.; Tuma, Roman

    2006-01-01

    Chlorosomes are the main light harvesting complexes of green photosynthetic bacteria. Recently, a lamellar model was proposed for the arrangement of pigment aggregates in Chlorobium tepidum chlorosomes, which contain bacteriochlorophyll (BChl) c as the main pigment. Here we demonstrate that the lamellar organization is also found in chlorosomes from two brown-colored species (Chl. phaeovibrioides and Chl. phaeobacteroides) containing BChl e as the main pigment. This suggests that the lamellar model is universal among green sulfur bacteria. In contrast to green-colored Chl. tepidum, chlorosomes from the brown-colored species often contain domains of lamellar aggregates that may help them to survive in extremely low light conditions. We suggest that carotenoids are localized between the lamellar planes and drive lamellar assembly by augmenting hydrophobic interactions. A model for chlorosome assembly, which accounts for the role of carotenoids and secondary BChl homologs, is presented. PMID:16731553

  17. Sulfide pulsing as the controlling factor of spinae production in Chlorobium limicola strain UdG 6038

    PubMed

    Pibernat; Abella

    1996-04-01

    Chlorobium limicola UdG 6038, a green sulfur bacterium, was isolated from anoxic sediments. Cells were gram-negative, non-motile, ovoid shaped, and contained chlorobactene and bacteriochlorophyll c as the main photosynthetic pigments. The DNA G+C content was 56.4 mol%. Ultrastructural studies revealed the presence of abundant spinae (45-110 spinae per cell) attached to the cell wall. India-ink-stained cells observed under the optical microscope were surrounded by a large capsule (5-11 &mgr;m total diameter). The presence of this capsule was coincident with the presence of a large number of spinae (> 30 spinae per cell). The mucilaginous capsule was attached to the spinae without penetrating it. In batch culture, the synthesis of spinae in strain UdG 6038 was not affected by changes in temperature, pH, salt concentration, or illumination at physiological ranges and hence, the cells remained spined. The control of spinae production was experimentally confirmed using a semicontinuous batch culture refed by sulfide pulsing. The culture remained at a low spination level (> 30 spinae per cell) only when the duration of sulfide starvation between pulses was less than 5 h. After longer sulfide starvation periods, the cells remained spined (more than 38 ± 6.3 spinae per cell). This observation supports the idea that the duration of sulfide limitation in the culture plays a key role in controlling the spination process in strain C. limicola UdG 6038. Chlorobium spinae may play an eco-physiological role in buoyancy capacity and adhesion of sulfur globules to the cells in natural environments where sulfide concentrations are expected to be highly variable. PMID:8952947

  18. Chlorobium limicola forma thiosulfatophilum: biocatalyst in the production of sulfur and organic carbon from a gas stream containing H/sub 2/O and CO/sub 2/

    SciTech Connect

    Cork, D.J.; Garunas, R.; Sajjad, A.

    1983-03-01

    Chlorobium limicola forma thiosulfatophilum (ATCC 17092) was grown in a 1-liter continuously stirred tank reactor (800-ml liquid volume) at pH 6.8, 30/sup 0/C, saturated light intensity, and gas flow rate of 23.6 ml/min from a gas cylinder blend consisting of 3.9 mol% H/sub 2/S, 9.2 mol% CO/sub 2/, 86.4 mol% N/sub 2/, and 0.5 mol% H/sub 2/. This is the first demonstration of photoautotrophic growth of a Chlorobium sp. on a continuous inorganic gas feed. A significant potential exists for applying this photoautotrophic process to desulfurization and CO/sub 2/ fixation of gases containing acidic components (H/sub 2/S and CO/sub 2/).

  19. Pyrite oxidation by thermophilic archaebacteria

    SciTech Connect

    Larsson, L.; Olsson, G.; Holst, O.; Karlsson, H.T. )

    1990-03-01

    Three species of thermophilic archaebacteria of the genera Sulfolobus (Sulfolobus acidocaldarius and S. solfataricus) and Acidianus (Acidianus brierleyi) were tested for their ability to oxidize pyrite and to grow autotropbically on pyrite, to explore their potential for use in coal desulfurization. Only A. brierleyi was able to oxidize and grow autotrophically on pyrite. Jarosite was formed during the pyrite oxidation, resulting in the precipitation of sulfate and iron. The medium composition affected the extent of jarosite formation.

  20. Finding extraterrestrial sites for thermophiles.

    PubMed

    Naylor, T

    2004-04-01

    Virtually our entire knowledge of the universe comes from two sorts of measurement of the electromagnetic radiation from the stars and galaxies within it; either their flux through relatively wide bandpasses (photometry), or measurements of the shape and wavelength of relatively narrow lines via spectroscopy. These techniques are now being used to discover planets outside our solar system, and perhaps in the next 10 years will begin to characterize them. If a serious search is to be made for extraterrestrial thermophiles, we need predictions for the effects of thermophiles on their host planets that are observable with these techniques. In this paper I shall outline what sorts of observation are likely to be used in the next 15 years for extra-solar planet work. All of the journal articles quoted here can be found through http://adsabs.harvard.edu/abstract_service.html, and often also accessed as preprints at http://uk.arxiv.org/form/astro%20ph?MULTI=form%20+/-%20interface. PMID:15046563

  1. Metabolic analysis of Chlorobium chlorochromatii CaD3 reveals clues of the symbiosis in ‘Chlorochromatium aggregatum'.

    PubMed Central

    Cerqueda-García, Daniel; Martínez-Castilla, León P; Falcón, Luisa I; Delaye, Luis

    2014-01-01

    A symbiotic association occurs in ‘Chlorochromatium aggregatum', a phototrophic consortium integrated by two species of phylogenetically distant bacteria composed by the green-sulfur Chlorobium chlorochromatii CaD3 epibiont that surrounds a central β-proteobacterium. The non-motile chlorobia can perform nitrogen and carbon fixation, using sulfide as electron donors for anoxygenic photosynthesis. The consortium can move due to the flagella present in the central β-protobacterium. Although Chl. chlorochromatii CaD3 is never found as free-living bacteria in nature, previous transcriptomic and proteomic studies have revealed that there are differential transcription patterns between the symbiotic and free-living status of Chl. chlorocromatii CaD3 when grown in laboratory conditions. The differences occur mainly in genes encoding the enzymatic reactions involved in nitrogen and amino acid metabolism. We performed a metabolic reconstruction of Chl. chlorochromatii CaD3 and an in silico analysis of its amino acid metabolism using an elementary flux modes approach (EFM). Our study suggests that in symbiosis, Chl. chlorochromatii CaD3 is under limited nitrogen conditions where the GS/GOGAT (glutamine synthetase/glutamate synthetase) pathway is actively assimilating ammonia obtained via N2 fixation. In contrast, when free-living, Chl. chlorochromatii CaD3 is in a condition of nitrogen excess and ammonia is assimilated by the alanine dehydrogenase (AlaDH) pathway. We postulate that ‘Chlorochromatium aggregatum' originated from a parasitic interaction where the N2 fixation capacity of the chlorobia would be enhanced by injection of 2-oxoglutarate from the β-proteobacterium via the periplasm. This consortium would have the advantage of motility, which is fundamental to a phototrophic bacterium, and the syntrophy of nitrogen and carbon sources. PMID:24285361

  2. Hexanol-induced order-disorder transitions in lamellar self-assembling aggregates of bacteriochlorophyll c in Chlorobium tepidum chlorosomes.

    PubMed

    Arellano, Juan B; Torkkeli, Mika; Tuma, Roman; Laurinmäki, Pasi; Melø, Thor B; Ikonen, Teemu P; Butcher, Sarah J; Serimaa, Ritva E; Psencík, Jakub

    2008-03-01

    Chlorosomes are light-harvesting complexes of green photosynthetic bacteria. Chlorosomes contain bacteriochlorophyll (BChl) c, d, or e aggregates that exhibit strong excitonic coupling. The short-range order, which is responsible for the coupling, has been proposed to be augmented by pigment arrangement into undulated lamellar structures with spacing between 2 and 3 nm. Treatment of chlorosomes with hexanol reversibly converts the aggregated chlorosome chlorophylls into a form with spectral properties very similar to that of the monomer. Although this transition has been extensively studied, the structural basis remains unclear due to variability in the obtained morphologies. Here we investigated hexanol-induced structural changes in the lamellar organization of BChl c in chlorosomes from Chlorobium tepidum by a combination of X-ray scattering, electron cryomicroscopy, and optical spectroscopy. At a low hexanol/pigment ratio, the lamellae persisted in the presence of hexanol while the short-range order and exciton interactions between chlorin rings were effectively eliminated, producing a monomer-like absorption. The result suggested that hexanol hydroxyls solvated the chlorin rings while the aliphatic tail partitioned into the hydrophobic part of the lamellar structure. This partitioning extended the chlorosome along its long axis. Further increase of the hexanol/pigment ratio produced round pigment-hexanol droplets, which lost all lamellar order. After hexanol removal the spectral properties were restored. In the samples treated under the high hexanol/pigment ratio, lamellae reassembled in small domains after hexanol removal while the shape and long-range order were irreversibly lost. Thus, all the interactions required for establishing the short-range order by self-assembly are provided by BChl c molecules alone. However, the long-range order and overall shape are imposed by an external structure, e.g., the proteinaceous chlorosome baseplate. PMID:18197717

  3. Consolidated bioprocessing method using thermophilic microorganisms

    DOEpatents

    Mielenz, Jonathan Richard

    2016-02-02

    The present invention is directed to a method of converting biomass to biofuel, and particularly to a consolidated bioprocessing method using a co-culture of thermophilic and extremely thermophilic microorganisms which collectively can ferment the hexose and pentose sugars produced by degradation of cellulose and hemicelluloses at high substrate conversion rates. A culture medium therefor is also provided as well as use of the methods to produce and recover cellulosic ethanol.

  4. Variations of culturable thermophilic microbe numbers and bacterial communities during the thermophilic phase of composting.

    PubMed

    Li, Rong; Li, Linzhi; Huang, Rong; Sun, Yifei; Mei, Xinlan; Shen, Biao; Shen, Qirong

    2014-06-01

    Composting is a process of stabilizing organic wastes through the degradation of biodegradable components by microbial communities under controlled conditions. In the present study, genera and species diversities, amylohydrolysis, protein and cellulose degradation abilities of culturable bacteria in the thermophilic phase of composting of cattle manure with plant ash and rice bran were investigated. The number of culturable thermophilic bacteria and actinomyces decreased with the increasing temperature. At the initiation and end of the thermophilic phase, genera and specie diversities and number of bacteria possessing degradation abilities were higher than during the middle phase. During the thermophilic composting phase, Bacillus, Geobacillus and Ureibacillus were the dominant genera, and Geobacillus thermodenitrificans was the dominant species. In later thermophilic phases, Geobacillus toebii and Ureibacillus terrenus were dominant. Bacillus, at the initiation, and Ureibacillus and Geobacillus, at the later phase, contributed the multiple degradation abilities. These data will facilitate the control of composting in the future. PMID:24415499

  5. Hemicellulases from anaerobic thermophiles. Progress report

    SciTech Connect

    Wiegel, J.

    1994-05-01

    The longterm goal of this research effort is to obtain an anaerobic thermophilic bacterium that efficiently converts various hemicellulose-containing biomass to ethanol over a broad pH range. The strategy is to modify the outfit and regulation of the rate-limiting xylanases, glycosidases and xylan esterases in the ethanologenic, anaerobic thermophile Thermoanaerobacter ethanolicus, which grows between pH 4.5 and 9.5. Although it utilizes xylans, the xylanase, acetyl(xylan) esterase and O-methylglucuronidase activities in T. ethanolicus are barely measurable and regarded as the rate limiting steps in its xylan utilization. Thus, and also due to the presently limited knowledge of hemicellulases in anaerobic thermophiles, we characterize the hemicellulolytic enzymes from this and other anaerobic thermophiles as enzyme donors. Beside the active xylosidase/arabinosidase from T. ethanolicus, exhibiting the two different activities, we characterized 2 xylosidases, two acetyl(xylan) esterases, and an O-methylglucuronidase from Thermoanaerobacterium spec. We will continue with the characterization of xylanases from novel isolated slightly acidophilic, neutrophilic and slightly alkalophilic thermophiles. We have cloned, subcloned and partially sequenced the 165,000 Da (2 x 85,000) xylosidase/arabinosidase from T. ethanolicus and started with the cloning of the esterases from Thermoanaerobacterium spec. Consequently, we will develop a shuttle vector and continue to apply electroporation of autoplasts as a method for cloning into T. ethanolicus.

  6. Effect of seeding during thermophilic composting of sewage sludge

    SciTech Connect

    Nakasaki, K.; Sasaki, M.; Shoda, M.; Kubota, H.

    1985-03-01

    The effect of seeding on the thermophilic composting of sewage sludge was examined by measuring the changes in CO/sub 2/ evolution rates and microbial numbers. Although the succession of thermophilic bacteria and thermophilic actinomycetes clearly reflected the effect of seeding, no clear difference was observed in the overall rate of composting or quality of the composted product. 7 references.

  7. Coal Depyritization by the Thermophilic Archaeon Metallosphaera sedula

    PubMed Central

    Clark, Thomas R.; Baldi, Franco; Olson, Gregory J.

    1993-01-01

    The kinetics of pyrite oxidation by Metallosphaera sedula were investigated with mineral pyrite and two coals with moderate (Pittsburgh no. 8) and high (New Brunswick, Canada) pyritic sulfur content. M. sedula oxidized mineral pyrite at a greater rate than did another thermophile, Acidianus brierleyi, or a mesophile, Thiobacillus ferrooxidans. Maximum rates of coal depyritization were also greater with M. sedula, although the magnitude of biological stimulation above abiotic rates was notably less than with mineral pyrite. Coal depyritization appears to be limited by the oxidation of pyrite with ferric ions and not by the rate of biotic oxidation of ferrous iron, as evidenced by the maintenance of a high ratio of ferric to ferrous iron in solution by M. sedula. Significant precipitation of hydronium jarosite at elevated temperature occurred only with New Brunswick coal. PMID:16349006

  8. Bioprospecting thermophiles for cellulase production: a review

    PubMed Central

    Acharya, Somen; Chaudhary, Anita

    2012-01-01

    Most of the potential bioprospecting is currently related to the study of the extremophiles and their potential use in industrial processes. Recently microbial cellulases find applications in various industries and constitute a major group of industrial enzymes. Considerable amount of work has been done on microbial cellulases, especially with resurgence of interest in biomass ethanol production employing cellulases and use of cellulases in textile and paper industry. Most efficient method of lignocellulosic biomass hydrolysis is through enzymatic saccharification using cellulases. Significant information has also been gained about the physiology of thermophilic cellulases producers and process development for enzyme production and biomass saccharification. The review discusses the current knowledge on cellulase producing thermophilic microorganisms, their physiological adaptations and control of cellulase gene expression. It discusses the industrial applications of thermophilic cellulases, their cost of production and challenges in cellulase research especially in the area of improving process economics of enzyme production. PMID:24031898

  9. Fate of selected emerging micropollutants during mesophilic, thermophilic and temperature co-phased anaerobic digestion of sewage sludge.

    PubMed

    Samaras, Vasilios G; Stasinakis, Athanasios S; Thomaidis, Nikolaos S; Mamais, Daniel; Lekkas, Themistokles D

    2014-06-01

    The removal of endocrine disrupting compounds (EDCs) and non-steroidal anti-inflammatory drugs (NSAIDs) was studied in three lab-scale anaerobic digestion (AD) systems; a single-stage mesophilic, a single-stage thermophilic and a two-stage thermophilic/mesophilic. All micropollutants underwent microbial degradation. High removal efficiency (>80%) was calculated for diclofenac, ibuprofen, naproxen and ketoprofen; whereas triclosan, bisphenol A and the sum of nonylphenol (NP), nonylphenol monoethoxylate (NP1EO) and nonylphenol diethoxylate were moderately removed (40-80%). NSAIDs removal was not affected by the type of AD system used; whereas slightly higher EDCs removal was observed in two-stage system. In this system, most microcontaminants were removed in thermophilic digester. Biotransformation of NP1EO and NP was affected by the temperature applied to bioreactors. Under mesophilic conditions, higher removal of NP1EO and accumulation of NP was noticed; whereas the opposite was observed under thermophilic conditions. For most analytes, higher specific removal rates were calculated under thermophilic conditions and 20 days SRT. PMID:24768891

  10. A thermophilic microbial fuel cell design

    NASA Astrophysics Data System (ADS)

    Carver, Sarah M.; Vuoriranta, Pertti; Tuovinen, Olli H.

    Microbial fuel cells (MFCs) are reactors able to generate electricity by capturing electrons from the anaerobic respiratory processes of microorganisms. While the majority of MFCs have been tested at ambient or mesophilic temperatures, thermophilic systems warrant evaluation because of the potential for increased microbial activity rates on the anode. MFC studies at elevated temperatures have been scattered, using designs that are already established, specifically air-cathode single chambers and two-chamber designs. This study was prompted by our previous attempts that showed an increased amount of evaporation in thermophilic MFCs, adding unnecessary technical difficulties and causing excessive maintenance. In this paper, we describe a thermophilic MFC design that prevents evaporation. The design was tested at 57 °C with an anaerobic, thermophilic consortium that respired with glucose to generate a power density of 375 mW m -2 after 590 h. Polarization and voltage data showed that the design works in the batch mode but the design allows for adoption to continuous operation.

  11. Are thermophilic microorganisms active in cold environments?

    NASA Astrophysics Data System (ADS)

    Cockell, Charles S.; Cousins, Claire; Wilkinson, Paul T.; Olsson-Francis, Karen; Rozitis, Ben

    2015-07-01

    The mean air temperature of the Icelandic interior is below 10 °C. However, we have previously observed 16S rDNA sequences associated with thermophilic lineages in Icelandic basalts. Measurements of the temperatures of igneous rocks in Iceland showed that solar insolation of these low albedo substrates achieved a peak surface temperature of 44.5 °C. We isolated seven thermophilic Geobacillus species from basalt with optimal growth temperatures of ~65 °C. The minimum growth temperature of these organisms was ~36 °C, suggesting that they could be active in the rock environment. Basalt dissolution rates at 40 °C were increased in the presence of one of the isolates compared to abiotic controls, showing its potential to be involved in active biogeochemistry at environmental temperatures. These data raise the possibility of transient active thermophilic growth in macroclimatically cold rocky environments, implying that the biogeographical distribution of active thermophiles might be greater than previously understood. These data show that temperatures measured or predicted over large scales on a planet are not in themselves adequate to assess niches available to extremophiles at micron scales.

  12. Systematic Underutilization of Glutamine In Thermophile Proteins

    NASA Technical Reports Server (NTRS)

    Liang, Shoudan; Weber, Arthur; Biegel, Bryan A. (Technical Monitor)

    2002-01-01

    Rapid racemization above 100 C of L-amino acids to Domino acids, as well as deamidation, is probably a hazard for high temperature life. For example, the half-life of some asparaginyl peptides can be as short as 10 minutes at 100 C. High temperature organisms could protect themselves by reducing usage of amino acids that are easily racemized/deamidazed, by having a rapid rate of protein turnover which requires energy, or by adapting special cis-peptide conformations. We have searched eight completely sequenced thermophile genomes, and compare them to mesophile genomes, in order to identify underutilized amino acids. To our surprise, asparagine, the most unstable amino acid to deamidation, is used at about the same level in thermophile proteins in comparison to mesophiles whereas it is the second most unstable amino acid, glutamine, that is underutilized in all of eight thermophile species. Glutamines are present at 2% level in a typical thermophile protein, instead of 4% in mesophile. We argue that it is easier to protect asparagines from deamidation by cis-peptide conformations. We discuss statistical as well as structural evidence in support of our conclusions.

  13. Draft Genome Sequence of Paenibacillus Strain P1XP2, a Polysaccharide-Degrading, Thermophilic, Facultative Anaerobic Bacterium Isolated from a Commercial Bioreactor Degrading Food Waste

    PubMed Central

    Adelskov, Joseph

    2015-01-01

    The analysis of the ~5.8-Mb draft genome sequence of a moderately thermophilic, heterotrophic, facultative anaerobic bacterium, Paenibacillus strain P1XP2, identified genes for enzymes with the potential for degrading complex food wastes, a property consistent with the ecological habitat of the isolate. PMID:25635015

  14. Cellulases from Thermophilic Fungi: Recent Insights and Biotechnological Potential

    PubMed Central

    Li, Duo-Chuan; Li, An-Na; Papageorgiou, Anastassios C.

    2011-01-01

    Thermophilic fungal cellulases are promising enzymes in protein engineering efforts aimed at optimizing industrial processes, such as biomass degradation and biofuel production. The cloning and expression in recent years of new cellulase genes from thermophilic fungi have led to a better understanding of cellulose degradation in these species. Moreover, crystal structures of thermophilic fungal cellulases are now available, providing insights into their function and stability. The present paper is focused on recent progress in cloning, expression, regulation, and structure of thermophilic fungal cellulases and the current research efforts to improve their properties for better use in biotechnological applications. PMID:22145076

  15. [Progress in the thermophilic and alkalophilic xylanases].

    PubMed

    Bai, Wenqin; Wang, Qinhong; Ma, Yanhe

    2014-06-01

    Xylanase is the key enzyme to degrade xylan that is a major component of hemicellulose. The enzyme has potential industrial applications in the food, feed, paper and flax degumming industries. The use of xylanases becomes more and more important in the paper industry for bleaching purposes. Xylanases used in the pulp bleaching process should be stable and active at high temperature and alkaline pH. Thermophilic and alkalophilic xylanases could be obtained by screening the wild type xylanases or engineering the mesophilic and neutral enzymes. In this paper, we reviewed recent progress of screening of the thermophilic and alkalophilic xylanases, molecular mechanism of thermal and alkaline adaptation and molecular engineering. Future research prospective was also discussed. PMID:25212001

  16. Thermophilic biohydrogen production: how far are we?

    PubMed

    Pawar, Sudhanshu S; van Niel, Ed W J

    2013-09-01

    Apart from being applied as an energy carrier, hydrogen is in increasing demand as a commodity. Currently, the majority of hydrogen (H2) is produced from fossil fuels, but from an environmental perspective, sustainable H2 production should be considered. One of the possible ways of hydrogen production is through fermentation, in particular, at elevated temperature, i.e. thermophilic biohydrogen production. This short review recapitulates the current status in thermophilic biohydrogen production through fermentation of commercially viable substrates produced from readily available renewable resources, such as agricultural residues. The route to commercially viable biohydrogen production is a multidisciplinary enterprise. Microbiological studies have pointed out certain desirable physiological characteristics in H2-producing microorganisms. More process-oriented research has identified best applicable reactor types and cultivation conditions. Techno-economic and life cycle analyses have identified key process bottlenecks with respect to economic feasibility and its environmental impact. The review has further identified current limitations and gaps in the knowledge, and also deliberates directions for future research and development of thermophilic biohydrogen production. PMID:23948723

  17. Thermophilic fungi in an aridland ecosystem.

    PubMed

    Powell, Amy J; Parchert, Kylea J; Bustamante, Joslyn M; Ricken, J Bryce; Hutchinson, Miriam I; Natvig, Donald O

    2012-01-01

    We report a comprehensive multi-year study of thermophilic fungi at the Sevilleta National Wildlife Refuge in central New Mexico. Recovery of thermophilic fungi from soils showed seasonal fluctuations, with greater abundance correlating with spring and summer precipitation peaks. In addition to grassland soils, we obtained and characterized isolates from grassland and riparian litter, herbivore dung and biological soil crusts. All strains belonged to either the Eurotiales or Sordariales (Chaetomiaceae). No particular substrate or microhabitat associations were detected. Molecular typing of strains revealed substantial phylogenetic diversity, eight ad hoc phylogroups across the two orders were identified and genetic diversity was present within each phylogroup. Growth tests over a range of temperatures showed substantial variation in maximum growth rates among strains and across phylogroups but consistency within phylogroups. Results demonstrated that 45-50 C represents the optimal temperature for growth of most isolates, with a dramatic decline at 60 C. Most strains grew at 60 C, albeit slowly, whereas none grew at 65 C, providing empirical confirmation that 60 C presents an evolutionary threshold for fungal growth. Our results support the hypothesis that fungal thermophily is an adaptation to transient seasonal and diurnal high temperatures, rather than simply an adaptation to specialized high-temperature environments. We note that the diversity observed among strains and the frequently confused taxonomy within these groups highlight the need for comprehensive biosystematic revision of thermophilic taxa in both orders. PMID:22505432

  18. X-Ray Scattering and Electron Cryomicroscopy Study on the Effect of Carotenoid Biosynthesis to the Structure of Chlorobium tepidum Chlorosomes

    PubMed Central

    Ikonen, T. P.; Li, H.; Pšenčík, J.; Laurinmäki, P. A.; Butcher, S. J.; Frigaard, N.-U.; Serimaa, R. E.; Bryant, D. A.; Tuma, R.

    2007-01-01

    Chlorosomes, the main antenna complexes of green photosynthetic bacteria, were isolated from null mutants of Chlorobium tepidum, each of which lacked one enzyme involved in the biosynthesis of carotenoids. The effects of the altered carotenoid composition on the structure of the chlorosomes were studied by means of x-ray scattering and electron cryomicroscopy. The chlorosomes from each mutant strain exhibited a lamellar arrangement of the bacteriochlorophyll c aggregates, which are the major constituents of the chlorosome interior. However, the carotenoid content and composition had a pronounced effect on chlorosome biogenesis and structure. The results indicate that carotenoids with a sufficiently long conjugated system are important for the biogenesis of the chlorosome baseplate. Defects in the baseplate structure affected the shape of the chlorosomes and were correlated with differences in the arrangement of lamellae and spacing between the lamellar planes of bacteriochlorophyll aggregates. In addition, comparisons among the various mutants enabled refinement of the assignments of the x-ray scattering peaks. While the main scattering peaks come from the lamellar structure of bacteriochlorophyll c aggregates, some minor peaks may originate from the paracrystalline arrangement of CsmA in the baseplate. PMID:17468163

  19. Role of Phe-99 and Trp-196 of sepiapterin reductase from Chlorobium tepidum in the production of L-threo-tetrahydrobiopterin.

    PubMed

    Supangat, Supangat; Park, Sun Ok; Seo, Kyung Hye; Lee, Sang Yeol; Park, Young Shik; Lee, Kon Ho

    2008-06-01

    Sepiapterin reductase from Chlorobium tepidum (cSR) catalyzes the synthesis of a distinct tetrahydrobiopterin (BH4), L-threo-BH4, different from the mammalian enzyme product. The 3-D crystal structure of cSR has revealed that the product configuration is determined solely by the substrate binding mode within the well-conserved catalytic triads. In cSR, the sepiapterin is stacked between two aromatic side chains of Phe-99 and Trp-196 and rotated approximately 180 degrees C around the active site from the position in mouse sepiapterin reductase. To confirm their roles in substrate binding, we mutated Phe-99 and/or Trp-196 to alanine (F99A, W196A) by site-directed mutagenesis and comparatively examined substrate binding of the purified proteins by kinetics analysis and differential scanning calorimetry. These mutants had higher Km values than the wild type. Remarkably, the W196A mutation resulted in a higher Km increase compared with the F99A mutation. Consistent with the results, the melting temperature (Tm) in the presence of sepiapterin was lower in the mutant proteins and the worst was W196A. These findings indicate that the two residues are indispensable for substrate binding in cSR, and Trp-196 is more important than Phe-99 for different stereoisomer production. PMID:18542834

  20. The hemicellulases from the ethanologenic thermophile, Thermoanaerobacter ethanolicus and similar anaerobic thermophiles. Annual technical progress report

    SciTech Connect

    Wiegel, J.

    1995-07-01

    A Xylanase was fractionated from Thermoanaerobacter ethanolicus, an ethanologenic thermophile, and the preparation so obtained was used to determined enzymatic parameters such as pH profile of enzyme activity. The ability of various mono- and di-saccharides as well as temperature variations to induce this enzyme activity were studied.

  1. Thermophiles in the genomic era: Biodiversity, science, and applications.

    PubMed

    Urbieta, M Sofía; Donati, Edgardo R; Chan, Kok-Gan; Shahar, Saleha; Sin, Lee Li; Goh, Kian Mau

    2015-11-01

    Thermophiles and hyperthermophiles are present in various regions of the Earth, including volcanic environments, hot springs, mud pots, fumaroles, geysers, coastal thermal springs, and even deep-sea hydrothermal vents. They are also found in man-made environments, such as heated compost facilities, reactors, and spray dryers. Thermophiles, hyperthermophiles, and their bioproducts facilitate various industrial, agricultural, and medicinal applications and offer potential solutions to environmental damages and the demand for biofuels. Intensified efforts to sequence the entire genome of hyperthermophiles and thermophiles are increasing rapidly, as evidenced by the fact that over 120 complete genome sequences of the hyperthermophiles Aquificae, Thermotogae, Crenarchaeota, and Euryarchaeota are now available. In this review, we summarise the major current applications of thermophiles and thermozymes. In addition, emphasis is placed on recent progress in understanding the biodiversity, genomes, transcriptomes, metagenomes, and single-cell sequencing of thermophiles in the genomic era. PMID:25911946

  2. Single gene insertion drives bioalcohol production by a thermophilic archaeon

    SciTech Connect

    Basen, M; Schut, GJ; Nguyen, DM; Lipscomb, GL; Benn, RA; Prybol, CJ; Vaccaro, BJ; Poole, FL; Kelly, RM; Adams, MWW

    2014-12-09

    Bioethanol production is achieved by only two metabolic pathways and only at moderate temperatures. Herein a fundamentally different synthetic pathway for bioalcohol production at 70 degrees C was constructed by insertion of the gene for bacterial alcohol dehydrogenase (AdhA) into the archaeon Pyrococcus furiosus. The engineered strain converted glucose to ethanol via acetate and acetaldehyde, catalyzed by the host-encoded aldehyde ferredoxin oxidoreductase (AOR) and heterologously expressed AdhA, in an energy-conserving, redox-balanced pathway. Furthermore, the AOR/AdhA pathway also converted exogenously added aliphatic and aromatic carboxylic acids to the corresponding alcohol using glucose, pyruvate, and/or hydrogen as the source of reductant. By heterologous coexpression of a membrane-bound carbon monoxide dehydrogenase, CO was used as a reductant for converting carboxylic acids to alcohols. Redirecting the fermentative metabolism of P. furiosus through strategic insertion of foreign genes creates unprecedented opportunities for thermophilic bioalcohol production. Moreover, the AOR/AdhA pathway is a potentially game-changing strategy for syngas fermentation, especially in combination with carbon chain elongation pathways.

  3. Single gene insertion drives bioalcohol production by a thermophilic archaeon

    PubMed Central

    Basen, Mirko; Schut, Gerrit J.; Nguyen, Diep M.; Lipscomb, Gina L.; Benn, Robert A.; Prybol, Cameron J.; Vaccaro, Brian J.; Poole, Farris L.; Kelly, Robert M.; Adams, Michael W. W.

    2014-01-01

    Bioethanol production is achieved by only two metabolic pathways and only at moderate temperatures. Herein a fundamentally different synthetic pathway for bioalcohol production at 70 °C was constructed by insertion of the gene for bacterial alcohol dehydrogenase (AdhA) into the archaeon Pyrococcus furiosus. The engineered strain converted glucose to ethanol via acetate and acetaldehyde, catalyzed by the host-encoded aldehyde ferredoxin oxidoreductase (AOR) and heterologously expressed AdhA, in an energy-conserving, redox-balanced pathway. Furthermore, the AOR/AdhA pathway also converted exogenously added aliphatic and aromatic carboxylic acids to the corresponding alcohol using glucose, pyruvate, and/or hydrogen as the source of reductant. By heterologous coexpression of a membrane-bound carbon monoxide dehydrogenase, CO was used as a reductant for converting carboxylic acids to alcohols. Redirecting the fermentative metabolism of P. furiosus through strategic insertion of foreign genes creates unprecedented opportunities for thermophilic bioalcohol production. Moreover, the AOR/AdhA pathway is a potentially game-changing strategy for syngas fermentation, especially in combination with carbon chain elongation pathways. PMID:25368184

  4. Thermophilic anaerobic digestion of high strength wastewaters

    SciTech Connect

    Wiegant, W.M.; Claassen, J.A.; Lettinga, G.

    1985-09-01

    Investigations on the thermophilic anaerobic treatment of high-strength wastewaters (14-65 kg COD/mT) are presented. Vinasse, the wastewater of alcohol distilleries, was used as an example of such wastewaters. Semicontinuously fed digestion experiments at high retention times revealed that the effluent quality of digestion at 55C is comparable with that at 30C at similar loading rates. The amount of methane formed per kilogram of vinasse drops almost linearly with increasing vinasse concentrations. The treatment of vinasse was also investigated using upflow anaerobic sludge blanket (UASB) reactors.

  5. Structure of Chlorobium tepidum sepiapterin reductase complex reveals the novel substrate binding mode for stereospecific production of L-threo-tetrahydrobiopterin.

    PubMed

    Supangat, Supangat; Seo, Kyung Hye; Choi, Yong Kee; Park, Young Shik; Son, Daeyoung; Han, Chang-deok; Lee, Kon Ho

    2006-01-27

    Sepiapterin reductase (SR) is involved in the last step of tetrahydrobiopterin (BH(4)) biosynthesis by reducing the di-keto group of 6-pyruvoyl tetrahydropterin. Chlorobium tepidum SR (cSR) generates a distinct BH(4) product, L-threo-BH(4) (6R-(1'S,2'S)-5,6,7,8-BH(4)), whereas animal enzymes produce L-erythro-BH(4) (6R-(1'R,2'S)-5,6,7,8-BH(4)) although it has high amino acid sequence similarities to the other animal enzymes. To elucidate the structural basis for the different reaction stereospecificities, we have determined the three-dimensional structures of cSR alone and complexed with NADP and sepiapterin at 2.1 and 1.7 A resolution, respectively. The overall folding of the cSR, the binding site for the cofactor NADP(H), and the positions of active site residues were quite similar to the mouse and the human SR. However, significant differences were found in the substrate binding region of the cSR. In comparison to the mouse SR complex, the sepiapterin in the cSR is rotated about 180 degrees around the active site and bound between two aromatic side chains of Trp-196 and Phe-99 so that its pterin ring is shifted to the opposite side, but its side chain position is not changed. The swiveled sepiapterin binding results in the conversion of the side chain configuration, exposing the opposite face for hydride transfer from NADPH. The different sepiapterin binding mode within the conserved catalytic architecture presents a novel strategy of switching the reaction stereospecificities in the same protein fold. PMID:16308317

  6. The crystal structure of ferritin from Chlorobium tepidum reveals a new conformation of the 4-fold channel for this protein family.

    PubMed

    Arenas-Salinas, Mauricio; Townsend, Philip D; Brito, Christian; Marquez, Valeria; Marabolli, Vanessa; Gonzalez-Nilo, Fernando; Matias, Cata; Watt, Richard K; López-Castro, Juan D; Domínguez-Vera, José; Pohl, Ehmke; Yévenes, Alejandro

    2014-11-01

    Ferritins are ubiquitous iron-storage proteins found in all kingdoms of life. They share a common architecture made of 24 subunits of five α-helices. The recombinant Chlorobium tepidum ferritin (rCtFtn) is a structurally interesting protein since sequence alignments with other ferritins show that this protein has a significantly extended C-terminus, which possesses 12 histidine residues as well as several aspartate and glutamic acid residues that are potential metal ion binding residues. We show that the macromolecular assembly of rCtFtn exhibits a cage-like hollow shell consisting of 24 monomers that are related by 4-3-2 symmetry; similar to the assembly of other ferritins. In all ferritins of known structure the short fifth α-helix adopts an acute angle with respect to the four-helix bundle. However, the crystal structure of the rCtFtn presented here shows that this helix adopts a new conformation defining a new assembly of the 4-fold channel of rCtFtn. This conformation allows the arrangement of the C-terminal region into the inner cavity of the protein shell. Furthermore, two Fe(III) ions were found in each ferroxidase center of rCtFtn, with an average FeA-FeB distance of 3 Å; corresponding to a diferric μ-oxo/hydroxo species. This is the first ferritin crystal structure with an isolated di-iron center in an iron-storage ferritin. The crystal structure of rCtFtn and the biochemical results presented here, suggests that rCtFtn presents similar biochemical properties reported for other members of this protein family albeit with distinct structural plasticity. PMID:25079050

  7. Thermophilic fungi in the new age of fungal taxonomy.

    PubMed

    de Oliveira, Tássio Brito; Gomes, Eleni; Rodrigues, Andre

    2015-01-01

    Thermophilic fungi are of wide interest due to their potential to produce heat-tolerant enzymes for biotechnological processes. However, the taxonomy of such organisms remains obscure, especially given new developments in the nomenclature of fungi. Here, we examine the taxonomy of the thermophilic fungi most commonly used in industry in light of the recent taxonomic changes following the adoption of the International Code of Nomenclature for Algae, Fungi and Plants and also based on the movement One Fungus = One Name. Despite the widespread use of these fungi in applied research, several thermotolerant fungi still remain classified as thermophiles. Furthermore, we found that while some thermophilic fungi have had their genomes sequenced, many taxa still do not have barcode sequences of reference strains available in public databases. This lack of basic information is a limiting factor for the species identification of thermophilic fungi and for metagenomic studies in this field. Based on next-generation sequencing, such studies generate large amounts of data, which may reveal new species of thermophilic fungi in different substrates (composting systems, geothermal areas, piles of plant material). As discussed in this study, there are intrinsic problems associated with this method, considering the actual state of the taxonomy of thermophilic fungi. To overcome such difficulties, the taxonomic classification of this group should move towards standardizing the commonly used species names in industry and to assess the possibility of including new systems for describing species based on environmental sequences. PMID:25399310

  8. Thermophilic slurry-phase treatment of petroleum hydrocarbon waste sludges

    SciTech Connect

    Castaldi, F.J.; Bombaugh, K.J.; McFarland, B.

    1995-12-31

    Chemoheterotrophic thermophilic bacteria were used to achieve enhanced hydrocarbon degradation during slurry-phase treatment of oily waste sludges from petroleum refinery operations. Aerobic and anaerobic bacterial cultures were examined under thermophilic conditions to assess the effects of mode of metabolism on the potential for petroleum hydrocarbon degradation. The study determined that both aerobic and anaerobic thermophilic bacteria are capable of growth on petroleum hydrocarbons. Thermophilic methanogenesis is feasible during the degradation of hydrocarbons when a strict anaerobic condition is achieved in a slurry bioreactor. Aerobic thermophilic bacteria achieved the largest apparent reduction in chemical oxygen demand, freon extractable oil, total and volatile solid,s and polycyclic aromatic hydrocarbons (PAHs) when treating oily waste sludges. The observed shift with time in the molecular weight distribution of hydrocarbon material was more pronounced under aerobic metabolic conditions than under strict anaerobic conditions. The changes in the hydrocarbon molecular weight distribution, infrared spectra, and PAH concentrations during slurry-phase treatment indicate that the aerobic thermophilic bioslurry achieved a higher degree of hydrocarbon degradation than the anaerobic thermophilic bioslurry during the same time period.

  9. Diversity of Thermophilic Microorganisms within Hawaiian Fumaroles

    NASA Astrophysics Data System (ADS)

    Ackerman, C. A.; Anderson, S.; Anderson, C.

    2007-12-01

    Fumaroles provide heat and moisture characteristic of an environment suitable for thermophilic microorganisms. On the Island of Hawaii, fumaroles are scattered across the southeastern portion of the island as a result of the volcanic activity from Kilauea Crater and Pu'u' O'o vent. We used metagenomics to detect 16S rDNA from archaeal and bacterial thermophilic microorganisms indicating their presence in Hawaiian fumaroles. The fumaroles sampled exist along elevation and precipitation gradients; varying from sea level to 4,012ft and annual rainfall from less than 20in to greater than 80in. To determine the effects of environmental gradients (including temperature, pH, elevation, and precipitation) on microbial diversity within and among fumaroles, we obtained 22 samples from 7 fumaroles over a three-day period in February of 2007. Temperature variations within individual fumaroles vary from 2.3oC to 35oC and the pH variances that range from 0.4 to 2.0. Temperatures of the different fumaroles range from 29.9oC to greater than 105oC, with pH values that vary from 2.55 to 6.93. Further data on the microbial diversity within fumaroles and among fumaroles will be determined once the sequencing of the microbial 16S rDNA regions is completed. We are currently assembling and sequencing clone libraries of bacterial and archaeal 16S rDNA fragments from fumaroles.

  10. Cellulosomics of the cellulolytic thermophile Clostridium clariflavum

    PubMed Central

    2014-01-01

    Background Clostridium clariflavum is an anaerobic, thermophilic, Gram-positive bacterium, capable of growth on crystalline cellulose as a single carbon source. The genome of C. clariflavum has been sequenced to completion, and numerous cellulosomal genes were identified, including putative scaffoldin and enzyme subunits. Results Bioinformatic analysis of the C. clariflavum genome revealed 49 cohesin modules distributed on 13 different scaffoldins and 79 dockerin-containing proteins, suggesting an abundance of putative cellulosome assemblies. The 13-scaffoldin system of C. clariflavum is highly reminiscent of the proposed cellulosome system of Acetivibrio cellulolyticus. Analysis of the C. clariflavum type I dockerin sequences indicated a very high level of conservation, wherein the putative recognition residues are remarkably similar to those of A. cellulolyticus. The numerous interactions among the cellulosomal components were elucidated using a standardized affinity ELISA-based fusion-protein system. The results revealed a rather simplistic recognition pattern of cohesin-dockerin interaction, whereby the type I and type II cohesins generally recognized the dockerins of the same type. The anticipated exception to this rule was the type I dockerin of the ScaB adaptor scaffoldin which bound selectively to the type I cohesins of ScaC and ScaJ. Conclusions The findings reveal an intricate picture of predicted cellulosome assemblies in C. clariflavum. The network of cohesin-dockerin pairs provides a thermophilic alternative to those of C. thermocellum and a basis for subsequent utilization of the C. clariflavum cellulosomal system for biotechnological application. PMID:26413154

  11. Extreme Thermophiles: Moving beyond single-enzyme biocatalysis

    PubMed Central

    Frock, Andrew D.; Kelly, Robert M.

    2013-01-01

    Extremely thermophilic microorganisms have been sources of thermostable and thermoactive enzymes for over 30 years. However, information and insights gained from genome sequences, in conjunction with new tools for molecular genetics, have opened up exciting new possibilities for biotechnological opportunities based on extreme thermophiles that go beyond single-step biotransformations. Although the pace for discovering novel microorganisms has slowed over the past two decades, genome sequence data have provided clues to novel biomolecules and metabolic pathways, which can be mined for a range of new applications. Furthermore, recent advances in molecular genetics for extreme thermophiles have made metabolic engineering for high temperature applications a reality. PMID:23413412

  12. Thermophilic fermentation of hydrolysates: the effect of inhibitors on growth of thermophilic bacteria.

    PubMed

    Thomasser, Christiane; Danner, Herbert; Neureiter, Markus; Saidi, Bamusi; Braun, Rudolf

    2002-01-01

    Lignocellulosic biomass has great potential as a cheap feedstock in biological processes to produce biofuels or chemicals; however, dilute acid pretreatment at high temperatures produces undesirable compounds. Toxicity tests were done with inhibitors in standard media, to predict the growth-limiting effects on thermophilic strains. The 22 inhibitors included furfural, levulinic acid, acetic acid, and cinnamaldehyde. Neutralizing reagents and additional treatment steps have been tested. PMID:12018300

  13. Global Association between Thermophilicity and Vancomycin Susceptibility in Bacteria

    PubMed Central

    Roy, Chayan; Alam, Masrure; Mandal, Subhrangshu; Haldar, Prabir K.; Bhattacharya, Sabyasachi; Mukherjee, Trinetra; Roy, Rimi; Rameez, Moidu J.; Misra, Anup K.; Chakraborty, Ranadhir; Nanda, Ashish K.; Mukhopadhyay, Subhra K.; Ghosh, Wriddhiman

    2016-01-01

    Exploration of the aquatic microbiota of several circum-neutral (6.0–8.5 pH) mid-temperature (55–85°C) springs revealed rich diversities of phylogenetic relatives of mesophilic bacteria, which surpassed the diversity of the truly-thermophilic taxa. To gain insight into the potentially-thermophilic adaptations of the phylogenetic relatives of Gram-negative mesophilic bacteria detected in culture-independent investigations we attempted pure-culture isolation by supplementing the enrichment media with 50 μg ml−1 vancomycin. Surprisingly, this Gram-positive-specific antibiotic eliminated the entire culturable-diversity of chemoorganotrophic and sulfur-chemolithotrophic bacteria present in the tested hot water inocula. Moreover, it also killed all the Gram-negative hot-spring isolates that were obtained in vancomycin-free media. Concurrent literature search for the description of Gram-negative thermophilic bacteria revealed that at least 16 of them were reportedly vancomycin-susceptible. While these data suggested that vancomycin-susceptibility could be a global trait of thermophilic bacteria (irrespective of their taxonomy, biogeography and Gram-character), MALDI Mass Spectroscopy of the peptidoglycans of a few Gram-negative thermophilic bacteria revealed that tandem alanines were present in the fourth and fifth positions of their muropeptide precursors (MPPs). Subsequent phylogenetic analyses revealed a close affinity between the D-alanine-D-alanine ligases (Ddl) of taxonomically-diverse Gram-negative thermophiles and the thermostable Ddl protein of Thermotoga maritima, which is well-known for its high specificity for alanine over other amino acids. The Ddl tree further illustrated a divergence between the homologs of Gram-negative thermophiles and mesophiles, which broadly coincided with vancomycin-susceptibility and vancomycin-resistance respectively. It was thus hypothesized that thermophilic Ddls have been evolutionarily selected to favor a D

  14. Global Association between Thermophilicity and Vancomycin Susceptibility in Bacteria.

    PubMed

    Roy, Chayan; Alam, Masrure; Mandal, Subhrangshu; Haldar, Prabir K; Bhattacharya, Sabyasachi; Mukherjee, Trinetra; Roy, Rimi; Rameez, Moidu J; Misra, Anup K; Chakraborty, Ranadhir; Nanda, Ashish K; Mukhopadhyay, Subhra K; Ghosh, Wriddhiman

    2016-01-01

    Exploration of the aquatic microbiota of several circum-neutral (6.0-8.5 pH) mid-temperature (55-85°C) springs revealed rich diversities of phylogenetic relatives of mesophilic bacteria, which surpassed the diversity of the truly-thermophilic taxa. To gain insight into the potentially-thermophilic adaptations of the phylogenetic relatives of Gram-negative mesophilic bacteria detected in culture-independent investigations we attempted pure-culture isolation by supplementing the enrichment media with 50 μg ml(-1) vancomycin. Surprisingly, this Gram-positive-specific antibiotic eliminated the entire culturable-diversity of chemoorganotrophic and sulfur-chemolithotrophic bacteria present in the tested hot water inocula. Moreover, it also killed all the Gram-negative hot-spring isolates that were obtained in vancomycin-free media. Concurrent literature search for the description of Gram-negative thermophilic bacteria revealed that at least 16 of them were reportedly vancomycin-susceptible. While these data suggested that vancomycin-susceptibility could be a global trait of thermophilic bacteria (irrespective of their taxonomy, biogeography and Gram-character), MALDI Mass Spectroscopy of the peptidoglycans of a few Gram-negative thermophilic bacteria revealed that tandem alanines were present in the fourth and fifth positions of their muropeptide precursors (MPPs). Subsequent phylogenetic analyses revealed a close affinity between the D-alanine-D-alanine ligases (Ddl) of taxonomically-diverse Gram-negative thermophiles and the thermostable Ddl protein of Thermotoga maritima, which is well-known for its high specificity for alanine over other amino acids. The Ddl tree further illustrated a divergence between the homologs of Gram-negative thermophiles and mesophiles, which broadly coincided with vancomycin-susceptibility and vancomycin-resistance respectively. It was thus hypothesized that thermophilic Ddls have been evolutionarily selected to favor a D-ala-D-ala bonding

  15. Performance of mesophilic biohydrogen-producing cultures at thermophilic conditions.

    PubMed

    Gupta, Medhavi; Gomez-Flores, Maritza; Nasr, Noha; Elbeshbishy, Elsayed; Hafez, Hisham; Hesham El Naggar, M; Nakhla, George

    2015-09-01

    In this study, batch tests were conducted to investigate the performance of mesophilic anaerobic digester sludge (ADS) at thermophilic conditions and estimate kinetic parameters for co-substrate fermentation. Starch and cellulose were used as mono-substrate and in combination as co-substrates (1:1 mass ratio) to conduct a comparative assessment between mesophilic (37 °C) and thermophilic (60 °C) biohydrogen production. Unacclimatized mesophilic ADS responded well to the temperature change. The highest hydrogen yield of 1.13 mol H2/mol hexose was observed in starch-only batches at thermophilic conditions. The thermophilic cellulose-only yield (0.42 mol H2/mol hexose) was three times the mesophilic yield (0.13 mol H2/mol hexose). Interestingly, co-fermentation of starch-cellulose at mesophilic conditions enhanced the hydrogen yield by 26% with respect to estimated mono-substrate yields, while under thermophilic conditions no enhancement in the overall yield was observed. Interestingly, the estimated overall Monod kinetic parameters showed higher rates at mesophilic than thermophilic conditions. PMID:26101964

  16. Use of an intelligent control system To evaluate multiparametric effects on iron oxidation by thermophilic bacteria

    PubMed

    Stoner; Miller; Fife; Larsen; Tolle; Johnson

    1998-11-01

    A learning-based intelligent control system, the BioExpert, was developed and applied to the evaluation of multiparametric effects on iron oxidation by enrichment cultures of moderately thermophilic, acidophilic mining bacteria. The control system acquired and analyzed the data and then selected and maintained the sets of conditions that were evaluated. Through multiple iterations, the BioExpert selected sets of conditions that resulted in improved iron oxidation rates. The results obtained with the BioExpert suggested that temperature and pH were coupled, or interactive, parameters. Elevated temperatures (51.5 degreesC) in combination with a moderately high pH (pH 1.84) impaired the growth of and iron oxidation by the enrichment culture. Moderate-to-high oxidation rates were achieved with a relatively high pH in combination with a relatively low temperature or, conversely, with a relatively low pH in combination with a relatively high temperature. The interactive effect of pH and temperature was not apparent from the results obtained in an experiment in which temperature was the only parameter that was varied. When the BioExpert was applied to a mixed culture containing mesophilic and thermophilic bacteria, the computer "learned" that pH 1.8, 45 degreesC, and an inlet iron concentration from 30 to 35 mM were most favorable for iron oxidation. In conclusion, this study demonstrated that the learning-based intelligent control system BioExpert was an effective experimental tool that can be used to examine multiparametric effects on the growth and metabolic activity of mining bacteria. PMID:9797322

  17. Use of an Intelligent Control System To Evaluate Multiparametric Effects on Iron Oxidation by Thermophilic Bacteria

    PubMed Central

    Stoner, Daphne L.; Miller, Karen S.; Fife, Dee Jay; Larsen, Eric D.; Tolle, Charles R.; Johnson, John A.

    1998-01-01

    A learning-based intelligent control system, the BioExpert, was developed and applied to the evaluation of multiparametric effects on iron oxidation by enrichment cultures of moderately thermophilic, acidophilic mining bacteria. The control system acquired and analyzed the data and then selected and maintained the sets of conditions that were evaluated. Through multiple iterations, the BioExpert selected sets of conditions that resulted in improved iron oxidation rates. The results obtained with the BioExpert suggested that temperature and pH were coupled, or interactive, parameters. Elevated temperatures (51.5°C) in combination with a moderately high pH (pH 1.84) impaired the growth of and iron oxidation by the enrichment culture. Moderate-to-high oxidation rates were achieved with a relatively high pH in combination with a relatively low temperature or, conversely, with a relatively low pH in combination with a relatively high temperature. The interactive effect of pH and temperature was not apparent from the results obtained in an experiment in which temperature was the only parameter that was varied. When the BioExpert was applied to a mixed culture containing mesophilic and thermophilic bacteria, the computer “learned” that pH 1.8, 45°C, and an inlet iron concentration from 30 to 35 mM were most favorable for iron oxidation. In conclusion, this study demonstrated that the learning-based intelligent control system BioExpert was an effective experimental tool that can be used to examine multiparametric effects on the growth and metabolic activity of mining bacteria. PMID:9797322

  18. Hydrolysis and acidification of dewatered sludge under mesophilic, thermophilic and extreme thermophilic conditions: effect of pH.

    PubMed

    Liu, Xiaoguang; Dong, Bin; Dai, Xiaohu

    2013-11-01

    This study investigated the effect of pH (uncontrolled, 8.0, 10.0 and 12.0) and temperature (mesophilic, thermophilic and extreme thermophilic) on hydrolysis and acidification of dewatered sludge in 7-day batch fermentation experiment. Solublization of COD, protein and carbohydrates as well as concentration and composition of VFAs were investigated. Sludge hydrolysis was enhanced with higher pH and temperature. The maximum SCOD, soluble protein and carbohydrates was observed at pH 12.0 at extreme thermophilic condition. The maximum VFAs yield was obtained at thermophilic and was 2.15 times that at mesophilic condition, but it took more time to reach the maximum. The VFAs consisted of acetic, propionic, iso-butyric, n-butyric, iso-valeric, and n-valeric acids, and acetic acid was the prevalent product in most cases except for uncontrolled pH and pH 8.0 at mesophilic condition. The methane production was as follows: pH 8.0>pH 10.0>uncontrolled (0.015)>pH 12.0; mesophilic>thermophilic>extreme thermophilic. PMID:24077155

  19. Thermophilic fermentation of acetoin and 2,3-butanediol by a novel Geobacillus strain

    PubMed Central

    2012-01-01

    Background Acetoin and 2,3-butanediol are two important biorefinery platform chemicals. They are currently fermented below 40°C using mesophilic strains, but the processes often suffer from bacterial contamination. Results This work reports the isolation and identification of a novel aerobic Geobacillus strain XT15 capable of producing both of these chemicals under elevated temperatures, thus reducing the risk of bacterial contamination. The optimum growth temperature was found to be between 45 and 55°C and the medium initial pH to be 8.0. In addition to glucose, galactose, mannitol, arabionose, and xylose were all acceptable substrates, enabling the potential use of cellulosic biomass as the feedstock. XT15 preferred organic nitrogen sources including corn steep liquor powder, a cheap by-product from corn wet-milling. At 55°C, 7.7 g/L of acetoin and 14.5 g/L of 2,3-butanediol could be obtained using corn steep liquor powder as a nitrogen source. Thirteen volatile products from the cultivation broth of XT15 were identified by gas chromatography–mass spectrometry. Acetoin, 2,3-butanediol, and their derivatives including a novel metabolite 2,3-dihydroxy-3-methylheptan-4-one, accounted for a total of about 96% of all the volatile products. In contrast, organic acids and other products were minor by-products. α-Acetolactate decarboxylase and acetoin:2,6-dichlorophenolindophenol oxidoreductase in XT15, the two key enzymes in acetoin metabolic pathway, were found to be both moderately thermophilic with the identical optimum temperature of 45°C. Conclusions Geobacillus sp. XT15 is the first naturally occurring thermophile excreting acetoin and/or 2,3-butanediol. This work has demonstrated the attractive prospect of developing it as an industrial strain in the thermophilic fermentation of acetoin and 2,3-butanediol with improved anti-contamination performance. The novel metabolites and enzymes identified in XT15 also indicated its strong promise as a precious

  20. Conductive iron oxides accelerate thermophilic methanogenesis from acetate and propionate.

    PubMed

    Yamada, Chihaya; Kato, Souichiro; Ueno, Yoshiyuki; Ishii, Masaharu; Igarashi, Yasuo

    2015-06-01

    Anaerobic digester is one of the attractive technologies for treatment of organic wastes and wastewater, while continuous development and improvements on their stable operation with efficient organic removal are required. Particles of conductive iron oxides (e.g., magnetite) are known to facilitate microbial interspecies electron transfer (termed as electric syntrophy). Electric syntrophy has been reported to enhance methanogenic degradation of organic acids by mesophilic communities in soil and anaerobic digester. Here we investigated the effects of supplementation of conductive iron oxides (magnetite) on thermophilic methanogenic microbial communities derived from a thermophilic anaerobic digester. Supplementation of magnetite accelerated methanogenesis from acetate and propionate under thermophilic conditions, while supplementation of ferrihydrite also accelerated methanogenesis from propionate. Microbial community analysis revealed that supplementation of magnetite drastically changed bacterial populations in the methanogenic acetate-degrading cultures, in which Tepidoanaerobacter sp. and Coprothermobacter sp. dominated. These results suggest that supplementation of magnetite induce electric syntrophy between organic acid-oxidizing bacteria and methanogenic archaea and accelerate methanogenesis even under thermophilic conditions. Findings from this study would provide a possibility for the achievement of stably operating thermophilic anaerobic digestion systems with high efficiency for removal of organics and generation of CH4. PMID:25488041

  1. Microbial influenced corrosion by thermophilic bacteria

    NASA Astrophysics Data System (ADS)

    Lata, Suman; Sharma, Chhaya; Singh, Ajay K.

    2012-03-01

    The present study was undertaken to investigate microbial influenced corrosion (MIC) on stainless steels due to thermophilic bacteria Desulfotomaculum nigrificans. The objective of the study was to measure the extent of corrosion and correlate it with the growth of the biofilm by monitoring the composition of its extracellular polymeric substances (EPS). The toxic effect of heavy metals on MIC was also observed. For this purpose, stainless steels 304L, 316L and 2205 were subjected to electrochemical polarization and immersion tests in the modified Baar's media, control and inoculated, in anaerobic conditions at room temperature. Scanning electron microscopy (SEM)/energy dispersive spectroscopy (EDS) were used to identify the chemicals present in/outside the pit. The results show maximum corrosive conditions when bacterial activity is highest, which in turn minimizes the amount of carbohydrate and protein along with the increase in the fraction of uronic acid in carbohydrate in EPS of the biofilm. However, although bacterial activity and corrosion rate decreases, the amount of biofilm components continue to increase. It is also observed that the toxicity of metals ions affect the bacterial activity and EPS production. It was observed that Desulfotomaculum sp. has the ability to biodegrade its own EPS.

  2. Protease Production by Different Thermophilic Fungi

    NASA Astrophysics Data System (ADS)

    Macchione, Mariana M.; Merheb, Carolina W.; Gomes, Eleni; da Silva, Roberto

    A comparative study was carried out to evaluate protease production in solid-state fermentation (SSF) and submerged fermentation (SmF) by nine different thermophilic fungi — Thermoascus aurantiacus Miehe, Thermomyces lanuginosus, T. lanuginosus TO.03, Aspergillus flavus 1.2, Aspergillus sp. 13.33, Aspergillus sp. 13.34, Aspergillus sp. 13.35, Rhizomucor pusillus 13.36 and Rhizomucor sp. 13.37 — using substrates containing proteins to induce enzyme secretion. Soybean extract (soybean milk), soybean flour, milk powder, rice, and wheat bran were tested. The most satisfactory results were obtained when using wheat bran in SSF. The fungi that stood out in SSF were T. lanuginosus, T. lanuginosus TO.03, Aspergillus sp. 13.34, Aspergillus sp. 13.35, and Rhizomucor sp. 13.37, and those in SmF were T. aurantiacus, T. lanuginosus TO.03, and 13.37. In both fermentation systems, A. flavus 1.2 and R. pusillus 13.36 presented the lowest levels of proteolytic activity.

  3. Phylogeography of the thermophilic cyanobacterium Mastigocladus laminosus.

    PubMed

    Miller, Scott R; Castenholz, Richard W; Pedersen, Deana

    2007-08-01

    We have taken a phylogeographic approach to investigate the demographic and evolutionary processes that have shaped the geographic patterns of genetic diversity for a sample of isolates of the cosmopolitan thermophilic cyanobacterial Mastigocladus laminosus morphotype collected from throughout most of its range. Although M. laminosus is found in thermal areas throughout the world, our observation that populations are typically genetically differentiated on local geographic scales suggests the existence of dispersal barriers, a conclusion corroborated by evidence for genetic isolation by distance. Genealogies inferred using nitrogen metabolism gene sequence data suggest that a significant amount of the extant global diversity of M. laminosus can be traced back to a common ancestor associated with the western North American hot spot currently located below Yellowstone National Park. Estimated intragenic recombination rates are comparable to those of pathogenic bacteria known for their capacity to exchange DNA, indicating that genetic exchange has played an important role in generating novel variation during M. laminosus diversification. Selection has constrained protein changes at loci involved in the assimilation of both dinitrogen and nitrate, suggesting the historic use of both nitrogen sources in this heterocystous cyanobacterium. Lineage-specific differences in thermal performance were also observed. PMID:17557856

  4. Experimental evidence for the thermophilicity of ancestral life

    PubMed Central

    Akanuma, Satoshi; Nakajima, Yoshiki; Yokobori, Shin-ichi; Kimura, Mitsuo; Nemoto, Naoki; Mase, Tomoko; Miyazono, Ken-ichi; Tanokura, Masaru; Yamagishi, Akihiko

    2013-01-01

    Theoretical studies have focused on the environmental temperature of the universal common ancestor of life with conflicting conclusions. Here we provide experimental support for the existence of a thermophilic universal common ancestor. We present the thermal stabilities and catalytic efficiencies of nucleoside diphosphate kinases (NDK), designed using the information contained in predictive phylogenetic trees, that seem to represent the last common ancestors of Archaea and of Bacteria. These enzymes display extreme thermal stabilities, suggesting thermophilic ancestries for Archaea and Bacteria. The results are robust to the uncertainties associated with the sequence predictions and to the tree topologies used to infer the ancestral sequences. Moreover, mutagenesis experiments suggest that the universal ancestor also possessed a very thermostable NDK. Because, as we show, the stability of an NDK is directly related to the environmental temperature of its host organism, our results indicate that the last common ancestor of extant life was a thermophile that flourished at a very high temperature. PMID:23776221

  5. Removal of fecal coliforms by thermophilic anaerobic digestion processes.

    PubMed

    De León, C; Jenkins, D

    2002-01-01

    Recent U.S. EPA regulations (40 CFR Section 503) specify maximum concentrations of pathogens and metals for Class A wastewater treatment plant sludges. The most common sludge process is mesophilic (35 degrees C) digestion which stabilizes the solids, produces a combustible gas but does not create an effluent that meets the 503 Class A pathogen requirements. This investigation was conducted to determine whether anaerobic digestion processes incorporating a thermophilic stage could achieve 503 Class A pathogen levels. The research reported here was a bench-scale screening study meant to identify the most promising process alternatives for further investigation. Fecal Coliform (FC) concentrations were used to assess disinfection efficiency. Digesters were 30 L capacity fed semi-continuously in draw-fill mode. Digester startup was rapid to produce true thermophiles. Temperature staging and pH were assessed in 3 sets of experiments: Set 1 were one stage ("acid phase"), Set 2 were one stage ("acid + methane phases") and Set 3 were two stage ("acid phase" then "methanogenic phase"). Feed was a 1:1 mixture of Thickened Waste Activated Sludge and Primary Sludge. The following anaerobic digestion configurations and operating parameters allowed the production of digested sludge with a mean FC concentration statistically less than 10(3) (the regulatory value for Class A sludge): thermophilic single stage acid phase at 52 and 62 degrees C; thermophilic single stage acid + methane phase at 48 degrees C, 52 degrees C and 62 degrees C; two-stage mesophilic acid phase followed by mesophilic methane phase; two stage mesophilic acid phase followed by thermophilic methane phase at 48 degrees C, 52 degrees C and 62 degrees C. If the maximum digested FC concentration must be below 10(3) MPN/g TS then the following digester configurations and operating conditions will be compliant: two stage mesophilic acid phase followed by thermophilic methane phase at 52 degrees C and 62 degrees C

  6. Cellulase production by a thermophilic clostridium species.

    PubMed

    Lee, B H; Blackburn, T H

    1975-09-01

    Strain M7, a thermophilic, anaerobic, terminally sporing bacterium (0.6 by 4.0 mum) was isolated from manure. It degraded filter paper in 1 to 2 days at 60 C in a minimal cellulose medium but was stimulated by yeast extract. It fermented a wide variety of sugars but produced cellulase only in cellulose or carboxymethyl-cellulose media. Cellulase synthesis not only was probably repressed by 0.4% glucose and 0.3% cellobiose, but also cellulase activity appeared to be inhibited by these sugars at these concentrations. Both C(1) cellulase (degrades native cellulose) and C(x) cellulase (beta-1,4-glucanase) activities in strain M7 cultures were assayed by measuring the liberation of reducing sugars with dinitrosalicylic acid. Both activities had optima at pH 6.5 and 67 C. One milliliter of a 48-h culture of strain M7 hydrolyzed 0.044-meq of glucose per min from cotton fibers. The cellulase(s) from strain M7 was extracellular, produced during exponential growth, but was not free in the growth medium until approximately 30% of the cellulose was hydrolyzed. Glucose and cellobiose were the major soluble products liberated from cellulose by the cellulase. ZnCl(2) precipitation appeared initially to be a good method for the concentration of cellulase activity, but subsequent purification was not successful. Isoelectric focusing indicated the presence of four C(x) cellulases (pI 4.5, 6.3, 6.8, and 8.7). The rapid production and high activity of cellulases from this organism strongly support the basic premise that increased hydrolysis of native cellulose is possible at elevated temperature. PMID:16350033

  7. THERMOPHILE ENDOSPORES HAVE RESPONSIVE EXOSPORIUM FOR ATTACHMENT

    SciTech Connect

    PANESSA-WARREN,B.; TORTORA,G.T.; WARREN,J.; SABATINI,R.

    1999-08-01

    Recently studies examining the colonization of Clostridial pathogens on agar and human tissue culture cells, demonstrated that (C. sporogenes ATCC 3584, C. difficile ATCC 43594 [patient isolate], C. difficile ATCC 9689 [non-clinical], C. clostridioforme [patient isolate]) bacterial spores (endospores) of the genus Clostridia have an outer membrane that becomes responsive at activation and exhibits extensions of the exosporial membrane that facilitate and maintain spore attachment to a nutritive substrate during germination and initial outgrowth of the newly developed bacterial cell. Therefore this attachment phenomenon plays an important role in insuring bacterial colonization of a surface and the initial stages of the infective process. To see if other non-clinical members of this genus also have this ability to attach to a substrate or food-source during spore germination, and how this attachment process in environmental thermophiles compares to the clinical paradigm (in relation to time sequence, exosporial membrane structure, type of attachment structures, composition of the membrane etc...), sediment samples were collected in sterile transport containers at 4 geothermal sites at Yellowstone National Park in Wyoming. Because spore forming bacteria will produce spores when conditions are unfavorable for growth, the samples were sealed and stored at 4 C. After 8 months the samples were screened for the presence of spores by light microscope examination using malachite green/safranin, and traditional endospores were identified in significant quantities from the Terrace Spring site (a 46 C lake with bacterial mats and a rapidly moving run-off channel leading to a traditional hot spring). The highest spore population was found in the top sediment and benthic water of the run-off channel, pH 8.1.

  8. Distribution of Thermophilic Marine Sulfate Reducers in North Sea Oil Field Waters and Oil Reservoirs

    PubMed Central

    Nilsen, R. K.; Beeder, J.; Thorstenson, T.; Torsvik, T.

    1996-01-01

    The distribution of thermophilic marine sulfate reducers in produced oil reservoir waters from the Gullfaks oil field in the Norwegian sector of the North Sea was investigated by using enrichment cultures and genus-specific fluorescent antibodies produced against the genera Archaeoglobus, Desulfotomaculum, and Thermodesulforhabdus. The thermophilic marine sulfate reducers in this environment could mainly be classified as species belonging to the genera Archaeoglobus and Thermodesulforhabdus. In addition, some unidentified sulfate reducers were present. Culturable thermophilic Desulfotomaculum strains were not detected. Specific strains of thermophilic sulfate reducers inhabited different parts of the oil reservoir. No correlation between the duration of seawater injection and the numbers of thermophilic sulfate reducers in the produced waters was observed. Neither was there any correlation between the concentration of hydrogen sulfide and the numbers of thermophilic sulfate reducers. The results indicate that thermophilic and hyperthermophilic sulfate reducers are indigenous to North Sea oil field reservoirs and that they belong to a deep subterranean biosphere. PMID:16535321

  9. [Biodegradation of organic pollutants by thermophiles and their applications: a review].

    PubMed

    Cui, Jing-Lan; Chen, Chen; Qin, Zhi-Hui; Yu, Chun-Na; Shen, Hui; Shen, Chao-Feng; Chen, Ying-Xu

    2012-11-01

    Persistent organic pollutants have increasingly become a critical environmental concern, while thermophiles have the high potential of degrading various kinds of environmental organic pollutants. At high temperatures, thermophiles have higher metabolic activity, and the competition by mesophiles is reduced, meanwhile, the solubility and bioavailability of some persistent organic pollutants are greatly increased, and thus, the degradation of the pollutants by thermophiles is more rapid and complete. Therefore, thermophils are of great significance for the bio-treatment of organic wastewater and the bioremediation of organic pollutants-contaminated sites. This paper introduced the research progress on the degradation of organic pollutants by thermophiles in terms of the characteristics of thermophiles in degrading organic pollutants, the effects of temperature on the degradation, the degradation pathways, the degradation enzymes, their coding genes, and practical engineering applications. The future research directions including the degradation mechanisms of thermophiles, their resources reserve, related technology strategies and their applications were also prospected. PMID:23431811

  10. Thermal adaptation of dihydrofolate reductase from the moderate thermophile Geobacillus stearothermophilus.

    PubMed

    Guo, Jiannan; Luk, Louis Y P; Loveridge, E Joel; Allemann, Rudolf K

    2014-05-01

    The thermal melting temperature of dihydrofolate reductase from Geobacillus stearothermophilus (BsDHFR) is ~30 °C higher than that of its homologue from the psychrophile Moritella profunda. Additional proline residues in the loop regions of BsDHFR have been proposed to enhance the thermostability of BsDHFR, but site-directed mutagenesis studies reveal that these proline residues contribute only minimally. Instead, the high thermal stability of BsDHFR is partly due to removal of water-accessible thermolabile residues such as glutamine and methionine, which are prone to hydrolysis or oxidation at high temperatures. The extra thermostability of BsDHFR can be obtained by ligand binding, or in the presence of salts or cosolvents such as glycerol and sucrose. The sum of all these incremental factors allows BsDHFR to function efficiently in the natural habitat of G. stearothermophilus, which is characterized by temperatures that can reach 75 °C. PMID:24730604

  11. Genome sequence of the moderately thermophilic halophile Flexistipes sinusarabici strain (MAS10T)

    SciTech Connect

    Lapidus, Alla L.; Chertkov, Olga; Nolan, Matt; Lucas, Susan; Hammon, Nancy; Deshpande, Shweta; Cheng, Jan-Fang; Tapia, Roxanne; Han, Cliff; Goodwin, Lynne A.; Pitluck, Sam; Liolios, Konstantinos; Pagani, Ioanna; Ivanova, N; Huntemann, Marcel; Mavromatis, K; Mikhailova, Natalia; Pati, Amrita; Chen, Amy; Palaniappan, Krishna; Land, Miriam L; Hauser, Loren John; Brambilla, Evelyne-Marie; Rohde, Manfred; Abt, Birte; Spring, Stefan; Goker, Markus; Bristow, James; Eisen, Jonathan; Markowitz, Victor; Hugenholtz, Philip; Kyrpides, Nikos C; Klenk, Hans-Peter; Woyke, Tanja

    2011-01-01

    Flexistipes sinusarabici Fiala et al. 2000 is the type species of the genus Flexistipes in the fami- ly Deferribacteraceae. The species is of interest because of its isolated phylogenetic location in a genomically under-characterized region of the tree of life, and because of its origin from a multiply extreme environment; the Atlantis Deep brines of the Red Sea, where it had to struggle with high temperatures, high salinity, and a high concentrations of heavy metals. This is the fourth completed genome sequence to be published of a type strain of the family Deferribacteraceae. The 2,526,590 bp long genome with its 2,346 protein-coding and 53 RNA genes is a part of the Genomic Encyclopedia of Bacteria and Archaea project.

  12. A Moderately Thermophilic Mixed Microbial Culture for Bioleaching of Chalcopyrite Concentrate at High Pulp Density

    PubMed Central

    Wang, Yuguang; Zeng, Weimin; Qiu, Guanzhou; Chen, Xinhua

    2014-01-01

    Three kinds of samples (acid mine drainage, coal mine wastewater, and thermal spring) derived from different sites were collected in China. Thereafter, these samples were combined and then inoculated into a basal salts solution in which different substrates (ferrous sulfate, elemental sulfur, and chalcopyrite) served as energy sources. After that, the mixed cultures growing on different substrates were pooled equally, resulting in a final mixed culture. After being adapted to gradually increasing pulp densities of chalcopyrite concentrate by serial subculturing for more than 2 years, the final culture was able to efficiently leach the chalcopyrite at a pulp density of 20% (wt/vol). At that pulp density, the culture extracted 60.4% of copper from the chalcopyrite in 25 days. The bacterial and archaeal diversities during adaptation were analyzed by denaturing gradient gel electrophoresis and constructing clone libraries of the 16S rRNA gene. The results show that the culture consisted mainly of four species, including Leptospirillum ferriphilum, Acidithiobacillus caldus, Sulfobacillus acidophilus, and Ferroplasma thermophilum, before adapting to a pulp density of 4%. However, L. ferriphilum could not be detected when the pulp density was greater than 4%. Real-time quantitative PCR was employed to monitor the microbial dynamics during bioleaching at a pulp density of 20%. The results show that A. caldus was the predominant species in the initial stage, while S. acidophilus rather than A. caldus became the predominant species in the middle stage. F. thermophilum accounted for the greatest proportion in the final stage. PMID:24242252

  13. Complete genome sequence of the moderate thermophile Anaerobaculum mobile type strain (NGAT)

    SciTech Connect

    Mavromatis, K; Stackebrandt, Erko; Held, Brittany; Lapidus, Alla L.; Nolan, Matt; Lucas, Susan; Hammon, Nancy; Deshpande, Shweta; Cheng, Jan-Fang; Tapia, Roxanne; Goodwin, Lynne A.; Pitluck, Sam; Liolios, Konstantinos; Pagani, Ioanna; Ivanova, N; Mikhailova, Natalia; Huntemann, Marcel; Pati, Amrita; Chen, Amy; Palaniappan, Krishna; Land, Miriam L; Rohde, Manfred; Spring, Stefan; Goker, Markus; Woyke, Tanja; Detter, J. Chris; Bristow, James; Eisen, Jonathan; Markowitz, Victor; Hugenholtz, Philip; Klenk, Hans-Peter; Kyrpides, Nikos C

    2013-01-01

    Anaerobaculum mobile Menes and Muxi 2002 is one of three described species of the genus Anaerobaculum, family Synergistaceae, phylum Synergistetes. This anaerobic and motile bac- terium ferments a range of carbohydrates and mono- and dicarboxylic acids with acetate, hy- drogen and CO2 as end products. A. mobile NGAT is the first member of the genus Anaerobaculum and the sixth member of the phylum Synergistetes with a completely se- quenced genome. Here we describe the features of this bacterium, together with the com- plete genome sequence, and annotation. The 2,160,700 bp long single replicon genome with its 2,053 protein-coding and 56 RNA genes is part of the Genomic Encyclopedia of Bacteria and Archaea project.

  14. A Novel Acidimicrobium Species in Continuous Cultures of Moderately Thermophilic, Mineral-Sulfide-Oxidizing Acidophiles▿

    PubMed Central

    Cleaver, Adam A.; Burton, Nicolas P.; Norris, Paul R.

    2007-01-01

    A novel species of Acidimicrobium appeared to be the predominant ferrous iron oxidizer in a mixed culture that effected the continuous, efficient extraction of nickel from a mineral concentrate at 49°C, but it was not isolated in pure culture. It outcompeted Acidimicrobium ferrooxidans, which was expected to have a major role in iron oxidation in reactors gassed with air, and was outnumbered at 49°C only by the sulfur-oxidizing Acidithiobacillus caldus. Sulfobacillus species were expected to compete with Acidimicrobium species when culture aeration was enriched with carbon dioxide, but they were a minor component of the populations with and without this enrichment. Sulfobacillus thermosulfidooxidans replaced the Acidimicrobium species and Acidithiobacillus caldus when the temperature was increased to 55°C. PMID:17468267

  15. Thermal Adaptation of Dihydrofolate Reductase from the Moderate Thermophile Geobacillus stearothermophilus

    PubMed Central

    2014-01-01

    The thermal melting temperature of dihydrofolate reductase from Geobacillus stearothermophilus (BsDHFR) is ∼30 °C higher than that of its homologue from the psychrophile Moritella profunda. Additional proline residues in the loop regions of BsDHFR have been proposed to enhance the thermostability of BsDHFR, but site-directed mutagenesis studies reveal that these proline residues contribute only minimally. Instead, the high thermal stability of BsDHFR is partly due to removal of water-accessible thermolabile residues such as glutamine and methionine, which are prone to hydrolysis or oxidation at high temperatures. The extra thermostability of BsDHFR can be obtained by ligand binding, or in the presence of salts or cosolvents such as glycerol and sucrose. The sum of all these incremental factors allows BsDHFR to function efficiently in the natural habitat of G. stearothermophilus, which is characterized by temperatures that can reach 75 °C. PMID:24730604

  16. Genome sequence of the moderately thermophilic halophile Flexistipes sinusarabici strain (MAS10T)

    PubMed Central

    Lapidus, Alla; Chertkov, Olga; Nolan, Matt; Lucas, Susan; Hammon, Nancy; Deshpande, Shweta; Cheng, Jan-Fang; Tapia, Roxanne; Han, Cliff; Goodwin, Lynne; Pitluck, Sam; Liolios, Konstantinos; Pagani, Ioanna; Ivanova, Natalia; Huntemann, Marcel; Mavromatis, Konstantinos; Mikhailova, Natalia; Pati, Amrita; Chen, Amy; Palaniappan, Krishna; Land, Miriam; Hauser, Loren; Brambilla, Evelyne-Marie; Rohde, Manfred; Abt, Birte; Spring, Stefan; Göker, Markus; Bristow, James; Eisen, Jonathan A.; Markowitz, Victor; Hugenholtz, Philip; Kyrpides, Nikos C.; Klenk, Hans-Peter; Woyke, Tanja

    2011-01-01

    Flexistipes sinusarabici Fiala et al. 2000 is the type species of the genus Flexistipes in the family Deferribacteraceae. The species is of interest because of its isolated phylogenetic location in a genomically under-characterized region of the tree of life, and because of its origin from a multiply extreme environment; the Atlantis Deep brines of the Red Sea, where it had to struggle with high temperatures, high salinity, and a high concentrations of heavy metals. This is the fourth completed genome sequence to be published of a type strain of the family Deferribacteraceae. The 2,526,590 bp long genome with its 2,346 protein-coding and 53 RNA genes is a part of the Genomic Encyclopedia of Bacteria and Archaea project. PMID:22180813

  17. Use of mesophilic and thermophilic bacteria for the improvement of copper extraction from a low-grade ore

    NASA Astrophysics Data System (ADS)

    Darezereshki, E.; Schaffie, M.; Lotfalian, M.; Seiedbaghery, S. A.; Ranjbar, M.

    2011-04-01

    Bioleaching was examined for copper extraction from a low grade ore using mesophilic and moderate thermophilic bacteria. Five equal size columns were used for the leaching of the ore. Sulfuric acid solution with a flow rate of 3.12 L·m-2·h-1 and pH 1.5 passed through each column continuously for 90 d. In the first and the second column, bioleaching was performed without agglomeration of the ore and on the agglomerated ore, respectively. 28wt% of the copper was extracted in the first column after 40 d, while this figure was 38wt% in the second column. After 90 d, however, the overall extractions were almost the same for both of them. Bioleaching with mesophilic bacteria was performed in the third column without agglomeration of the ore and in the fourth column on the agglomerated ore. After 40 d, copper extractions in the third and the fourth columns were 62wt% and 70wt%, respectively. Copper extractions were 75wt% for both the columns after 90 d. For the last column, bioleaching was performed with moderate thermophilic bacteria and agglomerated ore. Copper extractions were 80wt% and 85wt% after 40 and 90 d, respectively. It was concluded that crushing and agglomeration of the ore using bacteria could enhance the copper extraction considerably.

  18. [Conversion of acetic acid to methane by thermophiles: Progress report

    SciTech Connect

    Zinder, S.

    1991-12-31

    The objective of this project is to provide an understanding of thermophilic anaerobic microorganisms capable of breaking down acetic acid, the precursor of two-thirds of the methane produced by anaerobic bioreactors. Recent results include: (1) the isolation of Methanothrix strain CALLS-1, which grows much more rapidly than mesophilic strains; (2) the demonstration that thermophilic cultures of Methanosarcina and Methanothrix show minimum thresholds for acetate utilization of 1--2.5 mM and 10--20{mu}m respectively, in agreement with ecological data indicating that Methanothrix is favored by low acetate concentration; (3) the demonstration of high levels of thermostable acetyl-coA synthetase and carbon monoxide dehydrogenase in cell-free extracts of Methanothrix strains CALS-1; (4) the demonstration of methanogenesis from acetate and ATP in cell free extracts of strain CALS-1. (5) the demonstration that methanogenesis from acetate required 2 ATP/methane, and, in contrast to Methanosarcina, was independent of hydrogen and other electron donors; (6) the finding that entropy effects must be considered when predicting the level of hydrogen in thermophilic syntrophic cultures. (7) the isolation and characterization of the Desulfotomaculum thermoacetoxidans. Current research is centered on factors which allow thermophilic Methanothrix to compete with Methanosarcina.

  19. (Conversion of acetic acid to methane by thermophiles: Progress report)

    SciTech Connect

    Zinder, S.

    1991-01-01

    The objective of this project is to provide an understanding of thermophilic anaerobic microorganisms capable of breaking down acetic acid, the precursor of two-thirds of the methane produced by anaerobic bioreactors. Recent results include: (1) the isolation of Methanothrix strain CALLS-1, which grows much more rapidly than mesophilic strains; (2) the demonstration that thermophilic cultures of Methanosarcina and Methanothrix show minimum thresholds for acetate utilization of 1--2.5 mM and 10--20{mu}m respectively, in agreement with ecological data indicating that Methanothrix is favored by low acetate concentration; (3) the demonstration of high levels of thermostable acetyl-coA synthetase and carbon monoxide dehydrogenase in cell-free extracts of Methanothrix strains CALS-1; (4) the demonstration of methanogenesis from acetate and ATP in cell free extracts of strain CALS-1. (5) the demonstration that methanogenesis from acetate required 2 ATP/methane, and, in contrast to Methanosarcina, was independent of hydrogen and other electron donors; (6) the finding that entropy effects must be considered when predicting the level of hydrogen in thermophilic syntrophic cultures. (7) the isolation and characterization of the Desulfotomaculum thermoacetoxidans. Current research is centered on factors which allow thermophilic Methanothrix to compete with Methanosarcina.

  20. Bacteriostatic activities of monoacyl sugar alcohols against thermophilic sporeformers.

    PubMed

    Piao, Junkui; Kawahara-Aoyama, Yumiko; Inoue, Takashi; Adachi, Shuji

    2006-01-01

    The bacteriostatic activities of monoacyl sugar alcohols with different acyl chains and hydrophilic heads were examined against some thermophilic sporeformers. Monomyristoyl erythritol and xylitol efficaciously suppressed their spore development. The number and orientation of the hydroxyl groups also played important roles in this activity, and monomyristoyl xylitol exhibited the highest activity among the monomyristoyl sugar alcohols. PMID:16428845

  1. Phylogenetic analysis of anaerobic thermophilic bacteria: aid for their reclassification.

    PubMed Central

    Rainey, F A; Ward, N L; Morgan, H W; Toalster, R; Stackebrandt, E

    1993-01-01

    Small subunit rDNA sequences were determined for 20 species of the genera Acetogenium, Clostridium, Thermoanaerobacter, Thermoanaerobacterium, Thermoanaerobium, and Thermobacteroides, 3 non-validly described species, and 5 isolates of anaerobic thermophilic bacteria, providing a basis for a phylogenetic analysis of these organisms. Several species contain a version of the molecule significantly longer than that of Escherichia coli because of the presence of inserts. On the basis of normal evolutionary distances, the phylogenetic tree indicates that all bacteria investigated in this study with a maximum growth temperature above 65 degrees C form a supercluster within the subphylum of gram-positive bacteria that also contains Clostridium thermosaccharolyticum and Clostridium thermoaceticum, which have been previously sequenced. This supercluster appears to be equivalent in its phylogenetic depth to the supercluster of mesophilic clostridia and their nonspore-forming relatives. Several phylogenetically and phenotypically coherent clusters that are defined by sets of signature nucleotides emerge within the supercluster of thermophiles. Clostridium thermobutyricum and Clostridium thermopalmarium are members of Clostridium group I. A phylogenetic tree derived from transversion distances demonstrated the artificial clustering of some organisms with high rDNA G+C moles percent, i.e., Clostridium fervidus and the thermophilic, cellulolytic members of the genus Clostridium. The results of this study can be used as an aid for future taxonomic restructuring of anaerobic sporogenous and asporogenous thermophillic, gram-positive bacteria. PMID:7687600

  2. Thermophilic anaerobic digestion of industrial orange waste.

    PubMed

    Kaparaju, P L N; Rintala, J A

    2006-06-01

    Thermophilic anaerobic digestion of industrial orange waste (pulp and peel) with subsequent aerobic post-treatment of the digestate was evaluated. Methane production potential was first determined in batch assays and the effects of operational parameters such as hydraulic retention times (HRT) and organic loading rates (OLR) on process performance were studied through semi-continuous digestion. In batch assays, methane production potential of about 0.49 m(3) kg(-1) volatile solids (VS)(added waste) was achieved. In semi-continuous digestion, loading at 2.8 kgVS m(-3) d(-1) (2.9 kg total solids (TS) m(-3) d(-1)) and HRT of 26 d produced specific methane yields of 0.6 m(3) kg(-1) VS (added waste) (0.63 m(3) kg(-1) VS(added waste)). Operating at a higher OLR of 4.2 kgVS m(-3) d(-1) (4.4 kg TS m(-3) d(-1)) and 40 d HRT produced 0.5 m(3) of methane kg(-1) VS (added waste) (0.63-0.52 m(3) kg(-1) TS (added waste). Up to 70% of TS of industrial orange waste (11.6% TS) was methanised. Further increase in OLR to 5.6 kg VS m(-3) d(-1) (5.9 kg TS m(-3) d(-1); HRT of 20 d) resulted in an unstable and non-functional digester process shown directly through complete cessation of methanogenesis, drop in methane content, reduced pH and increase in volatile fatty acid (VFA) concentrations, especially acetate and soluble chemical oxygen demand. A pH adjustment (from an initial 3.2 to ca. 8) for the low pH orange waste was necessary and was found to be a crucial factor for stable digester operation as the process showed a tendency to be inhibited due to accumulation of VFAs and decrease in digester pH. Aerobic post-treatment of digestate resulted in removal of ammonia and VFAs. PMID:16865918

  3. Thermophilic hydrogen production from sludge pretreated by thermophilic bacteria: analysis of the advantages of microbial community and metabolism.

    PubMed

    Zheng, He-Shan; Guo, Wan-Qian; Yang, Shan-Shan; Feng, Xiao-Chi; Du, Juan-Shan; Zhou, Xian-Jiao; Chang, Jo-Shu; Ren, Nan-Qi

    2014-11-01

    In this study, the effects of thermophilic bacteria pretreatment and elevated fermentation temperature on hydrogen production from sludge were examined. The highest hydrogen yield of 19.9mlH2g(-1) VSS was achieved at 55°C by using pretreated sludge, which was 48.6% higher than raw sludge without pretreatment, and 28.39% higher than when fermented at 35°C. To explore the internal factors of this superior hydrogen production performance, the microbial community and the metabolism analysis were performed by using high-throughput sequencing and excitation-emission matrix. The pretreated sludge showed better utilization of dissolved organic matter and less inhibition of metabolism, especially at thermophilic condition. The 454 sequencing data indicated that microbial abundance was distinctly reduced and extremely high proportion of hydrogen-producing bacteria was found in the thermophilic community (Thermoanaerobacterium accounted for 93.75%). Thus, the pretreated sludge and thermophilic condition showed significant advantages in the hydrogen production using waste sludge as substrate. PMID:25260350

  4. [Conversion of acetic acid to methane by thermophiles

    SciTech Connect

    Zinder, S.H.

    1993-01-01

    The primary goal of this project is to obtain a better understanding of thermophilic microorganisms which convert acetic acid to CH[sub 4]. The previous funding period represents a departure from earlier research in this laboratory, which was more physiological and ecological. The present work is centered on the biochemistry of the thermophile Methanothrix sp. strain CALS-1. this organism presents a unique opportunity, with its purity and relatively rapid growth, to do comparative biochemical studies with the other major acetotrophic genus Methanosarcina. We previously found that Methanothrix is capable of using acetate at concentrations 100 fold lower than Methanosarcina. This finding suggests that there are significant differences in the pathways of methanogenesis from acetate in the two genera.

  5. Proteomic analysis of acetylation in thermophilic Geobacillus kaustophilus.

    PubMed

    Lee, Dong-Woo; Kim, Dooil; Lee, Yong-Jik; Kim, Jung-Ae; Choi, Ji Young; Kang, Sunghyun; Pan, Jae-Gu

    2013-08-01

    Recent analysis of prokaryotic N(ε)-lysine-acetylated proteins highlights the posttranslational regulation of a broad spectrum of cellular proteins. However, the exact role of acetylation remains unclear due to a lack of acetylated proteome data in prokaryotes. Here, we present the N(ε)-lysine-acetylated proteome of gram-positive thermophilic Geobacillus kaustophilus. Affinity enrichment using acetyl-lysine-specific antibodies followed by LC-MS/MS analysis revealed 253 acetylated peptides representing 114 proteins. These acetylated proteins include not only common orthologs from mesophilic Bacillus counterparts, but also unique G. kaustophilus proteins, indicating that lysine acetylation is pronounced in thermophilic bacteria. These data complement current knowledge of the bacterial acetylproteome and provide an expanded platform for better understanding of the function of acetylation in cellular metabolism. PMID:23696451

  6. Transformation of chenodeoxycholic acid by thermophilic Geobacillus stearothermophilus.

    PubMed

    Afzal, Mohammad; Oommen, Sosamma; Al-Awadi, Samira

    2011-01-01

    We performed a series of experiments with Geobacillus stearothermophilus, a thermophile isolated from oil-contaminated soil in the Kuwaiti desert. The organism has a good potential for the transformation of a broad spectrum of organic molecules such as steroids, amino acids, and aromatic hydrocarbons. In the present study, we tested its potential for the transformation of a bile component, chenodeoxycholic acid (CDCA). Five transformed products, namely, cholic acid, methylcholate, methylchenodeoxycholate, 3α-hydroxy-7-oxo-5β-cholanic acid, and 7α-hydroxy-3-oxo-5β-cholanic acid, were the major transformation products catalyzed by G. stearothermophilus. Under aerobic conditions, no evidence of side chain degradation, ring cleavage, or dehydrogenation was found among the metabolites of CDCA. CDCA transformation by a thermophile is reported for the first time. PMID:21838799

  7. Structure and comparative analysis of the genes encoding component C of methyl coenzyme M reductase in the extremely thermophilic archaebacterium Methanothermus fervidus.

    PubMed Central

    Weil, C F; Cram, D S; Sherf, B A; Reeve, J N

    1988-01-01

    A 6-kilobase-pair (kbp) region of the genome of the extremely thermophilic arachaebacterium Methanothermus fervidus which encodes the alpha, beta, and gamma subunit polypeptides of component C of methyl coenzyme M reductase was cloned and sequenced. Genes encoding the beta (mcrB) and gamma (mcrG) subunits were separated by two open reading frames (designated mcrC and mcrD) which encode unknown gene products. The M. fervidus genes were preceded by ribosome-binding sites, separated by short A + T-rich intergenic regions, contained unexpectedly few NNC codons, and exhibited inflexible codon usage at some locations. Sites of transcription initiation and termination flanking the mcrBDCGA cluster of genes in M. fervidus were identified. The sequences of the genes, the encoded polypeptides, and transcription regulatory signals in M. fervidus were compared with the functionally equivalent sequences from two mesophilic methanogens (Methanococcus vannielii and Methanosarcina barkeri) and from a moderate thermophile (Methanobacterium thermoautotrophicum Marburg). The amino acid sequences of the polypeptides encoded by the mcrBCGA genes in the two thermophiles were approximately 80% identical, whereas all other pairs of these gene products contained between 50 and 60% identical amino acid residues. The mcrD gene products have diverged more than the products of the other mcr genes. Identification of highly conserved regions within mcrA and mcrB suggested oligonucleotide sequences which might be developed as hybridization probes which could be used for identifying and quantifying all methanogens. Images PMID:3170483

  8. Isolation of full-length RNA from a thermophilic cyanobacterium.

    PubMed

    Luo, X Z; Stevens, S E

    1997-11-01

    Isolation of full-length mRNA without degradation is critical in the study of in vivo gene regulation and transcription, cDNA synthesis and reverse transcription (RT)-PCR. It is particularly difficult to isolate full-length mRNA from thermophiles, which have higher turnover rates of mRNA degradation. Mastigocladus laminosus is a thermophilic heterocystous cyanobacterium. The assay of M. laminosus cell lysates showed that RNase activity was high and was resistant to the conventional guanidine thiocyanate and 2-mercaptoethanol denaturation methods. The mRNA isolated by several conventional methods was completely degraded. A method was developed to purify full-length mRNA by a combination of fast cooling, vanadyl-ribonucleoside-complex inhibition, phenol-chloroform-isoamyl alcohol extraction, lithium chloride precipitation and the lysing of cells with the French Press. This method produced high-quality, full-length mRNA in high yield. Purified mRNA was suitable for Northern blotting, cDNA synthesis and RT-PCR. This method could be applicable to other thermophiles in which the RNase activity is high and/or is resistant to guanidine thiocyanate. PMID:9383558

  9. Potential and utilization of thermophiles and thermostable enzymes in biorefining

    PubMed Central

    Turner, Pernilla; Mamo, Gashaw; Karlsson, Eva Nordberg

    2007-01-01

    In today's world, there is an increasing trend towards the use of renewable, cheap and readily available biomass in the production of a wide variety of fine and bulk chemicals in different biorefineries. Biorefineries utilize the activities of microbial cells and their enzymes to convert biomass into target products. Many of these processes require enzymes which are operationally stable at high temperature thus allowing e.g. easy mixing, better substrate solubility, high mass transfer rate, and lowered risk of contamination. Thermophiles have often been proposed as sources of industrially relevant thermostable enzymes. Here we discuss existing and potential applications of thermophiles and thermostable enzymes with focus on conversion of carbohydrate containing raw materials. Their importance in biorefineries is explained using examples of lignocellulose and starch conversions to desired products. Strategies that enhance thermostablity of enzymes both in vivo and in vitro are also assessed. Moreover, this review deals with efforts made on developing vectors for expressing recombinant enzymes in thermophilic hosts. PMID:17359551

  10. Community dynamics of cellulose-adapted thermophilic bacterial consortia.

    PubMed

    Eichorst, Stephanie A; Varanasi, Patanjali; Stavila, Vatalie; Zemla, Marcin; Auer, Manfred; Singh, Seema; Simmons, Blake A; Singer, Steven W

    2013-09-01

    Enzymatic hydrolysis of cellulose is a key process in the global carbon cycle and the industrial conversion of biomass to biofuels. In natural environments, cellulose hydrolysis is predominately performed by microbial communities. However, detailed understanding of bacterial cellulose hydrolysis is primarily confined to a few model isolates. Developing models for cellulose hydrolysis by mixed microbial consortia will complement these isolate studies and may reveal new mechanisms for cellulose deconstruction. Microbial communities were adapted to microcrystalline cellulose under aerobic, thermophilic conditions using green waste compost as the inoculum to study cellulose hydrolysis in a microbial consortium. This adaptation selected for three dominant taxa--the Firmicutes, Bacteroidetes and Thermus. A high-resolution profile of community development during the enrichment demonstrated a community transition from Firmicutes to a novel Bacteroidetes population that clusters in the Chitinophagaceae family. A representative strain of this population, strain NYFB, was successfully isolated, and sequencing of a nearly full-length 16S rRNA gene demonstrated that it was only 86% identical compared with other validated strains in the phylum Bacteroidetes. Strain NYFB grew well on soluble polysaccharide substrates, but grew poorly on insoluble polysaccharide substrates. Similar communities were observed in companion thermophilic enrichments on insoluble wheat arabinoxylan, a hemicellulosic substrate, suggesting a common model for deconstruction of plant polysaccharides. Combining observations of community dynamics and the physiology of strain NYFB, a cooperative successional model for polysaccharide hydrolysis by the Firmicutes and Bacteroidetes in the thermophilic cellulolytic consortia is proposed. PMID:23763762

  11. Hemicellulases from the ethanologenic thermophile, Thermoanaerobacter ethanolicus and related anaerobic thermophiles. Final report, September 1992--June 1996

    SciTech Connect

    Wiegel, J.

    1998-09-01

    The short term goals of this application were to characterize hemicellulases from anaerobic thermophiles on the biochemical and molecular level to extend the presently limited knowledge of hemicellulases in anaerobic thermophilic bacteria. This objective includes the following tasks: (1) Traditional purification and biochemical/biophysical characterization of xylanases from the newly isolated, slightly alkalitolerant strain NDF190, and the slightly acid-tolerant strain YS485, both with high xylanolytic activities, and of the 4-O-methyl glucuronidase and arabinosidase from strain NDF190 and the acetyl (xylan) esterase from T. ethanolicus. This also includes determining the N-terminal sequences and obtaining gene probes. (2) Elucidation of the regulation of hemicellulolytic enzymes in anaerobic thermophiles. (3) To clone into E. coli and identify the multiplicity of the enzymes involved in hemicellulose degradation by T. ethanolicus and other suitable organisms. (4) To purify and characterize the recombinant enzymes with the goal of identifying the best enzymes for cloning into the ethanologenic T. ethanolicus to obtain an optimized hemicellulose utilization by this bacterium.

  12. Hemicellulases from the ethanologenic thermophile Thermoanaerobacter ethanolicus and related anaerobic thermophiles. Final report, September 1992--June 1996

    SciTech Connect

    Wiegel, J.

    1998-05-01

    The SHORT TERM GOALS of this application were to characterize hemicellulases from anaerobic thermophiles on the biochemical and molecular level to extend the presently limited knowledge of hemicellulases in anaerobic thermophilic bacteria. This objective includes the following TASKS: (1) Traditional purification and biochemical/biophysical characterization of xylanases from the newly isolated, slightly alkalitolerant strain NDF190, and the slightly acid-tolerant strain YS485, both with high xylanolytic activities, and of the 4-0-methyl glucuronidase and arabinosidase from strain NDF190 and the acetyl (xylan) esterase from T. ethanolicus. This also includes determining the N-terminal sequences and obtaining gene probes. (2) Elucidation of the regulation of hemicellulolytic enzymes in anaerobic thermophiles. (3) To clone into E. coli and identify the multiplicity of the enzymes involved in hemicellulose degradation by T. ethanolicus and other suitable organisms. (4) To purify and characterize the recombinant enzymes with the goal of identifying the best enzymes for cloning into the ethanologenic T. ethanolicus to obtain an optimized hemicellulose utilization by this bacterium (one of our long term goals).

  13. Draft Genome Sequence of the Cellulolytic and Xylanolytic Thermophile Clostridium clariflavum Strain 4-2a.

    PubMed

    Rooney, Elise A; Rowe, Kenneth T; Guseva, Anna; Huntemann, Marcel; Han, James K; Chen, Amy; Kyrpides, Nikos C; Mavromatis, Konstantinos; Markowitz, Victor M; Palaniappan, Krishna; Ivanova, Natalia; Pati, Amrita; Liolios, Konstantinos; Nordberg, Henrik P; Cantor, Michael N; Hua, Susan X; Shapiro, Nicole; Woyke, Tanja; Lynd, Lee R; Izquierdo, Javier A

    2015-01-01

    Clostridium clariflavum strain 4-2a, a novel strain isolated from a thermophilic biocompost pile, has demonstrated an extensive capability to utilize both cellulose and hemicellulose under thermophilic anaerobic conditions. Here, we report the draft genome of this strain. PMID:26205857

  14. ENVIRONMENTAL REGULATIONS AND TECHNOLOGY - AUTOTHERMAL THERMOPHILIC AEROBIC DIGESTION OF MUNICIPAL WASTEWATER SLUDGE

    EPA Science Inventory

    This document describes a promising technology — autothermal thermophilic aerobic digestion — for meeting the current and proposed U.S. federal requirements for pathogen controJ and land application of municipal wastewater sludge. Autothermal thermophilic aerobic digestion, or AT...

  15. REACTOR MODERATOR STRUCTURE

    DOEpatents

    Greenstreet, B.L.

    1963-12-31

    A system for maintaining the alignment of moderator block structures in reactors is presented. Integral restraining grids are placed between each layer of blocks in the moderator structure, at the top of the uppermost layer, and at the bottom of the lowermost layer. Slots are provided in the top and bottom surfaces of the moderator blocks so as to provide a keying action with the grids. The grids are maintained in alignment by vertical guiding members disposed about their peripheries. (AEC)

  16. Rapid start-up of thermophilic anaerobic digestion with the turf fraction of MSW as inoculum.

    PubMed

    Suwannoppadol, Suwat; Ho, Goen; Cord-Ruwisch, Ralf

    2011-09-01

    This study aims to determine suitable start-up conditions and inoculum sources for thermophilic anaerobic digestion. Within days of incubation MSW at 55°C, methane was produced at a high rate. In an attempt to narrow down which components of typical MSW contained the thermophilic methanogens, vacuum cleaner dust, banana peel, kitchen waste, and garden waste were tested as inoculum for thermophilic methanogenesis with acetate as the substrate. Results singled out grass turf as the key source of thermophilic acetate degrading methanogenic consortia. Within 4 days of anaerobic incubation (55°C), anaerobically incubated grass turf samples produced methane accompanied by acetate degradation enabling successful start-up of thermophilic anaerobic digestion. Other essential start-up conditions are specified. Stirring of the culture was not conducive for successful start-up as it resulted specifically in propionate accumulation. PMID:21723117

  17. Isolation from soil and properties of the extreme thermophile Clostridium thermohydrosulfuricum.

    PubMed Central

    Wiegel, J; Ljungdahl, L G; Rawson, J R

    1979-01-01

    Thirteen strains of a strict anaerobic, extreme thermophilic bacterium were isolated from soil samples of moderate temperature, from a sewage plant in Georgia, and from hot springs in Utah and Wyoming. They were identified as strains of Clostridium thermohydrosulfuricum. The guanosine + cytosine content (moles percent) was 37.6 (determined by buoyant density) and 34.1 (determined by melting temperature). All strains required a factor present in yeast extract or tryptone growth. Growth characteristics were as follows: a pH range of 5 to 9, with the optimum between 6.9 to 7.5, in a temperature range of 40 to 78 degrees C, with the optimum at 68 degrees C. The doubling time, when grown on glucose at temperature and pH optima, was 1.2 h. The main products of glucose fermentation were ethanol, lactate, acetate, CO2, and H2. The fermentation was inhibited by H2. Formation of spores occurred easily on glucose-agar medium or when cultures growing at temperatures above 65 degrees C were allowed to cool to temperature below 55 degrees C. C. thermohydrosulfuricum occurs widely distributed in the natural environment. PMID:39062

  18. Three novel halotolerant and thermophilic Geobacillus strains from shallow marine vents.

    PubMed

    Maugeri, Teresa L; Gugliandolo, Concetta; Caccamo, Daniela; Stackebrandt, Erko

    2002-10-01

    During a polyphasic taxonomic analysis performed on isolates from shallow marine hydrothermal vents of Eolian Islands (Italy), three thermophilic, halotolerant bacilli, designated as strain 1bw, strain 5-2 and strain 10-1, could not be affiliated to any described species. Physiological and biochemical characteristics, membrane lipids composition, mol % G+C content, and phylogenetic relationships determined on the basis of the 16S rRNA gene sequence analysis, placed these strains within the genus Geobacillus. The three strains were only moderately related to species of Geobacillus and their relatives, members of Saccharococcus. Determination of the relatedness among each other at a higher taxonomic level by DNA-DNA reassociation experiments demonstrated the three isolates to represent three different novel Geobacillus genomospecies. The taxonomic novelty of these three marine strains was substantiated by their physiological properties and by fatty acid patterns that did not match closely those of any Geobacillus type strain. These three novel strains could be of interest to biotechnology because of their ability to produce exopolysaccharides and to adhere on polystirene, characteristics undescribed so far for other Geobacillus species. They are also able to utilise hydrocarbons such as gas oil, kerosene and mineral lubricating oil. Strain 5-2 is tolerant to zinc. PMID:12421083

  19. Melghiribacillus thermohalophilus gen. nov., sp. nov., a novel filamentous, endospore-forming, thermophilic and halophilic bacterium.

    PubMed

    Addou, Nariman Ammara; Schumann, Peter; Spröer, Cathrin; Ben Hania, Wajdi; Hacene, Hocine; Fauque, Guy; Cayol, Jean-Luc; Fardeau, Marie-Laure

    2015-04-01

    A novel filamentous, endospore-forming, thermophilic and moderately halophilic bacterium designated strain Nari2A(T) was isolated from soil collected from an Algerian salt lake, Chott Melghir. The novel isolate was Gram-staining-positive, aerobic, catalase-negative and oxidase-positive. Optimum growth occurred at 50-55 °C, 7-10% (w/v) NaCl and pH 7-8. The strain exhibited 95.4, 95.4 and 95.2% 16S rRNA gene sequence similarity to Thalassobacillus devorans G19.1(T), Sediminibacillus halophilus EN8d(T) and Virgibacillus kekensis YIM-kkny16(T), respectively. The major menaquinone was MK-7. The polar lipid profile consisted of phosphatidylglycerol, diphosphatidylglycerol, three unknown phosphoglycolipids and two unknown phospholipids. The predominant cellular fatty acids were iso-C(15 : 0) and iso-C(17 : 0). The DNA G+C content was 41.9 mol%. Based on the phenotypic, chemotaxonomic and phylogenetic data, strain Nari2A(T) is considered to represent a novel species of a new genus in the family Bacillaceae , order Bacillales , for which the name Melghiribacillus thermohalophilus gen. nov., sp. nov. is proposed. The type strain of Melghiribacillus thermohalophilus is Nari2A(T) ( = DSM 25894(T) = CCUG 62543(T)). PMID:25604343

  20. Rapid establishment of thermophilic anaerobic microbial community during the one-step startup of thermophilic anaerobic digestion from a mesophilic digester.

    PubMed

    Tian, Zhe; Zhang, Yu; Li, Yuyou; Chi, Yongzhi; Yang, Min

    2015-02-01

    The purpose of this study was to explore how fast the thermophilic anaerobic microbial community could be established during the one-step startup of thermophilic anaerobic digestion from a mesophilic digester. Stable thermophilic anaerobic digestion was achieved within 20 days from a mesophilic digester treating sewage sludge by adopting the one-step startup strategy. The succession of archaeal and bacterial populations over a period of 60 days after the temperature increment was followed by using 454-pyrosequencing and quantitative PCR. After the increase of temperature, thermophilic methanogenic community was established within 11 days, which was characterized by the fast colonization of Methanosarcina thermophila and two hydrogenotrophic methanogens (Methanothermobacter spp. and Methanoculleus spp.). At the same time, the bacterial community was dominated by Fervidobacterium, whose relative abundance rapidly increased from 0 to 28.52 % in 18 days, followed by other potential thermophilic genera, such as Clostridium, Coprothermobacter, Anaerobaculum and EM3. The above result demonstrated that the one-step startup strategy could allow the rapid establishment of the thermophilic anaerobic microbial community. PMID:25463927

  1. Abiotic and microbiotic factors controlling biofilm formation by thermophilic sporeformers.

    PubMed

    Zhao, Yu; Caspers, Martien P M; Metselaar, Karin I; de Boer, Paulo; Roeselers, Guus; Moezelaar, Roy; Nierop Groot, Masja; Montijn, Roy C; Abee, Tjakko; Kort, Remco

    2013-09-01

    One of the major concerns in the production of dairy concentrates is the risk of contamination by heat-resistant spores from thermophilic bacteria. In order to acquire more insight in the composition of microbial communities occurring in the dairy concentrate industry, a bar-coded 16S amplicon sequencing analysis was carried out on milk, final products, and fouling samples taken from dairy concentrate production lines. The analysis of these samples revealed the presence of DNA from a broad range of bacterial taxa, including a majority of mesophiles and a minority of (thermophilic) spore-forming bacteria. Enrichments of fouling samples at 55°C showed the accumulation of predominantly Brevibacillus and Bacillus, whereas enrichments at 65°C led to the accumulation of Anoxybacillus and Geobacillus species. Bacterial population analysis of biofilms grown using fouling samples as an inoculum indicated that both Anoxybacillus and Geobacillus preferentially form biofilms on surfaces at air-liquid interfaces rather than on submerged surfaces. Three of the most potent biofilm-forming strains isolated from the dairy factory industrial samples, including Geobacillus thermoglucosidans, Geobacillus stearothermophilus, and Anoxybacillus flavithermus, have been characterized in detail with respect to their growth conditions and spore resistance. Strikingly, Geobacillus thermoglucosidans, which forms the most thermostable spores of these three species, is not able to grow in dairy intermediates as a pure culture but appears to be dependent for growth on other spoilage organisms present, probably as a result of their proteolytic activity. These results underscore the importance of abiotic and microbiotic factors in niche colonization in dairy factories, where the presence of thermophilic sporeformers can affect the quality of end products. PMID:23851093

  2. Abiotic and Microbiotic Factors Controlling Biofilm Formation by Thermophilic Sporeformers

    PubMed Central

    Zhao, Yu; Caspers, Martien P. M.; Metselaar, Karin I.; de Boer, Paulo; Roeselers, Guus; Moezelaar, Roy; Nierop Groot, Masja; Montijn, Roy C.; Abee, Tjakko

    2013-01-01

    One of the major concerns in the production of dairy concentrates is the risk of contamination by heat-resistant spores from thermophilic bacteria. In order to acquire more insight in the composition of microbial communities occurring in the dairy concentrate industry, a bar-coded 16S amplicon sequencing analysis was carried out on milk, final products, and fouling samples taken from dairy concentrate production lines. The analysis of these samples revealed the presence of DNA from a broad range of bacterial taxa, including a majority of mesophiles and a minority of (thermophilic) spore-forming bacteria. Enrichments of fouling samples at 55°C showed the accumulation of predominantly Brevibacillus and Bacillus, whereas enrichments at 65°C led to the accumulation of Anoxybacillus and Geobacillus species. Bacterial population analysis of biofilms grown using fouling samples as an inoculum indicated that both Anoxybacillus and Geobacillus preferentially form biofilms on surfaces at air-liquid interfaces rather than on submerged surfaces. Three of the most potent biofilm-forming strains isolated from the dairy factory industrial samples, including Geobacillus thermoglucosidans, Geobacillus stearothermophilus, and Anoxybacillus flavithermus, have been characterized in detail with respect to their growth conditions and spore resistance. Strikingly, Geobacillus thermoglucosidans, which forms the most thermostable spores of these three species, is not able to grow in dairy intermediates as a pure culture but appears to be dependent for growth on other spoilage organisms present, probably as a result of their proteolytic activity. These results underscore the importance of abiotic and microbiotic factors in niche colonization in dairy factories, where the presence of thermophilic sporeformers can affect the quality of end products. PMID:23851093

  3. Energetic and hydrogen limitations of thermophilic and hyperthermophilic methanogens

    NASA Astrophysics Data System (ADS)

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

    2013-12-01

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

  4. Thermophilic and alkaliphilic Actinobacteria: biology and potential applications

    PubMed Central

    Shivlata, L.; Satyanarayana, Tulasi

    2015-01-01

    Microbes belonging to the phylum Actinobacteria are prolific sources of antibiotics, clinically useful bioactive compounds and industrially important enzymes. The focus of the current review is on the diversity and potential applications of thermophilic and alkaliphilic actinobacteria, which are highly diverse in their taxonomy and morphology with a variety of adaptations for surviving and thriving in hostile environments. The specific metabolic pathways in these actinobacteria are activated for elaborating pharmaceutically, agriculturally, and biotechnologically relevant biomolecules/bioactive compounds, which find multifarious applications. PMID:26441937

  5. A novel thermophilic methane-oxidizing bacteria from thermal springs of Uzon volcano caldera, Kamchatka

    NASA Astrophysics Data System (ADS)

    Dvorianchikova, E.; Kizilova, A.; Kravchenko, I.; Galchenko, V.

    2012-04-01

    Methane is a radiatively active trace gas, contributing significantly to the greenhouse effect. It is 26 times more efficient in absorbing and re-emitting infrared radiation than carbon dioxide. Methanotrophs play an essential role in the global carbon cycle by oxidizing 50-75% of the biologically produced methane in situ, before it reaches the atmosphere. Methane-oxidizing bacteria are isolated from the various ecosystems and described at present. Their biology, processes of methane oxidation in fresh-water, marsh, soil and marine habitats are investigated quite well. Processes of methane oxidation in places with extreme physical and chemical conditions (high or low , salinity and temperature values) are studied in much smaller degree. Such ecosystems occupy a considerable part of the Earth's surface. The existence of aerobic methanotrophs inhabiting extreme environments has been verified so far by cultivation experiments and direct detection of methane monooxygenase genes specific to almost all aerobic methanotrophs. Thermophilic and thermotolerant methanotrophs have been isolated from such extreme environments and consist of the gammaproteobacterial (type I) genera Methylothermus, Methylocaldum, Methylococcus and the verrucomicrobial genus Methylacidiphilum. Uzon volcano caldera is a unique area, where volcanic processes still happen today. Hydrothermal springs of the area are extreme ecosystems which microbial communities represent considerable scientific interest of fundamental and applied character. A thermophilic aerobic methane-oxidising bacterium was isolated from a sediment sample from a hot spring (56.1; 5.3) of Uzon caldera. Strain S21 was isolated using mineral low salt medium. The headspace gas was composed of CH4, Ar, CO2, and O2 (40:40:15:5). The temperature of cultivation was 50, pH 5.5. Cells of strain S21 in exponential and early-stationary phase were coccoid bacilli, about 1 μm in diameter, and motile with a single polar flagellum. PCR and

  6. The Isis cold moderators

    SciTech Connect

    Allen, G. M.; Broome, T. A.; Burridge, R. A.; Cragg, D.; Hall, R.; Haynes, D.; Hirst, J.; Hogston, J. R.; Jones, H. H.; Sexton, J.; Wright, P.

    1997-09-01

    ISIS is a pulsed spallation neutron source where neutrons are produced by the interaction of a 160 kW proton beam of energy 800 MeV in a water-cooled Tantalum Target. The fast neutrons produced are thermalized in four moderators: two ambient water, one liquid methane operating at 100K and a liquid hydrogen moderator at 20 K. This paper gives a description of the construction of both cold moderator systems, details of the operating experience and a description of the current development program.

  7. Knowledge-based discovery for designing CRISPR-CAS systems against invading mobilomes in thermophiles.

    PubMed

    Chellapandi, P; Ranjani, J

    2015-09-01

    Clustered regularly interspaced short palindromic repeats (CRISPRs) are direct features of the prokaryotic genomes involved in resistance to their bacterial viruses and phages. Herein, we have identified CRISPR loci together with CRISPR-associated sequences (CAS) genes to reveal their immunity against genome invaders in the thermophilic archaea and bacteria. Genomic survey of this study implied that genomic distribution of CRISPR-CAS systems was varied from strain to strain, which was determined by the degree of invading mobiloms. Direct repeats found to be equal in some extent in many thermopiles, but their spacers were differed in each strain. Phylogenetic analyses of CAS superfamily revealed that genes cmr, csh, csx11, HD domain, devR were belonged to the subtypes of cas gene family. The members in cas gene family of thermophiles were functionally diverged within closely related genomes and may contribute to develop several defense strategies. Nevertheless, genome dynamics, geological variation and host defense mechanism were contributed to share their molecular functions across the thermophiles. A thermophilic archaean, Thermococcus gammotolerans and thermophilic bacteria, Petrotoga mobilis and Thermotoga lettingae have shown superoperons-like appearance to cluster cas genes, which were typically evolved for their defense pathways. A cmr operon was identified with a specific promoter in a thermophilic archaean, Caldivirga maquilingensis. Overall, we concluded that knowledge-based genomic survey and phylogeny-based functional assignment have suggested for designing a reliable genetic regulatory circuit naturally from CRISPR-CAS systems, acquired defense pathways, to thermophiles in future synthetic biology. PMID:26279704

  8. Aeration control of thermophilic aerobic digestion using fluorescence monitoring.

    PubMed

    Kim, Young-Kee; Oh, Byung-Keun

    2009-01-01

    The thermophilic aerobic digestion (TAD) process is recognized as an effective method for rapid waste activated sludge (WAS) degradation and the deactivation of pathogenic microorganisms. Yet, high energy costs due to heating and aeration have limited the commercialization of economical TAD processes. Previous research on autothermal thermophilic aerobic digestion (ATAD) has already reduced the heating cost. However, only a few studies have focused on reducing the aeration cost. Therefore, this study applied a two-step aeration control strategy to a fill-and-draw mode semicontinuous TAD process. The NADH-dependent fluorescence was monitored throughout the TAD experiment, and the aeration rate shifted according to the fluorescence intensity. As a result, the simple two-step aeration control operation achieved a 20.3% reduction in the total aeration, while maintaining an effective and stable operation. It is also expected that more savings can be achieved with a further reduction of the lower aeration rate or multisegmentation of the aeration rate. PMID:19190414

  9. Solubilization, solution equilibria, and biodegradation of PAH's under thermophilic conditions.

    PubMed

    Viamajala, Sridhar; Peyton, Brent M; Richards, Lee A; Petersen, James N

    2007-01-01

    Biodegradation rates of PAHs are typically low at mesophilic conditions and it is believed that the kinetics of degradation is controlled by PAH solubility and mass transfer rates. Solubility tests were performed on phenanthrene, fluorene and fluoranthene at 20 degrees C, 40 degrees C and 60 degrees C and, as expected, a significant increase in the equilibrium solubility concentration and of the rate of dissolution of these polycyclic aromatic hydrocarbons (PAHs) was observed with increasing temperature. A first-order model was used to describe the PAH dissolution kinetics and the thermodynamic property changes associated with the dissolution process (enthalpy, entropy and Gibb's free energy of solution) were evaluated. Further, other relevant thermodynamic properties for these PAHs, including the activity coefficients at infinite dilution, Henry's law constants and octanol-water partition coefficients, were calculated in the temperature range 20-60 degrees C. In parallel with the dissolution studies, three thermophilic Geobacilli were isolated from compost that grew on phenanthrene at 60 degrees C and degraded the PAH more rapidly than other reported mesophiles. Our results show that while solubilization rates of PAHs are significantly enhanced at elevated temperatures, the biodegradation of PAHs under thermophilic conditions is likely mass transfer limited due to enhanced degradation rates. PMID:16934313

  10. Global transport of thermophilic bacteria in atmospheric dust.

    PubMed

    Perfumo, Amedea; Marchant, Roger

    2010-04-01

    Aerosols from dust storms generated in the Sahara-Sahel desert area of Africa are transported north over Europe and periodically result in dry dust precipitation in the Mediterranean region. Samples of dust collected in Turkey and Greece following two distinct desert storm events contained viable thermophilic organisms of the genus Geobacillus, namely G. thermoglucosidasius and G. thermodenitrificans, and the recently reclassified Aeribacillus pallidus (formerly Geobacillus pallidus). We present here evidence that African dust storms create an atmospheric bridge between distant geographical regions and that they are also probably the source of thermophilic geobacilli later deposited over northern Europe by rainfall or dust plumes themselves. The same organisms (99% similarity in the 16S rDNA sequence) were found in dust collected in the Mediterranean region and inhabiting cool soils in Northern Ireland. This study also contributes new insights to the taxonomic identification of Geobacillus sp. Attempts to identify these organisms using 16S rRNA gene sequences have revealed that they contain multiple and diverse copies of the ribosomal RNA operon (up to 10 copies with nine different sequences), which dictates care in interpreting data about the systematics of this genus. PMID:23766086

  11. Hyper-thermophilic aerobic bacterial ecology for space agriculture

    NASA Astrophysics Data System (ADS)

    Oshima, T.; Kanazawa, S.; Moriya, T.; Ishikawa, Y.; Hashimoto, H.; Yamashita, M.; Space Agriculture Task Force, J.

    A material recycling is one of core issues in engineering for habitation on extraterrestrial bodies such as Mars A new composting system has been developed in Japan which utilizes some thermophilic bacteria to attain higher temperature than normally expected in the ordinary composting system Dead body of rat was found to be eaten up by the thermophilic bacteria under aerated condition and oxidized to carbon dioxide and few other inorganics within two hours Ecology of these composting bacteria is structured on the intensive symbiotic interactions among various species that participate in various reaction networks in a concert Complexity in the composting bacteria might be based on multiple interaction and interdependency among participating species and organisms Species identification and phylogeny of symbiotic bacteria and understanding of their ecology have been made Those bacterial systems are active and durable under temperature high in a range of 80 to 100 r C Biological combustion release heat and temperature goes up when air is fed through the reaction bed Since microbial activity decreases at exceeding temperature and release of heat decreases as well temperature in the reacting bed itself-regulated in the range Even though it should be verified composting bacteria themselves are presumed to be safe for human agricultural plant and animal species Their activity is restricted only to the condition under elevated temperature Their activities depend greatly on their symbiotic partners and extreme environment created by them The

  12. Solubilization, Solution Equilibria, and Biodegradation of PAH's under Thermophilic Conditions

    SciTech Connect

    Viamajala, S.; Peyton, B. M.; Richards, L. A.; Petersen, J. N.

    2007-01-01

    Biodegradation rates of PAHs are typically low at mesophilic conditions and it is believed that the kinetics of degradation is controlled by PAH solubility and mass transfer rates. Solubility tests were performed on phenanthrene, fluorene and fluoranthene at 20 C, 40 C and 60 C and, as expected, a significant increase in the equilibrium solubility concentration and of the rate of dissolution of these polycyclic aromatic hydrocarbons (PAHs) was observed with increasing temperature. A first-order model was used to describe the PAH dissolution kinetics and the thermodynamic property changes associated with the dissolution process (enthalpy, entropy and Gibb's free energy of solution) were evaluated. Further, other relevant thermodynamic properties for these PAHs, including the activity coefficients at infinite dilution, Henry's law constants and octanol-water partition coefficients, were calculated in the temperature range 20-60 C. In parallel with the dissolution studies, three thermophilic Geobacilli were isolated from compost that grew on phenanthrene at 60 C and degraded the PAH more rapidly than other reported mesophiles. Our results show that while solubilization rates of PAHs are significantly enhanced at elevated temperatures, the biodegradation of PAHs under thermophilic conditions is likely mass transfer limited due to enhanced degradation rates.

  13. Purification and physicochemical properties of lipase from thermophilic Bacillus aerius.

    PubMed

    Saun, Nitin Kumar; Mehta, Poonam; Gupta, Reena

    2014-01-01

    A thermophilic bacterial isolate producing lipase was isolated from soil of hot spring and identified as Bacillus aerius (MTCC 10978). Peak lipase activity was observed when 30 h old inoculum was used and incubated in shaking conditions for 48 h. The optimal temperature and pH for the bacterial growth and lipase production was found to be 55°C and 8.0 respectively with cottonseed oil as carbon source, yeast extract and beef extract as nitrogen source. The enzyme produced by thermophilic Bacillus aerius (MTCC 10978) was purified to 9-fold with 7.2% recovery by ammonium sulfate precipitation and DEAE-Cellulose Column Chromatography. The enzyme was found to be a protein having a molecular weight of 33 kDa on SDS-PAGE. The Km and Vmax value of lipase using p-nitrophenyl palmitate as calculated from Lineweaver-Burk plot was 2.13 mM and 0.66 µmol/ml/min respectively. PMID:25391687

  14. Report: antibiotic production by thermophilic Bacillus specie SAT-4.

    PubMed

    Muhammad, Syed Aun; Ahmad, Safia; Hameed, Abdul

    2009-07-01

    Production of antimicrobial compounds seems to be a general phenomenon for most bacteria. The prevalence of antimicrobial resistance among key microbial pathogens is increasing at an alarming rate worldwide. Current solutions involve development of a more rationale approach to antibiotic use and discover of new antimicrobials. Bacillus species produce a large number of biological compounds active against bacteria, fungi, protozoa and viruses. The process of production usually involves screening of wide range of microorganisms, testing and modification. Production is carried out using fermentation. Thermophilic spore-forming, gram positive, motile rod bacterial strains were isolated from the Thar Desserts, Sindh Province, Pakistan. These strains were screened and checked for antibacterial activity. The best activity was observed by SAT4 against Micrococcus luteus, Staphylococcus aureus and Pseudomonas aeroginosa. The activity was only observed against gram positive bacteria and no activity was seen against Pseudomonas aeroginosa. Thermophilic Bacillus specie SAT4 was found to be active in the fermentation process to produce the antimicrobial agents. Further optimizations of different conditions (time of incubation, media, pH, glucose concentrations, nitrogen concentrations, and temperature) for antimicrobial production by the selected bacterial strain was performed. Agar diffusion assay was performed to evaluate the antibacterial activity. Optimum conditions for the production of antimicrobials by selected isolate were observed to be 48 hour, pH 5, temperature 55 degrees C, 2% glucose and 1.5% nitrogen concentration. This newly isolated bacterial strain has great potential for antimicrobial production at industrial scale. PMID:19553186

  15. Conformational Preferences Underlying Reduced Activity of a Thermophilic Ribonuclease H

    PubMed Central

    Stafford, Kate A.; Trbovic, Nikola; Butterwick, Joel A.; Abel, Robert; Friesner, Richard A.; Palmer, Arthur G.

    2015-01-01

    The conformational basis for reduced activity of the thermophilic ribonuclease HI enzyme from Thermus thermophilus, compared to its mesophilic homolog from Escherichia coli, is elucidated using a combination of NMR spectroscopy and molecular dynamics (MD) simulations. Explicit-solvent all-atom MD simulations of the two wild-type proteins and an E. coli mutant in which a glycine residue is inserted after position 80 to mimic the T. thermophilus protein reproduce the differences in conformational dynamics determined from 15N spin-relaxation NMR spectroscopy of three loop regions that surround the active site and contain functionally important residues: the glycine-rich region, the handle region, and the β5/αE loop. Examination of the MD trajectories indicates that the thermophilic protein samples conformations productive for substrate binding and activity less frequently than the mesophilic enzyme, although these differences may manifest as either increased or decreased relative flexibility of the different regions. Additional MD simulations indicate that mutations increasing activity of the T. thermophilus enzyme at mesophilic temperatures do so by reconfiguring the local environments of the mutated sites to more closely resemble active conformations. Taken together, the results show that both locally increased and decreased flexibility contribute to an overall reduction in activity of T. thermophilus ribonuclease H compared to its mesophilic E. coli homolog. PMID:25550198

  16. Psychrophilic, mesophilic, and thermophilic triosephosphate isomerases from three clostridial species.

    PubMed Central

    Shing, Y W; Akagi, J M; Himes, R H

    1975-01-01

    Triosephosphate isomerase was purified to homogeneity as judged by analytical gel electrophoresis from clostridium sp. strain 69, clostridium pasteurianum, and C. thermosaccharolyticum, which grow optimally at 18, 37, and 55 C, respectively. Comparative studies on these purified proteins showed that they had the same molecular weight (53,000) and subunit molecular weight (26,500). They were equally susceptible to the active site-directed inhibitor, glycidol phosphate. However, their temperature and pH optima, as well as their stabilities to heat, urea, and sodium dodecyl sulfate, differed. The proteins also had different mobilities in acrylamide gel electrophoresis. This difference in ionic character was also reflected in the elution behavior of the enzymes from hydroxyapatite and in the isoelectric points determined by isoelectric focusing in acrylamide gel. The amino acid composition of these proteins showed that the thermophilic enzyme contains a greater amount of proline than the other enzymes. The ratio of acidic amino acids to basic amino acids was 1.79, 1.38, and 1.66 for the thermophilic mesophilic and psychrophilic enzymes, respectively. This is consistent with the relative isoelectric point values of these three enzymes. Images PMID:235509

  17. Microbial community dynamics in thermophilic undefined milk starter cultures.

    PubMed

    Parente, Eugenio; Guidone, Angela; Matera, Attilio; De Filippis, Francesca; Mauriello, Gianluigi; Ricciardi, Annamaria

    2016-01-18

    Model undefined thermophilic starter cultures were produced from raw milk of nine pasta-filata cheesemaking plants using a selective procedure based on pasteurization and incubation at high temperature with the objective of studying the microbial community dynamics and the variability in performances under repeated (7-13) reproduction cycles with backslopping. The traditional culture-dependent approach, based on random isolation and molecular characterization of isolates was coupled to the determination of pH and the evaluation of the ability to produce acid and fermentation metabolites. Moreover, a culture-independent approach based on amplicon-targeted next-generation sequencing was employed. The microbial diversity was evaluated by 16S rRNA gene sequencing (V1-V3 regions), while the microdiversity of Streptococcus thermophilus populations was explored by using novel approach based on sequencing of partial amplicons of the phosphoserine phosphatase gene (serB). In addition, the occurrence of bacteriophages was evaluated by qPCR and by multiplex PCR. Although it was relatively easy to select for a community dominated by thermophilic lactic acid bacteria (LAB) within a single reproduction cycle, final pH, LAB populations and acid production activity fluctuated over reproduction cycles. Both culture-dependent and -independent methods showed that the cultures were dominated by either S. thermophilus or Lactobacillus delbrueckii subsp. lactis or by both species. Nevertheless, subdominant mesophilic species, including lactococci and spoilage organisms, persisted at low levels. A limited number of serB sequence types (ST) were present in S. thermophilus populations. L. delbrueckii and Lactococcus lactis bacteriophages were below the detection limit of the method used and high titres of cos type S. thermophilus bacteriophages were detected in only two cases. In one case a high titre of bacteriophages was concurrent with a S. thermophilus biotype shift in the culture

  18. Monitoring of thermophilic adaptation of mesophilic anaerobe fermentation of sugar beet pressed pulp.

    PubMed

    Tukacs-Hájos, Annamária; Pap, Bernadett; Maróti, Gergely; Szendefy, Judit; Szabó, Piroska; Rétfalvi, Tamás

    2014-08-01

    Anaerobe fermentation of sugar beet pressed pulp was investigated in pilot-scale digesters. Thermophilic adaptation of mesophilic culture was monitored using chemical analysis and metagenomic characterization of the sludge. Temperature adaptation was achieved by increasing the temperature gradually (2 °C day(-1)) and by greatly decreasing the OLR. During stable run, the OLR was increased gradually to 11.29 kg VS m(-3)d(-1) and biogas yield was 5% higher in the thermophilic reactor. VFA levels increased in the thermophilic reactor with increased OLR (acetic acid 646 mg L(-1), propionic acid 596 mg L(-1)), then VFA decreased and the operation was manageable beside the relative high tVFA (1300-2000 mg L(-1)). The effect of thermophilic adaptation on the microbial communities was studied using a sequencing-based metagenomic approach. Connections between physico-chemical parameters and populations of bacteria and methanogen archaea were revealed. PMID:24926601

  19. ENGINEERING AND ECONOMIC ASSESSMENT OF AUTOHEATED THERMOPHILIC AEROBIC DIGESTION WITH AIR AERATION

    EPA Science Inventory

    A major disadvantage of aerobic digestion is that it requires long detention times, particularly in colder climates, to insure adequate stabilization. Autoheated thermophilic aerobic digestion (ATAD) offers the potential to decrease the required detention time. ATAD takes advanta...

  20. A novel membrane distillation-thermophilic bioreactor system: biological stability and trace organic compound removal.

    PubMed

    Wijekoon, Kaushalya C; Hai, Faisal I; Kang, Jinguo; Price, William E; Guo, Wenshan; Ngo, Hao H; Cath, Tzahi Y; Nghiem, Long D

    2014-05-01

    The removal of trace organic compounds (TrOCs) by a novel membrane distillation-thermophilic bioreactor (MDBR) system was examined. Salinity build-up and the thermophilic conditions to some extent adversely impacted the performance of the bioreactor, particularly the removal of total nitrogen and recalcitrant TrOCs. While most TrOCs were well removed by the thermophilic bioreactor, compounds containing electron withdrawing functional groups in their molecular structure were recalcitrant to biological treatment and their removal efficiency by the thermophilic bioreactor was low (0-53%). However, the overall performance of the novel MDBR system with respect to the removal of total organic carbon, total nitrogen, and TrOCs was high and was not significantly affected by the conditions of the bioreactor. All TrOCs investigated here were highly removed (>95%) by the MDBR system. Biodegradation, sludge adsorption, and rejection by MD contribute to the removal of TrOCs by MDBR treatment. PMID:24658107

  1. [Treatment of Flue Gas from Sludge Drying Process by A Thermophilic Biofilter].

    PubMed

    Chen, Wen-he; Deng, Ming-jia; Luo, Hui; Ding, Wen-iie; Li, Lin; Lin, Jian; Liu, Jun-xin

    2016-01-15

    A thermophilic biofilter was employed to treat the flue gas generated from sludge drying process, and the performance in both the start period and the stationary phase was studied under the gas flow rate of 2 700-3 100 m3 x h(-1) and retention time of 21.88-25.10 s. The results showed that the thermophilic biofilter could effectively treat gases containing sulfur dioxide, ammonia and volatile organic compounds (VOC). The removal efficiencies could reach 100%, 93.61% and 87.01%, respectively. Microbial analysis indicated that most of the population belonged to thermophilic bacteria. Paenibacillus sp., Chelatococcus sp., Bacillus sp., Clostridium thermosuccinogenes, Pseudoxanthomonas sp. and Geobacillus debilis which were abundant in the thermophilic biofilter, had the abilities of denitrification, desulfurization and degradation of volatile organic compounds. PMID:27078980

  2. REACTOR MODERATOR STRUCTURE

    DOEpatents

    Fraas, A.P.; Tudor, J.J.

    1963-08-01

    An improved moderator structure for nuclear reactors consists of moderator blocks arranged in horizontal layers to form a multiplicity of vertically stacked columns of blocks. The blocks in each vertical column are keyed together, and a ceramic grid is disposed between each horizontal layer of blocks. Pressure plates cover- the lateral surface of the moderator structure in abutting relationship with the peripheral terminal lengths of the ceramic grids. Tubular springs are disposed between the pressure plates and a rigid external support. The tubular springs have their axes vertically disposed to facilitate passage of coolant gas through the springs and are spaced apart a selected distance such that at sonae preselected point of spring deflection, the sides of the springs will contact adjacent springs thereby causing a large increase in resistance to further spring deflection. (AEC)

  3. Cold moderators at ORNL

    SciTech Connect

    Lucas, A. T.

    1997-09-01

    The Advanced Neutron Source (ANS) cold moderators were not an 'Oak Ridge first', but would have been the largest both physically and in terms of cold neutron flux. Two cold moderators were planned each 410 mm in diameter and containing about 30L of liquid deuterium. They were to be completely independent of each other. A modular system design was used to provide greater reliability and serviceability. When the ANS was terminated, up–grading of the resident High Flux Isotope Reactor (HFIR) was examined and an initial study was made into the feasibility of adding a cold source. Because the ANS design was modular, it was possible to use many identical design features. Sub-cooled liquid at 4 bar abs was initially chosen for the HFIR design concept, but this was subsequently changed to 15 bar abs to operate above the critical pressure. As in the ANS, the hydrogen will operate at a constant pressure throughout the temperature range and a completely closed loop with secondary containment was adopted. The heat load of 2 kW made the heat flux comparable with that of the ANS. Subsequent studies into the construction of cryogenic moderators for the proposed new Synchrotron Neutron source indicated that again many of the same design concepts could be used. By connecting the two cold sources together in series, the total heat load of 2 kW is very close to that of the HFIR allowing a very similar supercritical hydrogen system to be configured. The two hydrogen moderators of the SNS provide a comparable heat load to the HFIR moderator. It is subsequently planned to connect the two in series and operate from a single cold loop system, once again using supercritical hydrogen. The spallation source also provided an opportunity to re-examine a cold pellet solid methane moderator operating at 20K.

  4. Coupled moderator neutronics

    SciTech Connect

    Russell, G.J.; Pitcher, E.J.; Ferguson, P.D.

    1995-12-01

    Optimizing the neutronic performance of a coupled-moderator system for a Long-Pulse Spallation Source is a new and challenging area for the spallation target-system designer. For optimal performance of a neutron source, it is essential to have good communication with instrument scientists to obtain proper design criteria and continued interaction with mechanical, thermal-hydraulic, and materials engineers to attain a practical design. A good comprehension of the basics of coupled-moderator neutronics will aid in the proper design of a target system for a Long-Pulse Spallation Source.

  5. Genetics of thermophilic bacteria. [Bacillus stearothermophilus:a2

    SciTech Connect

    Welker, N.E.

    1991-01-01

    Organisms adapted to high temperature have evolved a variety of unique solutions to the biochemical problems imposed by this environment. Adaptation is commonly used to describe the biochemical properties of organisms which have become adapted to their environment (genetic adaptation). It can also mean the direct response-at the cellular level-of an organism to changes in temperature (physiological adaptation). Thermophilic bacilli (strains of Bacillus stearothermophilus) can exhibit a variety of biochemical adaptations in response to changes in temperature. These include changes in the composition and stability of the membrane, metabolic potential, the transport of amino acids, regulatory mechanisms, ribose methylation of tRNA, protein thermostability, and nutritional requirements. The objectives of the research were to develop efficient and reliable genetic systems to analyze and manipulate B. Stearothermophilus, and to use these systems initiate a biochemical, molecular, and genetic investigations of genes that are required for growth at high temperature.

  6. Biocorrosive Thermophilic Microbial Communities in Alaskan North Slope Oil Facilities

    SciTech Connect

    Duncan, Kathleen E.; Gieg, Lisa M.; Parisi, Victoria A.; Tanner, Ralph S.; Green Tringe, Susannah; Bristow, Jim; Suflita, Joseph M.

    2009-09-16

    Corrosion of metallic oilfield pipelines by microorganisms is a costly but poorly understood phenomenon, with standard treatment methods targeting mesophilic sulfatereducing bacteria. In assessing biocorrosion potential at an Alaskan North Slope oil field, we identified thermophilic hydrogen-using methanogens, syntrophic bacteria, peptideand amino acid-fermenting bacteria, iron reducers, sulfur/thiosulfate-reducing bacteria and sulfate-reducing archaea. These microbes can stimulate metal corrosion through production of organic acids, CO2, sulfur species, and via hydrogen oxidation and iron reduction, implicating many more types of organisms than are currently targeted. Micromolar quantities of putative anaerobic metabolites of C1-C4 n-alkanes in pipeline fluids were detected, implying that these low molecular weight hydrocarbons, routinely injected into reservoirs for oil recovery purposes, are biodegraded and provide biocorrosive microbial communities with an important source of nutrients.

  7. Pathway engineering to improve ethanol production by thermophilic bacteria

    SciTech Connect

    Lynd, L.R.

    1998-12-31

    Continuation of a research project jointly funded by the NSF and DOE is proposed. The primary project goal is to develop and characterize strains of C. thermocellum and C. thermosaccharolyticum having ethanol selectivity similar to more convenient ethanol-producing organisms. An additional goal is to document the maximum concentration of ethanol that can be produced by thermophiles. These goals build on results from the previous project, including development of most of the genetic tools required for pathway engineering in the target organisms. As well, we demonstrated that the tolerance of C. thermosaccharolyticum to added ethanol is sufficiently high to allow practical utilization should similar tolerance to produced ethanol be demonstrated, and that inhibition by neutralizing agents may explain the limited concentrations of ethanol produced in studies to date. Task 1 involves optimization of electrotransformation, using either modified conditions or alternative plasmids to improve upon the low but reproducible transformation, frequencies we have obtained thus far.

  8. The bioleaching of different sulfide concentrates using thermophilic bacteria

    NASA Astrophysics Data System (ADS)

    Torres, F.; Blázquez, M. L.; González, F.; Ballester, A.; Mier, J. L.

    1995-05-01

    The bioleaching of different mineral sulfide concentrates with thermophilic bacteria (genus Sulfolobus @#@) was studied. Since the use of this type of bacteria in leaching systems involves stirring and the control of temperature, the influence of the type of stirring and the pulp density on dissolution rates was studied in order to ascertain the optimum conditions for metal recovery. At low pulp densities, the dissolution kinetic was favored by pneumatic stirring, but for higher pulp densities, orbital stirring produced the best results. A comparative study of three differential concentrates, one mixed concentrate, and one global concentrate was made. Copper and iron extraction is directly influenced by bacterial activity, while zinc dissolution is basically due to an indirect mechanism that is activated in the presence of copper ions. Galvanic interactions between the different sulfides favors the selective bioleaching of some phases (sphalerite and chalcopyrite) and leads to high metal recovery rates. However, the formation of galvanic couples depends on the type of concentrate.

  9. A hexokinase with broad sugar specificity from a thermophilic bacterium.

    PubMed

    Bae, Jungdon; Kim, Dooil; Choi, Yongseok; Koh, Sukhoon; Park, Jung Eun; Su Kim, Joong; Moon, Seong Hoon; Park, Bo-Hyun; Park, Miri; Song, Hye-Eun; Hong, Suk-In; Lee, Dae-Sil

    2005-09-01

    A recombinant thermophilic Thermus caldophilus GK24 hexokinase, one of the ROK-type (repressor protein, open reading frames, and sugar kinase) proteins, exists uniquely as a 120 kDa molecule with four subunits (31 kDa), in contrast to eukaryotic and bacterial sugar kinases which are monomers or dimers. The optimal temperature and pH for the enzyme reaction are 70-80 degrees C and 7.5, respectively. This enzyme shows broad specificity toward glucose, mannose, glucosamine, allose, 2-deoxyglucose, and fructose. To understand the sugar specificity at a structural level, the enzyme-ATP/Mg2+-sugar binding complex models have been constructed. It has been shown that the sugar specificity is probably dependent on the interaction energy occurred by the positional proximity of sugars bound in the active site of the enzyme, which exhibits a tolerance to modification at C2 or C3 of glucose. PMID:16053915

  10. Draft Genome Sequences of Four Thermophilic Spore Formers Isolated from a Dairy-Processing Environment

    PubMed Central

    Caspers, Martien P. M.; Boekhorst, Jos; de Jong, Anne; Kort, Remco; Nierop Groot, Masja

    2016-01-01

    Spores of thermophilic spore-forming bacteria are a common cause of contamination in dairy products. Here, we report draft genome sequences of four thermophilic strains from a milk-processing plant or standard milk, namely, a Geobacillus thermoglucosidans isolate (TNO-09.023), Geobacillus stearothermophilus TNO-09.027, and two Anoxybacillus flavithermus isolates (TNO-09.014 and TNO-09.016). PMID:27516503

  11. Draft Genome Sequences of Four Thermophilic Spore Formers Isolated from a Dairy-Processing Environment.

    PubMed

    Caspers, Martien P M; Boekhorst, Jos; de Jong, Anne; Kort, Remco; Nierop Groot, Masja; Abee, Tjakko

    2016-01-01

    Spores of thermophilic spore-forming bacteria are a common cause of contamination in dairy products. Here, we report draft genome sequences of four thermophilic strains from a milk-processing plant or standard milk, namely, a Geobacillus thermoglucosidans isolate (TNO-09.023), Geobacillus stearothermophilus TNO-09.027, and two Anoxybacillus flavithermus isolates (TNO-09.014 and TNO-09.016). PMID:27516503

  12. Hydrophobic and Electrostatic Cell Surface Properties of Thermophilic Dairy Streptococci

    PubMed Central

    van der Mei, H. C.; de Vries, J.; Busscher, H. J.

    1993-01-01

    Microbial adhesion to hydrocarbons (MATH) and microelectrophoresis were done in 10 mM potassium phosphate solutions to characterize the surfaces of thermophilic dairy streptococci, isolated from pasteurizers. Regardless of whether they were grown (in M17 broth) with lactose, sucrose, or glucose added, strains were relatively hydrophilic (showing low initial removal rates by hexadecane) and slightly negatively charged. A tendency exists for cells grown with sucrose added to be more hydrophilic than cells grown with glucose or lactose added. Also, the lowest isoelectric points, i.e., the pH values for which the zeta potentials are zero, were measured for strains with glucose added to the growth medium. The isoelectric points for the strains were all rather high, between pH 3 and 5, indicative of protein-rich surfaces, although X-ray photoelectron spectroscopy did not measure excessively large amounts of nitrogen on the cell surfaces. Both MATH and microelectrophoresis were done as a function of pH. Maxima in hydrophobicity were observed at certain pH values. Usually these pH values were in the range of the isoelectric points of the cells. Thus it appears that MATH measures an interplay of hydrophobicity and electrostatic interactions. MATH measures solely hydrophobicity only when electrostatic interactions are absent, i.e., close to the isoelectric points of the cells. Considering that these thermophilic streptococci are all rather hydrophilic, a possible pathway to prevent fouling in the pasteurization process might be to render the heat exchanger plates of the pasteurizer more hydrophobic. PMID:16349127

  13. The metabolism of hydrogen by extremely thermophilic bacteria

    SciTech Connect

    Adams, M.W.W.

    1991-01-01

    The novel archaebacterium, Pyrococcus furiosus, grows optimally at 100{degree}C by a fermentative metabolism and produces hydrogen (H{sub 2}). We have shown that this organism appears to ferment glucose and evolve H{sub 2} by a novel pathway. The following metalloenzymes and proteins involved in H{sub 2} metabolism have been purified and characterized: hydrogenase (NiFeS), ferredoxin (FeS), pyruvate ferredoxin oxidoreductase (FeS), and an new enzyme which contains tungsten, glyceraldehyde ferredoxin oxidoreductase (WFeS). A rubredoxin was also purified, and it and the ferredoxin have been sequenced. In addition, a second new enzyme has been identified, glucose ferredoxin oxidoreductase. These represent the first enzymes and proteins to be purified from any organism able to grow optimally above 90{degree}C. All are remarkably thermostable and show maximal catalytic activity >95{degree}C. The ferredoxin has several unique properties and is potentially an extremely thermostable model for the catalytic sites of a variety of mesophilic metalloenzymes. In addition, the FeS-containing enzymes, hydrogenase and pyruvate ferredoxin oxidoreductase, and a ferredoxin, have been purified from the most thermophilic eubacterium currently known, Thermotoga maritima. This organism grows up to 90{degree}C, also by fermentation. The hydrogenases of T. maritima and P. furiosus each have many unique properties in comparison with mesophilic hydrogenases, and both appear to contain new types of metal centers that are specifically adapted to catalyze H{sub 2} production at the extreme temperatures. Hydrogenase activity has also been measured in four other extremely thermophilic organisms, one of which is capable of growth at 120{degree}C. 1 fig.

  14. Silicification of Thermophilic Biofilms: Do Aquificales Affect the Mineralisation Process?

    NASA Astrophysics Data System (ADS)

    Konhauser, K.; Lalonde, S.; Aguiar, P.; Reysenbach, A.

    2003-12-01

    In geothermal environments, biomineralisation is an inevitable consequence of microbes growing in solute-rich waters. The process of silicification is of particular interest due to (1) apparent discrepancies between natural and laboratory silicification rates and (2) siliceous microfossils currently serve as the earliest physical evidence for life on Earth. Although mesophilic microbe-silica interactions have been studied in great detail, there is a paucity of information on the role that thermophiles play in the silicification process, i.e., does their metabolism in any way facilitate silicification and do their cellular remains fossilise? To help resolve some of these uncertainties, a thermophilic, biofilm-forming member of the Aquificales order, Sulfurihydrogenobium azorense, was grown in the presence of various concentrations of silica, ranging from undersaturated to those extremely supersaturated with respect to amorphous silica. Since the chemolithoautotrophic Aquificales use of a wide range and combination of electron donors and acceptors, the bacteria cultured were grown in the presence of H2 with O2, S and Fe(III) as terminal electron acceptors. This study focused on the rates of pH-induced silica polymerisation during a 48 hour interval, when the soluble silica phase was at its most reactive stage, and when the greatest amount of silica immobilisation was likely to occur. S. azorense was found to have no detectable effect on the polymerisation rate of silica under any condition tested, nor did it cause silica to precipitate in undersaturated conditions. In addition, transmission electron microscopy showed that although silica did indeed precipitate from solution, there was no obvious association between solid-phase silica and the cells walls. This suggests that under high silica levels there is such a strong chemical driving force for silica polymerisation, homogeneous nucleation, and ultimately silica precipitation that there is no obvious need for

  15. Genomic characterization of Defluviitoga tunisiensis L3, a key hydrolytic bacterium in a thermophilic biogas plant and its abundance as determined by metagenome fragment recruitment.

    PubMed

    Maus, Irena; Cibis, Katharina Gabriela; Bremges, Andreas; Stolze, Yvonne; Wibberg, Daniel; Tomazetto, Geizecler; Blom, Jochen; Sczyrba, Alexander; König, Helmut; Pühler, Alfred; Schlüter, Andreas

    2016-08-20

    The genome sequence of Defluviitoga tunisiensis L3 originating from a thermophilic biogas-production plant was established and recently published as Genome Announcement by our group. The circular chromosome of D. tunisiensis L3 has a size of 2,053,097bp and a mean GC content of 31.38%. To analyze the D. tunisiensis L3 genome sequence in more detail, a phylogenetic analysis of completely sequenced Thermotogae strains based on shared core genes was performed. It appeared that Petrotoga mobilis DSM 10674(T), originally isolated from a North Sea oil-production well, is the closest relative of D. tunisiensis L3. Comparative genome analyses of P. mobilis DSM 10674(T) and D. tunisiensis L3 showed moderate similarities regarding occurrence of orthologous genes. Both genomes share a common set of 1351 core genes. Reconstruction of metabolic pathways important for the biogas production process revealed that the D. tunisiensis L3 genome encodes a large set of genes predicted to facilitate utilization of a variety of complex polysaccharides including cellulose, chitin and xylan. Ethanol, acetate, hydrogen (H2) and carbon dioxide (CO2) were found as possible end-products of the fermentation process. The latter three metabolites are considered to represent substrates for methanogenic Archaea, the key organisms in the final step of the anaerobic digestion process. To determine the degree of relatedness between D. tunisiensis L3 and dominant biogas community members within the thermophilic biogas-production plant, metagenome sequences obtained from the corresponding microbial community were mapped onto the L3 genome sequence. This fragment recruitment revealed that the D. tunisiensis L3 genome is almost completely covered with metagenome sequences featuring high matching accuracy. This result indicates that strains highly related or even identical to the reference strain D. tunisiensis L3 play a dominant role within the community of the thermophilic biogas-production plant. PMID

  16. Diversity of Cultured Thermophilic Anaerobes in Hot Springs of Yunnan Province, China

    NASA Astrophysics Data System (ADS)

    Lin, L.; Lu, Y.; Dong, X.; Liu, X.; Wei, Y.; Ji, X.; Zhang, C.

    2010-12-01

    Thermophilic anaerobes including Archaea and Bacteria refer to those growing optimally at temperatures above 50°C and do not use oxygen as the terminal electron acceptor for growth. Study on thermophilic anaerobes will help to understand how life thrives under extreme conditions. Meanwhile thermophilic anaerobes are of importance in potential application and development of thermophilic biotechnology. We have surveyed culturable thermophilic anaerobes in hot springs (pH6.5-7.5; 70 - 94°C) in Rehai of Tengchong, Bangnazhang of Longlin, Eryuan of Dali,Yunnan, China. 50 strains in total were cultured from the hot springs water using Hungate anaerobic technique, and 30 strains were selected based on phenotypic diversity for analysis of 16S rDNA sequences. Phylogenetic analysis showed that 28 strains belonged to the members of five genera: Caldanaerobacter, Calaramator, Thermoanaerobacter, Dictyoglomus and Fervidobacterium, which formed five branches on the phylogenetic tree. Besides, 2 strains of methanogenic archaea were obtained. The majority of the isolates were the known species, however, seven strains were identified as novel species affiliated to the five genera based on the lower 16S rDNA sequence similarities (less than 93 - 97%) with the described species. This work would provide the future study on their diversity, distribution among different regions and the potential application of thermophilic enzyme. Supported by State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences(SKLMR-080605)and the Foundation of State Natural Science (30660009, 30960022, 31081220175).

  17. Presence and potential role of thermophilic bacteria in temperate terrestrial environments.

    PubMed

    Portillo, M C; Santana, M; Gonzalez, J M

    2012-01-01

    Organic sulfur and nitrogen are major reservoirs of these elements in terrestrial systems, although their cycling remains to be fully understood. Both sulfur and nitrogen mineralization are directly related to microbial metabolism. Mesophiles and thermophiles were isolated from temperate environments. Thermophilic isolates were classified within the Firmicutes, belonging to the Geobacillus, Brevibacillus, and Ureibacillus genera, and showed optimum growth temperatures between 50°C and 60°C. Sulfate and ammonium produced were higher during growth of thermophiles both for isolated strains and natural bacterial assemblages. They were positively related to organic nutrient load. Temperature also affected the release of sulfate and ammonium by thermophiles. Quantitative, real-time reverse-transcription polymerase chain reaction on environmental samples indicated that the examined thermophilic Firmicutes represented up to 3.4% of the total bacterial community RNA. Temperature measurements during summer days showed values above 40°C for more than 10 h a day in soils from southern Spain. These results support a potential role of thermophilic bacteria in temperate terrestrial environments by mineralizing organic sulfur and nitrogen ruled by the existence and length of warm periods. PMID:22159635

  18. Moderate and Binge Drinking

    MedlinePlus

    ... here Home » Alcohol & Your Health » Overview of Alcohol Consumption » Drinking Levels Defined In this Section Alcohol Facts & Statistics What Is A Standard Drink? Drinking Levels Defined Drinking Levels Defined Moderate alcohol consumption: According to the "Dietary Guidelines for Americans 2015- ...

  19. Moderator Chemistry Program

    SciTech Connect

    Dewitt, L.V.; Gibbs, A.; Lambert, D.P.; Bohrer, S.R.; Fanning, R.L.; Houston, M.W.; Stinson, S.L.; Deible, R.W.; Abdel-Khalik, S.I.

    1990-11-01

    Over the past fifteen months, the Systems Chemistry Group of the Reactor Engineering Department has undertaken a comprehensive study of the Department`s moderator chemistry program at Savannah River Site (SRS). An internal review was developed to formalize and document this program. Objectives were as outlined in a mission statement and action plan. In addition to the mission statement and action plan, nine separate task reports have been issued during the course of this study. Each of these task reports is included in this document as a chapter. This document is an organized compilation of the individual reports issued by the Systems Chemistry Group in assessment of SRS moderator chemistry to determine if there were significant gaps in the program as ft existed in October, 1989. While these reviews found no significant gaps in that mode of operation, or any items that adversely affected safety, items were identified that could be improved. Many of the items have already been dear with or are in the process of completion under this Moderator Chemistry Program and other Reactor Restart programs. A complete list of the items of improvement found under this assessment is found in Chapter 9, along with a proposed time table for correcting remaining items that can be improved for the chemistry program of SRS reactors. An additional external review of the moderator chemistry processes, recommendations, and responses to/from the Reactor Corrosion Mitigation Committee is included as Appendix to this compilation.

  20. Moderator Chemistry Program

    SciTech Connect

    Dewitt, L.V.; Gibbs, A.; Lambert, D.P.; Bohrer, S.R.; Fanning, R.L.; Houston, M.W.; Stinson, S.L.; Deible, R.W.; Abdel-Khalik, S.I.

    1990-11-01

    Over the past fifteen months, the Systems Chemistry Group of the Reactor Engineering Department has undertaken a comprehensive study of the Department's moderator chemistry program at Savannah River Site (SRS). An internal review was developed to formalize and document this program. Objectives were as outlined in a mission statement and action plan. In addition to the mission statement and action plan, nine separate task reports have been issued during the course of this study. Each of these task reports is included in this document as a chapter. This document is an organized compilation of the individual reports issued by the Systems Chemistry Group in assessment of SRS moderator chemistry to determine if there were significant gaps in the program as ft existed in October, 1989. While these reviews found no significant gaps in that mode of operation, or any items that adversely affected safety, items were identified that could be improved. Many of the items have already been dear with or are in the process of completion under this Moderator Chemistry Program and other Reactor Restart programs. A complete list of the items of improvement found under this assessment is found in Chapter 9, along with a proposed time table for correcting remaining items that can be improved for the chemistry program of SRS reactors. An additional external review of the moderator chemistry processes, recommendations, and responses to/from the Reactor Corrosion Mitigation Committee is included as Appendix to this compilation.

  1. Management of moderate lymphedema.

    PubMed

    Shumacker, H B

    1981-08-01

    Moderate chronic lymphedema generally requires a different program of management than mild or massive lymphedema. It responds well to a special management regimen based on home use of an intermittent limb compressor and utilization of proper compression support when the patient is not in the recumbent position. PMID:7259517

  2. Extremely thermophilic microorganisms as metabolic engineering platforms for production of fuels and industrial chemicals

    PubMed Central

    Zeldes, Benjamin M.; Keller, Matthew W.; Loder, Andrew J.; Straub, Christopher T.; Adams, Michael W. W.; Kelly, Robert M.

    2015-01-01

    Enzymes from extremely thermophilic microorganisms have been of technological interest for some time because of their ability to catalyze reactions of industrial significance at elevated temperatures. Thermophilic enzymes are now routinely produced in recombinant mesophilic hosts for use as discrete biocatalysts. Genome and metagenome sequence data for extreme thermophiles provide useful information for putative biocatalysts for a wide range of biotransformations, albeit involving at most a few enzymatic steps. However, in the past several years, unprecedented progress has been made in establishing molecular genetics tools for extreme thermophiles to the point that the use of these microorganisms as metabolic engineering platforms has become possible. While in its early days, complex metabolic pathways have been altered or engineered into recombinant extreme thermophiles, such that the production of fuels and chemicals at elevated temperatures has become possible. Not only does this expand the thermal range for industrial biotechnology, it also potentially provides biodiverse options for specific biotransformations unique to these microorganisms. The list of extreme thermophiles growing optimally between 70 and 100°C with genetic toolkits currently available includes archaea and bacteria, aerobes and anaerobes, coming from genera such as Caldicellulosiruptor, Sulfolobus, Thermotoga, Thermococcus, and Pyrococcus. These organisms exhibit unusual and potentially useful native metabolic capabilities, including cellulose degradation, metal solubilization, and RuBisCO-free carbon fixation. Those looking to design a thermal bioprocess now have a host of potential candidates to choose from, each with its own advantages and challenges that will influence its appropriateness for specific applications. Here, the issues and opportunities for extremely thermophilic metabolic engineering platforms are considered with an eye toward potential technological advantages for high

  3. Trade-off between mesophilic and thermophilic denitrification: rates vs. sludge production, settleability and stability.

    PubMed

    Courtens, Emilie N P; Vlaeminck, Siegfried E; Vilchez-Vargas, Ramiro; Verliefde, Arne; Jauregui, Ruy; Pieper, Dietmar H; Boon, Nico

    2014-10-15

    The development of thermophilic nitrogen removal strategies will facilitate sustainable biological treatment of warm nitrogenous wastewaters. Thermophilic denitrification was extensively compared to mesophilic denitrification for the first time in this study. Two sequential batch reactors (SBR) at 34 °C and 55 °C were inoculated with mesophilic activated sludge (26 °C), fed with synthetic influent in a first phase. Subsequently, the carbon source was switched from acetate to molasses, whereas in a third phase, the nitrate source was fertilizer industry wastewater. The denitrifying sludge maintained its activity at 55 °C, resulting in an immediate process start-up, obtaining nitrogen removal rates higher than 500 mg N g(-1) VSS d(-1) in less than one week. Although the mesophilic SBR showed twice as high specific nitrogen removal rates, the maximum thermophilic denitrifying activity in this study was nearly 10 times higher than the activities reported thus far. The thermophilic SBR moreover had a 73% lower sludge volume index, a 45% lower sludge production and a higher resilience towards a change in carbon source compared with the mesophilic SBR. The higher resilience was potentially related to a higher microbial diversity and evenness of the thermophilic community at the end of the synthetic feeding period. The thermophilic microbial community showed a higher similarity over the different feeding periods implying a more stable community. Overall, this study showed the capability of mesophilic denitrifiers to maintain their activity after a large temperature increase. Existing mesophilic process systems with cooling for the treatment of warm wastewaters could thus efficiently be converted to thermophilic systems with low sludge production and good settling properties. PMID:25007305

  4. Evaluation of continuous mesophilic, thermophilic and temperature phased anaerobic digestion of microwaved activated sludge.

    PubMed

    Coelho, Nuno Miguel Gabriel; Droste, Ronald L; Kennedy, Kevin J

    2011-04-01

    The effects of microwave (MW) pretreatment, staging and digestion temperature on anaerobic digestion were investigated in a setup of ten reactors. A mesophilic reactor was used as a control. Its performance was compared to single-stage mesophilic and thermophilic reactors treating pretreated and non-pretreated sludge, temperature-phased (TPAD) thermophilic-mesophilic reactors treating pretreated and non-pretreated sludge and thermophilic-thermophilic reactors also treating pretreated and non-pretreated sludge. Four different sludge retention times (SRTs) (20, 15, 10 and 5 d) were tested for all reactors. Two-stage thermo-thermo reactors treating pretreated sludge produced more biogas than all other reactors and removed more volatile solids. Maximum volatile solids (VS) removal was 53.1% at an SRT of 15 d and maximum biogas increase relative to control was 106% at the shortest SRT tested. Both the maximum VS removal and biogas relative increase were measured for a system with thermophilic acidogenic reactor and thermophilic methanogenic reactor. All the two-stage systems treating microwaved sludge produced sludge free of pathogen indicator bacteria, at all tested conditions even at a total system SRT of only 5 d. MW pretreatment and staging reactors allowed the application of very short SRT (5 d) with no significant decrease in performance in terms of VS removal in comparison with the control reactor. MW pretreatment caused the solubilization of organic material in sludge but also allowed more extensive hydrolysis of organic material in downstream reactors. The association of MW pretreatment and thermophilic operation improves dewaterability of digested sludge. PMID:21470653

  5. Reduction of hexavalent chromium by the thermophilic methanogen Methanothermobacter thermautotrophicus

    NASA Astrophysics Data System (ADS)

    Singh, Rajesh; Dong, Hailiang; Liu, Deng; Zhao, Linduo; Marts, Amy R.; Farquhar, Erik; Tierney, David L.; Almquist, Catherine B.; Briggs, Brandon R.

    2015-01-01

    Despite significant progress on iron reduction by thermophilic microorganisms, studies on their ability to reduce toxic metals are still limited, despite their common co-existence in high temperature environments (up to 70 °C). In this study, Methanothermobacter thermautotrophicus, an obligate thermophilic methanogen, was used to reduce hexavalent chromium. Experiments were conducted in a growth medium with H2/CO2 as substrate with various Cr6+ concentrations (0.2, 0.4, 1, 3, and 5 mM) in the form of potassium dichromate (K2Cr2O7). Time-course measurements of aqueous Cr6+ concentrations using 1,5-diphenylcarbazide colorimetric method showed complete reduction of the 0.2 and 0.4 mM Cr6+ solutions by this methanogen. However, much lower reduction extents of 43.6%, 13.0%, and 3.7% were observed at higher Cr6+ concentrations of 1, 3 and 5 mM, respectively. These lower extents of bioreduction suggest a toxic effect of aqueous Cr6+ to cells at this concentration range. At these higher Cr6+ concentrations, methanogenesis was inhibited and cell growth was impaired as evidenced by decreased total cellular protein production and live/dead cell ratio. Likewise, Cr6+ bioreduction rates decreased with increased initial concentrations of Cr6+ from 13.3 to 1.9 μM h-1. X-ray absorption near-edge structure (XANES) spectroscopy revealed a progressive reduction of soluble Cr6+ to insoluble Cr3+ precipitates, which was confirmed as amorphous chromium hydroxide by selected area electron diffraction pattern. However, a small fraction of reduced Cr occurred as aqueous Cr3+. Scanning and transmission electron microscope observations of M. thermautotrophicus cells after Cr6+ exposure suggest both extra- and intracellular chromium reduction mechanisms. Results of this study demonstrate the ability of M. thermautotrophicus cells to reduce toxic Cr6+ to less toxic Cr3+ and its potential application in metal bioremediation, especially at high temperature subsurface radioactive waste disposal

  6. Reduction of hexavalent chromium by the thermophilic methanogen Methanothermobacter thermautotrophicus

    DOE PAGESBeta

    Singh, Rajesh; Dong, Hailiang; Liu, Deng; Zhao, Linduo; Marts, Amy R.; Farquhar, Erik; Tierney, David L.; Almquist, Catherine B.; Briggs, Brandon R.

    2014-10-22

    Despite the significant progress on iron reduction by thermophilic microorganisms, studies on their ability to reduce toxic metals are still limited, despite their common co-existence in high temperature environments (up to 70°C). In this study, Methanothermobacter thermautotrophicus, an obligate thermophilic methanogen, was used to reduce hexavalent chromium. Experiments were conducted in a growth medium with H2/CO2 as substrate with various Cr6+ concentrations (0.2, 0.4, 1, 3, and 5 mM) in the form of potassium dichromate (K2Cr2O7). Time-course measurements of aqueous Cr6+ concentrations with the 1, 5-diphenylcarbazide colorimetric method showed complete reduction of the 0.2 and 0.4 mM Cr6+ solutions bymore » this methanogen. However, much lower reduction extents of 43.6%, 13.0%, and 3.7% were observed at higher Cr6+ concentrations of 1, 3 and 5 mM, respectively. These lower extents of bioreduction suggest a toxic effect of aqueous Cr6+ to cells at this concentration range. At these higher Cr6+ concentrations, methanogenesis was inhibited and cell growth was impaired as evidenced by decreased total cellular protein production and live/dead cell ratio. Likewise, Cr6+ bioreduction rates decreased with increased initial concentrations of Cr6+ from 13.3 to1.9 μM h₋1. X-ray absorption near-edge structure (XANES) spectroscopy revealed a progressive reduction of soluble Cr6+ to insoluble Cr3+ precipitates, which was confirmed as amorphous chromium hydroxide by X-ray diffraction and selected area electron diffraction pattern. However, a small fraction of reduced Cr occurred as aqueous Cr3+. Scanning and transmission electron microscope observations of M. thermautotrophicus cells after Cr6+ exposure suggest both extra- and intracellular chromium reduction mechanisms. Results of this study demonstrate the ability of M. thermautotrophicus cells to reduce toxic Cr6+ to less toxic Cr3+ and its potential application in metal bioremediation, especially at high temperature

  7. Reduction of hexavalent chromium by the thermophilic methanogen Methanothermobacter thermautotrophicus

    PubMed Central

    Singh, Rajesh; Dong, Hailiang; Liu, Deng; Zhao, Linduo; Marts, Amy R.; Farquhar, Erik; Tierney, David L.; Almquist, Catherine B.; Briggs, Brandon R.

    2015-01-01

    Despite the significant progress on iron reduction by thermophilic microorganisms, studies on their ability to reduce toxic metals are still limited, despite their common co-existence in high temperature environments (up to 70°C). In this study, Methanothermobacter thermautotrophicus, an obligate thermophilic methanogen, was used to reduce hexavalent chromium. Experiments were conducted in a growth medium with H2/CO2 as substrate with various Cr6+ concentrations (0.2, 0.4, 1, 3, and 5 mM) in the form of potassium dichromate (K2Cr2O7). Time-course measurements of aqueous Cr6+ concentrations with the 1, 5-diphenylcarbazide colorimetric method showed complete reduction of the 0.2 and 0.4 mM Cr6+ solutions by this methanogen. However, much lower reduction extents of 43.6%, 13.0%, and 3.7% were observed at higher Cr6+ concentrations of 1, 3 and 5 mM, respectively. These lower extents of bioreduction suggest a toxic effect of aqueous Cr6+ to cells at this concentration range. At these higher Cr6+ concentrations, methanogenesis was inhibited and cell growth was impaired as evidenced by decreased total cellular protein production and live/dead cell ratio. Likewise, Cr6+ bioreduction rates decreased with increased initial concentrations of Cr6+ from 13.3 to1.9 µM h−1. X-ray absorption near-edge structure (XANES) spectroscopy revealed a progressive reduction of soluble Cr6+ to insoluble Cr3+ precipitates, which was confirmed as amorphous chromium hydroxide by X-ray diffraction and selected area electron diffraction pattern. However, a small fraction of reduced Cr occurred as aqueous Cr3+. Scanning and transmission electron microscope observations of M. thermautotrophicus cells after Cr6+ exposure suggest both extra- and intracellular chromium reduction mechanisms. Results of this study demonstrate the ability of M. thermautotrophicus cells to reduce toxic Cr6+ to less toxic Cr3+ and its potential application in metal bioremediation, especially at high temperature

  8. Reduction of hexavalent chromium by the thermophilic methanogen Methanothermobacter thermautotrophicus

    SciTech Connect

    Singh, Rajesh; Dong, Hailiang; Liu, Deng; Zhao, Linduo; Marts, Amy R.; Farquhar, Erik; Tierney, David L.; Almquist, Catherine B.; Briggs, Brandon R.

    2014-10-22

    Despite the significant progress on iron reduction by thermophilic microorganisms, studies on their ability to reduce toxic metals are still limited, despite their common co-existence in high temperature environments (up to 70°C). In this study, Methanothermobacter thermautotrophicus, an obligate thermophilic methanogen, was used to reduce hexavalent chromium. Experiments were conducted in a growth medium with H2/CO2 as substrate with various Cr6+ concentrations (0.2, 0.4, 1, 3, and 5 mM) in the form of potassium dichromate (K2Cr2O7). Time-course measurements of aqueous Cr6+ concentrations with the 1, 5-diphenylcarbazide colorimetric method showed complete reduction of the 0.2 and 0.4 mM Cr6+ solutions by this methanogen. However, much lower reduction extents of 43.6%, 13.0%, and 3.7% were observed at higher Cr6+ concentrations of 1, 3 and 5 mM, respectively. These lower extents of bioreduction suggest a toxic effect of aqueous Cr6+ to cells at this concentration range. At these higher Cr6+ concentrations, methanogenesis was inhibited and cell growth was impaired as evidenced by decreased total cellular protein production and live/dead cell ratio. Likewise, Cr6+ bioreduction rates decreased with increased initial concentrations of Cr6+ from 13.3 to1.9 μM h₋1. X-ray absorption near-edge structure (XANES) spectroscopy revealed a progressive reduction of soluble Cr6+ to insoluble Cr3+ precipitates, which was confirmed as amorphous chromium hydroxide by X-ray diffraction and selected area electron diffraction pattern. However, a small fraction of reduced Cr occurred as aqueous Cr3+. Scanning and transmission electron microscope observations of M. thermautotrophicus cells after Cr6+ exposure suggest both extra- and intracellular chromium reduction mechanisms. Results of

  9. Complete genome sequence of the moderately thermophilic mineral-sulfide-oxidizing firmicute Sulfobacillus acidophilus type strain (NAL(T)).

    PubMed

    Anderson, Iain; Chertkov, Olga; Chen, Amy; Saunders, Elizabeth; Lapidus, Alla; Nolan, Matt; Lucas, Susan; Hammon, Nancy; Deshpande, Shweta; Cheng, Jan-Fang; Han, Cliff; Tapia, Roxanne; Goodwin, Lynne A; Pitluck, Sam; Liolios, Konstantinos; Pagani, Ioanna; Ivanova, Natalia; Mikhailova, Natalia; Pati, Amrita; Palaniappan, Krishna; Land, Miriam; Pan, Chongle; Rohde, Manfred; Pukall, Rüdiger; Göker, Markus; Detter, John C; Woyke, Tanja; Bristow, James; Eisen, Jonathan A; Markowitz, Victor; Hugenholtz, Philip; Kyrpides, Nikos C; Klenk, Hans-Peter; Mavromatis, Konstantinos

    2012-07-30

    Sulfobacillus acidophilus Norris et al. 1996 is a member of the genus Sulfobacillus which comprises five species of the order Clostridiales. Sulfobacillus species are of interest for comparison to other sulfur and iron oxidizers and also have biomining applications. This is the first completed genome sequence of a type strain of the genus Sulfobacillus, and the second published genome of a member of the species S. acidophilus. The genome, which consists of one chromosome and one plasmid with a total size of 3,557,831 bp harbors 3,626 protein-coding and 69 RNA genes, and is a part of the GenomicEncyclopedia ofBacteria andArchaea project. PMID:23407703

  10. Complete genome sequence of the moderately thermophilic mineral-sulfide-oxidizing firmicute Sulfobacillus acidophilus type strain (NALT)

    SciTech Connect

    Anderson, Iain; Chertkov, Olga; Chen, Amy; Saunders, Elizabeth H; Lapidus, Alla L.; Nolan, Matt; Lucas, Susan; Hammon, Nancy; Deshpande, Shweta; Cheng, Jan-Fang; Han, Cliff; Tapia, Roxanne; Goodwin, Lynne A.; Pitluck, Sam; Liolios, Konstantinos; Pagani, Ioanna; Ivanova, N; Mikhailova, Natalia; Pati, Amrita; Palaniappan, Krishna; Land, Miriam L; Pan, Chongle; Rohde, Manfred; Pukall, Rudiger; Goker, Markus; Detter, J. Chris; Woyke, Tanja; Bristow, James; Eisen, Jonathan; Markowitz, Victor; Hugenholtz, Philip; Kyrpides, Nikos C; Klenk, Hans-Peter; Mavromatis, K

    2012-01-01

    Sulfobacillus acidophilus Norris et al. 1996 is a member of the genus Sulfobacillus which comprises five species of the order Clostridiales. Sulfobacillus species are of interest for comparison to other sulfur and iron oxidizers and also have biomining applications. This is the first completed genome sequence of a type strain of the genus Sulfobacillus, and the second published genome of a member of the species S. acidophilus. The genome, which consists of one chromosome and one plasmid with a total size of 3,557,831 bp, harbors 3,626 protein-coding and 69 RNA genes, and is a part of the Genomic Encyclopedia of Bacteria and Archaea project.

  11. Sulfobacillus thermosulfidooxidans strain Cutipay enhances chalcopyrite bioleaching under moderate thermophilic conditions in the presence of chloride ion.

    PubMed

    Bobadilla-Fazzini, Roberto A; Cortés, Maria Paz; Maass, Alejandro; Parada, Pilar

    2014-12-01

    Currently more than 90% of the world's copper is obtained through sulfide mineral processing. Among the copper sulfides, chalcopyrite is the most abundant and therefore economically relevant. However, primary copper sulfide bioleaching is restricted due to high ionic strength raffinate solutions and particularly chloride coming from the dissolution of ores. In this work we describe the chalcopyrite bioleaching capacity of Sulfobacillus thermosulfidooxidans strain Cutipay (DSM 27601) previously described at the genomic level (Travisany et al. (2012) Draft genome sequence of the Sulfobacillus thermosulfidooxidans Cutipay strain, an indigenous bacterium isolated from a naturally extreme mining environment in Northern Chile. J Bacteriol 194:6327-6328). Bioleaching assays with the mixotrophic strain Cutipay showed a strong increase in copper recovery from chalcopyrite concentrate at 50°C in the presence of chloride ion, a relevant inhibitory element present in copper bioleaching processes. Compared to the abiotic control and a test with Sulfobacillus acidophilus DSM 10332, strain Cutipay showed an increase of 42 and 69% in copper recovery, respectively, demonstrating its high potential for chalcopyrite bioleaching. Moreover, a genomic comparison highlights the presence of the 2-Haloacid dehalogenase predicted-protein related to a potential new mechanism of chloride resistance in acidophiles. This novel and industrially applicable strain is under patent application CL 2013-03335. PMID:26267113

  12. Kinetic characterization of thermophilic and mesophilic anaerobic digestion for coffee grounds and waste activated sludge.

    PubMed

    Li, Qian; Qiao, Wei; Wang, Xiaochang; Takayanagi, Kazuyuki; Shofie, Mohammad; Li, Yu-You

    2015-02-01

    This study was conducted to characterize the kinetics of an anaerobic process (hydrolysis, acetogenesis, acidogenesis and methanogenesis) under thermophilic (55 °C) and mesophilic (35 °C) conditions with coffee grounds and waste activated sludge (WAS) as the substrates. Special focus was given to the kinetics of propionic acid degradation to elucidate the accumulation of VFAs. Under the thermophilic condition, the methane production rate of all substrates (WAS, ground coffee and raw coffee) was about 1.5 times higher than that under the mesophilic condition. However, the effects on methane production of each substrate under the thermophilic condition differed: WAS increased by 35.8-48.2%, raw coffee decreased by 76.3-64.5% and ground coffee decreased by 74.0-57.9%. Based on the maximum reaction rate (Rmax) of each anaerobic stage obtained from the modified Gompertz model, acetogenesis was found to be the rate-limiting step for coffee grounds and WAS. This can be explained by the kinetics of propionate degradation under thermophilic condition in which a long lag-phase (more than 18 days) was observed, although the propionate concentration was only 500 mg/L. Under the mesophilic condition, acidogenesis and hydrolysis were found to be the rate-limiting step for coffee grounds and WAS, respectively. Even though reducing the particle size accelerated the methane production rate of coffee grounds, but did not change the rate-limiting step: acetogenesis in thermophilic and acidogenesis in mesophilic. PMID:25534040

  13. Comparing residue clusters from thermophilic and mesophilic enzymes reveals adaptive mechanisms

    DOE PAGESBeta

    Sammond, Deanne W.; Kastelowitz, Noah; Himmel, Michael E.; Yin, Hang; Crowley, Michael F.; Bomble, Yannick J.

    2016-01-07

    Understanding how proteins adapt to function at high temperatures is important for deciphering the energetics that dictate protein stability and folding. While multiple principles important for thermostability have been identified, we lack a unified understanding of how internal protein structural and chemical environment determine qualitative or quantitative impact of evolutionary mutations. In this work we compare equivalent clusters of spatially neighboring residues between paired thermophilic and mesophilic homologues to evaluate adaptations under the selective pressure of high temperature. We find the residue clusters in thermophilic enzymes generally display improved atomic packing compared to mesophilic enzymes, in agreement with previous research.more » Unlike residue clusters from mesophilic enzymes, however, thermophilic residue clusters do not have significant cavities. In addition, anchor residues found in many clusters are highly conserved with respect to atomic packing between both thermophilic and mesophilic enzymes. As a result, the improvements in atomic packing observed in thermophilic homologues are not derived from these anchor residues but from neighboring positions, which may serve to expand optimized protein core regions.« less

  14. Comparing Residue Clusters from Thermophilic and Mesophilic Enzymes Reveals Adaptive Mechanisms

    PubMed Central

    Sammond, Deanne W.; Kastelowitz, Noah; Himmel, Michael E.; Yin, Hang; Crowley, Michael F.; Bomble, Yannick J.

    2016-01-01

    Understanding how proteins adapt to function at high temperatures is important for deciphering the energetics that dictate protein stability and folding. While multiple principles important for thermostability have been identified, we lack a unified understanding of how internal protein structural and chemical environment determine qualitative or quantitative impact of evolutionary mutations. In this work we compare equivalent clusters of spatially neighboring residues between paired thermophilic and mesophilic homologues to evaluate adaptations under the selective pressure of high temperature. We find the residue clusters in thermophilic enzymes generally display improved atomic packing compared to mesophilic enzymes, in agreement with previous research. Unlike residue clusters from mesophilic enzymes, however, thermophilic residue clusters do not have significant cavities. In addition, anchor residues found in many clusters are highly conserved with respect to atomic packing between both thermophilic and mesophilic enzymes. Thus the improvements in atomic packing observed in thermophilic homologues are not derived from these anchor residues but from neighboring positions, which may serve to expand optimized protein core regions. PMID:26741367

  15. Probing the Folding-Unfolding Transition of a Thermophilic Protein, MTH1880

    PubMed Central

    Jung, Youngjin; Han, Jeongmin; Yun, Ji-Hye; Chang, Iksoo; Lee, Weontae

    2016-01-01

    The folding mechanism of typical proteins has been studied widely, while our understanding of the origin of the high stability of thermophilic proteins is still elusive. Of particular interest is how an atypical thermophilic protein with a novel fold maintains its structure and stability under extreme conditions. Folding-unfolding transitions of MTH1880, a thermophilic protein from Methanobacterium thermoautotrophicum, induced by heat, urea, and GdnHCl, were investigated using spectroscopic techniques including circular dichorism, fluorescence, NMR combined with molecular dynamics (MD) simulations. Our results suggest that MTH1880 undergoes a two-state N to D transition and it is extremely stable against temperature and denaturants. The reversibility of refolding was confirmed by spectroscopic methods and size exclusion chromatography. We found that the hyper-stability of the thermophilic MTH1880 protein originates from an extensive network of both electrostatic and hydrophobic interactions coordinated by the central β-sheet. Spectroscopic measurements, in combination with computational simulations, have helped to clarify the thermodynamic and structural basis for hyper-stability of the novel thermophilic protein MTH1880. PMID:26766214

  16. Community dynamics and glycoside hydrolase activities of thermophilic bacterial consortia adapted to switchgrass

    SciTech Connect

    Gladden, J.M.; Allgaier, M.; Miller, C.S.; Hazen, T.C.; VanderGheynst, J.S.; Hugenholtz, P.; Simmons, B.A.; Singer, S.W.

    2011-05-01

    Industrial-scale biofuel production requires robust enzymatic cocktails to produce fermentable sugars from lignocellulosic biomass. Thermophilic bacterial consortia are a potential source of cellulases and hemicellulases adapted to harsher reaction conditions than commercial fungal enzymes. Compost-derived microbial consortia were adapted to switchgrass at 60 C to develop thermophilic biomass-degrading consortia for detailed studies. Microbial community analysis using small-subunit rRNA gene amplicon pyrosequencing and short-read metagenomic sequencing demonstrated that thermophilic adaptation to switchgrass resulted in low-diversity bacterial consortia with a high abundance of bacteria related to thermophilic paenibacilli, Rhodothermus marinus, and Thermus thermophilus. At lower abundance, thermophilic Chloroflexi and an uncultivated lineage of the Gemmatimonadetes phylum were observed. Supernatants isolated from these consortia had high levels of xylanase and endoglucanase activities. Compared to commercial enzyme preparations, the endoglucanase enzymes had a higher thermotolerance and were more stable in the presence of 1-ethyl-3-methylimidazolium acetate ([C2mim][OAc]), an ionic liquid used for biomass pretreatment. The supernatants were used to saccharify [C2mim][OAc]-pretreated switchgrass at elevated temperatures (up to 80 C), demonstrating that these consortia are an excellent source of enzymes for the development of enzymatic cocktails tailored to more extreme reaction conditions.

  17. A comparative study on thermomechanical pulping pressate treatment using thermophilic and mesophilic sequencing batch reactors.

    PubMed

    Zheng, Meiru; Liao, B Q

    2014-01-01

    A comparative study on the treatment of thermomechanical pulping (TMP) pressate was conducted under thermophilic (55 degrees C) and mesophilic (30 degrees C) temperatures to explore in-mill biological treatment, with the intention to operate under heat-efficient conditions. The experimental study involved sequencing batch reactors (SBRs) operated over 114 days. Receiving a total influent chemical oxygen demand (COD) of 3700-4100 mg L(-1), the COD removal efficiencies of 80-90% and 75-85% were achieved for the mesophilic and thermophilic SBRs, respectively, at a hydraulic retention time (HRT) of 12 and 24h. Excellent sludge settleability (sludge volume index < 100 mL g(-1) mixed liquor suspended solids) was obtained at both thermophilic and mesophilic SBRs. A higher level of effluent suspended solids was observed under thermophilic conditions. The results support the feasibility of applying thermophilic biological treatment of TMP pressate. The treated effluent has the potential for subsequent reuse as process water after polishing, thus addressing the long-standing desire to develop water system closure for the pulp and paper mill operation. PMID:24701939

  18. Genetic tool development underpins recent advances in thermophilic whole‐cell biocatalysts

    PubMed Central

    Taylor, M. P.; van Zyl, L.; Tuffin, I. M.; Leak, D. J.; Cowan, D. A.

    2011-01-01

    Summary The environmental value of sustainably producing bioproducts from biomass is now widely appreciated, with a primary target being the economic production of fuels such as bioethanol from lignocellulose. The application of thermophilic prokaryotes is a rapidly developing niche in this field, driven by their known catabolic versatility with lignocellulose‐derived carbohydrates. Fundamental to the success of this work has been the development of reliable genetic and molecular systems. These technical tools are now available to assist in the development of other (hyper)thermophilic strains with diverse phenotypes such as hemicellulolytic and cellulolytic properties, branched chain alcohol production and other ‘valuable bioproduct’ synthetic capabilities. Here we present an insight into the historical limitations, recent developments and current status of a number of genetic systems for thermophiles. We also highlight the value of reliable genetic methods for increasing our knowledge of thermophile physiology. We argue that the development of robust genetic systems is paramount in the evolution of future thermophilic based bioprocesses and make suggestions for future approaches and genetic targets that will facilitate this process. PMID:21310009

  19. Microthrix parvicella and Gordona amarae in mesophilic and thermophilic anaerobic digestion systems.

    PubMed

    Marneri, Matina; Mamais, Daniel; Koutsiouki, Efi

    2009-04-14

    The scope of the study presented in this paper is to determine the fate of the filamentous bacteria Gordona amarae and Microthrix parvicella in anaerobic digestion operating under mesophilic and thermophilic conditions. In order to detect and quantify foaming bacteria in the anaerobic digesters, a fluorescent in situ hybridization (FISH) method was developed and applied. This paper presents the results of a laboratory-scale study that involved the operation of four lab-scale anaerobic digestion systems operating in the mesophilic (35 degrees C) and thermophilic (55 degrees C) temperature ranges at 20 days' detention time. According to the FISH counts of G. amarae and M. parvicella, it appears that thermophilic conditions resulted in a higher destruction of both filamentous bacteria, averaging approximately 97% and 94% for the single thermophilic digester and the dual thermophilic/mesophilic system, respectively. Within the context of this study, the overall performance of the four different anaerobic digestion systems was evaluated in terms of biogas production per mass of volatile solids destroyed, COD destruction, sludge dewaterability and foaming characteristics. The dual stage systems used in this study outperformed the single stage digesters. PMID:19507434

  20. CO Metabolism in the Thermophilic Acetogen Thermoanaerobacter kivui.

    PubMed

    Weghoff, Marie Charlotte; Müller, Volker

    2016-04-01

    The thermophilic acetogenic bacterium Thermoanaerobacter kivui, previously described not to use carbon monoxide as a carbon and energy source, was adapted to grow on CO. This was achieved by using a preculture grown on H2 plus CO2 and by increasing the CO concentration in small, 10% increments.T. kivui was finally able to grow within a 100% CO atmosphere. Growth on CO was found in complex and mineral media, and vitamins were not required. Carbon monoxide consumption was accompanied by acetate and hydrogen production. Cells also grew on synthesis gas (syngas) with the simultaneous use of CO and H2 coupled to acetate production. CO oxidation in resting cells was coupled to hydrogen and acetate production and accompanied by the synthesis of ATP. A protonophore abolished ATP synthesis but stimulated H2 production, which is consistent with a chemiosmotic mechanism of ATP synthesis. Hydrogenase activity was highest in crude extracts of CO-grown cells, and carbon monoxide dehydrogenase (CODH) activity was highest in H2-plus-CO2- or CO-grown cells. The genome of T. kivui harbors two CODH gene clusters, and both CODH proteins were present in crude extracts, but one CODH was more prevalent in crude extracts from CO-grown cells. PMID:26850300

  1. Role of Polyphosphate in Thermophilic Synechococcus sp. from Microbial Mats

    PubMed Central

    Fazeli, Fariba; Grote, Alexandra; Grossman, Arthur R.; Bhaya, Devaki

    2013-01-01

    Synechococcus OS-B′, a thermophilic unicellular cyanobacterium, recently isolated from the microbial mats in Octopus Spring (Yellowstone National Park), induces a suite of genes, including phosphatases and transporters, in response to phosphorus (P) starvation. Here we describe two different approaches to examine the ability of Synechococcus OS-B′ to synthesize and break down polyphosphate (poly P), a key storage compound in many prokaryotes. First, we developed a transformation protocol to create mutants in the polyphosphate kinase (ppk), the major enzyme responsible for the synthesis of poly P. The ppk mutant exhibited a pleiotropic phenotype with defects in poly P accumulation, aberrant levels of Pho regulon transcripts, growth defects, and changes in cell size and exopolysaccharide levels, among others. Second, we measured transcripts of ppk and ppx (encoding the polyphosphatase) directly from mat samples and found that the levels varied dramatically over a diel cycle. We also used Western blot analysis to quantify levels of PPK and PPX and found that these enzymes differentially accumulated during the diel cycle. Levels of polyphosphate kinase peaked at night, while polyphosphatase levels were highest during the early morning hours. We hypothesize that the opposing activities of these two enzymes allow cells to store and utilize poly P to optimize growth over a diel cycle. PMID:23687278

  2. Kinetics of inactivation of indicator pathogens during thermophilic anaerobic digestion.

    PubMed

    Popat, Sudeep C; Yates, Marylynn V; Deshusses, Marc A

    2010-12-01

    Thermophilic anaerobic sludge digestion is a promising process to divert waste to beneficial use, but an important question is the required temperature and holding time to achieve a given degree of pathogen inactivation. In this study, the kinetics of inactivation of Ascaris suum and vaccine strain poliovirus type 1 (PVS-1), selected as indicators for helminth ova and enteric viruses respectively, were determined during anaerobic digestion at temperatures ranging from 51 to 56 °C. Inactivation of both indicator organisms was fast with greater than two log reductions achieved within 2 h for A. suum and three log reductions for PVS-1, suggesting that the current U.S. regulations are largely conservative. The first-order inactivation rate constants k followed Arrhenius relationship with activation energies of 105 and 39 KJ mol(-1) for A. suum and PVS-1, respectively indicating that A. suum was more sensitive to temperature. Although inactivation was fast, the presence of compounds in the sludge that are known to be protective of pathogen inactivation was observed, suggesting that composition-dependent time-temperature relationships are necessary. PMID:20692678

  3. Biogeography of thermophilic phototrophic bacteria belonging to Roseiflexus genus.

    PubMed

    Gaisin, Vasil A; Grouzdev, Denis S; Namsaraev, Zorigto B; Sukhacheva, Marina V; Gorlenko, Vladimir M; Kuznetsov, Boris B

    2016-03-01

    Isolated environments such as hot springs are particularly interesting for studying the microbial biogeography. These environments create an 'island effect' leading to genetic divergence. We studied the phylogeographic pattern of thermophilic anoxygenic phototrophic bacteria, belonging to the Roseiflexus genus. The main characteristic of the observed pattern was geographic and geochronologic fidelity to the hot springs within Circum-Pacific and Alpine-Himalayan-Indonesian orogenic belts. Mantel test revealed a correlation between genetic divergence and geographic distance among the phylotypes. Cluster analysis revealed a regional differentiation of the global phylogenetic pattern. The phylogeographic pattern is in correlation with geochronologic events during the break up of Pangaea that led to the modern configuration of continents. To our knowledge this is the first geochronological scenario of intercontinental prokaryotic taxon divergence. The existence of the modern phylogeographic pattern contradicts with the existence of the ancient evolutionary history of the Roseiflexus group proposed on the basis of its deep-branching phylogenetic position. These facts indicate that evolutionary rates in Roseiflexus varied over a wide range. PMID:26826142

  4. Carbohydrate Transport by the Anaerobic Thermophile Clostridium thermocellum LQRI

    PubMed Central

    Strobel, H. J.; Caldwell, F. C.; Dawson, K. A.

    1995-01-01

    Clostridium thermocellum is an anaerobic thermophilic bacterium which degrades cellulose and ferments the resulting glucose, cellobiose, and cellodextrins predominantly to ethanol. However, relatively little information was available on carbohydrate uptake by this bacterium. Washed cells internalized intact oligomers as large as cellopentaose. Since cellobiose and cellodextrin phosphorylase activities were detected in the cytosol and were not associated with cell membranes, phosphorylation of carbohydrates occurred intracellularly. Kinetic studies indicated that cellobiose and larger cellodextrins were taken up by a common uptake system while glucose entered via a separate mechanism. When cells were treated with metabolic inhibitors including iodoacetate and arsenate, the uptake of radiolabeled glucose or cellobiose was reduced by as much as 90%, and this reduction was associated with a 95% decline in intracellular ATP content. A combination of the ionophores nigericin and valinomycin abolished the proton-motive force but only slightly decreased transport and ATP. These results suggested that the two modes of carbohydrate transport in C. thermocellum were ATP dependent. This work is the first demonstration of cellodextrin transport by a cellulolytic bacterium. PMID:16535164

  5. Isolation and characterization of novel thermophilic lipase-secreting bacteria

    PubMed Central

    Rabbani, Mohammed; Bagherinejad, Mohammad Reza; Sadeghi, Hamid MirMohammad; Shariat, Ziaedin Samsam; Etemadifar, Zahra; Moazen, Fatemeh; Rahbari, Manizheh; Mafakher, Ladan; Zaghian, Saeideh

    2013-01-01

    The purpose of the present study was to screen and identify the lipase-producing microorganisms from various regions of Iran. Samples collected from hot spring, Persian Gulf, desert area and oil-contaminated soil, were analyzed for thermophilic extracellular-lipase producing organisms. Six strains with high activity on rhodamine B plates were selected for chemical identification and further study. Among these isolated bacteria, four strains show higher activity in pH-Stat method at 55 °C. These strains were identified by PCR amplification of 16s rRNA genes using universal primers. Fermentation increased the activity up to 50%. The growth medium, designed for lipase production, increased the activity up to 4.55 folds. The crude supernatant of ZR-5 after fermentation and separation the cells, was lyophilized and the activity was measured. Total activity of this strain was 12 kU/g that shows its potential for industrial uses. Further study is required for purification of enzyme and calculation its specific activity. Immobilization is another approach should be considered. PMID:24688500

  6. Isolation and characterization of novel thermophilic lipase-secreting bacteria.

    PubMed

    Rabbani, Mohammed; Bagherinejad, Mohammad Reza; Sadeghi, Hamid MirMohammad; Shariat, Ziaedin Samsam; Etemadifar, Zahra; Moazen, Fatemeh; Rahbari, Manizheh; Mafakher, Ladan; Zaghian, Saeideh

    2013-12-01

    The purpose of the present study was to screen and identify the lipase-producing microorganisms from various regions of Iran. Samples collected from hot spring, Persian Gulf, desert area and oil-contaminated soil, were analyzed for thermophilic extracellular-lipase producing organisms. Six strains with high activity on rhodamine B plates were selected for chemical identification and further study. Among these isolated bacteria, four strains show higher activity in pH-Stat method at 55 °C. These strains were identified by PCR amplification of 16s rRNA genes using universal primers. Fermentation increased the activity up to 50%. The growth medium, designed for lipase production, increased the activity up to 4.55 folds. The crude supernatant of ZR-5 after fermentation and separation the cells, was lyophilized and the activity was measured. Total activity of this strain was 12 kU/g that shows its potential for industrial uses. Further study is required for purification of enzyme and calculation its specific activity. Immobilization is another approach should be considered. PMID:24688500

  7. Allosteric regulation in phosphofructokinase from the extreme thermophile Thermus thermophilus.

    PubMed

    McGresham, Maria S; Lovingshimer, Michelle; Reinhart, Gregory D

    2014-01-14

    An investigation into the kinetics and regulatory properties of the type-1 phosphofructokinase (PFK) from the extreme thermophile Thermus thermophilus (TtPFK) reveals an enzyme that is inhibited by PEP and activated by ADP by modifying the affinity exhibited for the substrate fructose 6-phosphate (Fru-6-P) in a manner analogous to other prokaryotic PFKs. However, TtPFK binds both of these allosteric ligands significantly more tightly than other bacterial PFKs while effecting a substantially more modest extent of inhibition or activation at 25 °C, reinforcing the principle that binding affinity and effectiveness can be both independent and uncorrelated to one another. These properties have allowed us to establish rigorously that PEP only inhibits by antagonizing the binding of Fru-6-P and not by influencing turnover, a conclusion that requires kcat to be determined under conditions in which both inhibitor and substrate are saturating simultaneously. In addition, the temperature dependence of the allosteric effects on Fru-6-P binding indicate that the coupling free energies are entropy-dominated, as observed previously for PFK from Bacillus stearothermophilus but not for PFK from Escherichia coli , supporting the hypothesis that entropy-dominated allosteric effects may be a characteristic of enzymes derived from thermostable organisms. For such enzymes, the root cause of the allosteric effect may not be easily discerned from static structural information such as that obtained from X-ray crystallography. PMID:24328040

  8. Hydrogenomics of the Extremely Thermophilic Bacterium Caldicellulosiruptor saccharolyticus▿ †

    PubMed Central

    van de Werken, Harmen J. G.; Verhaart, Marcel R. A.; VanFossen, Amy L.; Willquist, Karin; Lewis, Derrick L.; Nichols, Jason D.; Goorissen, Heleen P.; Mongodin, Emmanuel F.; Nelson, Karen E.; van Niel, Ed W. J.; Stams, Alfons J. M.; Ward, Donald E.; de Vos, Willem M.; van der Oost, John; Kelly, Robert M.; Kengen, Servé W. M.

    2008-01-01

    Caldicellulosiruptor saccharolyticus is an extremely thermophilic, gram-positive anaerobe which ferments cellulose-, hemicellulose- and pectin-containing biomass to acetate, CO2, and hydrogen. Its broad substrate range, high hydrogen-producing capacity, and ability to coutilize glucose and xylose make this bacterium an attractive candidate for microbial bioenergy production. Here, the complete genome sequence of C. saccharolyticus, consisting of a 2,970,275-bp circular chromosome encoding 2,679 predicted proteins, is described. Analysis of the genome revealed that C. saccharolyticus has an extensive polysaccharide-hydrolyzing capacity for cellulose, hemicellulose, pectin, and starch, coupled to a large number of ABC transporters for monomeric and oligomeric sugar uptake. The components of the Embden-Meyerhof and nonoxidative pentose phosphate pathways are all present; however, there is no evidence that an Entner-Doudoroff pathway is present. Catabolic pathways for a range of sugars, including rhamnose, fucose, arabinose, glucuronate, fructose, and galactose, were identified. These pathways lead to the production of NADH and reduced ferredoxin. NADH and reduced ferredoxin are subsequently used by two distinct hydrogenases to generate hydrogen. Whole-genome transcriptome analysis revealed that there is significant upregulation of the glycolytic pathway and an ABC-type sugar transporter during growth on glucose and xylose, indicating that C. saccharolyticus coferments these sugars unimpeded by glucose-based catabolite repression. The capacity to simultaneously process and utilize a range of carbohydrates associated with biomass feedstocks is a highly desirable feature of this lignocellulose-utilizing, biofuel-producing bacterium. PMID:18776029

  9. Continuous culture of Methanococcus jannaschii, an extremely thermophilic methanogen

    SciTech Connect

    Jiahuey Tsao; Kaneshiro, S.M.; Shusan Yu; Clark, D.S. . Dept. of Chemical Engineering)

    1994-02-05

    Methanococcus jannaschii, an extremely thermophilic methanogen isolated from a deep-sea hydrothermal vent was grown at 80 C in continuous culture on a mineral salts medium gassed with H[sub 2] and CO[sub 2] at three different flow rates. The maximum specific growth rate was 0.56 h[sup [minus]1], and the maximum specific methane productivity was 0.32 (mol g[sup [minus]1] h[sup [minus]1]). Uncoupling of growth and methane production was evidenced by an increase in the non-growth-associated rate of methane formation, [Beta], with increasing gaseous input. The specific hydrogenase activity exhibited growth-associated behavior at low growth rates, but showed no dependence on growth at higher growth rates. The growth dependence of hydrogenase activity is consistent with the pressure dependence of hydrogenase activity measured in previous experiments. In contrast, the specific protease activity was independent of the growth rate over the entire range of dilution rates studied.

  10. How do people define moderation?

    PubMed

    vanDellen, Michelle R; Isherwood, Jennifer C; Delose, Julie E

    2016-06-01

    Eating in moderation is considered to be sound and practical advice for weight maintenance or prevention of weight gain. However, the concept of moderation is ambiguous, and the effect of moderation messages on consumption has yet to be empirically examined. The present manuscript examines how people define moderate consumption. We expected that people would define moderate consumption in ways that justified their current or desired consumption rather than view moderation as an objective standard. In Studies 1 and 2, moderate consumption was perceived to involve greater quantities of an unhealthy food (chocolate chip cookies, gummy candies) than perceptions of how much one should consume. In Study 3, participants generally perceived themselves to eat in moderation and defined moderate consumption as greater than their personal consumption. Furthermore, definitions of moderate consumption were related to personal consumption behaviors. Results suggest that the endorsement of moderation messages allows for a wide range of interpretations of moderate consumption. Thus, we conclude that moderation messages are unlikely to be effective messages for helping people maintain or lose weight. PMID:26964691

  11. Effect of temperature on solids reductions and on degradation kinetics during thermophilic aerobic digestion of a simulated sludge.

    PubMed

    Toki, C J

    2008-07-01

    Laboratory-scale experiments were conducted to determine the influence of higher thermophilic temperatures on thermophilic aerobic digestion treatment of a simulated sludge. The efficiency of the process was evaluated in respect of solids removal and degradation rate constants at four thermophilic temperatures. Batch runs were operated at a retention time of one day and temperatures of 65, 70, 72 and 75 degrees C. The results indicated that temperature increase did not impart any significant benefits to the digestion operation in terms of suspended solids and biochemichal oxygen demand reduction. The findings from this research also suggested that the treatment would not appear to benefit from temperatures higher than 65 degrees C, as classically suggested by Van't Hoff-Arrhenius. Therefore, increase of thermophilic temperature in the tested 65-75 degrees C range does not enhance the efficiency of thermophilic, aerobic sludge digestion treatment. PMID:18697516

  12. Characterization of Melioribacter roseus gen. nov., sp. nov., a novel facultatively anaerobic thermophilic cellulolytic bacterium from the class Ignavibacteria, and a proposal of a novel bacterial phylum Ignavibacteriae.

    PubMed

    Podosokorskaya, Olga A; Kadnikov, Vitaly V; Gavrilov, Sergey N; Mardanov, Andrey V; Merkel, Alexander Y; Karnachuk, Olga V; Ravin, Nikolay V; Bonch-Osmolovskaya, Elizaveta A; Kublanov, Ilya V

    2013-06-01

    A novel moderately thermophilic, facultatively anaerobic chemoorganotrophic bacterium strain P3M-2(T) was isolated from a microbial mat developing on the wooden surface of a chute under the flow of hot water (46°C) coming out of a 2775-m-deep oil exploration well (Tomsk region, Russia). Strain P3M-2(T) is a moderate thermophile and facultative anaerobe growing on mono-, di- or polysaccharides by aerobic respiration, fermentation or by reducing diverse electron acceptors [nitrite, Fe(III), As(V)]. Its closest cultivated relative (90.8% rRNA gene sequence identity) is Ignavibacterium album, the only chemoorganotrophic member of the phylum Chlorobi. New genus and species Melioribacter roseus are proposed for isolate P3M-2(T) . Together with I. album, the new organism represents the class Ignavibacteria assigned to the phylum Chlorobi. The revealed group includes a variety of uncultured environmental clones, the 16S rRNA gene sequences of some of which have been previously attributed to the candidate division ZB1. Phylogenetic analysis of M. roseus and I. album based on their 23S rRNA and RecA sequences confirmed that these two organisms could represent an even deeper, phylum-level lineage. Hence, we propose a new phylum Ignavibacteriae within the Bacteroidetes-Chlorobi group with a sole class Ignavibacteria, two families Ignavibacteriaceae and Melioribacteraceae and two species I. album and M. roseus. This proposal correlates with chemotaxonomic data and phenotypic differences of both organisms from other cultured representatives of Chlorobi. The most essential differences, supported by the analyses of complete genomes of both organisms, are motility, facultatively anaerobic and obligately organotrophic mode of life, the absence of chlorosomes and the apparent inability to grow phototrophically. PMID:23297868

  13. Herbinix hemicellulosilytica gen. nov., sp. nov., a thermophilic cellulose-degrading bacterium isolated from a thermophilic biogas reactor.

    PubMed

    Koeck, Daniela E; Ludwig, Wolfgang; Wanner, Gerhard; Zverlov, Vladimir V; Liebl, Wolfgang; Schwarz, Wolfgang H

    2015-08-01

    Phenotypic and phylogenetic studies were performed on new isolates of a novel Gram-stain-positive, anaerobic, non-sporulating, rod-shaped bacterium isolated from a thermophilic biogas plant. The novel organisms were able to degrade crystalline cellulose. 16S rRNA gene comparative sequence analysis demonstrated that the isolates formed a hitherto unknown subline within the family Lachnospiraceae. As a representative of the whole group of isolates, strain T3/55T was further characterized. The closest relative of T3/55T among the taxa with validly published names is Mobilitalea sibirica, sharing 93.9% 16S rRNA gene sequence similarity. Strain T3/55T was catalase-negative, indole-negative, and produced acetate, ethanol and propionic acid as major end products from cellulose metabolism. The major cellular fatty acids (>1%) were 16 : 0 dimethyl acetal, 16 : 0 fatty acid methyl ester and 16 : 0 aldehyde. The DNA G+C content was 36.6 mol%. A novel genus and species, Herbinix hemicellulosilytica gen. nov., sp. nov., is proposed based on phylogenetic analysis and physiological properties of the novel isolate. Strain T3/55T ( = DSM 29228T = CECT 8801T), represents the type strain of Herbinix hemicellulosilytica gen. nov., sp. nov. PMID:25872956

  14. Characteristics of Recombinant Phytase (rSt-Phy) of the Thermophilic mold Sporotrichum thermophile and its applicability in dephytinizing foods.

    PubMed

    Ranjan, Bibhuti; Singh, Bijender; Satyanarayana, T

    2015-12-01

    Sporotrichum thermophile produces very low titres of phytase (St-Phy) extracellularly, which is acidstable, thermostable, and protease insensitive with broad substrate specificity, and therefore, the gene encoding phytase (St-Phy) has been cloned and expressed in E. coli. The purified recombinant phytase (rSt-Phy) has the molecular mass of 55 kDa with Km and Vmax (calcium phytate), kcat and kcat/Km of 0.143 mM, 185.05 nmoles mg(-1)  s(-1), 5.1 × 10(3) s(-1), and 3.5 × 10(7) M(-1) s(-1), respectively. Mg(2+) and Ba(2+) display slight stimulatory effect on the enzyme, while it is inhibited by other ions to a varied extent. The enzyme is also inhibited by chaotropic agents (guanidinium hydrochloride, potassium iodide, and urea), Woodward's reagent K, and 2,3-butanedione but resistant to both pepsin and trypsin. The rSt-Phy is useful in dephytinization of tandoori and naan (unleavened flat Indian breads), and bread, liberating soluble inorganic phosphate that mitigates anti-nutrient effects of phytic acid. PMID:26433599

  15. Comparison of mesophilic and thermophilic anaerobic digestion of sugar beet pulp: performance, dewaterability and foam control.

    PubMed

    Suhartini, Sri; Heaven, Sonia; Banks, Charles J

    2014-01-01

    Digestion of sugar beet pulp was assessed in relation to biogas and methane production, foaming potential, and digestate dewaterability. Four 4-litre working volume digesters were operated mesophilically (37±0.5 °C) and four thermophilically (55±0.5 °C) over three hydraulic retention times. Digesters were operated in duplicate at organic loading rates (OLR) of 4 and 5 g volatile solids l(-1) day(-1) without water addition. Thermophilic digestion gave higher biogas and methane productivity than mesophilic and was able to operate at the higher OLR, where mesophilic digestion showed signs of instability. Digestate dewaterability was assessed using capillary suction time and frozen image centrifugation. The occurrence of, or potential for, stable foam formation was assessed using a foaming potential test. Thermophilic operation allowed higher loadings to be applied without loss of performance, and gave a digestate with superior dewatering characteristics and very little foaming potential. PMID:24291796

  16. Anaerobic digestion of mixed microalgae cultivated in secondary effluent under mesophilic and thermophilic conditions.

    PubMed

    Cea-Barcia, Glenda; Moreno, Gloria; Buitrón, Germán

    2015-01-01

    The anaerobic digestion of mixed indigenous microalgae, grown in a secondary effluent, was evaluated in batch tests at mesophilic (35°C) and thermophilic (50°C) conditions. Under mesophilic conditions, specific methane production varied from 178 to 207 mL CH4/g volatile solids (VS) and the maximum production rate varied from 8.8 to 26.1 mL CH4/(gVS day), depending on the type of microalgae culture. Lower methane parameters were observed in those cultures where Scenedesmus represents more than 95% of the microalge. The culture with the lowest digestion performances under mesophilic conditions was studied under thermophilic conditions. The increase in the incubation temperature significantly increased the specific methane production (390 mL CH4/g VS) and rate (26.0 mL CH4/(gVS day)). However, under thermophilic conditions a lag period of 30 days was observed. PMID:26465311

  17. Comparison of multi-enzyme and thermophilic bacteria on the hydrolysis of mariculture organic waste (MOW).

    PubMed

    Guo, Liang; Sun, Mei; Zong, Yan; Zhao, Yangguo; Gao, Mengchun; She, Zonglian

    2016-01-01

    Mariculture organic waste (MOW) is rich in organic matter, which is a potential energy resource for anaerobic digestion. In order to enhance the anaerobic fermentation, the MOW was hydrolyzed by multi-enzyme and thermophilic bacteria. It was advantageous for soluble chemical oxygen demand (SCOD) release at MOW concentrations of 6 and 10 g/L with multi-enzyme and thermophilic bacteria pretreatments. For multi-enzyme, the hydrolysis was not obvious at substrate concentrations of 1 and 3 g/L, and the protein and carbohydrate increased with hydrolysis time at substrate concentrations of 6 and 10 g/L. For thermophilic bacteria, the carbohydrate was first released at 2-4 h and then consumed, and the protein increased with hydrolysis time. The optimal enzyme hydrolysis for MOW was determined by measuring the changes of SCOD, protein, carbohydrate, ammonia and total phosphorus, and comparing with acid and alkaline pretreatments. PMID:27120653

  18. FLUID MODERATED REACTOR

    DOEpatents

    Wigner, E.P.; Ohlinger, L.A.; Young, G.J.; Weinberg, A.M.

    1957-10-22

    A reactor which utilizes fissionable fuel elements in rod form immersed in a moderator or heavy water and a means of circulating the heavy water so that it may also function as a coolant to remove the heat generated by the fission of the fuel are described. In this design, the clad fuel elements are held in vertical tubes immersed in heavy water in a tank. The water is circulated in a closed system by entering near the tops of the tubes, passing downward through the tubes over the fuel elements and out into the tank, where it is drawn off at the bottom, passed through heat exchangers to give up its heat and then returned to the tops of the tubes for recirculation.

  19. Dispersal of thermophilic Desulfotomaculum endospores into Baltic Sea sediments over thousands of years

    PubMed Central

    de Rezende, Júlia Rosa; Kjeldsen, Kasper Urup; Hubert, Casey R J; Finster, Kai; Loy, Alexander; Jørgensen, Bo Barker

    2013-01-01

    Patterns of microbial biogeography result from a combination of dispersal, speciation and extinction, yet individual contributions exerted by each of these mechanisms are difficult to isolate and distinguish. The influx of endospores of thermophilic microorganisms to cold marine sediments offers a natural model for investigating passive dispersal in the ocean. We investigated the activity, diversity and abundance of thermophilic endospore-forming sulfate-reducing bacteria (SRB) in Aarhus Bay by incubating pasteurized sediment between 28 and 85 °C, and by subsequent molecular diversity analyses of 16S rRNA and of the dissimilatory (bi)sulfite reductase (dsrAB) genes within the endospore-forming SRB genus Desulfotomaculum. The thermophilic Desulfotomaculum community in Aarhus Bay sediments consisted of at least 23 species-level 16S rRNA sequence phylotypes. In two cases, pairs of identical 16S rRNA and dsrAB sequences in Arctic surface sediment 3000 km away showed that the same phylotypes are present in both locations. Radiotracer-enhanced most probable number analysis revealed that the abundance of endospores of thermophilic SRB in Aarhus Bay sediment was ca. 104 per cm3 at the surface and decreased exponentially to 100 per cm3 at 6.5 m depth, corresponding to 4500 years of sediment age. Thus, a half-life of ca. 300 years was estimated for the thermophilic SRB endospores deposited in Aarhus Bay sediments. These endospores were similarly detected in the overlying water column, indicative of passive dispersal in water masses preceding sedimentation. The sources of these thermophiles remain enigmatic, but at least one source may be common to both Aarhus Bay and Arctic sediments. PMID:22832348

  20. Biogas production and methanogenic archaeal community in mesophilic and thermophilic anaerobic co-digestion processes.

    PubMed

    Yu, D; Kurola, J M; Lähde, K; Kymäläinen, M; Sinkkonen, A; Romantschuk, M

    2014-10-01

    Over 258 Mt of solid waste are generated annually in Europe, a large fraction of which is biowaste. Sewage sludge is another major waste fraction. In this study, biowaste and sewage sludge were co-digested in an anaerobic digestion reactor (30% and 70% of total wet weight, respectively). The purpose was to investigate the biogas production and methanogenic archaeal community composition in the anaerobic digestion reactor under meso- (35-37 °C) and thermophilic (55-57 °C) processes and an increasing organic loading rate (OLR, 1-10 kg VS m(-3) d(-1)), and also to find a feasible compromise between waste treatment capacity and biogas production without causing process instability. In summary, more biogas was produced with all OLRs by the thermophilic process. Both processes showed a limited diversity of the methanogenic archaeal community which was dominated by Methanobacteriales and Methanosarcinales (e.g. Methanosarcina) in both meso- and thermophilic processes. Methanothermobacter was detected as an additional dominant genus in the thermophilic process. In addition to operating temperatures, the OLRs, the acetate concentration, and the presence of key substrates like propionate also affected the methanogenic archaeal community composition. A bacterial cell count 6.25 times higher than archaeal cell count was observed throughout the thermophilic process, while the cell count ratio varied between 0.2 and 8.5 in the mesophilic process. This suggests that the thermophilic process is more stable, but also that the relative abundance between bacteria and archaea can vary without seriously affecting biogas production. PMID:24837280

  1. Caenibacillus caldisaponilyticus gen. nov., sp. nov., a thermophilic, spore-forming and phospholipid-degrading bacterium isolated from acidulocompost.

    PubMed

    Tsujimoto, Yoshiyuki; Saito, Ryo; Furuya, Hiroto; Ishihara, Daisuke; Sahara, Takehiko; Kimura, Nobutada; Nishino, Tokuzo; Tsuruoka, Naoki; Shigeri, Yasushi; Watanabe, Kunihiko

    2016-07-01

    A thermophilic and phospholipid-degrading bacterium, designated strain B157T, was isolated from acidulocompost, a garbage compost processed under acidic conditions at moderately high temperature. The organism was Gram-stain-positive, aerobic, spore-forming and rod-shaped. Growth was observed to occur at 40-65 °C and pH 4.8-8.1 (optimum growth: 50-60 °C, pH 6.2). The strain was catalase- and oxidase-positive. The cell wall contained meso-diaminopimelic acid, alanine, glutamic acid and galactose. The predominant respiratory quinone was menaquinone-7 (MK-7) and the major fatty acids were anteiso-C17 : 0 and iso-C17 : 0. Comparative 16S rRNA gene sequence analysis showed that strain B157T was related most closely to Tuberibacillus calidus 607T (94.8 % identity), and the phylogenetic analysis revealed that it belonged to the family Sporolactobacillaceae. The DNA G+C content was determined as 51.8 mol%. In spite of many similarities with the type strains of members of the family Sporolactobacillaceae, genotypic analyses suggest that strain B157T represents a novel species of a new genus, Caenibacilluscaldisaponilyticus gen. nov., sp. nov. The type strain of Caenibacilluscaldisaponilyticus is B157T (=NBRC 111400T=DSM 101100T). PMID:27117268

  2. Isolation and characterization of a fast-growing, thermophilic Methanobacterium species

    SciTech Connect

    Zhao, Y.; Zhang, H.; Boone, D.R.; Mah, R.A.

    1986-11-01

    A thermophilic, autotrophic methanogen (strain CB12, DSM 3664) was isolated from a mesophilic biogas digestor. This bacterium used H/sub 2/-CO/sub 2/ or formate as a substrate and grew as short rods, sometimes in pairs and in crooked filaments. Motility was not observed. Its optimum temperature (56/sup 0/C) was lower than that of other thermophilic members of the genus Methanobacterium. The maximum observed specific growth rate was 0.564 h/sup -1/ (74-min doubling time).

  3. Chromosome map of the thermophilic archaebacterium Thermococcus celer

    NASA Technical Reports Server (NTRS)

    Noll, K. M.; Woese, C. R. (Principal Investigator)

    1989-01-01

    A physical map for the chromosome of the thermophilic archaebacterium Thermococcus celer Vu13 has been constructed. Thirty-four restriction endonucleases were tested for their ability to generate large restriction fragments from the chromosome of T. celer. Of these, the enzymes NheI, SpeI, and XbaI yielded the fewest fragments when analyzed by pulsed-field electrophoresis. NheI and SpeI each gave 5 fragments, while XbaI gave 12. The size of the T. celer chromosome was determined from the sum of the apparent sizes of restriction fragments derived from single and double digests by using these enzymes and was found to be 1,890 +/- 27 kilobase pairs. Partial and complete digests allowed the order of all but three small (less than 15 kilobase pairs) fragments to be deduced. These three fragments were assigned positions by using hybridization probes derived from these restriction fragments. The positions of the other fragments were confirmed by using hybridization probes derived in the same manner. The positions of the 5S, 16S, and 23S rRNA genes as well as the 7S RNA gene were located on this map by using cloned portions of these genes as hybridization probes. The 5S rRNA gene was localized 48 to 196 kilobases from the 5' end of the 16S gene. The 7S RNA gene was localized 190 to 504 kilobases from the 3' end of the 23S gene. These analyses demonstrated that the chromosome of T. celer is a single, circular DNA molecule. This is the first such demonstration of the structure of an archaebacterial chromosome.

  4. Kinetics of thermophilic anaerobes in fixed-bed reactors.

    PubMed

    Perez, M; Romero, L I; Sales, D

    2001-08-01

    The main objective of this study is to estimate growth kinetic constants and the concentration of "active" attached biomass in two anaerobic thermophilic reactors which contain different initial sizes of immobilized anaerobic mixed cultures and decompose distillery wastewater. This paper studies the substrate decomposition in two lab-scale fixed-bed reactors operating at batch conditions with corrugated tubes as support media. It can be demonstrated that high micro-organisms-substrate ratios favor the degradation activity of the different anaerobic cultures, allowing the stable operation without lag-phases and giving better quality in effluent. The kinetic parameters obtained--maximum specific growth rates (mu(max)), non-biodegradable substrate (S(NB)) and "active or viable biomass" concentrations (X(V0))--were obtained by applying the Romero kinetic model [L.I. Romero, 1991. Desarrollo de un modelo matemático general para los procesos fermentativos, Cinética de la degradación anaerobia, Ph.D. Thesis, University of Cádiz (Spain), Serv. Pub. Univ. Cádiz], with COD as substrate and methane (CH4) as the main product of the anaerobic process. This method is suitable to calculate and to differentiate the main kinetic parameters of both the total anaerobic mixed culture and the methanogenic population. Comparison of experimental measured concentration of volatile attached solids (VS(att)) in both reactors with the estimated "active" biomass concentrations obtained by applying Romero kinetic model [L.I. Romero, 1991. Desarrollo de un modelo matemático general para los procesos fermentativos, Cinética de la degradación anaerobia, Ph.D. Thesis, University of Cádiz (Spain), Serv. Pub. Univ. Cádiz] shows that a large amount of inert matter is present in the fixed-bed reactor. PMID:11513409

  5. Optimum conditions for aerobic thermophilic pretreatment of municipal sludge

    SciTech Connect

    Cheunbarn, T.; Pagilla, K.R.

    1998-07-01

    Lab scale experiments were conducted to determine optimum sludge residence time (SRT) and temperature of aerobic thermophilic pretreatment (ATP) of mixed (thickened waste activated and primary) sludge to achieve maximum pathogen destruction and best process performance. 4L lab scale ATP reactors were operated at SRT of 0.6, 1.0, and 1.5 days, and at temperature of 55, 58, 62 and 65 C. ATP at temperature {ge} 62 C and SRT {ge} 0.6 days reduced the feed sludge fecal coliform density by at least 4-logs from 10{sup 7} MPN/g total solids to < 10{sup 4} MPN/g total solids. Salmonella in the feed sludge was reduced to < 1 MPN/g total solids from 2 to 18 MPN/4 g total solids by ATP at temperature {ge} 55 C and SRT {ge} 0.6 days. ATP was able to increase sludge volatile acids concentration by 100--200% over the feed sludge volatile acid concentration, and reduce the supernatant COD from 20,000--22,000 mg/L in the feed to 13,000--17,000 mg/L in ATP reactor sludge. Volatile solids destruction by ATP was increased from 25% to 40% when SRT was increased from 0.6 days to 1.5 days, and only 5% increase in volatile solids destruction was seen at each SRT of 0.6, 1.0, and 1.5 days when ATP temperature was increased from 55 to 65 C.

  6. Experimental evolution of a facultative thermophile from a mesophilic ancestor.

    PubMed

    Blaby, Ian K; Lyons, Benjamin J; Wroclawska-Hughes, Ewa; Phillips, Grier C F; Pyle, Tyler P; Chamberlin, Stephen G; Benner, Steven A; Lyons, Thomas J; Crécy-Lagard, Valérie de; Crécy, Eudes de

    2012-01-01

    Experimental evolution via continuous culture is a powerful approach to the alteration of complex phenotypes, such as optimal/maximal growth temperatures. The benefit of this approach is that phenotypic selection is tied to growth rate, allowing the production of optimized strains. Herein, we demonstrate the use of a recently described long-term culture apparatus called the Evolugator for the generation of a thermophilic descendant from a mesophilic ancestor (Escherichia coli MG1655). In addition, we used whole-genome sequencing of sequentially isolated strains throughout the thermal adaptation process to characterize the evolutionary history of the resultant genotype, identifying 31 genetic alterations that may contribute to thermotolerance, although some of these mutations may be adaptive for off-target environmental parameters, such as rich medium. We undertook preliminary phenotypic analysis of mutations identified in the glpF and fabA genes. Deletion of glpF in a mesophilic wild-type background conferred significantly improved growth rates in the 43-to-48°C temperature range and altered optimal growth temperature from 37°C to 43°C. In addition, transforming our evolved thermotolerant strain (EVG1064) with a wild-type allele of glpF reduced fitness at high temperatures. On the other hand, the mutation in fabA predictably increased the degree of saturation in membrane lipids, which is a known adaptation to elevated temperature. However, transforming EVG1064 with a wild-type fabA allele had only modest effects on fitness at intermediate temperatures. The Evolugator is fully automated and demonstrates the potential to accelerate the selection for complex traits by experimental evolution and significantly decrease development time for new industrial strains. PMID:22020511

  7. Characterization of Thermophilic Consortia from Two Souring Oil Reservoirs

    PubMed Central

    Mueller, R. F.; Nielsen, P. H.

    1996-01-01

    The microbial consortia from produced water at two different oil fields in Alaska (Kuparuk) and the North Sea (Ninian) were investigated for sulfate-reducing and methanogenic activity over a range of temperatures and for a variety of substrates. The consortia were sampled on site, and samples were either incubated on site at 60(deg)C with various substrates or frozen for later incubation and analyses. Temperature influenced the rates of sulfate reduction, hydrogen sulfide production, and substrate oxidation, as well as the cell morphology. The highest rates of sulfate reduction and substrate oxidation were found between 50 and 60(deg)C. Formate and n-butyrate were the most favorable electron donors at any tested temperature. Acetate was utilized at 35(deg)C but not at 50 or 70(deg)C and was produced at 60(deg)C. This indicates that the high levels of acetate found in produced water from souring oil formations are due mainly to an incomplete oxidation of volatile fatty acids to acetate. The cell size distribution of the microbial consortium indicated a nonuniform microbial composition in the original sample from the Kuparuk field. At different temperatures, different microbial morphologies and physiologies were observed. Methane-producing activity at thermophilic temperatures (60(deg)C) was found only for the Kuparuk consortium when hydrogen and carbon dioxide were present. No methane production from acetate was observed. Suppression of methanogenic activity in the presence of sulfate indicated a competition with sulfate-reducing bacteria for hydrogen. PMID:16535394

  8. Geobacillus sp., a thermophilic soil bacterium producing volatile antibiotics.

    PubMed

    Ren, Yuhao; Strobel, Gary; Sears, Joe; Park, Melina

    2010-07-01

    Geobacillus, a bacterial genus, is represented by over 25 species of Gram-positive isolates from various man-made and natural thermophilic areas around the world. An isolate of this genus (M-7) has been acquired from a thermal area near Yellowstone National Park, MT and partially characterized. The cells of this organism are globose (ca. 0.5 mu diameter), and they are covered in a matrix capsule which gives rise to elongate multicelled bacilliform structures (ranging from 3 to 12 mum) as seen by light and atomic force microscopy, respectively. The organism produces unique petal-shaped colonies (undulating margins) on nutrient agar, and it has an optimum pH of 7.0 and an optimum temperature range of 55-65 degrees C. The partial 16S rRNA sequence of this organism has 97% similarity with Geobacillus stearothermophilus, one of its closest relatives genetically. However, uniquely among all members of this genus, Geobacillus sp. (M-7) produces volatile organic substances (VOCs) that possess potent antibiotic activities. Some of the more notable components of the VOCs are benzaldehyde, acetic acid, butanal, 3-methyl-butanoic acid, 2-methyl-butanoic acid, propanoic acid, 2-methyl-, and benzeneacetaldehyde. An exposure of test organisms such as Aspergillus fumigatus, Botrytis cinerea, Verticillium dahliae, and Geotrichum candidum produced total inhibition of growth on a 48-h exposure to Geobacillus sp.(M-7) cells (ca.10(7)) and killing at a 72-h exposure at higher bacterial cell concentrations. A synthetic mixture of those available volatile compounds, at the ratios occurring in Geobacillus sp. (M-7), mimicked the bioactivity of this organism. PMID:20091406

  9. Treatment of sewage sludge in a thermophilic membrane reactor (TMR) with alternate aeration cycles.

    PubMed

    Collivignarelli, Maria Cristina; Castagnola, Federico; Sordi, Marco; Bertanza, Giorgio

    2015-10-01

    The management of sewage sludge is becoming a more and more important issue, both at national and international level, in particular due to the uncertain recovery/disposal future options. Therefore, it is clear that the development of new technologies that can mitigate the problem at the source by reducing sludge production is necessary, such as the European Directive 2008/98/EC prescribes. This work shows the results obtained with a thermophilic membrane reactor, for processing a biological sludge derived from a wastewater treatment plant (WWTP) that treats urban and industrial wastewater. Sewage sludge was treated in a thermophilic membrane reactor (TMR), at pilot-scale (1 m(3) volume), with alternate aeration cycles. The experimentation was divided into two phases: a "startup phase" during which, starting with a psychrophilic/mesophilic biomass, thermophilic conditions were progressively reached, while feeding a highly biodegradable substrate; the obtained thermophilic biomass was then used, in the "regime phase", to digest biological sludge which was fed to the plant. Good removal yields were observed: 64% and 57% for volatile solids (VS) and total COD (CODtot), respectively, with an average hydraulic retention time (HRT) equal to 20 d, an organic loading rate (OLR) of about 1.4-1.8 kg COD m(-3) d(-1) and aeration/non aeration cycles alternated every 4 h. PMID:26233586

  10. Thermal adaptation analyzed by comparison of protein sequences from mesophilic and extremely thermophilic Methanococcus species

    NASA Technical Reports Server (NTRS)

    Haney, P. J.; Badger, J. H.; Buldak, G. L.; Reich, C. I.; Woese, C. R.; Olsen, G. J.

    1999-01-01

    The genome sequence of the extremely thermophilic archaeon Methanococcus jannaschii provides a wealth of data on proteins from a thermophile. In this paper, sequences of 115 proteins from M. jannaschii are compared with their homologs from mesophilic Methanococcus species. Although the growth temperatures of the mesophiles are about 50 degrees C below that of M. jannaschii, their genomic G+C contents are nearly identical. The properties most correlated with the proteins of the thermophile include higher residue volume, higher residue hydrophobicity, more charged amino acids (especially Glu, Arg, and Lys), and fewer uncharged polar residues (Ser, Thr, Asn, and Gln). These are recurring themes, with all trends applying to 83-92% of the proteins for which complete sequences were available. Nearly all of the amino acid replacements most significantly correlated with the temperature change are the same relatively conservative changes observed in all proteins, but in the case of the mesophile/thermophile comparison there is a directional bias. We identify 26 specific pairs of amino acids with a statistically significant (P < 0.01) preferred direction of replacement.

  11. Endospores of thermophilic bacteria as tracers of microbial dispersal by ocean currents.

    PubMed

    Müller, Albert Leopold; de Rezende, Júlia Rosa; Hubert, Casey R J; Kjeldsen, Kasper Urup; Lagkouvardos, Ilias; Berry, David; Jørgensen, Bo Barker; Loy, Alexander

    2014-06-01

    Microbial biogeography is influenced by the combined effects of passive dispersal and environmental selection, but the contribution of either factor can be difficult to discern. As thermophilic bacteria cannot grow in the cold seabed, their inactive spores are not subject to environmental selection. We therefore conducted a global experimental survey using thermophilic endospores that are passively deposited by sedimentation to the cold seafloor as tracers to study the effect of dispersal by ocean currents on the biogeography of marine microorganisms. Our analysis of 81 different marine sediments from around the world identified 146 species-level 16S rRNA phylotypes of endospore-forming, thermophilic Firmicutes. Phylotypes showed various patterns of spatial distribution in the world oceans and were dispersal-limited to different degrees. Co-occurrence of several phylotypes in locations separated by great distances (west of Svalbard, the Baltic Sea and the Gulf of California) demonstrated a widespread but not ubiquitous distribution. In contrast, Arctic regions with water masses that are relatively isolated from global ocean circulation (Baffin Bay and east of Svalbard) were characterized by low phylotype richness and different compositions of phylotypes. The observed distribution pattern of thermophilic endospores in marine sediments suggests that the impact of passive dispersal on marine microbial biogeography is controlled by the connectivity of local water masses to ocean circulation. PMID:24351936

  12. Draft Genome Sequence of a Thermophilic Desulfurization Bacterium, Geobacillus thermoglucosidasius Strain W-2

    PubMed Central

    Zhu, Lin; Li, Mingchang; Guo, Shuyi

    2016-01-01

    Geobacillus thermoglucosidasius strain W-2 is a thermophilic bacterium isolated from a deep-subsurface oil reservoir in northern China, which is capable of degrading organosulfur compounds. Here, we report the draft genome sequence of G. thermoglucosidasius strain W-2, which may help to elucidate the genetic basis of biodegradation of organosulfur pollutants under heated conditions. PMID:27491977

  13. Microbial diversity in innovative mesophilic/thermophilic temperature-phased anaerobic digestion of sludge.

    PubMed

    Gagliano, M C; Braguglia, C M; Gallipoli, A; Gianico, A; Rossetti, S

    2015-05-01

    Anaerobic digestion (AD) is one of the few sustainable technologies that both produce energy and treat waste streams. Driven by a complex and diverse community of microbes, AD may be affected by different factors, many of which also influence the composition and activity of the microbial community. In this study, the biodiversity of microbial populations in innovative mesophilic/thermophilic temperature-phased AD of sludge was evaluated by means of fluorescence in situ hybridization (FISH). The increase of digestion temperature drastically affected the microbial composition and selected specialized biomass. Hydrogenotrophic Methanobacteriales and the protein fermentative bacterium Coprothermobacter spp. were identified in the thermophilic anaerobic biomass. Shannon-Weaver diversity (H') and evenness (E) indices were calculated using FISH data. Species richness was lower under thermophilic conditions compared with the values estimated in mesophilic samples, and it was flanked by similar trend of the evenness indicating that thermophilic communities may be therefore more susceptible to sudden changes and less prompt to adapting to operative variations. PMID:24875310

  14. COMPARATIVE EVALUATION OF MESOPHILIC AND THERMOPHILIC ANAEROBIC DIGESTION. PHASE 2. STEADY STATE STUDIES

    EPA Science Inventory

    A study was conducted of the relative performance of anaerobic digestion systems under mesophilic and thermophilic conditions. Fifty liter laboratory scale digesters were fed primary sludge from the Allentown, PA Waste Water Treatment Plant. Long-term, steady-state performance da...

  15. COMPARATIVE EVALUATION OF MESOPHILIC AND THERMOPHILIC DIGESTION - PHASE II. STEADY STATE STUDIES

    EPA Science Inventory

    A study of the relative performance of anaerobic digestion systems under mesophilic and thermophilic conditions was conducted. Fifty liter laboratory scale digesters were fed primary sludge from the Allentown, PA Waste Water Treatment Plant. Long-term, steady-state performance da...

  16. Proteogenomic Analysis of a Thermophilic Bacterial Consortium Adapted to Deconstruct Switchgrass

    PubMed Central

    D'haeseleer, Patrik; Gladden, John M.; Allgaier, Martin; Chain, Patrik S. G.; Tringe, Susannah G.; Malfatti, Stephanie A.; Aldrich, Joshua T.; Nicora, Carrie D.; Robinson, Errol W.; Paša-Tolić, Ljiljana; Hugenholtz, Philip; Simmons, Blake A.; Singer, Steven W.

    2013-01-01

    Thermophilic bacteria are a potential source of enzymes for the deconstruction of lignocellulosic biomass. However, the complement of proteins used to deconstruct biomass and the specific roles of different microbial groups in thermophilic biomass deconstruction are not well-explored. Here we report on the metagenomic and proteogenomic analyses of a compost-derived bacterial consortium adapted to switchgrass at elevated temperature with high levels of glycoside hydrolase activities. Near-complete genomes were reconstructed for the most abundant populations, which included composite genomes for populations closely related to sequenced strains of Thermus thermophilus and Rhodothermus marinus, and for novel populations that are related to thermophilic Paenibacilli and an uncultivated subdivision of the little-studied Gemmatimonadetes phylum. Partial genomes were also reconstructed for a number of lower abundance thermophilic Chloroflexi populations. Identification of genes for lignocellulose processing and metabolic reconstructions suggested Rhodothermus, Paenibacillus and Gemmatimonadetes as key groups for deconstructing biomass, and Thermus as a group that may primarily metabolize low molecular weight compounds. Mass spectrometry-based proteomic analysis of the consortium was used to identify >3000 proteins in fractionated samples from the cultures, and confirmed the importance of Paenibacillus and Gemmatimonadetes to biomass deconstruction. These studies also indicate that there are unexplored proteins with important roles in bacterial lignocellulose deconstruction. PMID:23894306

  17. Thermal and Spectroscopic Characterization of a Proton Pumping Rhodopsin from an Extreme Thermophile*

    PubMed Central

    Tsukamoto, Takashi; Inoue, Keiichi; Kandori, Hideki; Sudo, Yuki

    2013-01-01

    So far retinylidene proteins (∼rhodopsin) have not been discovered in thermophilic organisms. In this study we investigated and characterized a microbial rhodopsin derived from the extreme thermophilic bacterium Thermus thermophilus, which lives in a hot spring at around 75 °C. The gene for the retinylidene protein, named thermophilic rhodopsin (TR), was chemically synthesized with codon optimization. The codon optimized TR protein was functionally expressed in the cell membranes of Escherichia coli cells and showed active proton transport upon photoillumination. Spectroscopic measurements revealed that the purified TR bound only all-trans-retinal as a chromophore and showed an absorption maximum at 530 nm. In addition, TR exhibited both photocycle kinetics and pH-dependent absorption changes, which are characteristic of rhodopsins. Of note, time-dependent thermal denaturation experiments revealed that TR maintained its absorption even at 75 °C, and the denaturation rate constant of TR was much lower than those of other proton pumping rhodopsins such as archaerhodopsin-3 (200 ×), Haloquadratum walsbyi bacteriorhodopsin (by 10-times), and Gloeobacter rhodopsin (100 ×). Thus, these results suggest that microbial rhodopsins are also distributed among thermophilic organisms and have high stability. TR should allow the investigation of the molecular mechanisms of ion transport and protein folding. PMID:23740255

  18. Genome Sequence of Anoxybacillus geothermalis Strain GSsed3, a Novel Thermophilic Endospore-Forming Species

    PubMed Central

    Filippidou, Sevasti; Jaussi, Marion; Junier, Thomas; Wunderlin, Tina; Roussel-Delif, Ludovic; Jeanneret, Nicole; Vieth-Hillebrand, Andrea; Vetter, Alexandra; Regenspurg, Simona; McMurry, Kim; Gleasner, Cheryl D.; Lo, Chien-Chi; Li, Paul; Vuyisich, Momchilo; Chain, Patrick S.

    2015-01-01

    Anoxybacillus geothermalis strain GSsed3 is an endospore-forming thermophilic bacterium isolated from filter deposits in a geothermal site. This novel species has a larger genome size (7.2 Mb) than that of any other Anoxybacillus species, and it possesses genes that support its phenotypic metabolic characterization and suggest an intriguing link to metals. PMID:26067952

  19. Dissecting and engineering metabolic and regulatory networks of thermophilic bacteria for biofuel production.

    PubMed

    Lin, Lu; Xu, Jian

    2013-11-01

    Interest in thermophilic bacteria as live-cell catalysts in biofuel and biochemical industry has surged in recent years, due to their tolerance of high temperature and wide spectrum of carbon-sources that include cellulose. However their direct employment as microbial cellular factories in the highly demanding industrial conditions has been hindered by uncompetitive biofuel productivity, relatively low tolerance to solvent and osmic stresses, and limitation in genome engineering tools. In this work we review recent advances in dissecting and engineering the metabolic and regulatory networks of thermophilic bacteria for improving the traits of key interest in biofuel industry: cellulose degradation, pentose-hexose co-utilization, and tolerance of thermal, osmotic, and solvent stresses. Moreover, new technologies enabling more efficient genetic engineering of thermophiles were discussed, such as improved electroporation, ultrasound-mediated DNA delivery, as well as thermo-stable plasmids and functional selection systems. Expanded applications of such technological advancements in thermophilic microbes promise to substantiate a synthetic biology perspective, where functional parts, module, chassis, cells and consortia were modularly designed and rationally assembled for the many missions at industry and nature that demand the extraordinary talents of these extremophiles. PMID:23510903

  20. Characterization of thermophilic fungal community associated with pile fermentation of Pu-erh tea.

    PubMed

    Zhang, Wei; Yang, Ruijuan; Fang, Wenjun; Yan, Liang; Lu, Jun; Sheng, Jun; Lv, Jie

    2016-06-16

    This study aimed to characterize the thermophilic fungi in pile-fermentation process of Pu-erh tea. Physicochemical analyses showed that the high temperature and low pH provided optimal conditions for propagation of fungi. A number of fungi, including Blastobotrys adeninivorans, Thermomyces lanuginosus, Rasamsonia emersonii, Aspergillus fumigatus, Rhizomucor pusillus, Rasamsonia byssochlamydoides, Rasamsonia cylindrospora, Aspergillus tubingensis, Aspergillus niger, Candida tropicalis and Fusarium graminearum were isolated as thermophilic fungi under combination of high temperature and acid culture conditions from Pu-erh tea pile-fermentation. The fungal communities were analyzed by PCR-DGGE. Results revealed that those fungi are closely related to Debaryomyces hansenii, Cladosporium cladosporioides, A. tubingensis, R. emersonii, R. pusillus, A. fumigatus and A. niger, and the last four presented as dominant species in the pile process. These four preponderant thermophilic fungi reached the order of magnitude of 10(7), 10(7), 10(7) and 10(6)copies/g dry tea, respectively, measured by real-time quantitative PCR (q-PCR). The results indicate that the thermophilic fungi play an important role in Pu-erh tea pile fermentation. PMID:27046629

  1. Complete Genome Sequence of the Cellulolytic Thermophile Clostridium thermocellum DSM1313

    SciTech Connect

    Feinberg, Lawrence F; Foden, Justine; Barrett, Trisha; Davenport, Karen W.; Bruce, David; Detter, J. Chris; Tapia, Roxanne; Han, Cliff; Lapidus, Alla L.; Lucas, Susan; Cheng, Jan-Fang; Pitluck, Sam; Woyke, Tanja; Ivanova, N; Mikhailova, Natalia; Land, Miriam L; Hauser, Loren John; Argyros, Aaron; Goodwin, Lynne A.; Hogsett, David; Caiazza, Nicky

    2011-01-01

    Clostridium thermocellum DSM1313 is a thermophilic, anaerobic bacterium with some of the highest rates of cellulose hydrolysis reported. The complete genome sequence reveals a suite of carbohydrate-active enzymes and demonstrates a level of diversity at the species level distinguishing it from the type strain ATCC27405.

  2. Complete Genome Sequence of the Cellulolytic Thermophile Caldicellulosiruptor obsidiansis OB47T

    SciTech Connect

    Elkins, James G; Lochner, Adriane; Hamilton-Brehm, Scott; Walston Davenport, Karen; Podar, Mircea; Brown, Steven D; Land, Miriam L; Hauser, Loren John; Klingeman, Dawn Marie; Raman, Babu; Goodwin, Lynne A.; Tapia, Roxanne; Meincke, Linda; Detter, J C; Bruce, David; Han, Cliff; Palumbo, Anthony Vito; Cottingham, Robert W; Keller, Martin; Graham, David E

    2010-01-01

    Caldicellulosiruptor obsidiansis OB47T (ATCC BAA-2073; JCM 16842) is an extremely thermophilic, anaerobic bacterium capable of hydrolyzing plant-derived polymers through the expression of multidomain/multifunctional hydrolases. The complete genome sequence reveals a diverse set of carbohydrate-active enzymes and provides further insight into lignocellulosic biomass hydrolysis at high temperatures.

  3. Inactivation of Clostridium difficile in sewage sludge by anaerobic thermophilic digestion.

    PubMed

    Xu, Changyun; Salsali, Hamidreza; Weese, Scott; Warriner, Keith

    2016-01-01

    There has been an increase in community-associated Clostridium difficile infections with biosolids derived from wastewater treatment being identified as one potential source. The current study evaluated the efficacy of thermophilic digestion in decreasing levels of C. difficile ribotype 078 associated with sewage sludge. Five isolates of C. difficile 078 were introduced (final density of 5 log CFU/g) into digested sludge and subjected to anaerobic digestion at mesophilic (36 or 42 °C) or thermophilic (55 °C) temperatures for up to 60 days. It was found that mesophilic digestion at 36 °C did not result in a significant reduction in C. difficile spore levels. In contrast, thermophilic sludge digestion reduced endospore levels at a rate of 0.19-2.68 log CFU/day, depending on the strain tested. The mechanism of lethality was indirect - by stimulating germination then inactivating the resultant vegetative cells. Acidification of sludge by adding acetic acid (6 g/L) inhibited the germination of spores regardless of the sludge digestion temperature. In conclusion, thermophilic digestion can be applied to reduce C. difficile in biosolids, thereby reducing the environmental burden of the enteric pathogen. PMID:26564276

  4. Leg allometry in ants: extreme long-leggedness in thermophilic species.

    PubMed

    Sommer, Stefan; Wehner, Rüdiger

    2012-01-01

    The thermophilic ant genera Cataglyphis and Ocymyrmex share a variety of specialisations that enable them to engage in high-speed foraging at considerably higher temperatures than less heat-tolerant species. In the present account we test the hypothesis that thermophilic ants have longer legs than closely related species from more mesic habitats. By comparing large-sized, medium-sized, and small-sized species of Cataglyphis and Ocymyrmex with size-matched species of the closely related non-thermophilic genera Formica (Formicinae) and Messor (Myrmicinae), respectively, we show that the thermophilic species are equipped with considerably longer legs than their less heat-tolerant relatives. Hence phylogenetically, extreme long-leggedness has evolved at least twice in desert ants: in the Formicinae and the Myrmicinae. Functionally, this morphological trait is adaptive for a number of reasons. The long legs raise the body into cooler layers of air and enable higher running speeds, which increase convective cooling and reduce foraging time. These are important adaptations all the more as due to the low food density prevailing in desert habitats foraging Cataglyphis and Ocymyrmex ants have to cover large distances within their physically demanding foraging grounds. PMID:21992805

  5. Draft Genome of Thermanaerothrix daxensis GNS-1, a Thermophilic Facultative Anaerobe from the Chloroflexi Class Anaerolineae

    PubMed Central

    Pace, Laura A.; Ward, Lewis M.; Fischer, Woodward W.

    2015-01-01

    We present the draft genome of Thermanaerothrix daxensis GNS-1, a thermophilic member of the Chloroflexi phylum. This organism was initially characterized as a nonmotile, strictly anaerobic fermenter; however, genome analysis demonstrates that it encodes genes for a flagellum and multiple pathways for aerobic and anaerobic respiration. PMID:26586891

  6. Draft Genome of Thermanaerothrix daxensis GNS-1, a Thermophilic Facultative Anaerobe from the Chloroflexi Class Anaerolineae.

    PubMed

    Pace, Laura A; Hemp, James; Ward, Lewis M; Fischer, Woodward W

    2015-01-01

    We present the draft genome of Thermanaerothrix daxensis GNS-1, a thermophilic member of the Chloroflexi phylum. This organism was initially characterized as a nonmotile, strictly anaerobic fermenter; however, genome analysis demonstrates that it encodes genes for a flagellum and multiple pathways for aerobic and anaerobic respiration. PMID:26586891

  7. Anaerobic High-Throughput Cultivation Method for Isolation of Thermophiles Using Biomass-Derived Substrates

    SciTech Connect

    Hamilton-Brehm, Scott; Vishnivetskaya, Tatiana A; Allman, Steve L; Mielenz, Jonathan R; Elkins, James G

    2012-01-01

    Flow cytometry (FCM) techniques have been developed for sorting mesophilic organisms, but the difficulty increases if the target microbes are thermophilic anaerobes. We demonstrate a reliable, high-throughput method of screening thermophilic anaerobic organisms using FCM and 96-well plates for growth on biomass-relevant substrates. The method was tested using the cellulolytic thermophiles Clostridium ther- mocellum (Topt = 55 C), Caldicellulosiruptor obsidiansis (Topt = 78 C) and the fermentative hyperthermo- philes, Pyrococcus furiosus (Topt = 100 C) and Thermotoga maritima (Topt = 80 C). Multi-well plates were incubated at various temperatures for approximately 72 120 h and then tested for growth. Positive growth resulting from single cells sorted into individual wells containing an anaerobic medium was verified by OD600. Depending on the growth substrate, up to 80 % of the wells contained viable cultures, which could be transferred to fresh media. This method was used to isolate thermophilic microbes from Rabbit Creek, Yellowstone National Park (YNP), Wyoming. Substrates for enrichment cultures including crystalline cellulose (Avicel), xylan (from Birchwood), pretreated switchgrass and Populus were used to cultivate organisms that may be of interest to lignocellulosic biofuel production.

  8. Proteogenomic Analysis of a Thermophilic Bacterial Consortium Adapted to Deconstruct Switchgrass

    SciTech Connect

    D'haeseleer, Patrik; Gladden, John M.; Allgaier, Martin; Chain, Patrick; Tringe, Susannah G.; Malfatti, Stephanie; Aldrich, Joshua T.; Nicora, Carrie D.; Robinson, Errol W.; Pasa-Tolic, Ljiljana; Hugenholtz, Philip; Simmons, Blake A.; Singer, Steven W.

    2013-07-19

    Thermophilic bacteria are a potential source of enzymes for the deconstruction of lignocellulosic biomass. However, the complement of proteins used to deconstruct biomass and the specific roles of different microbial groups in thermophilic biomass deconstruction are not well-explored. Here we report on the metagenomic and proteogenomic analyses of a compost-derived bacterial consortium adapted to switchgrass at elevated temperature with high levels of glycoside hydrolase activities. Near-complete genomes were reconstructed for the most abundant populations, which included composite genomes for populations closely related to sequenced strains of Thermus thermophilus and Rhodothermus marinus, and for novel populations that are related to thermophilic Paenibacilli and an uncultivated subdivision of the littlestudied Gemmatimonadetes phylum. Partial genomes were also reconstructed for a number of lower abundance thermophilic Chloroflexi populations. Identification of genes for lignocellulose processing and metabolic reconstructions suggested Rhodothermus, Paenibacillus and Gemmatimonadetes as key groups for deconstructing biomass, and Thermus as a group that may primarily metabolize low molecular weight compounds. Mass spectrometry-based proteomic analysis of the consortium was used to identify .3000 proteins in fractionated samples from the cultures, and confirmed the importance of Paenibacillus and Gemmatimonadetes to biomass deconstruction. These studies also indicate that there are unexplored proteins with important roles in bacterial lignocellulose deconstruction.

  9. Polysaccharide-degrading thermophiles generated by heterologous gene expression in Geobacillus kaustophilus HTA426.

    PubMed

    Suzuki, Hirokazu; Yoshida, Ken-ichi; Ohshima, Toshihisa

    2013-09-01

    Thermophiles have important advantages over mesophiles as host organisms for high-temperature bioprocesses, functional production of thermostable enzymes, and efficient expression of enzymatic activities in vivo. To capitalize on these advantages of thermophiles, we describe here a new inducible gene expression system in the thermophile Geobacillus kaustophilus HTA426. Six promoter regions in the HTA426 genome were identified and analyzed for expression profiles using β-galactosidase reporter assay. This analysis identified a promoter region upstream of a putative amylose-metabolizing gene cluster that directed high-level expression of the reporter gene. The expression was >280-fold that without a promoter and was further enhanced 12-fold by maltose addition. In association with a multicopy plasmid, this promoter region was used to express heterologous genes. Several genes, including a gene whose product was insoluble when expressed in Escherichia coli, were successfully expressed as soluble proteins, with yields of 0.16 to 59 mg/liter, and conferred new functions to G. kaustophilus strains. Remarkably, cellulase and α-amylase genes conferred the ability to degrade cellulose paper and insoluble starch at high temperatures, respectively, generating thermophiles with the potential to degrade plant biomass. Our results demonstrate that this novel expression system expands the potential applications of G. kaustophilus. PMID:23793634

  10. Polysaccharide-Degrading Thermophiles Generated by Heterologous Gene Expression in Geobacillus kaustophilus HTA426

    PubMed Central

    Yoshida, Ken-ichi; Ohshima, Toshihisa

    2013-01-01

    Thermophiles have important advantages over mesophiles as host organisms for high-temperature bioprocesses, functional production of thermostable enzymes, and efficient expression of enzymatic activities in vivo. To capitalize on these advantages of thermophiles, we describe here a new inducible gene expression system in the thermophile Geobacillus kaustophilus HTA426. Six promoter regions in the HTA426 genome were identified and analyzed for expression profiles using β-galactosidase reporter assay. This analysis identified a promoter region upstream of a putative amylose-metabolizing gene cluster that directed high-level expression of the reporter gene. The expression was >280-fold that without a promoter and was further enhanced 12-fold by maltose addition. In association with a multicopy plasmid, this promoter region was used to express heterologous genes. Several genes, including a gene whose product was insoluble when expressed in Escherichia coli, were successfully expressed as soluble proteins, with yields of 0.16 to 59 mg/liter, and conferred new functions to G. kaustophilus strains. Remarkably, cellulase and α-amylase genes conferred the ability to degrade cellulose paper and insoluble starch at high temperatures, respectively, generating thermophiles with the potential to degrade plant biomass. Our results demonstrate that this novel expression system expands the potential applications of G. kaustophilus. PMID:23793634

  11. Complete Genome Sequence of the Cellulolytic Thermophile Caldicellulosiruptor obsidiansis OB47T▿

    PubMed Central

    Elkins, James G.; Lochner, Adriane; Hamilton-Brehm, Scott D.; Davenport, Karen Walston; Podar, Mircea; Brown, Steven D.; Land, Miriam L.; Hauser, Loren J.; Klingeman, Dawn M.; Raman, Babu; Goodwin, Lynne A.; Tapia, Roxanne; Meincke, Linda J.; Detter, J. Chris; Bruce, David C.; Han, Cliff S.; Palumbo, Anthony V.; Cottingham, Robert W.; Keller, Martin; Graham, David E.

    2010-01-01

    Caldicellulosiruptor obsidiansis OB47T (ATCC BAA-2073, JCM 16842) is an extremely thermophilic, anaerobic bacterium capable of hydrolyzing plant-derived polymers through the expression of multidomain/multifunctional hydrolases. The complete genome sequence reveals a diverse set of carbohydrate-active enzymes and provides further insight into lignocellulosic biomass hydrolysis at high temperatures. PMID:20851897

  12. Genome Sequence of Anoxybacillus geothermalis Strain GSsed3, a Novel Thermophilic Endospore-Forming Species.

    PubMed

    Filippidou, Sevasti; Jaussi, Marion; Junier, Thomas; Wunderlin, Tina; Roussel-Delif, Ludovic; Jeanneret, Nicole; Vieth-Hillebrand, Andrea; Vetter, Alexandra; Regenspurg, Simona; Johnson, Shannon L; McMurry, Kim; Gleasner, Cheryl D; Lo, Chien-Chi; Li, Paul; Vuyisich, Momchilo; Chain, Patrick S; Junier, Pilar

    2015-01-01

    Anoxybacillus geothermalis strain GSsed3 is an endospore-forming thermophilic bacterium isolated from filter deposits in a geothermal site. This novel species has a larger genome size (7.2 Mb) than that of any other Anoxybacillus species, and it possesses genes that support its phenotypic metabolic characterization and suggest an intriguing link to metals. PMID:26067952

  13. Effect of Temperature on the Respiration and Cytochromes of an Extreme Thermophile

    PubMed Central

    McFeters, Gordon A.; Ulrich, J. Terry

    1972-01-01

    The rate of oxygen uptake in an extreme thermophile at 70 C was three times greater than at 50 C. Cytochromes a, b, and c were present in cells grown at 50, 60, and 70 C. The content of these electron transport system elements remained relatively constant as the growth temperature was raised. PMID:5022175

  14. Thermophilic anaerobic digestion of Lurgi coal gasification wastewater in a UASB reactor.

    PubMed

    Wang, Wei; Ma, Wencheng; Han, Hongjun; Li, Huiqiang; Yuan, Min

    2011-02-01

    Lurgi coal gasification wastewater (LCGW) is a refractory wastewater, whose anaerobic treatment has been a severe problem due to its toxicity and poor biodegradability. Using a mesophilic (35±2°C) reactor as a control, thermophilic anaerobic digestion (55±2°C) of LCGW was investigated in a UASB reactor. After 120 days of operation, the removal of COD and total phenols by the thermophilic reactor could reach 50-55% and 50-60% respectively, at an organic loading rate of 2.5 kg COD/(m(3) d) and HRT of 24 h; the corresponding efficiencies were both only 20-30% in the mesophilic reactor. After thermophilic digestion, the wastewater concentrations of the aerobic effluent COD could reach below 200 mg/L compared with around 294 mg/L if mesophilic digestion was done and around 375 mg/L if sole aerobic pretreatment was done. The results suggested that thermophilic anaerobic digestion improved significantly both anaerobic and aerobic biodegradation of LCGW. PMID:21112778

  15. Discrimination among thermophilic Campylobacter species by polymerase chain reaction amplification of 23S rRNA gene fragments.

    PubMed Central

    Eyers, M; Chapelle, S; Van Camp, G; Goossens, H; De Wachter, R

    1993-01-01

    By comparing nucleic acid sequences determined for one of the most variable areas of 23S rRNA genes of 23 Campylobacter strains, we were able to identify regions specific for thermophilic Campylobacter strains. Oligonucleotide primers corresponding to these unique regions were synthesized and used in the polymerase chain reaction. One primer pair selectively detected all thermophilic Campylobacter species, while four other primer pairs allowed discrimination among the thermophilic species Campylobacter coli, Campylobacter jejuni subsp. jejuni, Campylobacter lari, and Campylobacter upsaliensis. All primer sets were tested successfully on a large number of clinical isolates. Images PMID:7508460

  16. Properties of an amylase from thermophilic Bacillus SP

    PubMed Central

    de Carvalho, Raquel Vieira; Côrrea, Thamy Lívia Ribeiro; da Silva, Júlia Caroline Matos; de Oliveira Mansur, Luciana Ribeiro Coutinho; Martins, Meire Lelis Leal

    2008-01-01

    α-Amylase production by thermophilic Bacillus sp strain SMIA-2 cultivated in liquid cultures containing soluble starch as a carbon source and supplemented with 0.05% whey protein and 0.2% peptone reached a maximum activity at 32 h, with levels of 37 U/mL. Studies on the amylase characterization revealed that the optimum temperature of this enzyme was 90°C. The enzyme was stable for 1 h at temperatures ranging from 40-50°C while at 90°C, 66% of its maximum activity was lost. However, in the presence of 5 mM CaCl2, the enzyme was stable at 90°C for 30 min and retained about 58% residual activity after 1 h. The optimum pH of the enzyme was found to be 8.5. After incubation of enzyme for 2 h at pH 9.5 and 11.0 was observed a decrease of about 6.3% and 16.5% of its original activity. At pH 6.0 the enzyme lost about 36% of its original activity. The enzyme was strongly inhibited by Co2+, Cu2+ and Ba2+, but less affected by Mg2+, Na+ and K+. In the presence of 2.0 M NaCl, 63% of amylase activity was retained after 2 h incubation at 45°C. The amylase exhibited more than 70% activity when incubated for 1 h at 50°C with sodium dodecyl sulphate. However, very little residual activity was obtained with sodium hypochlorite and with hydrogen peroxide the enzyme was completely inhibited. The compatibility of Bacillus sp SMIA-2 amylase with certain commercial detergents was shown to be good as the enzyme retained 86%, 85% and 75% of its activity after 20 min incubation at 50°C in the presence of the detergent brands Omo®, Campeiro® and Tide®, respectively. PMID:24031188

  17. Stability and activity of mesophilic subtilisin E and its thermophilic homolog: Insights from molecular dynamics simulations

    SciTech Connect

    Colombo, G.; Merz, K.M. Jr.

    1999-07-28

    This report examines the origin of the high-temperature (250 K) behavior of a thermophilic mutant enzyme (labeled at 5-3H5; see Zhao and Arnold Prot. Eng. 1999, 12, 47--53) derived from subtilisin E by eight amino acid substitutions. Through the use of molecular dynamics (MD) simulations, the authors have provided molecular-level insights into how point mutations can affect protein structure and dynamics. From simulations the authors observed a reduced rmsd in several key regions, an increased overall flexibility, an increase in the number of hydrogen bonds, and an increase in the number of stabilizing interactions in the thermophilic system. It was shown that it is not a necessary requirement that thermophilic enzymes be less flexible than their mesophilic counterparts at low temperatures. However, thermophilic enzymes must retain their three-dimensional structures and flexibility at high temperatures in order to retain activity. Furthermore, the authors have been able to point out the effects of some of the single substitutions. Even if it is not possible yet to give general rules for rational protein design, the authors are able to make some predictions on how a protein should be stabilized in order to be thermophilic. In particular, the authors suggest that a promising strategy toward speeding up the design of thermally stable proteins would be to identify fluxional regions within a protein through the use of MD simulations (or suitable experiments). Presumably these regions allow for autocatalytic reactions to occur and are also involved in allowing water to gain access to the interior of the protein and initiate protein unfolding. These fluxional regions could also adversely affect the positioning of the catalytic machinery, thereby decreasing catalytic efficiency. Thus, once these locations have been identified, focused directed evolution studies could be designed that stabilize these fluxional regions.

  18. Conventional mesophilic vs. thermophilic anaerobic digestion: a trade-off between performance and stability?

    PubMed

    Labatut, Rodrigo A; Angenent, Largus T; Scott, Norman R

    2014-04-15

    A long-term comparative study using continuously-stirred anaerobic digesters (CSADs) operated at mesophilic and thermophilic temperatures was conducted to evaluate the influence of the organic loading rate (OLR) and chemical composition on process performance and stability. Cow manure was co-digested with dog food, a model substrate to simulate a generic, multi-component food-like waste and to produce non-substrate specific, composition-based results. Cow manure and dog food were mixed at a lower - and an upper co-digestion ratio to produce a low-fiber, high-strength substrate, and a more recalcitrant, lower-strength substrate, respectively. Three increasing OLRs were evaluated by decreasing the CSADs hydraulic retention time (HRT) from 20 to 10 days. At longer HRTs and lower manure-to-dog food ratio, the thermophilic CSAD was not stable and eventually failed as a result of long-chain fatty acid (LCFA) accumulation/degradation, which was triggered by the compounded effects of temperature on reaction rates, mixing intensity, and physical state of LCFAs. At shorter HRTs and upper manure-to-dog food ratio, the thermophilic CSAD marginally outperformed the biomethane production rates and substrate stabilization of the mesophilic CSAD. The increased fiber content relative to lipids at upper manure-to-dog food ratios improved the stability and performance of the thermophilic process by decreasing the concentration of LCFAs in solution, likely adsorbed onto the manure fibers. Overall, results of this study show that stability of the thermophilic co-digestion process is highly dependent on the influent substrate composition, and particularly for this study, on the proportion of manure to lipids in the influent stream. In contrast, mesophilic co-digestion provided a more robust and stable process regardless of the influent composition, only with marginally lower biomethane production rates (i.e., 7%) for HRTs as short as 10 days (OLR = 3 g VS/L-d). PMID:24530545

  19. Novel thermophilic hemicellulases for the conversion of lignocellulose for second generation biorefineries.

    PubMed

    Cobucci-Ponzano, Beatrice; Strazzulli, Andrea; Iacono, Roberta; Masturzo, Giuseppe; Giglio, Rosa; Rossi, Mosè; Moracci, Marco

    2015-10-01

    The biotransformation of lignocellulose biomasses into fermentable sugars is a very complex procedure including, as one of the most critical steps, the (hemi) cellulose hydrolysis by specific enzymatic cocktails. We explored here, the potential of stable glycoside hydrolases from thermophilic organisms, so far not used in commercial enzymatic preparations, for the conversion of glucuronoxylan, the major hemicellulose of several energy crops. Searches in the genomes of thermophilic bacteria led to the identification, efficient production, and detailed characterization of novel xylanase and α-glucuronidase from Alicyclobacillus acidocaldarius (GH10-XA and GH67-GA, respectively) and a α-glucuronidase from Caldicellulosiruptor saccharolyticus (GH67-GC). Remarkably, GH10-XA, if compared to other thermophilic xylanases from this family, coupled good specificity on beechwood xylan and the best stability at 65 °C (3.5 days). In addition, GH67-GC was the most stable α-glucuronidases from this family and the first able to hydrolyse both aldouronic acid and aryl-α-glucuronic acid substrates. These enzymes, led to the very efficient hydrolysis of beechwood xylan by using 7- to 9-fold less protein (concentrations <0.3 μM) and in much less reaction time (2h vs 12h) if compared to other known biotransformations catalyzed by thermophilic enzymes. In addition, remarkably, together with a thermophilic β-xylosidase, they catalyzed the production of xylose from the smart cooking pre-treated biomass of one of the most promising energy crops for second generation biorefineries. We demonstrated that search by the CAZy Data Bank of currently available genomes and detailed enzymatic characterization of recombinant enzymes allow the identification of glycoside hydrolases with novel and interesting properties and applications. PMID:26215346

  20. Thermophilic Sulfate Reduction in Hydrothermal Sediment of Lake Tanganyika, East Africa

    PubMed Central

    Elsgaard, Lars; Prieur, Daniel; Mukwaya, Gashagaza M.; Jørgensen, Bo B.

    1994-01-01

    In environments with temperatures above 60°C, thermophilic prokaryotes are the only metabolically active life-forms. By using the 35SO42- tracer technique, we studied the activity of sulfate-reducing microorganisms (SRM) in hot sediment from a hydrothermal vent site in the northern part of freshwater Lake Tanganyika (East Africa). Incubation of slurry samples at 8 to 90°C demonstrated meso- and thermophilic sulfate reduction with optimum temperatures of 34 to 45°C and 56 to 65°C, respectively, and with an upper temperature limit of 80°C. Sulfate reduction was stimulated at all temperatures by the addition of short-chain fatty acids and benzoate or complex substrates (yeast extract and peptone). A time course experiment showed that linear thermophilic sulfate consumption occurred after a lag phase (12 h) and indicated the presence of a large population of SRM in the hydrothermal sediment. Thermophilic sulfate reduction had a pH optimum of about 7 and was completely inhibited at pH 8.8 to 9.2. SRM could be enriched from hydrothermal chimney and sediment samples at 60 and 75°C. In lactate-grown enrichments, sulfide production occurred at up to 70 and 75°C, with optima at 63 and 71°C, respectively. Several sporulating thermophilic enrichments were morphologically similar to Desulfotomaculum spp. Dissimilatory sulfate reduction in the studied hydrothermal area of Lake Tanganyika apparently has an upper temperature limit of 80°C. PMID:16349249

  1. Defluviitalea phaphyphila sp. nov., a Novel Thermophilic Bacterium That Degrades Brown Algae

    PubMed Central

    Ji, Shi-Qi; Wang, Bing; Lu, Ming

    2015-01-01

    Brown algae are one of the largest groups of oceanic primary producers for CO2 removal and carbon sinks for coastal regions. However, the mechanism for brown alga assimilation remains largely unknown in thermophilic microorganisms. In this work, a thermophilic alginolytic community was enriched from coastal sediment, from which an obligate anaerobic and thermophilic bacterial strain, designated Alg1, was isolated. Alg1 shared a 16S rRNA gene identity of 94.6% with Defluviitalea saccharophila LIND6LT2T. Phenotypic, chemotaxonomic, and phylogenetic studies suggested strain Alg1 represented a novel species of the genus Defluviitalea, for which the name Defluviitalea phaphyphila sp. nov. is proposed. Alg1 exhibited an intriguing ability to convert carbohydrates of brown algae, including alginate, laminarin, and mannitol, to ethanol and acetic acid. Three gene clusters participating in this process were predicted to be in the genome, and candidate enzymes were successfully expressed, purified, and characterized. Six alginate lyases were demonstrated to synergistically deconstruct alginate into unsaturated monosaccharide, followed by one uronic acid reductase and two 2-keto-3-deoxy-d-gluconate (KDG) kinases to produce pyruvate. A nonclassical mannitol 1-phosphate dehydrogenase, catalyzing d-mannitol 1-phosphate to fructose 1-phosphate in the presence of NAD+, and one laminarase also were disclosed. This work revealed that a thermophilic brown alga-decomposing system containing numerous novel thermophilic alginate lyases and a unique mannitol 1-phosphate dehydrogenase was adopted by the natural ethanologenic strain Alg1 during the process of evolution in hostile habitats. PMID:26590273

  2. Defluviitalea phaphyphila sp. nov., a Novel Thermophilic Bacterium That Degrades Brown Algae.

    PubMed

    Ji, Shi-Qi; Wang, Bing; Lu, Ming; Li, Fu-Li

    2016-02-01

    Brown algae are one of the largest groups of oceanic primary producers for CO2 removal and carbon sinks for coastal regions. However, the mechanism for brown alga assimilation remains largely unknown in thermophilic microorganisms. In this work, a thermophilic alginolytic community was enriched from coastal sediment, from which an obligate anaerobic and thermophilic bacterial strain, designated Alg1, was isolated. Alg1 shared a 16S rRNA gene identity of 94.6% with Defluviitalea saccharophila LIND6LT2(T). Phenotypic, chemotaxonomic, and phylogenetic studies suggested strain Alg1 represented a novel species of the genus Defluviitalea, for which the name Defluviitalea phaphyphila sp. nov. is proposed. Alg1 exhibited an intriguing ability to convert carbohydrates of brown algae, including alginate, laminarin, and mannitol, to ethanol and acetic acid. Three gene clusters participating in this process were predicted to be in the genome, and candidate enzymes were successfully expressed, purified, and characterized. Six alginate lyases were demonstrated to synergistically deconstruct alginate into unsaturated monosaccharide, followed by one uronic acid reductase and two 2-keto-3-deoxy-d-gluconate (KDG) kinases to produce pyruvate. A nonclassical mannitol 1-phosphate dehydrogenase, catalyzing D-mannitol 1-phosphate to fructose 1-phosphate in the presence of NAD(+), and one laminarase also were disclosed. This work revealed that a thermophilic brown alga-decomposing system containing numerous novel thermophilic alginate lyases and a unique mannitol 1-phosphate dehydrogenase was adopted by the natural ethanologenic strain Alg1 during the process of evolution in hostile habitats. PMID:26590273

  3. Single-step ethanol production from lignocellulose using novel extremely thermophilic bacteria

    PubMed Central

    2013-01-01

    Background Consolidated bioprocessing (CBP) of lignocellulosic biomass to ethanol using thermophilic bacteria provides a promising solution for efficient lignocellulose conversion without the need for additional cellulolytic enzymes. Most studies on the thermophilic CBP concentrate on co-cultivation of the thermophilic cellulolytic bacterium Clostridium thermocellum with non-cellulolytic thermophilic anaerobes at temperatures of 55°C-60°C. Results We have specifically screened for cellulolytic bacteria growing at temperatures >70°C to enable direct conversion of lignocellulosic materials into ethanol. Seven new strains of extremely thermophilic anaerobic cellulolytic bacteria of the genus Caldicellulosiruptor and eight new strains of extremely thermophilic xylanolytic/saccharolytic bacteria of the genus Thermoanaerobacter isolated from environmental samples exhibited fast growth at 72°C, extensive lignocellulose degradation and high yield ethanol production on cellulose and pretreated lignocellulosic biomass. Monocultures of Caldicellulosiruptor strains degraded up to 89-97% of the cellulose and hemicellulose polymers in pretreated biomass and produced up to 72 mM ethanol on cellulose without addition of exogenous enzymes. In dual co-cultures of Caldicellulosiruptor strains with Thermoanaerobacter strains the ethanol concentrations rose 2- to 8.2-fold compared to cellulolytic monocultures. A co-culture of Caldicellulosiruptor DIB 087C and Thermoanaerobacter DIB 097X was particularly effective in the conversion of cellulose to ethanol, ethanol comprising 34.8 mol% of the total organic products. In contrast, a co-culture of Caldicellulosiruptor saccharolyticus DSM 8903 and Thermoanaerobacter mathranii subsp. mathranii DSM 11426 produced only low amounts of ethanol. Conclusions The newly discovered Caldicellulosiruptor sp. strain DIB 004C was capable of producing unexpectedly large amounts of ethanol from lignocellulose in fermentors. The established co

  4. Reactor performance and microbial community dynamics during solid-state anaerobic digestion of corn stover at mesophilic and thermophilic conditions.

    PubMed

    Shi, Jian; Wang, Zhongjiang; Stiverson, Jill A; Yu, Zhongtang; Li, Yebo

    2013-05-01

    Reactor performance and microbial community dynamics were investigated during solid state anaerobic digestion (SS-AD) of corn stover at mesophilic and thermophilic conditions. Thermophilic SS-AD led to faster and greater reductions of cellulose and hemicelluloses during the first 12 days compared to mesophilic SS-AD. However, accumulation of volatile fatty acids (VFAs) was 5-fold higher at thermophilic than mesophilic temperatures, resulting in a large pH drop during days 6-12 in the thermophilic reactors. Culture-based enumeration revealed 10-50 times greater populations of cellulolytic and xylanolytic microbes during thermophilic SS-AD than mesophilic SS-AD. DGGE analysis of PCR amplified 16S rRNA genes showed dynamic shifts, especially during the thermophilic SS-AD, of bacterial and archaeal communities over the 38 days of SS-AD as a result of acclimation of the initial seed microbial consortia to the lignocellulosic feedstock. The findings of this study can guide future studies to improve efficiency and stability of SS-AD. PMID:23567733

  5. Decontamination of heavy metal laden sewage sludge with simultaneous solids reduction using thermophilic sulfur and ferrous oxidizing species.

    PubMed

    Mehrotra, A; Kundu, K; Sreekrishnan, T R

    2016-02-01

    A possibility of using simultaneous sewage sludge digestion and metal leaching (SSDML) process at the thermophilic temperature to remove heavy metals and suspended solids from sewage sludge is explored in this study. Though thermophilic sludge digestion efficiently produces a stable sludge, its inability to remove heavy metals requires it to be used in tandem with another process like bioleaching for metal reduction. Previously, different temperature optima were known for the heterotrophs (thermophilic) responsible for the sludge digestion and the autotrophs involved in bioleaching (mesophilic), because of which the metal concentration was brought down separately in a different reactor. In our study, SSDML process was carried out at 50 °C (thermophilic) by using ferrous sulfate (batch-1) and sulfur (batch-2) as the energy source in two reactors. The concentration of volatile suspended solids reduced by >40% in both batches, while that of heavy metals zinc, copper, chromium, cadmium and nickel decreased by >50% in both batch-1 and batch-2. Lead got leached out only in batch-1. Using 16S rRNA gene-based PCR-denaturing gradient gel electrophoresis analysis, Alicyclobacillus tolerans was found to be the microorganism responsible for lowering the pH in both the reactors at thermophilic temperature. The indicator organism count was also below the maximum permissible limit making sludge suitable for agricultural use. Our results indicate that SSDML at thermophilic temperature can be effectively used for reduction of heavy metals and suspended solids from sewage sludge. PMID:26686075

  6. Alicyclobacillus dauci sp. nov., a slightly thermophilic, acidophilic bacterium isolated from a spoiled mixed vegetable and fruit juice product.

    PubMed

    Nakano, Chisa; Takahashi, Naoto; Tanaka, Naoto; Okada, Sanae

    2015-02-01

    A novel, moderately thermophilic, acidophilic, Gram-variable, rod-shaped, endospore-forming bacterium was isolated from a spoiled mixed vegetable and fruit juice product that had the off-flavour of guaiacol. The bacterium, strain 4F(T), grew aerobically at 20-50 °C (optimum 40 °C) and pH 3.0-6.0 (optimum pH 4.0) and produced acid from glycerol, d-galactose and d-glucose. It contained menaquinone-7 (MK-7) as the major isoprenoid quinone and the DNA G+C content was 49.6 mol%. The predominant cellular fatty acids of strain 4F(T) were ω-alicyclic (ω-cyclohexane fatty acids), which are characteristic of the genus Alicyclobacillus. Phylogenetic analyses based on 16S rRNA gene sequences showed that the strain belongs to the Alicyclobacillus cluster, and is related most closely to the type strains of Alicyclobacillus acidoterrestris (97.4 % similarity) and Alicyclobacillus fastidiosus (97.3 %). Strain 4F(T) produced guaiacol from vanillic acid. It can be distinguished from related species by its acid production type and guaiacol production. On the basis of phenotypic characteristics, phylogenetic analysis and DNA-DNA relatedness values, it can be concluded that the strain represents a novel species of the genus Alicyclobacillus, for which the name Alicyclobacillus dauci sp. nov. is proposed; the type strain is 4F(T) ( = DSM 28700(T) = NBRC 108949(T) = NRIC 0938(T)). PMID:25505343

  7. [Conversion of acetic acid to methane by thermophiles]. Progress report, May 15, 1989--May 14, 1993

    SciTech Connect

    Zinder, S.H.

    1993-06-01

    The primary goal of this project is to obtain a better understanding of thermophilic microorganisms which convert acetic acid to CH{sub 4}. The previous funding period represents a departure from earlier research in this laboratory, which was more physiological and ecological. The present work is centered on the biochemistry of the thermophile Methanothrix sp. strain CALS-1. this organism presents a unique opportunity, with its purity and relatively rapid growth, to do comparative biochemical studies with the other major acetotrophic genus Methanosarcina. We previously found that Methanothrix is capable of using acetate at concentrations 100 fold lower than Methanosarcina. This finding suggests that there are significant differences in the pathways of methanogenesis from acetate in the two genera.

  8. Distribution and Diversity of Symbiotic Thermophiles, Symbiobacterium thermophilum and Related Bacteria, in Natural Environments

    PubMed Central

    Ueda, Kenji; Ohno, Michiyo; Yamamoto, Kaori; Nara, Hanae; Mori, Yujiro; Shimada, Masafumi; Hayashi, Masahiko; Oida, Hanako; Terashima, Yuko; Nagata, Mitsuyo; Beppu, Teruhiko

    2001-01-01

    Symbiobacterium thermophilum is a tryptophanase-positive thermophile which shows normal growth only in coculture with its supporting bacteria. Analysis of the 16S rRNA gene (rDNA) indicated that the bacterium belongs to a novel phylogenetic branch at the outermost position of the gram-positive bacterial group without clustering to any other known genus. Here we describe the distribution and diversity of S. thermophilum and related bacteria in the environment. Thermostable tryptophanase activity and amplification of the specific 16S rDNA fragment were effectively employed to detect the presence of Symbiobacterium. Enrichment with kanamycin raised detection sensitivity. Mixed cultures of thermophiles containing Symbiobacterium species were frequently obtained from compost, soil, animal feces, and contents in the intestinal tracts, as well as feeds. Phylogenetic analysis and denaturing gradient gel electrophoresis of the specific 16S rDNA amplicons revealed a diversity of this group of bacteria in the environment. PMID:11525967

  9. [Effect of substrate concentration on pathogen indicators inactivation during thermophilic anaerobic digestion].

    PubMed

    Cao, Hong-Qing; Zhang, Fei-Fei; Li, Jian; Tong, Zi-Lin; Hu, Zhen-Hu

    2014-03-01

    Because excess sludge contains high density of pathogens, it has to be treated to reduce pathogens before being disposed for land application. In this study, the effect of substrate concentration on pathogen inactivation during thermophilic anaerobic digestion was investigated. The results show that, with the increase of substrate concentration, VFAs and cumulative methane production increased. The density of total coliforms in the suspension liquid has a 2.0-3.0 orders of magnitude decline and fecal coliforms has 1.8-3.3 orders of magnitude decline after 28 days thermophilic anaerobic digestion at substrate concentration of 28-84 g x L(-1) and temperature of 55 degrees C. More than 99% of total coliforms and fecal coliforms have been killed after 28 days digestion. Salmonella spp. was not detected in the suspension and solid after anaerobic digestion. When substrate concentration was higher than 45 g x L(-1), the inactivation of total coliforms and fecal coliforms declined. PMID:24881391

  10. Thermophilic anaerobic co-digestion of poultry litter and thin stillage.

    PubMed

    Sharma, Deepak; Espinosa-Solares, Teodoro; Huber, David H

    2013-05-01

    The purpose of this study was to test whether the performance of a thermophilic CSTR digester that has been stabilized on poultry litter will be enhanced or diminished by the addition of thin stillage as co-substrate. Replicate laboratory digesters, derived from a stable pilot-scale digester, were operated with increasing ratios (w/w) of thin stillage/poultry litter feedstock. After a period of adaptation to 20% and 40% thin stillage, digester performance showed increases in biogas, percent methane and COD removal, as well as a decrease in volatile acids. Peak performance occurred with 60% thin stillage. However, 80% thin stillage caused significant reduction of performance, including declines of methanogenic activity and COD removal. In conclusion, supplementing the thermophilic digestion of poultry litter with thin stillage improved the bioenergy (methane) output, but thin stillage became inhibitory at high concentrations. PMID:23567688

  11. Sulfur oxidation activities of pure and mixed thermophiles and sulfur speciation in bioleaching of chalcopyrite.

    PubMed

    Zhu, Wei; Xia, Jin-Lan; Yang, Yi; Nie, Zhen-yuan; Zheng, Lei; Ma, Chen-yan; Zhang, Rui-yong; Peng, An-an; Tang, Lu; Qiu, Guan-zhou

    2011-02-01

    The sulfur oxidation activities of four pure thermophilic archaea Acidianus brierleyi (JCM 8954), Metallosphaera sedula (YN 23), Acidianus manzaensis (YN 25) and Sulfolobus metallicus (YN 24) and their mixture in bioleaching chalcopyrite were compared. Meanwhile, the relevant surface sulfur speciation of chalcopyrite leached with the mixed thermophilic archaea was investigated. The results showed that the mixed culture, with contributing significantly to the raising of leaching rate and accelerating the formation of leaching products, may have a higher sulfur oxidation activity than the pure cultures, and jarosite was the main passivation component hindering the dissolution of chalcopyrite, while elemental sulfur seemed to have no influence on the dissolution of chalcopyrite. In addition, the present results supported the former speculation, i.e., covellite might be converted from chalcocite during the leaching experiments, and the elemental sulfur may partially be the derivation of covellite and chalcocite. PMID:21194927

  12. Biodegradation of crude oil by thermophilic bacteria isolated from a volcano island.

    PubMed

    Meintanis, Christos; Chalkou, Kalliopi I; Kormas, Konstantinos Ar; Karagouni, Amalia D

    2006-03-01

    One-hundred and fifty different thermophilic bacteria isolated from a volcanic island were screened for detection of an alkane hydroxylase gene using degenerated primers developed to amplify genes related to the Pseudomonas putida and Pseudomonas oleovorans alkane hydroxylases. Ten isolates carrying the alkJ gene were further characterized by 16s rDNA gene sequencing. Nine out of ten isolates were phylogenetically affiliated with Geobacillus species and one isolate with Bacillus species. These isolates were able to grow in liquid cultures with crude oil as the sole carbon source and were found to degrade long chain crude oil alkanes in a range between 46.64% and 87.68%. Results indicated that indigenous thermophilic hydrocarbon degraders of Bacillus and Geobacillus species are of special significance as they could be efficiently used for bioremediation of oil-polluted soil and composting processes. PMID:16456612

  13. Why use a thermophilic aerobic membrane reactor for the treatment of industrial wastewater/liquid waste?

    PubMed

    Collivignarelli, Maria Cristina; Abbà, Alessandro; Bertanza, Giorgio

    2015-01-01

    This paper describes the advantages of thermophilic aerobic membrane reactor (TAMR) for the treatment of high strength wastewaters. The results were obtained from the monitoring of an industrial and a pilot scale plant. The average chemical oxygen demand (COD) removal yield was equal to 78% with an organic loading rate (OLR) up to 8-10 kgCOD m(-3) d(-1) despite significant scattering of the influent wastewater composition. Total phosphorus (TP) was removed with a rate of 90%, the most important removal mechanism being chemical precipitation (as hydroxyapatite, especially), which is improved by the continuous aeration that promotes phosphorus crystallization. Moreover, surfactants were removed with efficiency between 93% and 97%. Finally, the experimental work showed that thermophilic processes (TPPs) are complementary with respect to mesophilic treatments. PMID:25704477

  14. Redirection of the Reaction Specificity of a Thermophilic Acetolactate Synthase toward Acetaldehyde Formation

    PubMed Central

    Cheng, Maria; Yoshiyasu, Hayato; Okano, Kenji; Ohtake, Hisao; Honda, Kohsuke

    2016-01-01

    Acetolactate synthase and pyruvate decarboxylase are thiamine pyrophosphate-dependent enzymes that convert pyruvate into acetolactate and acetaldehyde, respectively. Although the former are encoded in the genomes of many thermophiles and hyperthermophiles, the latter has been found only in mesophilic organisms. In this study, the reaction specificity of acetolactate synthase from Thermus thermophilus was redirected to catalyze acetaldehyde formation to develop a thermophilic pyruvate decarboxylase. Error-prone PCR and mutant library screening led to the identification of a quadruple mutant with 3.1-fold higher acetaldehyde-forming activity than the wild-type. Site-directed mutagenesis experiments revealed that the increased activity of the mutant was due to H474R amino acid substitution, which likely generated two new hydrogen bonds near the thiamine pyrophosphate-binding site. These hydrogen bonds might result in the better accessibility of H+ to the substrate-cofactor-enzyme intermediate and a shift in the reaction specificity of the enzyme. PMID:26731734

  15. Effects of total solids content on waste activated sludge thermophilic anaerobic digestion and its sludge dewaterability.

    PubMed

    Wang, Tianfeng; Chen, Jie; Shen, Honglang; An, Dong

    2016-10-01

    The role of total solids content on sludge thermophilic anaerobic digestion was investigated in batch reactors. A range of total solids content from 2% to 10% was evaluated with two replicates. The lowest inhibitory concentration for free ammonia and total ammonia of sludge thermophilic anaerobic digestion was 110.9-171.4mg/L and 1313.1-1806.7mg/L, respectively. The volumetric biogas production rate increased with increasing of total solids content, but the corresponding biogas yield per gram volatile solid decreased. The result of normalized capillary suction time indicated that the dewaterability of digested sludge at high total solids content was poor, while solid content of sediment obtained by centrifuging sludge at 2000g for 10min increased with increasing of total solids content of sludge. The results suggest that thickened sludge mixed with dewatered sludge at an appropriate ratio could get high organic loading rate, high biogas yield and adequate dewatering effort. PMID:26897469

  16. Biocatalytic synthesis of poly(δ-valerolactone) using a thermophilic esterase from archaeoglobus fulgidus as catalyst.

    PubMed

    Cao, Hong; Han, Haobo; Li, Guangquan; Yang, Jiebing; Zhang, Lingfei; Yang, Yan; Fang, Xuedong; Li, Quanshun

    2012-01-01

    The ring-opening polymerization of δ-valerolactone catalyzed by a thermophilic esterase from the archaeon Archaeoglobus fulgidus was successfully conducted in organic solvents. The effects of enzyme concentration, temperature, reaction time and reaction medium on monomer conversion and product molecular weight were systematically evaluated. Through the optimization of reaction conditions, poly(δ-valerolactone) was produced in 97% monomer conversion, with a number-average molecular weight of 2225 g/mol, in toluene at 70 °C for 72 h. This paper has produced a new biocatalyst for the synthesis of poly(δ-valerolactone), and also deeper insight has been gained into the mechanism of thermophilic esterase-catalyzed ring-opening polymerization. PMID:23202895

  17. Genome sequence of a native-feather degrading extremely thermophilic Eubacterium, Fervidobacterium islandicum AW-1.

    PubMed

    Lee, Yong-Jik; Jeong, Haeyoung; Park, Gun-Seok; Kwak, Yunyoung; Lee, Sang-Jae; Lee, Sang Jun; Park, Min-Kyu; Kim, Ji-Yeon; Kang, Hwan Ku; Shin, Jae-Ho; Lee, Dong-Woo

    2015-01-01

    Fervidobacterium islandicum AW-1 (KCTC 4680) is an extremely thermophilic anaerobe isolated from a hot spring in Indonesia. This bacterium could degrade native chicken feathers completely at 70 °C within 48 h, which is of potential importance on the basis of relevant environmental and agricultural issues in bioremediation and development of eco-friendly bioprocesses for the treatment of native feathers. However, its genomic and phylogenetic analysis remains unclear. Here, we report the high-quality draft genome sequence of an extremely thermophilic anaerobe, F. islandicum AW-1. The genome consists of 2,359,755 bp, which encodes 2,184 protein-coding genes and 64 RNA-encoding genes. This may reveal insights into anaerobic metabolism for keratin degradation and also provide a biological option for poultry waste treatments. PMID:26421103

  18. Enumeration of Thermophilic Heterotrophs in Geothermally Heated Soils from Mount Erebus, Ross Island, Antarctica

    PubMed Central

    Hudson, J. Andrew; Daniel, Roy M.

    1988-01-01

    Soil samples with temperatures up to 64°C were collected from Mount Erebus, an active volcano located on Ross Island, Antarctica. Acridine orange direct counts and most probable number counts of soil samples stored at 4°C for 2 months showed a wide variation in the number of thermophilic microorganisms in different soils. Organisms similar to Clostridium thermohydrosulfuricum, Bacillus schlegelii, and Bacillus acidocaldarius, as well as neutrophilic Bacillus strains, were isolated. PMID:16347573

  19. Enumeration of thermophilic heterotrophs in geothermally heated soils from mount erebus, ross island, antarctica.

    PubMed

    Hudson, J A; Daniel, R M

    1988-02-01

    Soil samples with temperatures up to 64 degrees C were collected from Mount Erebus, an active volcano located on Ross Island, Antarctica. Acridine orange direct counts and most probable number counts of soil samples stored at 4 degrees C for 2 months showed a wide variation in the number of thermophilic microorganisms in different soils. Organisms similar to Clostridium thermohydrosulfuricum, Bacillus schlegelii, and Bacillus acidocaldarius, as well as neutrophilic Bacillus strains, were isolated. PMID:16347573

  20. The F- or V-type Na(+)-ATPase of the thermophilic bacterium Clostridium fervidus.

    PubMed Central

    Speelmans, G; Poolman, B; Abee, T; Konings, W N

    1994-01-01

    Clostridium fervidus is a thermophilic, anaerobic bacterium which uses solely Na+ as a coupling ion for energy transduction. Important features of the primary Na+ pump (ATPase) that generates the sodium motive force are presented. The advantage of using a sodium rather than a proton motive force at high temperatures becomes apparent from the effect of temperature on H+ and Na+ permeation in liposomes. PMID:8051034

  1. Comparing mesophilic and thermophilic anaerobic digestion of chicken manure: Microbial community dynamics and process resilience

    SciTech Connect

    Niu, Qigui; Takemura, Yasuyuki; Kubota, Kengo; Li, Yu-You

    2015-09-15

    Highlights: • Microbial community dynamics and process functional resilience were investigated. • The threshold of TAN in mesophilic reactor was higher than the thermophilic reactor. • The recoverable archaeal community dynamic sustained the process resilience. • Methanosarcina was more sensitive than Methanoculleus on ammonia inhibition. • TAN and FA effects the dynamic of hydrolytic and acidogenic bacteria obviously. - Abstract: While methane fermentation is considered as the most successful bioenergy treatment for chicken manure, the relationship between operational performance and the dynamic transition of archaeal and bacterial communities remains poorly understood. Two continuous stirred-tank reactors were investigated under thermophilic and mesophilic conditions feeding with 10%TS. The tolerance of thermophilic reactor on total ammonia nitrogen (TAN) was found to be 8000 mg/L with free ammonia (FA) 2000 mg/L compared to 16,000 mg/L (FA1500 mg/L) of mesophilic reactor. Biomethane production was 0.29 L/gV S{sub in} in the steady stage and decreased following TAN increase. After serious inhibition, the mesophilic reactor was recovered successfully by dilution and washing stratagem compared to the unrecoverable of thermophilic reactor. The relationship between the microbial community structure, the bioreactor performance and inhibitors such as TAN, FA, and volatile fatty acid was evaluated by canonical correspondence analysis. The performance of methanogenic activity and substrate removal efficiency were changed significantly correlating with the community evenness and phylogenetic structure. The resilient archaeal community was found even after serious inhibition in both reactors. Obvious dynamics of bacterial communities were observed in acidogenic and hydrolytic functional bacteria following TAN variation in the different stages.

  2. Complete Genome Sequence of the Thermophilic, Piezophilic, Heterotrophic Bacterium Marinitoga piezophila KA3

    SciTech Connect

    Lucas, Susan; Han, James; Lapidus, Alla L.; Cheng, Jan-Fang; Goodwin, Lynne A.; Pitluck, Sam; Peters, Lin; Mikhailova, Natalia; Teshima, Hazuki; Detter, J. Chris; Han, Cliff; Tapia, Roxanne; Land, Miriam L; Hauser, Loren John; Kyrpides, Nikos C; Ivanova, N; Pagani, Ioanna; Vannier, Pauline; Oger, Phil; Bartlett, Douglas; Noll, Kenneth M; Woyke, Tanja; Jebbar, Mohamed

    2012-01-01

    Marinitoga piezophila KA3 is a thermophilic, anaerobic, chemoorganotrophic, sulfur-reducing bacterium isolated from the Grandbonum deep-sea hydrothermal vent site at the East Pacific Rise (13 degrees N, 2,630-m depth). The genome of M. piezophila KA3 comprises a 2,231,407-bp circular chromosome and a 13,386-bp circular plasmid. This genome was sequenced within Department of Energy Joint Genome Institute CSP 2010.

  3. Hydrogen Production by the Thermophilic Alga Mastigocladus laminosus: Effects of Nitrogen, Temperature, and Inhibition of Photosynthesis

    PubMed Central

    Miyamoto, Kazuhisa; Hallenbeck, Patrick C.; Benemann, John R.

    1979-01-01

    Hydrogen production by nitrogen-limited cultures of a thermophilic blue-green alga (cyanobacterium), Mastigocladus laminosus, was studied to develop the concept of a high-temperature biophotolysis system. Biophotolytic production of hydrogen by solar radiation was also demonstrated. Hydrogen consumption activity in these cultures was relatively high and is the present limiting factor on both the net rate and duration of hydrogen production. PMID:16345432

  4. Behavior of cellulose-degrading bacteria in thermophilic anaerobic digestion process.

    PubMed

    Syutsubo, K; Nagaya, Y; Sakai, S; Miya, A

    2005-01-01

    Previously, we found that the newly isolated Clostridium sp. strain JC3 became the dominant cellulose-degrading bacterium in thermophilic methanogenic sludge. In the present study, the behavior of strain JC3 in the thermophilic anaerobic digestion process was investigated quantitatively by molecular biological techniques. A cellulose-degrading experiment was conducted at 55 degrees C with a 9.5 L of anaerobic baffled reactor having three compartments (Nos. 1, 2, 3). Over 80% of the COD input was converted into methane when 2.5 kgCOD m(-3) d(-1) was loaded for an HRT of 27 days. A FISH probe specific for strain JC3 was applied to sludge samples harvested from the baffled reactor. Consequently, the ratio of JC3 cells to DAPI-stained cells increased from below 0.5% (undetectable) to 9.4% (compartment 1), 13.1% (compartment 2) and 21.6% (compartment 3) at day 84 (2.5 kgCOD m(-3)d(-1)). The strain JC3 cell numbers determined by FISH correlated closely with the cellulose-degrading methanogenic activities of retained sludge. A specific primer set targeting the cellulase gene (cellobiohydrolaseA: cbhA) of strain JC3 was designed and applied to digested sludge for treating solid waste such as coffee grounds, wastepaper, garbage, cellulose and so on. The strain JC3 cell numbers determined by quantitative PCR correlated closely with the cellulose-sludge loading of the thermophilic digester. Strain JC3 is thus important in the anaerobic hydrolysis of cellulose in thermophilic anaerobic digestion processes. PMID:16180412

  5. Catalytic Biomineralization of Fluorescent Calcite by the Thermophilic Bacterium Geobacillus thermoglucosidasius▿

    PubMed Central

    Yoshida, Naoto; Higashimura, Eiji; Saeki, Yuichi

    2010-01-01

    The thermophilic Geobacillus bacterium catalyzed the formation of 100-μm hexagonal crystals at 60°C in a hydrogel containing sodium acetate, calcium chloride, and magnesium sulfate. Under fluorescence microscopy, crystals fluoresced upon excitation at 365 ± 5, 480 ± 20, or 545 ± 15 nm. X-ray diffraction indicated that the crystals were magnesium-calcite in calcite-type calcium carbonate. PMID:20851984

  6. PCR detection of thermophilic spore-forming bacteria involved in canned food spoilage.

    PubMed

    Prevost, S; Andre, S; Remize, F

    2010-12-01

    Thermophilic bacteria that form highly heat-resistant spores constitute an important group of spoilage bacteria of low-acid canned food. A PCR assay was developed in order to rapidly trace these bacteria. Three PCR primer pairs were designed from rRNA gene sequences. These primers were evaluated for the specificity and the sensitivity of detection. Two primer pairs allowed detection at the species level of Geobacillus stearothermophilus and Moorella thermoacetica/thermoautrophica. The other pair allowed group-specific detection of anaerobic thermophilic bacteria of the genera Thermoanaerobacterium, Thermoanaerobacter, Caldanerobium and Caldanaerobacter. After a single enrichment step, these PCR assays allowed the detection of 28 thermophiles from 34 cans of spoiled low-acid food. In addition, 13 ingredients were screened for the presence of these bacteria. This PCR assay serves as a detection method for strains able to spoil low-acid canned food treated at 55°C. It will lead to better reactivity in the canning industry. Raw materials and ingredients might be qualified not only for quantitative spore contamination, but also for qualitative contamination by highly heat-resistant spores. PMID:20397018

  7. Thermophilic bacteria in Moroccan hot springs, salt marshes and desert soils

    PubMed Central

    Aanniz, Tarik; Ouadghiri, Mouna; Melloul, Marouane; Swings, Jean; Elfahime, Elmostafa; Ibijbijen, Jamal; Ismaili, Mohamed; Amar, Mohamed

    2015-01-01

    The diversity of thermophilic bacteria was investigated in four hot springs, three salt marshes and 12 desert sites in Morocco. Two hundred and forty (240) thermophilic bacteria were recovered, identified and characterized. All isolates were Gram positive, rod-shaped, spore forming and halotolerant. Based on BOXA1R-PCR and 16S rRNA gene sequencing, the recovered isolates were dominated by the genus Bacillus (97.5%) represented by B. licheniformis (119), B. aerius (44), B. sonorensis (33), B. subtilis (subsp. spizizenii (2) and subsp. inaquosurum (6)), B. amyloliquefaciens (subsp. amyloliquefaciens (4) and subsp. plantarum (4)), B. tequilensis (3), B. pumilus (3) and Bacillus sp. (19). Only six isolates (2.5%) belonged to the genus Aeribacillus represented by A. pallidus (4) and Aeribacillus sp. (2). In this study, B. aerius and B. tequilensis are described for the first time as thermophilic bacteria. Moreover, 71.25%, 50.41% and 5.41% of total strains exhibited high amylolytic, proteolytic or cellulolytic activity respectively. PMID:26273259

  8. Electron transfer kinetics of caa3 oxidase from Bacillus stearothermophilus: a hypothesis for thermophilicity.

    PubMed Central

    Giuffrè, A; Watmough, N J; Giannini, S; Brunori, M; Konings, W N; Greenwood, C

    1999-01-01

    The O2 reaction and the reverse electron transfer of the thermophilic caa3 terminal oxidase of Bacillus stearothermophilus have been studied by laser flash-photolysis. The results show that both reactions, although studied at a temperature of 20 degreesC, far from the optimal temperature of > 60 degreesC for caa3, follow a kinetic behavior essentially identical to that observed with the electrostatic complex between mammalian cyt c and cyt c oxidase. In the O2 reaction cyt a and cyt a3 are very quickly oxidized; cyt a is then re-reduced via CuA, whereas cyt c oxidation is apparently rate-limited by the oxidation of CuA. Upon photodissociation of the mixed valence-CO caa3, reverse electron transfer from the binuclear center to cyt a3+ (tau1 = 3 micros) and CuA2+ (tau2 = 64 micros) is observed, while cyt c is not reduced by any detectable level. These results seem to rule out accounting for enzymatic thermophilicity by altered kinetics of intramolecular electron transfer involving the cyt center in the reduced configuration, which is very fast. On the basis of these results and previous data, we propose that thermophilicity involves an increased activation barrier for the reduction of cyt a3-CuB in the configuration typical of the oxidized site. PMID:9876155

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

    NASA Astrophysics Data System (ADS)

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

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

  10. Development of a Continuous Bioconversion System Using a Thermophilic Whole-Cell Biocatalyst

    PubMed Central

    Ninh, Pham Huynh; Yokohigashi, Yukako; Okano, Kenji; Omasa, Takeshi; Ohtake, Hisao

    2013-01-01

    The heat treatment of recombinant mesophilic cells having heterologous thermophilic enzymes results in the denaturation of indigenous mesophilic enzymes and the elimination of undesired side reactions; therefore, highly selective whole-cell catalysts comparable to purified enzymes can be readily prepared. However, the thermolysis of host cells leads to the heat-induced leakage of thermophilic enzymes, which are produced as soluble proteins, limiting the exploitation of their excellent stability in repeated and continuous reactions. In this study, Escherichia coli cells having the thermophilic fumarase from Thermus thermophilus (TtFTA) were treated with glutaraldehyde to prevent the heat-induced leakage of the enzyme, and the resulting cells were used as a whole-cell catalyst in repeated and continuous reactions. Interestingly, although electron microscopic observations revealed that the cellular structure of glutaraldehyde-treated E. coli was not apparently changed by the heat treatment, the membrane permeability of the heated cells to relatively small molecules (up to at least 3 kDa) was significantly improved. By applying the glutaraldehyde-treated E. coli having TtFTA to a continuous reactor equipped with a cell-separation membrane filter, the enzymatic hydration of fumarate to malate could be operated for more than 600 min with a molar conversion yield of 60% or higher. PMID:23335777

  11. Keratin Degradation by Fervidobacterium pennavorans, a Novel Thermophilic Anaerobic Species of the Order Thermotogales

    PubMed Central

    Friedrich, A. B.; Antranikian, G.

    1996-01-01

    From a hot spring of the Azores islands a novel thermophilic bacterium belonging to the Thermotogales order was isolated. This strain, which grows optimally at 70(deg)C and pH 6.5, is the first known extreme thermophile that is able to degrade native feathers at high temperatures. The enzyme system converts feather meal to amino acids and peptides. On the basis of physiological, morphological, and 16S rDNA studies the new isolate was found to be a member of the Thermotogales order and was identified as Fervidobacterium pennavorans. The strain was highly related to Fervidobacterium islandicum and Fervidobacterium pullulanolyticum. The cell-bound keratinolytic enzyme system was purified 32-fold by detergent treatment with CHAPS (3-[(3-cholamidopropyl)-dimethyl-ammonio]-1-propanesulfonate) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme was characterized as a serine protease with a molecular mass of 130 kDa and an isoelectric point of 3.8. Optimal activity was measured at 80(deg)C and pH 10.0. Furthermore, 19 anaerobic thermophilic archaea and bacteria belonging to the orders Thermococcales, Thermoproteales, Thermotogales, and Clostridiales (growth temperatures between 60 and 105(deg)C) were tested for their abilities to grow on feathers and produce heat-stable keratinolytic enzymes. None of the tested extremophilic microorganisms was able to attack the substrate in a native form. PMID:16535379

  12. Growth of extreme thermophile Sulfolobus acidocaldarius in a hyperbaric helium bioreactor

    SciTech Connect

    Sturm, F.J.; Hurwitz, S.A.; Deming, J.W.; Kelly, R.M.

    1987-01-01

    The relationship between pressure and temperature as it affects microbial growth and metabolism has been examined only for a limited number of bacterial species. Because many newly-discovered, extremely thermophilic bacteria have been isolated from pressurized environments, this relationship merits closer scrutiny. In this study, the extremely thermophilic bacterium, Sulfolobus acidocaldarius, was cultured successfully in a hyperbaric chamber containing helium and air enriched with 5% carbon dioxide. Over a pressure range of approximately 1-120 bar and a temperature range of 67-80/sup 0/C, growth was achieved in a heterotrophic medium with the air mixture at partial pressures up to 3.5 bar. Helium was used to obtain the final, desired incubation pressure. No significant growth was noted above 80/sup 0/C over the same range of hyperbaric pressures, or at 70/sup 0/C when pressure was applied hydrostatically. Growth experiments conducted under hyperbaric conditions may provide a means to study these bacteria under simulated in situ conditions and simultaneously avoid the complications associated with hydrostatic experiments. Results indicate that hyperbaric helium bioreactors will be important in the study of extremely thermophilic bacteria that are isolated from pressurized environments.

  13. Evaluation of thermochemical pretreatment and continuous thermophilic condition in rice straw composting process enhancement.

    PubMed

    Hosseini, Seyed Mohammad; Abdul Aziz, Hamidi

    2013-04-01

    The effects of thermochemical pretreatment and continuous thermophilic conditions on the composting of a mixture of rice straw residue and cattle manure were investigated using a laboratory-scale composting reactor. Results indicate that the composting period of rice straw can be shortened to less than 10 days by applying alkali pre-treatment and continuous thermophilic composting conditions. The parameters obtained on day 9 of this study are similar to the criteria level published by the Canadian Council of Ministers of the Environment. The moisture content, organic matter reduction, pH level, electrical conductivity, total organic carbon reduction, soluble chemical oxygen demand reduction, total Kjeldahl nitrogen, carbon-to-nitrogen ratio, and germination index were 62.07%, 16.99%, 7.30%, 1058 μS/cm, 17.00%, 83.43%, 2.06%, 16.75%, and 90.33%, respectively. The results of this study suggest that the application of chemical-biological integrated processes under thermophilic conditions is a novel method for the rapid degradation and maturation of rice straw residue. PMID:23428821

  14. Genetics of thermophilic bacteria: Progress report, May 1, 1986--June 30, 1988

    SciTech Connect

    Welker, N.E.

    1988-01-01

    Efficient and reliable protoplasting, regeneration and fusion techniques have been established for a prototrophic strain of Bacillus stearothermophilus. A variety of auxotrophic mutants and a restriction-deficient mutant were isolated and protoplast fusion was used to construct isogenic strains and for chromosomal mapping. Two linkage groups (hom thr and his gly pur-1) were established using this system. The order of the markers is similar to analagous markers on the Bacillus subtilis chromosome. We have evidence that this genetic exchange system can be used for linear chromosomal DNA transformation of protoplasts. Investigations have recently been initiated to develop a transducing system in this strain. Attempts to introduce a thermophile plasmid into strain NUB36 by transformation of protoplasts or protoplast fusion were unsuccessful. We think that the lack of success encountered in these studies is because this plasmid is not stably maintained at high temperature and/or is incompatible with the cryptic plasmids contained in this strain. This is the first report of a reliable genetic exchange system in B. stearothermophilus. This system can be used for the genetic analysis of this organism and for molecular cloning. The development of a host-vector system for cloning in B. stearothermophilus will make it possible to clone genes encoding cellulose hydrolysis, alcohol formation, or methane production. In addition, information gained from the genetic analysis of thermophilic bacilli may be of value in developing genetic systems in thermophilic eubacteria and archaebacteria. 3 tabs.

  15. Upflow anaerobic solid-state (UASS) digestion of horse manure: Thermophilic vs. mesophilic performance.

    PubMed

    Böske, Janina; Wirth, Benjamin; Garlipp, Felix; Mumme, Jan; Van den Weghe, Herman

    2015-01-01

    Energetic use of complex lignocellulosic wastes has gained global interest. Thermophilic digestion of horse manure based on straw was investigated using the upflow anaerobic solid-state (UASS) process. Increasing the organic loading rate from 2.5 to 5.5gvsL(-)(1)d(-)(1) enhanced the average methane production rate from 0.387 to 0.687LCH4L(-)(1)d(-)(1), whereas the yield decreased from 154.8 to 124.8LCH4kgvs(-)(1). A single-stage and two-stage process design showed almost the same performance. Compared to prior experiments at mesophilic conditions, thermophilic conditions showed a significantly higher efficiency with an increase of 59.8% in methane yield and 58.1% in methane production rate. Additional biochemical methane potential (BMP) tests with two types of horse manure and four different bedding materials showed that wheat straw obtained the highest BMP. The results show that the thermophilic UASS process can be the key to an efficient energy recovery from straw-based manures. PMID:25459798

  16. High-solids enrichment of thermophilic microbial communities and their enzymes on bioenergy feedstocks

    SciTech Connect

    Reddy, A. P.; Allgaier, M.; Singer, S.W.; Hazen, T.C.; Simmons, B.A.; Hugenholtz, P.; VanderGheynst, J.S.

    2011-04-01

    Thermophilic microbial communities that are active in a high-solids environment offer great potential for the discovery of industrially relevant enzymes that efficiently deconstruct bioenergy feedstocks. In this study, finished green waste compost was used as an inoculum source to enrich microbial communities and associated enzymes that hydrolyze cellulose and hemicellulose during thermophilic high-solids fermentation of the bioenergy feedstocks switchgrass and corn stover. Methods involving the disruption of enzyme and plant cell wall polysaccharide interactions were developed to recover xylanase and endoglucanase activity from deconstructed solids. Xylanase and endoglucanase activity increased by more than a factor of 5, upon four successive enrichments on switchgrass. Overall, the changes for switchgrass were more pronounced than for corn stover; solids reduction between the first and second enrichments increased by a factor of four for switchgrass while solids reduction remained relatively constant for corn stover. Amplicon pyrosequencing analysis of small-subunit ribosomal RNA genes recovered from enriched samples indicated rapid changes in the microbial communities between the first and second enrichment with the simplified communities achieved by the third enrichment. The results demonstrate a successful approach for enrichment of unique microbial communities and enzymes active in a thermophilic high-solids environment.

  17. Biofilms of thermophilic bacilli isolated from dairy processing plants and efficacy of sanitizers.

    PubMed

    Burgess, Sara A; Lindsay, Denise; Flint, Steve H

    2014-01-01

    In many environments, bacteria can attach to a surface and grow into multicellular structures, otherwise known as biofilms. Many systems for studying these biofilms in the laboratory are available. To study biofilms of the thermophilic bacilli in milk powder-manufacturing plants, standard laboratory biofilm techniques need to be adapted. The focus of this chapter is on techniques that can be used for growing and analyzing biofilms of thermophilic bacilli that are isolated from dairy processing plants. These techniques include laboratory methods as well as how to set up a pilot-scale experiment. The laboratory methods consist of a microtiter plate assay, which is used for strain selection, and the CDC reactor, which is used for testing sanitizers and antimicrobial surfaces. In dairy processing, if a new sanitizer or antimicrobial surface appears to be promising, it is useful to carry out pilot-scale experiments before introducing it to a manufacturing plant. We describe how to set up a pilot-scale experiment for testing the efficacy of sanitizers against the thermophilic bacilli. PMID:24664846

  18. Bioelectrochemical analyses of the development of a thermophilic biocathode catalyzing electromethanogenesis.

    PubMed

    Fu, Qian; Kuramochi, Yoshihiro; Fukushima, Naoya; Maeda, Haruo; Sato, Kozo; Kobayashi, Hajime

    2015-01-20

    The use of thermophilic microorganisms as biocatalysts for electromethanogenesis was investigated. Single-chamber reactors inoculated with thermophiles and operated at 55 °C showed high CH4 production rates (max. 1103 mmol m(–2) day(–1) at an applied voltage of 0.8 V) with current-capture efficiencies >90%, indicating that thermophiles have high potential as biocatalysts. To improve the electromethanogenic activity, the developed biocathode was transferred to a two-chamber reactor and operated at a poised potential of −0.5 V vs SHE. The CH4 production rates of the biocathode were enhanced approximately 6-fold in 160 h of poised-potential incubation, indicating that the acclimation of the biocathode resulted in performance improvement. Compositional alteration of the cathodic microbiota suggested that a Methanothermobacter-related methanogen and synergistetes- and thermotogae-related bacteria were selected during the acclimation. Cyclic voltammetry of the “acclimated” biocathode showed an augmented cathodic catalytic wave with a midpoint potential at ca. −0.35 V vs SHE. Moreover, the biocathode was able to catalyze electromethanogenesis at −0.35 V vs SHE. These results suggested that the ability of the biocathode to catalyze electromethanogenesis via direct electron transfer was enhanced by the acclimation. This study provides new technological and fundamental information on electromethanogenic bioelectrochemical systems (BESs) that may be extended to other BESs. PMID:25544349

  19. Screening of thermotolerant and thermophilic fungi aiming β-xylosidase and arabinanase production

    PubMed Central

    Benassi, Vivian Machado; de Lucas, Rosymar Coutinho; Jorge, João Atílio; Polizeli, Maria de Lourdes Teixeira de Moraes

    2014-01-01

    Plant cell wall is mainly composed by cellulose, hemicellulose and lignin. The heterogeneous structure and composition of the hemicellulose are key impediments to its depolymerization and subsequent use in fermentation processes. Thus, this study aimed to perform a screening of thermophilic and thermotolerant filamentous fungi collected from different regions of the São Paulo state, and analyze the production of β-xylosidase and arabinanase at different temperatures. These enzymes are important to cell wall degradation and synthesis of end products as xylose and arabinose, respectively, which are significant sugars to fermentation and ethanol production. A total of 12 fungal species were analyzed and 9 of them grew at 45 °C, suggesting a thermophilic or thermotolerant character. Additionally Aspergillus thermomutatus anamorph of Neosartorya and A. parasiticus grew at 50 °C. Aspergillus niger and Aspergillus thermomutatus were the filamentous fungi with the most expressive production of β-xylosidase and arabinanase, respectively. In general for most of the tested microorganisms, β-xylosidase and arabinanase activities from mycelial extract (intracellular form) were higher in cultures grown at high temperatures (35–40 °C), while the correspondent extracellular activities were favorably secreted from cultures at 30 °C. This study contributes to catalogue isolated fungi of the state of São Paulo, and these findings could be promising sources for thermophilic and thermotolerant microorganisms, which are industrially important due to their enzymes. PMID:25763055

  20. Mesophilic-hydrothermal-thermophilic (M-H-T) digestion of green corn straw.

    PubMed

    Li, Dong; Wang, Qingjing; Li, Jiang; Li, Zhidong; Yuan, Yuexiang; Yan, Zhiying; Mei, Zili; Liu, Xiaofeng

    2016-02-01

    Mesophilic-hydrothermal (80-160 °C, 30 min)-thermophilic (M-H-T) digestion and control tests of mesophilic (M), thermophilic (T), hydrothermal-mesophilic (H-M), and mesophilic-thermophilic digestion (M-T) of green corn straw were conducted for a 20-day fermentation period. The results indicate that M-H-T is an efficient method to improve methane production. A maximum methane yield of 371.74 mL/g volatile solid was obtained by the M (3 days)-H (140 °C)-T (17 days) process, which was 20.44%, 16.55%, 31.44%, and 14.31% higher than the yields of the M, T, 140-M, and M-T processes. The enhanced methane production was attributed to (1) the improved hemicellulose degradation and lignin disorganization; (2) prevention of the degradation of soluble sugar, easily hydrolyzed hemicellulose and cellulose into furfural and methylfurfural; and (3) lack of formation of Maillard reaction products during initial hydrothermal treatment. PMID:26700755

  1. Highly Thermostable Xylanase Production from A Thermophilic Geobacillus sp. Strain WSUCF1 Utilizing Lignocellulosic Biomass

    PubMed Central

    Bhalla, Aditya; Bischoff, Kenneth M.; Sani, Rajesh Kumar

    2015-01-01

    Efficient enzymatic hydrolysis of lignocellulose to fermentable sugars requires a complete repertoire of biomass deconstruction enzymes. Hemicellulases play an important role in hydrolyzing hemicellulose component of lignocellulose to xylooligosaccharides and xylose. Thermostable xylanases have been a focus of attention as industrially important enzymes due to their long shelf life at high temperatures. Geobacillus sp. strain WSUCF1 produced thermostable xylanase activity (crude xylanase cocktail) when grown on xylan or various inexpensive untreated and pretreated lignocellulosic biomasses such as prairie cord grass and corn stover. The optimum pH and temperature for the crude xylanase cocktail were 6.5 and 70°C, respectively. The WSUCF1 crude xylanase was found to be highly thermostable with half-lives of 18 and 12 days at 60 and 70°C, respectively. At 70°C, rates of xylan hydrolysis were also found to be better with the WSUCF1 secretome than those with commercial enzymes, i.e., for WSUCF1 crude xylanase, Cellic-HTec2, and AccelleraseXY, the percent xylan conversions were 68.9, 49.4, and 28.92, respectively. To the best of our knowledge, WSUCF1 crude xylanase cocktail is among the most thermostable xylanases produced by thermophilic Geobacillus spp. and other thermophilic microbes (optimum growth temperature ≤70°C). High thermostability, activity over wide range of temperatures, and better xylan hydrolysis than commercial enzymes make WSUCF1 crude xylanase suitable for thermophilic lignocellulose bioconversion processes. PMID:26137456

  2. The HPr Proteins from the Thermophile Bacillus stearothermophilus Can Form Domain-swapped Dimers

    SciTech Connect

    Sridharan, Sudharsan; Razvi, Abbas; Scholtz, J. Martin; Sacchettini, James C.

    2010-07-20

    The study of proteins from extremophilic organisms continues to generate interest in the field of protein folding because paradigms explaining the enhanced stability of these proteins still elude us and such studies have the potential to further our knowledge of the forces stabilizing proteins. We have undertaken such a study with our model protein HPr from a mesophile, Bacillus subtilis, and a thermophile, Bacillus stearothermophilus. We report here the high-resolution structures of the wild-type HPr protein from the thermophile and a variant, F29W. The variant proved to crystallize in two forms: a monomeric form with a structure very similar to the wild-type protein as well as a domain-swapped dimer. Interestingly, the structure of the domain-swapped dimer for HPr is very different from that observed for a homologous protein, Crh, from B. subtilis. The existence of a domain-swapped dimer has implications for amyloid formation and is consistent with recent results showing that the HPr proteins can form amyloid fibrils. We also characterized the conformational stability of the thermophilic HPr proteins using thermal and solvent denaturation methods and have used the high-resolution structures in an attempt to explain the differences in stability between the different HPr proteins. Finally, we present a detailed analysis of the solution properties of the HPr proteins using a variety of biochemical and biophysical methods.

  3. A Numerical Evaluation on the Viability of Heap Thermophilic Bioleaching of Chalcopyrite

    NASA Astrophysics Data System (ADS)

    Vilcaez, J.; Suto, K.; Inoue, C.

    2007-03-01

    The present numerical evaluation explores into the interactions among the many variables governing the mass and heat transport processes that take place in a heap thermophilic bioleaching system. The necessity of using mesophiles together with thermophiles is proved by tracing the activity of both microorganisms individually at each point throughout the heap. The role of key variables such as the fraction of FeS2 per CuFeS2 leached was quantified and its importance highlighted. In this evaluation, the heat transfer process plays the main role because of the heat accumulation required to maintain the heap temperature within the range of 60 °C to 80 °C where thermophilic microorganisms are capable of completing the unfinished dissolution of copper started by mesophilic microorganisms at 30 °C. The evaluation was done taking into consideration: biological activity as function of the temperature in the heap, heat loss due to conduction and advection from the top and bottom of the heap, and mass transfer between the gas and liquid phases as a function of temperature. The exothermic nature of the leaching reactions of CuFeS2 and FeS2 makes the system auto-thermal.

  4. Cadmium Ion Biosorption by the Thermophilic Bacteria Geobacillus stearothermophilus and G. thermocatenulatus

    PubMed Central

    Hetzer, Adrian; Daughney, Christopher J.; Morgan, Hugh W.

    2006-01-01

    This study reports surface complexation models (SCMs) for quantifying metal ion adsorption by thermophilic microorganisms. In initial cadmium ion toxicity tests, members of the genus Geobacillus displayed the highest tolerance to CdCl2 (as high as 400 to 3,200 μM). The thermophilic, gram-positive bacteria Geobacillus stearothermophilus and G. thermocatenulatus were selected for further electrophoretic mobility, potentiometric titration, and Cd2+ adsorption experiments to characterize Cd2+ complexation by functional groups within and on the cell wall. Distinct one-site SCMs described the extent of cadmium ion adsorption by both studied Geobacillus sp. strains over a range of pH values and metal/bacteria concentration ratios. The results indicate that a functional group with a deprotonation constant pK value of approximately 3.8 accounts for 66% and 80% of all titratable sites for G. thermocatenulatus and G. stearothermophilus, respectively, and is dominant in Cd2+ adsorption reactions. The results suggest a different type of functional group may be involved in cadmium biosorption for both thermophilic strains investigated here, compared to previous reports for mesophilic bacteria. PMID:16751511

  5. Enhancing ethanol production from thermophilic and mesophilic solid digestate using ozone combined with aqueous ammonia pretreatment.

    PubMed

    Wang, Dianlong; Xi, Jiang; Ai, Ping; Yu, Liang; Zhai, Hong; Yan, Shuiping; Zhang, Yanlin

    2016-05-01

    Pretreatment with ozone combined with aqueous ammonia was used to recover residual organic carbon from recalcitrant solid digestate for ethanol production after anaerobic digestion (AD) of rice straw. Methane yield of AD at mesophilic and thermophilic conditions, and ethanol production of solid digestate were investigated. The results showed that the methane yield at thermophilic temperature was 72.2% higher than that at mesophilic temperature under the same conditions of 24days and 17% solid concentration. And also the ethanol production efficiency of solid digestate after thermophilic process was 24.3% higher than that of solid digestate after mesophilic process. In this study, the optimal conditions for integrated methane and ethanol processes were determined as 55°C, 17% solid concentration and 24days. 58.6% of glucose conversion, 142.8g/kg of methane yield and 65.2g/kg of ethanol yield were achieved, and the highest net energy balance was calculated as 6416kJ/kg. PMID:26868156

  6. Innovative two-stage mesophilic/thermophilic anaerobic degradation of sonicated sludge: performances and energy balance.

    PubMed

    Gianico, A; Braguglia, C M; Gallipoli, A; Mininni, G

    2015-05-01

    This study investigates for the first time, on laboratory scale, the possible application of an innovative enhanced stabilization process based on sequential mesophilic/thermophilic anaerobic digestion of waste-activated sludge, with low-energy sonication pretreatment. The first mesophilic digestion step was conducted at short hydraulic retention time (3-5 days), in order to favor volatile fatty acid production, followed by a longer thermophilic step of 10 days to enhance the bioconversion kinetics, assuring a complete pathogen removal. The high volatile solid removals, up to 55%, noticeably higher compared to the performances of a single-stage process carried out in same conditions, can guarantee the stability of the final digestate for land application. The ultrasonic pretreatment influenced significantly the fatty acid formation and composition during the first mesophilic step, improving consequently the thermophilic conversion of these compounds into methane. Methane yield from sonicated sludge digestion reached values up to 0.2 Nm(3)/kgVSfed. Positive energy balances highlighted the possible exploitation of this innovative two-stage digestion in place of conventional single-stage processes. PMID:24906832

  7. Thermophilic microbial cellulose decomposition and methanogenesis pathways recharacterized by metatranscriptomic and metagenomic analysis

    PubMed Central

    Xia, Yu; Wang, Yubo; Fang, Herbert H. P.; Jin, Tao; Zhong, Huanzi; Zhang, Tong

    2014-01-01

    The metatranscriptomic recharacterization in the present study captured microbial enzymes at the unprecedented scale of 40,000 active genes belonged to 2,269 KEGG functions were identified. The novel information obtained herein revealed interesting patterns and provides an initial transcriptional insight into the thermophilic cellulose methanization process. Synergistic beta-sugar consumption by Thermotogales is crucial for cellulose hydrolysis in the thermophilic cellulose-degrading consortium because the primary cellulose degraders Clostridiales showed metabolic incompetence in subsequent beta-sugar pathways. Additionally, comparable transcription of putative Sus-like polysaccharide utilization loci (PULs) was observed in an unclassified order of Bacteroidetes suggesting the importance of PULs mechanism for polysaccharides breakdown in thermophilic systems. Despite the abundance of acetate as a fermentation product, the acetate-utilizing Methanosarcinales were less prevalent by 60% than the hydrogenotrophic Methanobacteriales. Whereas the aceticlastic methanogenesis pathway was markedly more active in terms of transcriptional activities in key genes, indicating that the less dominant Methanosarcinales are more active than their hydrogenotrophic counterparts in methane metabolism. These findings suggest that the minority of aceticlastic methanogens are not necessarily associated with repressed metabolism, in a pattern that was commonly observed in the cellulose-based methanization consortium, and thus challenge the causal likelihood proposed by previous studies. PMID:25330991

  8. Temporal variation of microbial population in a thermophilic biofilter for SO₂ removal.

    PubMed

    Zhang, Jingying; Li, Lin; Liu, Junxin

    2016-01-01

    The performance of a biofilter relies on the activity of microorganisms during the gas contaminant treatment process. In this study, SO2 was treated using a laboratory-scale biofilter packed with polyurethane foam cubes (PUFC), on which thermophilic desulfurization bacteria were attached. The thermophilic biofilter effectively reduced SO2 within 10months of operation time, with a maximum elimination capacity of 48.29 g/m(3)/hr. Temporal shifts in the microbial population in the thermophilic biofilter were determined through polymerase chain reaction-denaturing gradient gel electrophoresis and deoxyribonucleic acid (DNA) sequence analysis. The substrate species and environmental conditions in the biofilter influenced the microbial population. Oxygen distribution in the PUFC was analyzed using a microelectrode. When the water-containing rate in PUFC was over 98%, the oxygen distribution presented aerobic-anoxic-aerobic states along the test route on the PUFC. The appearance of sulfate-reducing bacteria was caused by the anaerobic conditions and sulfate formation after 4months of operation. PMID:26899638

  9. Syntrophic degradation of proteinaceous materials by the thermophilic strains Coprothermobacter proteolyticus and Methanothermobacter thermautotrophicus.

    PubMed

    Sasaki, Kengo; Morita, Masahiko; Sasaki, Daisuke; Nagaoka, Jun; Matsumoto, Norio; Ohmura, Naoya; Shinozaki, Hiraku

    2011-11-01

    Protein is a major component of organic solid wastes, and therefore, it is necessary to further elucidate thermophilic protein degradation process. The effects of hydrogenotrophic methanogens on protein degradation were investigated using the proteolytic bacterial strain CT-1 that was isolated from a methanogenic thermophilic (55°C) packed-bed reactor degrading artificial garbage slurry. Strain CT-1 was closely related to Coprothermobacter proteolyticus, which is frequently found in methanogenic reactors degrading organic solid wastes. Strain CT-1 was cultivated in the absence or presence of Methanothermobacter thermautotrophicus by using 3 kinds of proteinaceous substrates. Degradation rates of casein, gelatin, and bovine serum albumin were higher in co-cultures than in monocultures. Strain CT-1 showed faster growth in co-cultures than in monocultures. M. thermautotrophicus comprised 5.5-6.0% of the total cells in co-culture. Increased production of ammonia and acetate was observed in co-cultures than in monocultures, suggesting that addition of M. thermautotrophicus increases the products of protein degradation. Hydrogen produced in the monocultures was converted to methane in co-cultures. These results suggest that thermophilic proteolytic bacteria find it favorable to syntrophically degrade protein in a methanogenic environment, and that it is important to retain hydrogen-scavenging methanogens within the reactor. PMID:21802987

  10. Proteins responsible for lysogeny of deep-sea thermophilic bacteriophage GVE2 at high temperature.

    PubMed

    Song, Qing; Ye, Ting; Zhang, Xiaobo

    2011-06-15

    The lytic and lysogenic life cycle switch of bacteriophages plays very important roles in virus-host interactions. However, the lysogeny of thermophilic bacteriophage infecting thermophile at high temperatures has not been addressed. In this study, two lysogeny-related genes encoding the CI protein and recombinase of GVE2, a thermophilic bacteriophage obtained from a deep-sea hydrothermal vent, were characterized. Temporal analyses showed that the two genes were expressed at early stages of GVE2 infection. Based on chromatin immunoprecipitation (ChIP) assay and electrophoretic mobility shift assay (EMSA), the GVE2 CI protein was bound with only one DNA fragment located at 24264-24036 bp in the GVE2 genome. This location might be the original transcription site and the lysis-lysogeny switch site, which was very different from mesophilic bacteriophages. The GVE2 CI and recombinase proteins could function only at high temperatures. Therefore our study improved our understanding of the lysogeny process of bacteriophages at high temperatures. PMID:21303688

  11. Comparing mesophilic and thermophilic anaerobic digestion of chicken manure: Microbial community dynamics and process resilience.

    PubMed

    Niu, Qigui; Takemura, Yasuyuki; Kubota, Kengo; Li, Yu-You

    2015-09-01

    While methane fermentation is considered as the most successful bioenergy treatment for chicken manure, the relationship between operational performance and the dynamic transition of archaeal and bacterial communities remains poorly understood. Two continuous stirred-tank reactors were investigated under thermophilic and mesophilic conditions feeding with 10%TS. The tolerance of thermophilic reactor on total ammonia nitrogen (TAN) was found to be 8000mg/L with free ammonia (FA) 2000mg/L compared to 16,000mg/L (FA1500mg/L) of mesophilic reactor. Biomethane production was 0.29 L/gVSin in the steady stage and decreased following TAN increase. After serious inhibition, the mesophilic reactor was recovered successfully by dilution and washing stratagem compared to the unrecoverable of thermophilic reactor. The relationship between the microbial community structure, the bioreactor performance and inhibitors such as TAN, FA, and volatile fatty acid was evaluated by canonical correspondence analysis. The performance of methanogenic activity and substrate removal efficiency were changed significantly correlating with the community evenness and phylogenetic structure. The resilient archaeal community was found even after serious inhibition in both reactors. Obvious dynamics of bacterial communities were observed in acidogenic and hydrolytic functional bacteria following TAN variation in the different stages. PMID:26054964

  12. Thermophilic bacteria in Moroccan hot springs, salt marshes and desert soils.

    PubMed

    Aanniz, Tarik; Ouadghiri, Mouna; Melloul, Marouane; Swings, Jean; Elfahime, Elmostafa; Ibijbijen, Jamal; Ismaili, Mohamed; Amar, Mohamed

    2015-06-01

    The diversity of thermophilic bacteria was investigated in four hot springs, three salt marshes and 12 desert sites in Morocco. Two hundred and forty (240) thermophilic bacteria were recovered, identified and characterized. All isolates were Gram positive, rod-shaped, spore forming and halotolerant. Based on BOXA1R-PCR and 16S rRNA gene sequencing, the recovered isolates were dominated by the genus Bacillus (97.5%) represented by B. licheniformis (119), B. aerius (44), B. sonorensis (33), B. subtilis (subsp. spizizenii (2) and subsp. inaquosurum (6)), B. amyloliquefaciens (subsp. amyloliquefaciens (4) and subsp. plantarum (4)), B. tequilensis (3), B. pumilus (3) and Bacillus sp. (19). Only six isolates (2.5%) belonged to the genus Aeribacillus represented by A. pallidus (4) and Aeribacillus sp. (2). In this study, B. aerius and B. tequilensis are described for the first time as thermophilic bacteria. Moreover, 71.25%, 50.41% and 5.41% of total strains exhibited high amylolytic, proteolytic or cellulolytic activity respectively. PMID:26273259

  13. The Distribution of Thermophilic Sulfate-reducing Bacteria Along an Estuarine Gradient Reveals Multiple Origins of Endospores in Estuarine Sediments

    NASA Astrophysics Data System (ADS)

    Bell, E.

    2015-12-01

    Cold marine sediments harbour inactive spores of thermophilic bacteria. These misplaced thermophiles are genetically similar to microorganisms detected in deep biosphere environments, leading to the hypothesis that seabed fluid flow transports thermophiles out of warm subsurface environments and into the ocean. Estuaries form the transition between the marine and the terrestrial biosphere and are influenced by tidal currents, surface run-off and groundwater seepage. Endospores from thermophilic bacteria present in estuarine sediments could therefore originate from a number of sources that may influence the estuary differently. We have therefore tested the hypothesis that this will lead to a gradient in the composition of thermophilic endospore populations in estuarine sediments. The distribution of thermophilic spore-forming sulfate-reducing bacteria along an estuarine gradient from freshwater (River Tyne, UK) to marine (North Sea) was investigated. Microbial community analysis by 16S rRNA gene amplicon sequencing revealed changes in the thermophilic population enriched at different locations within the estuary. Certain species were only detected at the marine end, highlighting possible links to deep marine biosphere habitats such as oil reservoirs that harbour closely related Desulfotomaculum spp. Conversely, other taxa were predominantly observed in the freshwater reaches of the estuary indicating dispersal from an upstream or terrestrial source. Different endospore populations were enriched dependent on incubation temperature and spore heat-resistance. Microcosms incubated at 50, 60 or 70°C showed a shift in the dominant species of Desulfotomaculum enriched as the temperature increased. Microcosms triple-autoclaved at 121°C prior to incubation still showed rapid and reproducible sulfate-reduction and some Desulfotomaculum spp. remained active after autoclaving at 130°C. These results show that temperature physiology and biogeographic patterns can be used to

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

    NASA Astrophysics Data System (ADS)

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

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

  15. Graphite moderated (252)Cf source.

    PubMed

    Sajo-Bohus, Laszlo; Barros, Haydn; Greaves, Eduardo D; Vega-Carrillo, Hector Rene

    2015-06-01

    The Thorium molten-salt reactor is an attractive and affordable nuclear power option for developing countries with insufficient infrastructure and limited technological capability. In the aim of personnel training and experience gathering at the Universidad Simon Bolivar there is in progress a project of developing a subcritical thorium liquid-fuel reactor. The neutron source to run this subcritical reactor is a (252)Cf source and the reactor will use high-purity graphite as moderator. Using the MCNP5 code the neutron spectra of the (252)Cf in the center of the graphite moderator has been estimated along the channel where the liquid thorium salt will be inserted; also the ambient dose equivalent due to the source has been determined around the moderator. PMID:25770393

  16. Moderated ruthenium fischer-tropsch synthesis catalyst

    DOEpatents

    Abrevaya, Hayim

    1991-01-01

    The subject Fischer-Tropsch catalyst comprises moderated ruthenium on an inorganic oxide support. The preferred moderator is silicon. Preferably the moderator is effectively positioned in relationship to ruthenium particles through simultaneous placement on the support using reverse micelle impregnation.

  17. Two-stage thermophilic-mesophilic anaerobic digestion of waste activated sludge from a biological nutrient removal plant.

    PubMed

    Watts, S; Hamilton, G; Keller, J

    2006-01-01

    A two-stage thermophilic-mesophilic anaerobic digestion pilot-plant was operated solely on waste activated sludge (WAS) from a biological nutrient removal (BNR) plant. The first-stage thermophilic reactor (HRT 2 days) was operated at 47, 54 and 60 degrees C. The second-stage mesophilic digester (HRT 15 days) was held at a constant temperature of 36-37 degrees C. For comparison with a single-stage mesophilic process, the mesophilic digester was also operated separately with an HRT of 17 days and temperature of 36-37 degrees C. The results showed a truly thermophilic stage (60 degrees C) was essential to achieve good WAS degradation. The lower thermophilic temperatures examined did not offer advantages over single-stage mesophilic treatment in terms of COD and VS removal. At a thermophilic temperature of 60 degrees C, the plant achieved 35% VS reduction, representing a 46% increase compared to the single-stage mesophilic digester. This is a significant level of degradation which could make such a process viable in situations where there is no primary sludge generated. The fate of the biologically stored phosphorus in this BNR sludge was also investigated. Over 80% of the incoming phosphorus remained bound up with the solids and was not released into solution during the WAS digestion. Therefore only a small fraction of phosphorus would be recycled to the main treatment plant with the dewatering stream. PMID:16784199

  18. Effect of thermal additions on the density and distribution of thermophilic amoebae and pathogenic Naegleria fowleri in a newly created cooling lake

    SciTech Connect

    Tyndall, R.L.; Ironside, K.S.; Metler, P.L.; Tan, E.L. ); Hazen, T.C.; Fliermans, C.B. )

    1989-03-01

    Pathogenic Naegleria fowleri is the causative agent of fatal human amoebic meningoencephalitis. The protozoan is ubiquitous in nature, and its presence is enhanced by thermal additions. In this investigation, water and sediments from a newly created cooling lake were quantitatively analyzed for the presence of thermophilic amoebae, thermophilic Naegleria spp., and the pathogen Naegleria fowleri. During periods of thermal additions, the concentrations of thermophilic amoebae and thermophilic Naegleria spp. increased as much as 5 orders of magnitude, and the concentration of the pathogen N. fowleri increased as much as 2 orders of magnitude. Concentrations of amoebae returned to prior thermal perturbation levels within 30 to 60 days after cessation of thermal additions. Increases in the thermophilic amoeba concentrations were noted in Savannah River oxbows downriver from the Savannah River plant discharge streams as compared with oxbows upriver from the discharges. Concentrations of thermophilic amoebae and thermophilic Naegleria spp. correlated significantly with temperature and conductivity. Air samples taken proximal to the lade during periods of thermal addition showed no evidence of thermophilic Naegleria spp. Isoenzyme patterns of the N. fowleri isolated from the cooling lake were identical to patterns of N. fowleri isolated from other sites in the United States and Belgium.

  19. Draft Genome Sequence of Geobacillus subterraneus Strain K, a Hydrocarbon-Oxidizing Thermophilic Bacterium Isolated from a Petroleum Reservoir in Kazakhstan

    PubMed Central

    Poltaraus, Andrey B.; Sokolova, Diyana S.; Grouzdev, Denis S.; Ivanov, Timophey M.; Malakho, Sophia G.; Korshunova, Alena V.; Tourova, Tatiyana P.

    2016-01-01

    The draft genome sequence of Geobacillus subterraneus strain K, a thermophilic aerobic oil-oxidizing bacterium isolated from production water of the Uzen high-temperature oil field in Kazakhstan, is presented here. The genome is annotated for elucidation of the genomic and phenotypic diversity of thermophilic alkane-oxidizing bacteria. PMID:27491973

  20. Draft Genome Sequence of Geobacillus subterraneus Strain K, a Hydrocarbon-Oxidizing Thermophilic Bacterium Isolated from a Petroleum Reservoir in Kazakhstan.

    PubMed

    Poltaraus, Andrey B; Sokolova, Diyana S; Grouzdev, Denis S; Ivanov, Timophey M; Malakho, Sophia G; Korshunova, Alena V; Tourova, Tatiyana P; Nazina, Tamara N

    2016-01-01

    The draft genome sequence of Geobacillus subterraneus strain K, a thermophilic aerobic oil-oxidizing bacterium isolated from production water of the Uzen high-temperature oil field in Kazakhstan, is presented here. The genome is annotated for elucidation of the genomic and phenotypic diversity of thermophilic alkane-oxidizing bacteria. PMID:27491973

  1. THERMICANUS AEGYPTIUS GEN. NOV., SP. NOV., ISOLATED FROM OXIC SOIL, A FERMENTATIVE MICROAEROPHILE THAT GROWS COMMENSALLY WITH THE THERMOPHILIC ACETOGEN MOORELLA THERMOACETICA

    EPA Science Inventory

    A thermophilic, fermentative microaerophile (ET-5b) and a thermophilic acetogen (ET-5a) were coisolated from oxic soil obtained from Egypt. The 16S rRNA gene sequence of ET-5a was 99.8% identical to that of the classic acetogen Moorella thermoacetica. Further analyses confirmed t...

  2. Characteristic Features in the Structure and Collagen-Binding Ability of a Thermophilic Collagenolytic Protease from the Thermophile Geobacillus collagenovorans MO-1

    PubMed Central

    Itoi, Yuichi; Horinaka, Mano; Tsujimoto, Yoshiyuki; Matsui, Hiroshi; Watanabe, Kunihiko

    2006-01-01

    A collagen-degrading thermophile, Geobacillus collagenovorans MO-1, extracellularly produces a collagenolytic protease with a large molecular mass. Complete nucleotide sequencing of this gene after gene cloning revealed that the collagenolytic protease is a member of the subtilisin family of serine proteases and consists of a signal sequence for secretion, a prosequence for maturation, a catalytic region, 14 direct repeats of 20 amino acids at the C terminus, and a region with unknown function intervening between the catalytic region and the numerous repeats. Since the unusual repeats are most likely to be cleaved in the secreted form of the enzyme, the intervening region was investigated to determine whether it participates in collagen binding to facilitate collagen degradation. It was found that the mature collagenolytic protease containing the intervening region at the C terminus bound collagen but not the other insoluble proteins, elastin and keratin. Furthermore, the intervening region fused with glutathione S-transferase showed a collagen-binding ability comparable to that of the mature collagenolytic protease. The collagen-binding ability was finally attributed to two-thirds of the intervening region which is rich in β-strands and is approximately 35 kDa in molecular mass. In the collagenolytic protease from strain MO-1, hydrogen bonds most likely predominate over the hydrophobic interaction for collagen binding, since a higher concentration of NaCl released collagen from the enzyme surface but a nonionic detergent could not. To the best of our knowledge, this is the first report of a thermophilic collagenolytic protease containing the collagen-binding segment. PMID:16952949

  3. Influence of organics and silica on Fe(II) oxidation rates and cell-mineral aggregate formation by the green-sulfur Fe(II)-oxidizing bacterium Chlorobium ferrooxidans KoFox - Implications for Fe(II) oxidation in ancient oceans

    NASA Astrophysics Data System (ADS)

    Gauger, Tina; Byrne, James M.; Konhauser, Kurt O.; Obst, Martin; Crowe, Sean; Kappler, Andreas

    2016-06-01

    Most studies on microbial phototrophic Fe(II) oxidation (photoferrotrophy) have focused on purple bacteria, but recent evidence points to the importance of green-sulfur bacteria (GSB). Their recovery from modern ferruginous environments suggests that these photoferrotrophs can offer insights into how their ancient counterparts grew in Archean oceans at the time of banded iron formation (BIF) deposition. It is unknown, however, how Fe(II) oxidation rates, cell-mineral aggregate formation, and Fe-mineralogy vary under environmental conditions reminiscent of the geological past. To address this, we studied the Fe(II)-oxidizer Chlorobium ferrooxidans KoFox, a GSB living in co-culture with the heterotrophic Geospirillum strain KoFum. We investigated the mineralogy of Fe(III) metabolic products at low/high light intensity, and in the presence of dissolved silica and/or fumarate. Silica and fumarate influenced the crystallinity and particle size of the produced Fe(III) minerals. The presence of silica also enhanced Fe(II) oxidation rates, especially at high light intensities, potentially by lowering Fe(II)-toxicity to the cells. Electron microscopic imaging showed no encrustation of either KoFox or KoFum cells with Fe(III)-minerals, though weak associations were observed suggesting co-sedimentation of Fe(III) with at least some biomass via these aggregates, which could support diagenetic Fe(III)-reduction. Given that GSB are presumably one of the most ancient photosynthetic organisms, and pre-date cyanobacteria, our findings, on the one hand, strengthen arguments for photoferrotrophic activity as a likely mechanism for BIF deposition on a predominantly anoxic early Earth, but, on the other hand, also suggest that preservation of remnants of Fe(II)-oxidizing GSB as microfossils in the rock record is unlikely.

  4. Long-term stability of thermophilic co-digestion submerged anaerobic membrane reactor encountering high organic loading rate, persistent propionate and detectable hydrogen in biogas.

    PubMed

    Qiao, Wei; Takayanagi, Kazuyuki; Niu, Qigui; Shofie, Mohammad; Li, Yu You

    2013-12-01

    The performance of thermophilic anaerobic co-digestion of coffee grounds and sludge using membrane reactor was investigated for 148 days, out of a total research duration of 263 days. The OLR was increased from 2.2 to 33.7 kg-COD/m(3)d and HRT was shortened from 70 to 7 days. A significant irreversible drop in pH confirmed the overload of reactor. Under a moderately high OLR of 23.6 kg-COD/m(3)d, and with HRT and influent total solids of 10 days and 150 g/L, respectively, the COD removal efficiency was 44.5%. Hydrogen in biogas was around 100-200 ppm, which resulted in the persistent propionate of 1.0-3.2g/L. The VFA consumed approximately 60% of the total alkalinity. NH4HCO3 was supplemented to maintain alkalinity. The stability of system relied on pH management under steady state. The 16SrDNA results showed that hydrogen-utilizing methanogens dominates the archaeal community. The propionate-oxidizing bacteria in bacterial community was insufficient. PMID:24090872

  5. The Structure of a Novel Thermophilic Esterase from the Planctomycetes Species, Thermogutta terrifontis Reveals an Open Active Site Due to a Minimal ‘Cap’ Domain

    PubMed Central

    Sayer, Christopher; Szabo, Zalan; Isupov, Michail N.; Ingham, Colin; Littlechild, Jennifer A.

    2015-01-01

    A carboxyl esterase (TtEst2) has been identified in a novel thermophilic bacterium, Thermogutta terrifontis from the phylum Planctomycetes and has been cloned and over-expressed in Escherichia coli. The enzyme has been characterized biochemically and shown to have activity toward small p-nitrophenyl (pNP) carboxylic esters with optimal activity for pNP-acetate. The enzyme shows moderate thermostability retaining 75% activity after incubation for 30 min at 70°C. The crystal structures have been determined for the native TtEst2 and its complexes with the carboxylic acid products propionate, butyrate, and valerate. TtEst2 differs from most enzymes of the α/β-hydrolase family 3 as it lacks the majority of the ‘cap’ domain and its active site cavity is exposed to the solvent. The bound ligands have allowed the identification of the carboxyl pocket in the enzyme active site. Comparison of TtEst2 with structurally related enzymes has given insight into how differences in their substrate preference can be rationalized based upon the properties of their active site pockets. PMID:26635762

  6. HEAVY WATER MODERATED NEUTRONIC REACTOR

    DOEpatents

    Szilard, L.

    1958-04-29

    A nuclear reactor of the type which utilizes uranium fuel elements and a liquid coolant is described. The fuel elements are in the form of elongated tubes and are disposed within outer tubes extending through a tank containing heavy water, which acts as a moderator. The ends of the fuel tubes are connected by inlet and discharge headers, and liquid bismuth is circulated between the headers and through the fuel tubes for cooling. Helium is circulated through the annular space between the outer tubes in the tank and the fuel tubes to cool the water moderator to prevent boiling. The fuel tubes are covered with a steel lining, and suitable control means, heat exchange means, and pumping means for the coolants are provided to complete the reactor assembly.

  7. Cryogenic moderator simulations : confronting reality.

    SciTech Connect

    Iverson, E. B.

    1999-01-06

    The Intense Pulsed Neutron Source (IPNS) at Argonne National Laboratory is a spallation neutron source dedicated to materials research. Its three cryogenic methane moderators provide twelve neutron beams to fourteen instruments and test facilities. This report concerns ongoing activities for benchmarking our Monte Carlo model of the IPNS neutron generation system. This paper concentrates on the techniques (both experimental and calculational) used in such benchmarking activities.

  8. Net energy production associated with pathogen inactivation during mesophilic and thermophilic anaerobic digestion of sewage sludge.

    PubMed

    Ziemba, Christopher; Peccia, Jordan

    2011-10-15

    The potential for anaerobic digester energy production must be balanced with the sustainability of reusing the resultant biosolids for land application. Mesophilic, thermophilic, temperature-phased, and high temperature (60 or 70 °C) batch pre-treatment digester configurations have been systematically evaluated for net energy production and pathogen inactivation potential. Energy input requirements and net energy production were modeled for each digester scheme. First-order inactivation rate coefficients for Escherichia coli, Enterococcus faecalis and bacteriophage MS-2 were measured at each digester temperature and full-scale pathogen inactivation performance was estimated for each indicator organism and each digester configuration. Inactivation rates were found to increase dramatically at temperatures above 55 °C. Modeling full-scale performance using retention times based on U.S. EPA time and temperature constraints predicts a 1-2 log inactivation in mesophilic treatment, and a 2-5 log inactivation in 50-55 °C thermophilic and temperature-phased treatments. Incorporating a 60 or 70 °C batch pre-treatment phase resulted in dramatically higher potency, achieving MS-2 inactivation of 14 and 16 logs respectively, and complete inactivation (over 100 log reduction) of E. coli and E. faecalis. For temperatures less than 70 °C, viability staining of thermally-treated E. coli showed significantly reduced inactivation relative to standard culture enumeration. Due to shorter residence times in thermophilic reactors, the net energy production for all digesters was similar (less than 20% difference) with the 60 or 70 °C batch treatment configurations producing the most net energy and the mesophilic treatment producing the least. Incorporating a 60 or 70 °C pre-treatment phase can dramatically increase pathogen inactivation performance without decreasing net energy capture from anaerobic digestion. Energy consumption is not a significant barrier against

  9. Characterization and kinetic mechanism of mono- and bifunctional ornithine acetyltransferases from thermophilic microorganisms.

    PubMed

    Marc, F; Weigel, P; Legrain, C; Almeras, Y; Santrot, M; Glansdorff, N; Sakanyan, V

    2000-08-01

    The argJ gene coding for N2-acetyl-L-ornithine: L-glutamate N-acetyltransferase, the key enzyme involved in the acetyl cycle of L-arginine biosynthesis, has been cloned from thermophilic procaryotes: the archaeon Methanoccocus jannaschii, and the bacteria Thermotoga neapolitana and Bacillus stearothermophilus. Archaeal argJ only complements an Escherichia coli argE mutant (deficient in acetylornithinase, which catalyzes the fifth step in the linear biosynthetic pathway), whereas bacterial genes additionally complement an argA mutant (deficient in N-acetylglutamate synthetase, the first enzyme of the pathway). In keeping with these in vivo data the purified His-tagged ArgJ enzyme of M. jannaschii only catalyzes N2-acetylornithine conversion to ornithine, whereas T. neapolitana and B. stearothermophilus ArgJ also catalyze the conversion of glutamate to N-acetylglutamate using acetylCoA as the acetyl donor. M. jannaschii ArgJ is therefore a monofunctional enzyme, whereas T. neapolitana and B. stearothermophilus encoded ArgJ are bifunctional. Kinetic data demonstrate that in all three thermophilic organisms ArgJ-mediated catalysis follows ping-pong bi-bi kinetic mechanism. Acetylated ArgJ intermediates were detected in semireactions using [14C]acetylCoA or [14C]N2-acetyl-L-glutamate as acetyl donors. In this catalysis L-ornithine acts as an inhibitor; this amino acid therefore appears to be a key regulatory molecule in the acetyl cycle of L-arginine synthesis. Thermophilic ArgJ are synthesized as protein precursors undergoing internal cleavage to generate alpha and beta subunits which appear to assemble to alpha2beta2 heterotetramers in E. coli. The cleavage occurs between alanine and threonine residues within the highly conserved PXM-ATML motif detected in all available ArgJ sequences. PMID:10931207

  10. Spore-Forming Thermophilic Sulfate-Reducing Bacteria Isolated from North Sea Oil Field Waters

    PubMed Central

    Rosnes, Jan Thomas; Torsvik, Terje; Lien, Torleiv

    1991-01-01

    Thermophilic sulfate-reducing bacteria were isolated from oil field waters from oil production platforms in the Norwegian sector of the North Sea. Spore-forming rods dominated in the enrichments when lactate, propionate, butyrate, or a mixture of aliphatic fatty acids (C4 through C6) was added as a carbon source and electron donor. Representative strains were isolated and characterized. The isolates grew autotrophically on H2-CO2 and heterotrophically on fatty acids such as formate, propionate, butyrate, caproate, valerate, pyruvate, and lactate and on alcohols such as methanol, ethanol, and propanol. Sulfate, sulfite, and thiosulfate but not nitrate could be used as an electron acceptor. The temperature range for growth was 43 to 78°C; the spores were extremely heat resistant and survived 131°C for 20 min. The optimum pH was 7.0. The isolates grew well in salt concentrations ranging from 0 to 800 mmol of NaCl per liter. Sulfite reductase P582 was present, but cytochrome c and desulfoviridin were not found. Electron micrographs revealed a gram-positive cell organization. The isolates were classified as a Desulfotomaculum sp. on the basis of spore formation, general physiological characteristics, and submicroscopic organization. To detect thermophilic spore-forming sulfate-reducing bacteria in oil field water, polyvalent antisera raised against antigens from two isolates were used. These bacteria were shown to be widespread in oil field water from different platforms. The origin of thermophilic sulfate-reducing bacteria in the pore water of oil reservoirs is discussed. Images PMID:16348538

  11. Thermophilization of adult and juvenile tree communities in the northern tropical Andes

    PubMed Central

    Duque, Alvaro; Stevenson, Pablo R.; Feeley, Kenneth J.

    2015-01-01

    Climate change is expected to cause shifts in the composition of tropical montane forests towards increased relative abundances of species whose ranges were previously centered at lower, hotter elevations. To investigate this process of “thermophilization,” we analyzed patterns of compositional change over the last decade using recensus data from a network of 16 adult and juvenile tree plots in the tropical forests of northern Andes Mountains and adjacent lowlands in northwestern Colombia. Analyses show evidence that tree species composition is strongly linked to temperature and that composition is changing directionally through time, potentially in response to climate change and increasing temperatures. Mean rates of thermophilization [thermal migration rate (TMR), °C⋅y−1] across all censuses were 0.011 °C⋅y−1 (95% confidence interval = 0.002–0.022 °C⋅y−1) for adult trees and 0.027 °C⋅y−1 (95% confidence interval = 0.009–0.050 °C⋅y−1) for juvenile trees. The fact that thermophilization is occurring in both the adult and juvenile trees and at rates consistent with concurrent warming supports the hypothesis that the observed compositional changes are part of a long-term process, such as global warming, and are not a response to any single episodic event. The observed changes in composition were driven primarily by patterns of tree mortality, indicating that the changes in composition are mostly via range retractions, rather than range shifts or expansions. These results all indicate that tropical forests are being strongly affected by climate change and suggest that many species will be at elevated risk for extinction as warming continues. PMID:26261350

  12. Preservation of microbial communities enriched on lignocellulose under thermophilic and high-solid conditions

    DOE PAGESBeta

    Yu, Chaowei; Reddy, Amitha P.; Simmons, Christopher W.; Simmons, Blake A.; Singer, Steven W.; VanderGheynst, Jean S.

    2015-12-02

    Microbial communities enriched from diverse environments have shown considerable promise for the targeted discovery of microorganisms and enzymes for bioconversion of lignocellulose to liquid fuels. While preservation of microbial communities is important for commercialization and research, few studies have examined storage conditions ideal for preservation. The goal of this study was to evaluate the impact of preservation method on composition of microbial communities enriched on switchgrass before and after storage. The enrichments were completed in a high-solid and aerobic environment at 55 °C. Community composition was examined for each enrichment to determine when a stable community was achieved. Preservation methodsmore » included cryopreservation with the cryoprotective agents DMSO and glycerol, and cryopreservation without cryoprotective agents. Revived communities were examined for their ability to decompose switchgrass under high-solid and thermophilic conditions. High-throughput 16S rRNA gene sequencing of DNA extracted from enrichment samples showed that the majority of the shift in composition of the switchgrass-degrading community occurred during the initial three 2-week enrichments. Shifts in community structure upon storage occurred in all cryopreserved samples. Storage in liquid nitrogen in the absence of cryoprotectant resulted in variable preservation of dominant microorganisms in enriched samples. Cryopreservation with either DMSO or glycerol provided consistent and equivalent preservation of dominant organisms. In conclusion, a stable switchgrass-degrading microbial community was achieved after three 2-week enrichments. Dominant microorganisms were preserved equally well with DMSO and glycerol. DMSO-preserved communities required more incubation time upon revival to achieve pre-storage activity levels during high-solid thermophilic cultivation on switchgrass. Despite shifts in the community with storage, the samples were active upon revival

  13. Ultrasound-Mediated DNA Transformation in Thermophilic Gram-Positive Anaerobes

    PubMed Central

    Ji, Yuetong; He, Zhili; Pu, Yunting; Zhou, Jizhong; Xu, Jian

    2010-01-01

    Background Thermophilic, Gram-positive, anaerobic bacteria (TGPAs) are generally recalcitrant to chemical and electrotransformation due to their special cell-wall structure and the low intrinsic permeability of plasma membranes. Methodology/Principal Findings Here we established for any Gram-positive or thermophiles an ultrasound-based sonoporation as a simple, rapid, and minimally invasive method to genetically transform TGPAs. We showed that by applying a 40 kHz ultrasound frequency over a 20-second exposure, Texas red-conjugated dextran was delivered with 27% efficiency into Thermoanaerobacter sp. X514, a TGPA that can utilize both pentose and hexose for ethanol production. Experiments that delivered plasmids showed that host-cell viability and plasmid DNA integrity were not compromised. Via sonoporation, shuttle vectors pHL015 harboring a jellyfish gfp gene and pIKM2 encoding a Clostridium thermocellum β-1,4-glucanase gene were delivered into X514 with an efficiency of 6×102 transformants/µg of methylated DNA. Delivery into X514 cells was confirmed via detecting the kanamycin-resistance gene for pIKM2, while confirmation of pHL015 was detected by visualization of fluorescence signals of secondary host-cells following a plasmid-rescue experiment. Furthermore, the foreign β-1,4-glucanase gene was functionally expressed in X514, converting the host into a prototypic thermophilic consolidated bioprocessing organism that is not only ethanologenic but cellulolytic. Conclusions/Significance In this study, we developed an ultrasound-based sonoporation method in TGPAs. This new DNA-delivery method could significantly improve the throughput in developing genetic systems for TGPAs, many of which are of industrial interest yet remain difficult to manipulate genetically. PMID:20838444

  14. Preservation of microbial communities enriched on lignocellulose under thermophilic and high-solid conditions

    SciTech Connect

    Yu, Chaowei; Reddy, Amitha P.; Simmons, Christopher W.; Simmons, Blake A.; Singer, Steven W.; VanderGheynst, Jean S.

    2015-12-02

    Microbial communities enriched from diverse environments have shown considerable promise for the targeted discovery of microorganisms and enzymes for bioconversion of lignocellulose to liquid fuels. While preservation of microbial communities is important for commercialization and research, few studies have examined storage conditions ideal for preservation. The goal of this study was to evaluate the impact of preservation method on composition of microbial communities enriched on switchgrass before and after storage. The enrichments were completed in a high-solid and aerobic environment at 55 °C. Community composition was examined for each enrichment to determine when a stable community was achieved. Preservation methods included cryopreservation with the cryoprotective agents DMSO and glycerol, and cryopreservation without cryoprotective agents. Revived communities were examined for their ability to decompose switchgrass under high-solid and thermophilic conditions. High-throughput 16S rRNA gene sequencing of DNA extracted from enrichment samples showed that the majority of the shift in composition of the switchgrass-degrading community occurred during the initial three 2-week enrichments. Shifts in community structure upon storage occurred in all cryopreserved samples. Storage in liquid nitrogen in the absence of cryoprotectant resulted in variable preservation of dominant microorganisms in enriched samples. Cryopreservation with either DMSO or glycerol provided consistent and equivalent preservation of dominant organisms. In conclusion, a stable switchgrass-degrading microbial community was achieved after three 2-week enrichments. Dominant microorganisms were preserved equally well with DMSO and glycerol. DMSO-preserved communities required more incubation time upon revival to achieve pre-storage activity levels during high-solid thermophilic cultivation on switchgrass. Despite shifts in the community with storage, the samples were active upon revival under

  15. Thermophilization of adult and juvenile tree communities in the northern tropical Andes.

    PubMed

    Duque, Alvaro; Stevenson, Pablo R; Feeley, Kenneth J

    2015-08-25

    Climate change is expected to cause shifts in the composition of tropical montane forests towards increased relative abundances of species whose ranges were previously centered at lower, hotter elevations. To investigate this process of "thermophilization," we analyzed patterns of compositional change over the last decade using recensus data from a network of 16 adult and juvenile tree plots in the tropical forests of northern Andes Mountains and adjacent lowlands in northwestern Colombia. Analyses show evidence that tree species composition is strongly linked to temperature and that composition is changing directionally through time, potentially in response to climate change and increasing temperatures. Mean rates of thermophilization [thermal migration rate (TMR), °C ⋅ y(-1)] across all censuses were 0.011 °C ⋅ y(-1) (95% confidence interval = 0.002-0.022 °C ⋅ y(-1)) for adult trees and 0.027 °C ⋅ y(-1) (95% confidence interval = 0.009-0.050 °C ⋅ y(-1)) for juvenile trees. The fact that thermophilization is occurring in both the adult and juvenile trees and at rates consistent with concurrent warming supports the hypothesis that the observed compositional changes are part of a long-term process, such as global warming, and are not a response to any single episodic event. The observed changes in composition were driven primarily by patterns of tree mortality, indicating that the changes in composition are mostly via range retractions, rather than range shifts or expansions. These results all indicate that tropical forests are being strongly affected by climate change and suggest that many species will be at elevated risk for extinction as warming continues. PMID:26261350

  16. Effects of selected thermophilic microorganisms on crude oils at elevated temperatures and pressures

    SciTech Connect

    Premuzic, E.T.; Lin, M.S.

    1992-01-01

    The objective of this program is to determine the chemical and physical effects of thermophilic and thermoadapted organisms on crude oils and cores at elevated temperatures and pressures. Ultimately a data base will be generated which will be used in technical and economic feasibility studies leading to field applications. Progress to date are described for: construction of core-flooding systems; studies of trends in biochemical interactions between different microorganisms and crude oils; and comparative studies of interaction between different crude oils and microorganisms.

  17. Effects of selected thermophilic microorganisms on crude oils at elevated temperatures and pressures

    SciTech Connect

    Premuzic, E.T.; Lin, M.S.

    1992-10-01

    The objective of this program is to determine the chemical and physical effects of thermophilic and thermoadapted organisms on crude oils and cores at elevated temperatures and pressures. Ultimately a data base will be generated which will be used in technical and economic feasibility studies leading to field applications. Progress reports are presented for the following tasks: duration of biotreatment and media effects where comparative studies of microbial effects on different types of heavy oils are discussed; construction of core-flooding systems; and microscopic comparison of reaction mixtures.

  18. Draft Genome Sequence of Brevibacillus borstelensis cifa_chp40, a Thermophilic Strain Having Biotechnological Importance

    PubMed Central

    Tripathy, Swetaleena; Padhi, Soumesh Kumar; Sen, Rinil; Maji, Ushajyoti; Samanta, Mrinal; Mohanty, Sriprakash; Maiti, Nikhil Kumar

    2016-01-01

    Brevibacillus borstelensis cifa_chp40 is a thermophilic, strictly aerobic gram positive motile bacteria isolated from the alkaline hot water spring located in the Eastern Ghats zone of India. It could grow in a wide range of temperature and degrade low-density polythene at 37°C. The strain cifa_chp40 produces essential enzymes like protease, lipase, esterase and amidase at 50°C. Here, we report the draft genome sequence of B. borstelensis cifa_chp40 which will provide further insight into the metabolic capabilities, function and evolution of this important organism. PMID:26958091

  19. Complete Genome Sequence of the Thermophilic Bacterium Exiguobacterium sp. AT1b

    SciTech Connect

    Vishnivetskaya, T.; Lucas, Susan; Copeland, A; Lapidus, Alla L.; Glavina Del Rio, Tijana; Dalin, Eileen; Tice, Hope; Bruce, David; Goodwin, Lynne A.; Pitluck, Sam; Saunders, Elizabeth H; Brettin, Thomas S; Detter, J. Chris; Han, Cliff; Larimer, Frank W; Land, Miriam L; Hauser, Loren John; Kyrpides, Nikos C; Ovchinnikova, Galina; Kathariou, Sophia; Ramaley, Robert F.; Rodrigues, Debora F.; Hendrix, Christie; Richardson, Paul; Tiedje, James M.

    2011-01-01

    Here we present the genome of strain Exiguobacterium sp. AT1b, a thermophilic member of the genus Exiguobacterium whose representatives were isolated from various environments along a thermal and physico-chemical gradient. This genome was sequenced to be a comparative resource for study of thermal adaptation with a psychroactive representative of the genus, Exiguobacterium sibiricum strain 255-15, that was previously sequenced by the U.S. Department of Energy's (DOE) Joint Genome Institute (JGI) (http://genome.ornl.gov/microbial/exig/).

  20. Complete Genome Sequence of the Thermophilic Bacterium Exiguobacterium sp. AT1b

    SciTech Connect

    Vishnivetskaya, T.; Lucas, Susan; Copeland, A; Lapidus, Alla L; Glavina Del Rio, Tijana; Dalin, Eileen; Tice, Hope; Bruce, David; Goodwin, Lynne A.; Pitluck, Sam; Saunders, Elizabeth H; Brettin, Tom; Detter, J. Chris; Han, Cliff; Larimer, Frank W; Land, Miriam L; Hauser, Loren John; Kyrpides, Nikos C; Ovchinnikova, Galina; Kathariou, Sophia; Ramaley, Robert F.; Rodrigues, Debora F.; Hendrix, Christie; Richardson, Paul; Tiedje, James M.

    2011-01-01

    Here we present the genome of strain Exiguobacterium sp. AT1b, a thermophilic member of the genus Exiguobacterium whose representatives were isolated from various environments along a thermal and physicochemical gradient. This genome was sequenced to be a comparative resource for the study of thermal adaptation with a psychroactive representative of the genus, Exiguobacterium sibiricum strain 255-15, that was previously sequenced by the U.S. Department of Energy s (DOE s) Joint Genome Institute (JGI) (http://genome.ornl.gov/microbial/exig/).

  1. Temperature-Jump Fluorescence Provides Evidence for Fully Reversible Microsecond Dynamics in a Thermophilic Alcohol Dehydrogenase

    PubMed Central

    2015-01-01

    Protein dynamics on the microsecond (μs) time scale were investigated by temperature-jump fluorescence spectroscopy as a function of temperature in two variants of a thermophilic alcohol dehydrogenase: W87F and W87F:H43A. Both mutants exhibit a fast, temperature-independent μs decrease in fluorescence followed by a slower full recovery of the initial fluorescence. The results, which rule out an ionizing histidine as the origin of the fluorescence quenching, are discussed in the context of a Trp49-containing dimer interface that acts as a conduit for thermally activated structural change within the protein interior. PMID:26223665

  2. Pyrinomonas methylaliphatogenes gen. nov., sp. nov., a novel group 4 thermophilic member of the phylum Acidobacteria from geothermal soils.

    PubMed

    Crowe, M A; Power, J F; Morgan, X C; Dunfield, P F; Lagutin, K; Rijpstra, W I C; Rijpstra, I C; Vyssotski, G N S; Sinninghe Damste, J S; Houghton, K M; Ryan, J L J; Stott, M B

    2014-01-01

    An aerobic, thermophilic, moderately acidophilic non-spore-forming bacterium, strain K22(T), was isolated from geothermally heated soil at Mount Ngauruhoe, New Zealand. On the basis of 16S rRNA gene sequence similarity, K22(T) was shown to belong to subdivision 4 of the phylum Acidobacteria and to be most closely related to 'Candidatus Chloracidobacterium thermophilum' (86 %) and Blastocatella fastidiosa (86 %). Cells stained Gram-negative and were catalase and oxidase-positive. The major fatty acids detected were iso-C15 : 0, iso-C17 : 0, iso-C19 : 0 and iso-C21 : 0 when standard lipid extraction protocols were employed. Analysis of the total cell lipid acid hydrolysate also detected membrane-spanning and ether lipids, which made up approximately 40 % of the total membrane composition. These lipids included dicarboxylic (iso-diabolic) acid and the glyceryl ether of alkyl analogues of iso-C15 : 0 and iso-diabolic acid. The G+C content of the genomic DNA was 59.6 mol% and the primary respiratory quinone was MK-8. Strain K22(T) grew at 50-69 °C with an optimum temperature of 65 °C and at pH 4.1-7.8 with an optimum growth pH of 6.5. NaCl tolerance was up to 1 % (w/v). Cells displayed a chemoheterotrophic and obligately aerobic metabolism. Cells grew on nutrient broth, alginate, arabinose, Casamino acids, glucose, lactate, formate, mannose, sodium alginate, peptone, sucrose, tryptone, xanthan, xylan, xylose and yeast extract. Nitrogen sources included nitrate, ammonium, urea, yeast extract and Casamino acids, but not dinitrogen gas. The distinct phylogenetic position and the phenotypic characteristics separate strain K22(T) from all other members of the class Acidobacteria and indicate that it represents a novel species and genus, for which the name Pyrinomonas methylaliphatogenes gen. nov., sp. nov. is proposed. The type strain of the type species is K22(T) ( = DSM 25857(T) = ICMP 18710(T)). PMID:24048862

  3. Influence of mode of storage and drying of fodder on thermophilic actinomycete aerocontamination in dairy farms of the Doubs region of France.

    PubMed Central

    Dalphin, J C; Pernet, D; Reboux, G; Martinez, J; Dubiez, A; Barale, T; Depierre, A

    1991-01-01

    Airborne contamination by thermophilic actinomycetes, micromycetes and Gram negative bacteria was determined on 34 dairy farms and related to fodder drying and storage methods. Eighteen farms had a barn drying system, eight with additional heating; the remaining 16 had traditional fodder storage methods. Three air samples were obtained for each farm with a six stage Andersen sampler. The thermophilic actinomycetes were identified as Streptomyces and the dominant micromycetes as Aspergillus spp; there was no relation between the levels of these organisms. There were fewer thermophilic actinomycete colonies per Petri dish (stage 5 on the Anderson sampler) on farms with barn drying than on those with traditional storage (median (range) 7 (0-2628) and 56 (4-2628) respectively). The three farms where no thermophilic actinomycetes were found had barn drying with heating and the four most modern farms had lower thermophilic actinomycete colony counts than the others (median (range) 3 (0-10) and 48 (0-2628)). The level of thermophilic actinomycetes and, to a lesser degree, of micromycetes was higher where the farmer had farmer's lung. Thermophilic actinomycetes of the genus Streptomyces are probably the antigens associated with farmer's lung in the Doubs, and modern farms with barn drying and heating furnish some protection against this disease. PMID:1948788

  4. Changes of resistome, mobilome and potential hosts of antibiotic resistance genes during the transformation of anaerobic digestion from mesophilic to thermophilic.

    PubMed

    Tian, Zhe; Zhang, Yu; Yu, Bo; Yang, Min

    2016-07-01

    This study aimed to reveal how antibiotic resistance genes (ARGs) and their horizontal and vertical transfer-related items (mobilome and bacterial hosts) respond to the transformation of anaerobic digestion (AD) from mesophilic to thermophilic using one-step temperature increase. The resistomes and mobilomes of mesophilic and thermophilic sludge were investigated using metagenome sequencing, and the changes in 24 representative ARGs belonging to three categories, class 1 integron and bacterial genera during the transition period were further followed using quantitative PCR and 454-pyrosequencing. After the temperature increase, resistome abundance in the digested sludge decreased from 125.97 ppm (day 0, mesophilic) to 50.65 ppm (day 57, thermophilic) with the reduction of most ARG types except for the aminoglycoside resistance genes. Thermophilic sludge also had a smaller mobilome, including plasmids, insertion sequences and integrons, than that of mesophilic sludge, suggesting the lower horizontal transfer potential of ARGs under thermophilic conditions. On the other hand, the total abundance of 18 bacterial genera, which were suggested as the possible hosts for 13 ARGs through network analysis, decreased from 23.27% in mesophilic sludge to 11.92% in thermophilic sludge, indicating fewer hosts for the vertical expansion of ARGs after the increase in temperature. These results indicate that the better reduction of resistome abundance by thermophilic AD might be associated with the decrease of both the horizontal and vertical transferability of ARGs. PMID:27108212

  5. Molecular Characterization and Expression of a Phytase Gene from the Thermophilic Fungus Thermomyces lanuginosus

    PubMed Central

    Berka, Randy M.; Rey, Michael W.; Brown, Kimberly M.; Byun, Tony; Klotz, Alan V.

    1998-01-01

    The phyA gene encoding an extracellular phytase from the thermophilic fungus Thermomyces lanuginosus was cloned and heterologously expressed, and the recombinant gene product was biochemically characterized. The phyA gene encodes a primary translation product (PhyA) of 475 amino acids (aa) which includes a putative signal peptide (23 aa) and propeptide (10 aa). The deduced amino acid sequence of PhyA has limited sequence identity (ca. 47%) with Aspergillus niger phytase. The phyA gene was inserted into an expression vector under transcriptional control of the Fusarium oxysporum trypsin gene promoter and used to transform a Fusarium venenatum recipient strain. The secreted recombinant phytase protein was enzymatically active between pHs 3 and 7.5, with a specific activity of 110 μmol of inorganic phosphate released per min per mg of protein at pH 6 and 37°C. The Thermomyces phytase retained activity at assay temperatures up to 75°C and demonstrated superior catalytic efficiency to any known fungal phytase at 65°C (the temperature optimum). Comparison of this new Thermomyces catalyst with the well-known Aspergillus niger phytase reveals other favorable properties for the enzyme derived from the thermophilic gene donor, including catalytic activity over an expanded pH range. PMID:9797301

  6. Population dynamics during startup of thermophilic anaerobic digesters: the mixing factor.

    PubMed

    Ghanimeh, Sophia A; Saikaly, Pascal E; Li, Dong; El-Fadel, Mutasem

    2013-11-01

    Two thermophilic digesters were inoculated with manure and started-up under mixed and stagnant conditions. The Archaea in the mixed digester (A) were dominated by hydrogenotrophic Methanobateriaceae (61%) with most of the methane being produced via syntrophic pathways. Methanosarcinales (35%) were the only acetoclastic methanogens present. Acetate dissipation seems to depend on balanced hydrogenotrophic-to-acetotrophic abundance, which in turn was statistically correlated to free ammonia levels. Relative abundance of bacterial community was associated with the loading rate. However, in the absence of mixing (digester B), the relationship between microbial composition and operating parameters was not discernible. This was attributed to the development of microenvironments where environmental conditions are significantly different from average measured parameters. The impact of microenvironments was accentuated by the use of a non-acclimated seed that lacks adequate propionate degraders. Failure to disperse the accumulated propionate, and other organics, created high concentration niches where competitive and inhibiting conditions developed and favored undesired genera, such as Halobacteria (65% in B). As a result, digester B experienced higher acid levels and lower allowable loading rate. Mixing was found necessary to dissipate potential inhibitors, and improve stability and loading capacity, particularly when a non-acclimated seed, often lacking balanced thermophilic microflora, is used. PMID:23830181

  7. Microbial population dynamics in a thermophilic methane digester fed with garbage.

    PubMed

    Cheon, J; Hong, F; Hidaka, T; Koshikawa, H; Tsuno, H

    2007-01-01

    The diversity of microbial communities in three full-scale thermophilic anaerobic digesters which treated garbage, sewage sludge and livestock wastes (hereafter called TGD, TSD and TLD, respectively) was investigated using 16S rDNA clone libraries in triplicate. The population dynamics of TGD were also studied. The purposes were to show the microbial diversity in each reactor and to suggest which key microbes in a thermophilic methane digester fed with garbage, including a check of reproducibility and the suggestion of an error range in this molecular biology method. 736 clones were identified, and the maximum error was estimated to be around +/-10% for the same OTU (operational taxonomic unit) and for most detected OTUs. The most frequently detected OTU shows a close relationship to Uncultured bacterium clone MBA08, Unidentified bacterium clone TUG22 and Uncultured archaeal symbiont PA204 in TGD, TSD and TLD, respectively. The microbial population dynamics in TGD were studied over a period of 90 days, and the occupying ratios of Bacillus infernus and Methanothermobacter wolfeii were shown to change with the change in VFA concentration. From the dynamic change and characteristics of the microbes, it is concluded that Bacillus infernus and Methanothermobacter wolfeii played an important role and were recommended as key microbes in TGD. PMID:17564383

  8. Methanogenic pathway and community structure in a thermophilic anaerobic digestion process of organic solid waste.

    PubMed

    Sasaki, Daisuke; Hori, Tomoyuki; Haruta, Shin; Ueno, Yoshiyuki; Ishii, Masaharu; Igarashi, Yasuo

    2011-01-01

    The methanogenic pathway and microbial community in a thermophilic anaerobic digestion process of organic solid waste were investigated in a continuous-flow stirred-tank reactor using artificial garbage slurry as a feedstock. The decomposition pathway of acetate, a significant precursor of CH(4) and a key intermediate metabolite in the anaerobic digestion process, was analyzed by using stable isotopes. A tracer experiment using (13)C-labeled acetate revealed that approximately 80% of the acetate was decomposed via a non-aceticlastic oxidative pathway, whereas the remainder was converted to methane via an aceticlastic pathway. Archaeal 16S rRNA analyses demonstrated that the hydrogenotrophic methanogens Methanoculleus spp. accounted for >90% of detected methanogens, and the aceticlastic methanogens Methanosarcina spp. were the minor constituents. The clone library targeting bacterial 16S rRNA indicated the predominance of the novel Thermotogales bacterium (relative abundance: ~53%), which is related to anaerobic acetate oxidizer Thermotoga lettingae TMO, although the sequence similarity was low. Uncultured bacteria that phylogenetically belong to municipal solid waste cluster I were also predominant in the microflora (~30%). These results imply that the microbial community in the thermophilic degrading process of organic solid waste consists exclusively of unidentified bacteria, which efficiently remove acetate through a non-aceticlastic oxidative pathway. PMID:20851673

  9. Continuous thermophilic composting (CTC) for rapid biodegradation and maturation of organic municipal solid waste.

    PubMed

    Xiao, Yong; Zeng, Guang-Ming; Yang, Zhao-Hui; Shi, Wen-Jun; Huang, Cui; Fan, Chang-Zheng; Xu, Zheng-Yong

    2009-10-01

    Fewer and fewer municipal solid wastes are treated by composting in China because of the disadvantages of enormous investment, long processing cycle and unstable products in a conventional composting treatment. In this study, a continuous thermophilic composting (CTC) method, only a thermophilic phase within the process, has been applied to four bench-scale composting runs, and further compared with a conventional composting run by assessing the indexes of pH, total organic carbon (TOC), total Kjeldahl nitrogen (TKN), C/N ratio, germination index (GI), specific oxygen uptake rate (SOUR), dissolved organic carbon (DOC) and dehydrogenase activity. After composting for 14 days, 16 days, 18 days and 19 days in the four CTC runs, respectively, mature compost products were obtained, with quality similar to or better than which had been stabilized for 28 days in run A. The products from the CTC runs also showed favorable stability in room temperature environment after the short-term composting at high temperature. The study suggested CTC as a novel method for rapid degradation and maturation of organic municipal solid wastes. PMID:19487122

  10. Performance comparison between mesophilic and thermophilic anaerobic reactors for treatment of palm oil mill effluent.

    PubMed

    Jeong, Joo-Young; Son, Sung-Min; Pyon, Jun-Hyeon; Park, Joo-Yang

    2014-08-01

    The anaerobic digestion of palm oil mill effluent (POME) was carried out under mesophilic (37°C) and thermophilic (55°C) conditions without long-time POME storage in order to compare the performance of each condition in the field of Sumatra Island, Indonesia. The anaerobic treatment system was composed of anaerobic hybrid reactor and anaerobic baffled filter. Raw POME was pretreated by screw decanter to reduce suspended solids and residual oil. The total COD removal rate of 90-95% was achieved in both conditions at the OLR of 15kg[COD]/m(3)/d. The COD removal in thermophilic conditions was slightly better, however the biogas production was much higher than that in the mesophilic one at high OLR. The organic contents in pretreated POME were highly biodegradable in mesophilic under the lower OLRs. The biogas production was 13.5-20.0l/d at the 15kg[COD]/m(3)/d OLR, and the average content of carbon dioxide was 5-35% in both conditions. PMID:24797939

  11. Mesophilic versus thermophilic anaerobic digestion of cattle manure: methane productivity and microbial ecology.

    PubMed

    Moset, Veronica; Poulsen, Morten; Wahid, Radziah; Højberg, Ole; Møller, Henrik Bjarne

    2015-09-01

    In this study, productivity and physicochemical and microbiological (454 sequencing) parameters, as well as environmental criteria, were investigated in anaerobic reactors to contribute to the ongoing debate about the optimal temperature range for treating animal manure, and expand the general knowledge on the relation between microbiological and physicochemical process indicators. For this purpose, two reactor sizes were used (10 m(3) and 16 l), in which two temperature conditions (35°C and 50°C) were tested. In addition, the effect of the hydraulic retention time was evaluated (16 versus 20 days). Thermophilic anaerobic digestion showed higher organic matter degradation (especially fiber), higher pH and higher methane (CH₄) yield, as well as better percentage of ultimate CH₄ yield retrieved and lower residual CH₄ emission, when compared with mesophilic conditions. In addition, lower microbial diversity was found in the thermophilic reactors, especially for Bacteria, where a clear intensification towards Clostridia class members was evident. Independent of temperature, some similarities were found in digestates when comparing with animal manure, including low volatile fatty acids concentrations and a high fraction of Euryarchaeota in the total microbial community, in which members of Methanosarcinales dominated for both temperature conditions; these indicators could be considered a sign of process stability. PMID:25737010

  12. Comparative performance of mesophilic and thermophilic anaerobic digestion for high-solid sewage sludge.

    PubMed

    Hidaka, Taira; Wang, Feng; Togari, Taketo; Uchida, Tsutomu; Suzuki, Yutaka

    2013-12-01

    In local cities, many small sewage and waste treatment facilities are operated independently. To encourage processing by anaerobic digestion at a centralized sewage treatment plant (STP), high-solid sewage sludge is helpful because it reduces the energy and cost required for transporting the sludge from other STPs. Mesophilic and thermophilic anaerobic digestion of sewage sludge at total solids concentrations (TS) of 7.5% and 10% were evaluated using laboratory-scale continuous reactors. Under the mesophilic condition, sewage sludge of 10% TS was successfully treated. Under the thermophilic condition, sewage sludge of 7.5% TS was not successfully treated when the total ammonia concentration was over 2000 mg N/L. Batch experiments showed that it takes a few weeks for the methane fermentation activity to recover after being inhibited. The effectiveness of adding easily biodegradable organic matter was confirmed. These results show that high-solid sewage sludge is suitable for small facilities by controlling the operating conditions. PMID:24096284

  13. Aerobic moving bed biofilm reactor treating thermomechanical pulping whitewater under thermophilic conditions.

    PubMed

    Jahren, Sigrun J; Rintala, Jukka A; Odegaard, Hallvard

    2002-02-01

    The continuously operated laboratory scale Kaldnes moving bed biofilm reactor (MBBR) was used for thermophilic (55 degrees C) aerobic treatment of TMP whitewater. In the MBBR, the biomass is grown on carrier elements that move along with the water in the reactor. Inoculation with mesophilic activated sludge gave 60-65% SCOD removal from the first day onwards. During the 107 days of experiment, the 60-65% SCOD removals were achieved at organic loading rates of 2.5-3.5 kg SCODm(-3) d(-1), the highest loading rates applied during the run and HRT of 13-22h. Carbohydrates, which contributed to 50-60% of the influent SCOD. were removed by 90-95%, while less than 15% of the lignin-like material (30-35% of SCODin) was removed. The sludge yield was 0.23g VSSg SCOD(-1)removed. The results show that the aerobic biofilm process can be successfully operated under thermophilic conditions. PMID:11848344

  14. Effects of lipids on thermophilic anaerobic digestion and reduction of lipid inhibition upon addition of bentonite.

    PubMed

    Angelidaki, I; Petersen, S P; Ahring, B K

    1990-07-01

    The effect of bentonite-bound oil on thermophilic anaerobic digestion of cattle manure was investigated. In digestor experiments, addition of oil was found to be inhibitory during start-up and the inhibitory effect was less pronounced when the oil was added in the form of bentonite-bound oil compared to when the oil was added alone. After adaptation of the digestors, very rapid degradation of oil was observed and more than 80% of the oil was degraded within a few hours after daily feeding. In batch experiments, glyceride trioleate was found to be inhibitory to thermophilic anaerobic digestion when the concentrations were higher than 2.0 g/l. However, addition of bentonite (a clay mineral) at concentrations of 0.15% and 0.45% was found to partly overcome this inhibition. Addition of calcium chloride in concentration of 3 mM (0.033% w/v) showed a similar positive effect on the utilization of oil, but the effect was lower than with bentonite. PMID:1366749

  15. [Numerical taxonomy of a thermophilic "Bacillus" species isolated from West African rice soils (author's transl)].

    PubMed

    Garcia, J L; Roussos, S; Bensoussan, M; Bianchi, A; Mandel, M

    1982-01-01

    Fifty-seven strains of endospore-forming thermophilic bacteria, 37 of which were capable of denitrification, were isolated from rice soils of West Africa. They were compared with 17 strains of similar bacteria from culture collections, utilizing a total of 123 morphological, physiological and biochemical characteristics. A numerical analysis was performed using the complete linkage-clustering method and the Khi2 test. Seventy-five percent (55 strains) could be included in 12 groups at a taxonomic distance of 0.015. Wild strains of denitrifiers issued in phenons 8 to 12 and strains of phenon 4 (not denitrifying) were related to the named strains of phenons 1 and 7 (Bacillus stearothermophilus). Twenty-two wild strains, and 5 strains from culture collections, were only thermotolerating without growth at 65 degrees C. The strains of phenon 3 were related to the 3 named strains of B. coagulans. Phenons 5 and 6 were composed of strains related to B. circulans. The strains of phenon 2 denitrified and showed a swollen central endospore; they were closely related to B. brevis. The denitrifying thermophilic strains isolated from rice soils (phenons 8 to 12) were related to the first group (B. kaustophilus) of Walker and Wolf but their base compositions of DNA were significantly different from those found for the reference strains. PMID:7103312

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

    PubMed

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

    2016-11-01

    Proof of principle of biohythane and potential energy production from garden waste (GW) is demonstrated in this study in a two-step process coupling dark fermentation and anaerobic digestion. The synergistic effect of using co-cultures of extreme thermophiles to intensify biohydrogen dark fermentation is demonstrated using xylose, cellobiose and GW. Co-culture of Caldicellulosiruptor saccharolyticus and Thermotoga maritima showed higher hydrogen production yields from xylose (2.7±0.1molmol(-1) total sugar) and cellobiose (4.8±0.3molmol(-1) total sugar) compared to individual cultures. Co-culture of extreme thermophiles C. saccharolyticus and Caldicellulosiruptor bescii increased synergistically the hydrogen production yield from GW (98.3±6.9Lkg(-1) (VS)) compared to individual cultures and co-culture of T. maritima and C. saccharolyticus. The biochemical methane potential of the fermentation end-products was 322±10Lkg(-1) (CODt). Biohythane, a biogas enriched with 15% hydrogen could be obtained from GW, yielding a potential energy generation of 22.2MJkg(-1) (VS). PMID:27484669

  17. Mesophilic versus thermophilic anaerobic digestion of cattle manure: methane productivity and microbial ecology

    PubMed Central

    Moset, Veronica; Poulsen, Morten; Wahid, Radziah; Højberg, Ole; Møller, Henrik Bjarne

    2015-01-01

    In this study, productivity and physicochemical and microbiological (454 sequencing) parameters, as well as environmental criteria, were investigated in anaerobic reactors to contribute to the ongoing debate about the optimal temperature range for treating animal manure, and expand the general knowledge on the relation between microbiological and physicochemical process indicators. For this purpose, two reactor sizes were used (10 m3 and 16 l), in which two temperature conditions (35°C and 50°C) were tested. In addition, the effect of the hydraulic retention time was evaluated (16 versus 20 days). Thermophilic anaerobic digestion showed higher organic matter degradation (especially fiber), higher pH and higher methane (CH4) yield, as well as better percentage of ultimate CH4 yield retrieved and lower residual CH4 emission, when compared with mesophilic conditions. In addition, lower microbial diversity was found in the thermophilic reactors, especially for Bacteria, where a clear intensification towards Clostridia class members was evident. Independent of temperature, some similarities were found in digestates when comparing with animal manure, including low volatile fatty acids concentrations and a high fraction of Euryarchaeota in the total microbial community, in which members of Methanosarcinales dominated for both temperature conditions; these indicators could be considered a sign of process stability. PMID:25737010

  18. Antimicrobial Protein Candidates from the Thermophilic Geobacillus sp. Strain ZGt-1: Production, Proteomics, and Bioinformatics Analysis

    PubMed Central

    Alkhalili, Rawana N.; Bernfur, Katja; Dishisha, Tarek; Mamo, Gashaw; Schelin, Jenny; Canbäck, Björn; Emanuelsson, Cecilia; Hatti-Kaul, Rajni

    2016-01-01

    A thermophilic bacterial strain, Geobacillus sp. ZGt-1, isolated from Zara hot spring in Jordan, was capable of inhibiting the growth of the thermophilic G. stearothermophilus and the mesophilic Bacillus subtilis and Salmonella typhimurium on a solid cultivation medium. Antibacterial activity was not observed when ZGt-1 was cultivated in a liquid medium; however, immobilization of the cells in agar beads that were subjected to sequential batch cultivation in the liquid medium at 60 °C showed increasing antibacterial activity up to 14 cycles. The antibacterial activity was lost on protease treatment of the culture supernatant. Concentration of the protein fraction by ammonium sulphate precipitation followed by denaturing polyacrylamide gel electrophoresis separation and analysis of the gel for antibacterial activity against G. stearothermophilus showed a distinct inhibition zone in 15–20 kDa range, suggesting that the active molecule(s) are resistant to denaturation by SDS. Mass spectrometric analysis of the protein bands around the active region resulted in identification of 22 proteins with molecular weight in the range of interest, three of which were new and are here proposed as potential antimicrobial protein candidates by in silico analysis of their amino acid sequences. Mass spectrometric analysis also indicated the presence of partial sequences of antimicrobial enzymes, amidase and dd-carboxypeptidase. PMID:27548162

  19. Role of Mn2+ and compatible solutes in the radiation resistance of thermophilic bacteria and archaea.

    PubMed

    Webb, Kimberly M; DiRuggiero, Jocelyne

    2012-01-01

    Radiation-resistant bacteria have garnered a great deal of attention from scientists seeking to expose the mechanisms underlying their incredible survival abilities. Recent analyses showed that the resistance to ionizing radiation (IR) in the archaeon Halobacterium salinarum is dependent upon Mn-antioxidant complexes responsible for the scavenging of reactive oxygen species (ROS) generated by radiation. Here we examined the role of the compatible solutes trehalose, mannosylglycerate, and di-myo-inositol phosphate in the radiation resistance of aerobic and anaerobic thermophiles. We found that the IR resistance of the thermophilic bacteria Rubrobacter xylanophilus and Rubrobacter radiotolerans was highly correlated to the accumulation of high intracellular concentration of trehalose in association with Mn, supporting the model of Mn(2+)-dependent ROS scavenging in the aerobes. In contrast, the hyperthermophilic archaea Thermococcus gammatolerans and Pyrococcus furiosus did not contain significant amounts of intracellular Mn, and we found no significant antioxidant activity from mannosylglycerate and di-myo-inositol phosphate in vitro. We therefore propose that the low levels of IR-generated ROS under anaerobic conditions combined with highly constitutively expressed detoxification systems in these anaerobes are key to their radiation resistance and circumvent the need for the accumulation of Mn-antioxidant complexes in the cell. PMID:23209374

  20. Acquired thermotolerance and heat shock in the extremely thermophilic archaebacterium Sulfolobus sp. strain B12

    SciTech Connect

    Trent, J.D.; Osipiuk, J.; Pinkau, T. )

    1990-03-01

    The extreme thermophile Sulfolobus sp. strain B12 exhibits an acquired thermotolerance response. Thus, survival of cells from a 70{degrees}C culture at the lethal temperature of 92{degrees}C was enhanced by as much as 6 orders of magnitude over a 2-h period if the culture was preheated to 88{degrees}C for 60 min or longer before being exposed to the lethal temperature. In eubacteria and eucaryotes, acquired thermotolerance correlates with the induced synthesis of a dozen or so proteins known as heat shock proteins. In this Sulfolobus species, it correlates with the preferential synthesis of primarily one major protein (55 kilodaltons) and, to a much lesser extent, two minor proteins (28 and 35 kilodaltons). Since the synthesis of all other proteins was radically reduced and these proteins were apparently not degraded or exported, their relative abundance within the cell increased during the time the cells were becoming thermotolerant. They could not yet be related to known heat shock proteins. In immunoassays, they were not cross-reactive with antibodies against heat shock proteins from Escherichia coli (DnaK and GroE), which are highly conserved between eubacteria and eucaryotes. However, it appears that if acquired thermotolerance depends on the synthesis of protective proteins, then in this extremely thermophilic archaebacterium it depends primarily on one protein.

  1. [Screening and Enzyme Production Characteristics of Thermophilic Cellulase-producing Strains].

    PubMed

    Feng, Hong-mei; Qin, Yong-sheng; Li, Xiao-fan; Zhou, Jin-xing; Peng, Xia-wei

    2016-04-15

    A total of 6 thermophilic cellulase-producing strainswere isolated from organic garden waste mixed chicken composting at thermophilic period. These isolates were identified as Streptomyces thermoviolaceus, S. thermodiastaticus, S. thermocarboxydus, S. albidoflavus, S. thermovulgaris and Brevibacillus borstelensis through 16S rRNA gene sequence alignment and phylogenetic tree analysis. The cellulose-degrading microbial community has been investigated in few researches so far both at home and abroad. In this study, the mixed strains M-1 was made up of the 6 cellulose-decomposing microorganisms. The CMCase activity of the mixed strains M- 1 was stronger than any of the 6 single strains. Production of CMCase from mixed strains M-1 was studied by optimizing different physico-chemical parameters. The Maximum CMCase production (135.9 U · mL⁻¹) of strains M-1 was achieved at 45 °C in a liquid medium (pH 4) inoculated with 1% (volume fraction), containing a mixture of wheat bran and starch, corn flour and KNO₃. After optimization of separation conditions, CMCase production capacity was improved by 1.8 times. PMID:27548981

  2. Cloning and expression of an actinokinase gene from a thermophilic Streptomyces in Escherechia coli.

    PubMed

    Chitte, R R; Dey, S

    2001-05-01

    A thermophilic Streptomyces megasporus strain SD5, could secrete a new fibrinolytic (actinokinase) at 55 degrees C. The gene (ackS) encoding actinokinase was isolated from the chromosomal DNA of S. megasporus SD5 and cloned in different hosts and vectors. The expression was obtained in E. coli JM109 using Cla I linearized pBR322 as vector (pSR 500). The recombinant E. coli containing pSR 500 expressed active actinokinase but the expression was low and the recombinant was unstable in liquid culture. Deletion analysis revealed that removal of Bam H I-Sal I fragment from down stream and Cla I-EcoRI from upsream enhanced the stability and expression of ackS in both solid and liquid media. For over expresion, the ackS gene was cloned in E. coli C 600 using Bam HI linearized pT7-7. This seemed to be the most suitable host vector system. The recombinant and native form of actinokinase exhibited similar characteristics. Actinokinase was the first thrombolytic enzyme from a thermophile to be cloned and over expressed in a mesophilic heterologous expression system. PMID:11510122

  3. Biochemical and Structural Characterization of Enolase from Chloroflexus aurantiacus: Evidence for a Thermophilic Origin

    PubMed Central

    Zadvornyy, Oleg A.; Boyd, Eric S.; Posewitz, Matthew C.; Zorin, Nikolay A.; Peters, John W.

    2015-01-01

    Enolase catalyzes the conversion of 2-phosphoglycerate to phosphoenolpyruvate during both glycolysis and gluconeogenesis, and is required by all three domains of life. Here, we report the purification and biochemical and structural characterization of enolase from Chloroflexus aurantiacus, a thermophilic anoxygenic phototroph affiliated with the green non-sulfur bacteria. The protein was purified as a homodimer with a subunit molecular weight of 46 kDa. The temperature optimum for enolase catalysis was 80°C, close to the measured thermal stability of the protein which was determined to be 75°C, while the pH optimum for enzyme activity was 6.5. The specific activities of purified enolase determined at 25 and 80°C were 147 and 300 U mg−1 of protein, respectively. Km values for the 2-phosphoglycerate/phosphoenolpyruvate reaction determined at 25 and 80°C were 0.16 and 0.03 mM, respectively. The Km values for Mg2+ binding at these temperatures were 2.5 and 1.9 mM, respectively. When compared to enolase from mesophiles, the biochemical and structural properties of enolase from C. aurantiacus are consistent with this being thermally adapted. These data are consistent with the results of our phylogenetic analysis of enolase, which reveal that enolase has a thermophilic origin. PMID:26082925

  4. Thermophilic campylobacters in surface waters around Lancaster, UK: negative correlation with Campylobacter infections in the community.

    PubMed

    Jones, K; Betaieb, M; Telford, D R

    1990-11-01

    The incidence of campylobacter enteritis in Lancaster City Health Authority is three times the UK average for similar sizes of population and has marked seasonal peaks in May and June. Environmental monitoring of surface waters around Lancaster showed that thermophilic campylobacters were absent from drinking water from the fells and from the clean upper reaches of the River Conder but were present in the main rivers entering Morecambe Bay, the lower reaches of the River Conder, the Lancaster canal, and seawater from the Lune estuary and Morecambe Bay. All the surface waters tested showed the same seasonality, namely, higher numbers in the winter months and low numbers or none in May, June and July. The absence of thermophilic campylobacters in the summer months may be due to high sunshine levels because experiments on the effects of light showed that campylobacters in sewage effluent and seawater were eliminated within 60 and 30 min of daylight respectively but survived for 24 h in darkness. As the concentrations of campylobacters in surface waters were at their lowest precisely at the time of peak infections in the community it is unlikely that surface waters form Lancaster's reservoir of campylobacter infection for the community. PMID:2276990

  5. Substrate-Specific Development of Thermophilic Bacterial Consortia by Using Chemically Pretreated Switchgrass

    PubMed Central

    Eichorst, Stephanie A.; Joshua, Chijioke; Sathitsuksanoh, Noppadon; Singh, Seema; Simmons, Blake A.

    2014-01-01

    Microbial communities that deconstruct plant biomass have broad relevance in biofuel production and global carbon cycling. Biomass pretreatments reduce plant biomass recalcitrance for increased efficiency of enzymatic hydrolysis. We exploited these chemical pretreatments to study how thermophilic bacterial consortia adapt to deconstruct switchgrass (SG) biomass of various compositions. Microbial communities were adapted to untreated, ammonium fiber expansion (AFEX)-pretreated, and ionic-liquid (IL)-pretreated SG under aerobic, thermophilic conditions using green waste compost as the inoculum to study biomass deconstruction by microbial consortia. After microbial cultivation, gravimetric analysis of the residual biomass demonstrated that both AFEX and IL pretreatment enhanced the deconstruction of the SG biomass approximately 2-fold. Two-dimensional nuclear magnetic resonance (2D-NMR) experiments and acetyl bromide-reactive-lignin analysis indicated that polysaccharide hydrolysis was the dominant process occurring during microbial biomass deconstruction, and lignin remaining in the residual biomass was largely unmodified. Small-subunit (SSU) rRNA gene amplicon libraries revealed that although the dominant taxa across these chemical pretreatments were consistently represented by members of the Firmicutes, the Bacteroidetes, and Deinococcus-Thermus, the abundance of selected operational taxonomic units (OTUs) varied, suggesting adaptations to the different substrates. Combining the observations of differences in the community structure and the chemical and physical structure of the biomass, we hypothesize specific roles for individual community members in biomass deconstruction. PMID:25261509

  6. Treatment of high strength pharmaceutical wastewaters in a Thermophilic Aerobic Membrane Reactor (TAMR).

    PubMed

    Collivignarelli, M C; Abbà, A; Bertanza, G

    2014-10-15

    In the present work we studied the thermophilic biological treatability of high strength liquid wastes from a pharmaceutical industry (rich in organic matter - COD: Chemical Oxygen Demand, nutrients and salinity). Different mixtures (with concentrations of COD, phosphorus and chloride up to 57,000 mg L(-1), 2000 mg L(-1) and 9000 mg L(-1), respectively) were tested. The pilot plant used in this work was designed and built with dimensions comparable to a semi-industrial unit. The results are therefore representative for full-scale applications. During four months of experimentation, the pilot plant (TAMR - Thermophilic Aerobic Membrane Reactor) was operated at 49 ± 1 °C and the organic loading rate was 1.5-5.5 kgCOD m(-3) d(-1) with a hydraulic retention time of 5-10 days. Main results are the following: a) extremely high COD removal rate (up to 98%); b) very low sludge production (∼0.016 kgVSS produced kgCOD removed(-1)); c) suitability as a pre-treatment to a conventional (e.g. activated sludge) biological treatment (the studied pharmaceutical industrial wastewaters are discharged into the sewer system for final polishing in a centralized municipal wastewater treatment plant) and d) high phosphorus removal (up to 99%). PMID:25000201

  7. Immunomagnetically Captured Thermophilic Sulfate-Reducing Bacteria from North Sea Oil Field Waters

    PubMed Central

    Christensen, Bjørn; Torsvik, Terje; Lien, Torleiv

    1992-01-01

    Immunomagnetic beads (IMB) were used to recover thermophilic sulfate-reducing bacteria from oil field waters from oil production platforms in the Norwegian sector of the North Sea. IMB coated with polyclonal antibodies against whole-cell antigens of the thermophilic Thermodesulfobacterium mobile captured strains GFA1, GFA2, and GFA3. GFA1 was serologically and morphologically identical to T. mobile. GFA2 and GFA3 were spore forming and similar to the Desulfotomaculum strains T90A and T93B previously isolated from North Sea oil field waters by a classical enrichment procedure. Western blots (immunoblots) of whole cells showed that GFA2, GFA3, T90A, and T93B are different serotypes of the same Desulfotomaculum species. Monoclonal antibodies (MAb) against T. mobile type strain cells were produced and used as capture agents on IMB. These MAb, named A4F4, were immunoglobulin M; they were specific to T. mobile and directed against lipopolysaccharides. The prevailing cells immunocaptured with MAb A4F4 were morphologically and serologically similar to T. mobile type strain cells. T. mobile was not detected in these oil field waters by classical enrichment procedures. Furthermore, extraction with antibody-coated IMB allowed pure strains to be isolated directly from primary enrichment cultures without prior time-consuming subculturing and consecutive transfers to selective media. Images PMID:16348693

  8. Thermophilic production of polyhydroxyalkanoates by a novel Aneurinibacillus strain isolated from Gudao oilfield, China.

    PubMed

    Xiao, Zijun; Zhang, Yu; Xi, Lijun; Huo, Fangfang; Zhao, Jing-yi; Li, Jing

    2015-09-01

    Polyhydroxyalkanoates (PHAs) are usually biosynthesized using mesophilic strains, but the fermentation processes often suffer from bacterial contamination. This work reports the screening of thermophilic bacteria capable of producing PHAs under elevated temperatures to reduce the contamination risk. Strain XH2 was isolated from an oilfield and identified as Aneurinibacillus sp. by morphology, physiological-biochemical characterization, and 16S rDNA phylogenetic analysis. This strain can produce PHA granules, which was detected by Nile red staining and transmission electron microscopic imaging. At 55 °C, 111.6 mg l(-1) of PHA was produced in a fermentation medium containing glucose, peptone, and yeast extract. If peptone was removed from the medium, the yield of PHA would be enhanced by 2.4 times. The main monomers of the PHA product were identified to be 3-hydroxybutyrate and 3-hydroxyvalerate with a molar ratio of 17.2:1 by gas chromatography-mass spectroscopy (GC-MS) and nuclear magnetic resonance analyses. Two minor homologues, 3-hydroxyoctanoate, and 3-hydroxy-4-phenylbutanoate, were tentatively identified by GC-MS as well. This is the first report of thermophilic PHA bacterial producer from the Firmicutes phylum. PMID:25832555

  9. Effect of fillers on key characteristics of sludge thermophilic anaerobic digestion.

    PubMed

    Shao, Liming; Xu, Yuanshun; Wang, Tianfeng; Lü, Fan; He, Pinjing

    2015-10-01

    In anaerobic digestion (AD) of sludge, AD efficiency and digested sludge (DS) dewaterability are critical factors. In this study, polyester non-woven fabric fillers were integrated into a sludge digester. The effect of such fillers on digestion was investigated in thermophilic temperature range in semi-continuous mode. Methane production of filler system and control reactor were significantly different (P < 0.05, paired t-test). At hydraulic retention times of 18 days and 12 days, the corresponding methane yields from filler system were 140% and 161%, respectively, of the yields from control digester without filler. Improvement of DS dewaterability was uncertain during 110 days of operation. While after a longer period of digestion, filler system resulted in a lower normalized capillary suction time of DS (76.5 ± 21.6 s L/g total suspended solids) than control reactor (118.7 ± 32.9 s L/g total suspended solids). The results showed that the filler could improve thermophilic AD performance, except at too short hydraulic retention times. PMID:26151853

  10. Isolation and Characterization of Thermophilic Bacilli Degrading Cinnamic, 4-Coumaric, and Ferulic Acids

    PubMed Central

    Peng, Xue; Misawa, Norihiko; Harayama, Shigeaki

    2003-01-01

    Thirty-four thermophilic Bacillus sp. strains were isolated from decayed wood bark and a hot spring water sample based on their ability to degrade vanillic acid under thermophilic conditions. It was found that these bacteria were able to degrade a wide range of aromatic acids such as cinnamic, 4-coumaric, 3-phenylpropionic, 3-(p-hydroxyphenyl)propionic, ferulic, benzoic, and 4-hydroxybenzoic acids. The metabolic pathways for the degradation of these aromatic acids at 60°C were examined by using one of the isolates, strain B1. Benzoic and 4-hydroxybenzoic acids were detected as breakdown products from cinnamic and 4-coumaric acids, respectively. The β-oxidative mechanism was proposed to be responsible for these conversions. The degradation of benzoic and 4-hydroxybenzoic acids was determined to proceed through catechol and gentisic acid, respectively, for their ring fission. It is likely that a non-β-oxidative mechanism is the case in the ferulic acid catabolism, which involved 4-hydroxy-3-methoxyphenyl-β-hydroxypropionic acid, vanillin, and vanillic acid as the intermediates. Other strains examined, which are V0, D1, E1, G2, ZI3, and H4, were found to have the same pathways as those of strain B1, except that strains V0, D1, and H4 had the ability to transform 3-hydroxybenzoic acid to gentisic acid, which strain B1 could not do. PMID:12620824

  11. Some Properties of Acid Protease from the Thermophilic Fungus, Penicillium duponti K1014

    PubMed Central

    Hashimoto, Hikotaka; Iwaasa, Takashi; Yokotsuka, Tamotsu

    1973-01-01

    A purified acid protease from a true thermophilic fungus, Penicillium duponti K1014, was most active at pH 2.5 for milk casein and at pH 3.0 for hemoglobin. The enzyme was stable at a pH range of 2.5 to 6.0 at 30 C for 20 h. The acid protease retained full activity after 1 h at 60 C at a pH range between 3.5 and 5.5. At the most stable pH of 4.5, more than 65% of its activity remained after heat treatment for 1 h at 70 C. These thermal properties show the enzyme as a thermophilic protein. The enzyme activity was strongly inhibited by sodium lauryl sulfate and oxidizing reagents such as potassium permanganate and N-bromosuccinimide. No inhibition was caused by chelating reagents, potato inhibitor, and those reagents which convert sulfhydryl groups to mercaptides. Reducing reagents showed an activating effect. The enzyme showed the trypsinogen-activating property at an acidic pH range; optimal trypsinogen activation was obtained at a pH of approximately 3.0. The isoelectric point of the enzyme was estimated to be pH 3.89 by disk electrofocusing. By using gel filtration, an approximate value of 41,000 was estimated for the molecular weight. PMID:4699217

  12. [Effects of Thermophilic Composting on Antibiotic Resistance Genes (ARGs) of Swine Manure Source].

    PubMed

    Zheng, Ning-guo; Huang, Nan; Wang, Wei-wei; Yu, Man; Chen, Xiao-yang; Yao, Yan-lai; Wang, Wei-ping; Hong, Chun-lai

    2016-05-15

    To investigate the effects of thermophilic composting process on antibiotic resistance genes (ARGs) of swine manure source at a field scale, the abundance of four erythromycin resistance genes (ermA, ermB, ermC and ermF), three β-lactam resistance genes (blaTEM, blaCTX and blaSHV) and two quinolone resistance genes (qnrA and qnrS) were quantified by quantitative PCR ( qPCR) during the composting process. The results suggested that the erm genes' copy numbers were significantly higher than those of the bla and qnr genes in the early stage of composting (P < 0.01). The maximum abundance of erm genes was ermB (9.88 x 10⁸ copies · g⁻¹), following by ermF (9.4 x 10⁸ copies · g⁻¹). At the end of the composting process, bla and qnr genes were at low levels, while erm genes were still at high levels. Even through ermF was proliferated comparing with the initial copies. These results indicated that thermophilic composting process could not effectively remove all ARGs. For some ARGs, compost may be a good bioreactor resulting in their proliferation. Application of composting products on farmland may cause transference of ARGs. PMID:27506057

  13. Characterization of DNA transport in the thermophilic bacterium Thermus thermophilus HB27.

    PubMed

    Schwarzenlander, Cornelia; Averhoff, Beate

    2006-09-01

    Horizontal gene transfer has been a major force for genome plasticity over evolutionary history, and is largely responsible for fitness-enhancing traits, including antibiotic resistance and virulence factors. In particular, for adaptation of prokaryotes to extreme environments, lateral gene transfer seems to have played a crucial role. Recently, by performing a genome-wide mutagenesis approach with Thermus thermophilus HB27, we identified the first genes in a thermophilic bacterium for the uptake of free DNA, a process called natural transformation. Here, we present the first data on the biochemistry and bioenergetics of the DNA transport process in this thermophile. We report that linear and circular plasmid DNA are equally well taken up with a high maximal velocity of 1.5 microg DNA.(mg protein)(-1).min(-1), demonstrating an extremely efficient binding and uptake rate of 40 kb.s(-1).cell(-1). Uncouplers and ATPase inhibitors immediately inhibited DNA uptake, providing clear evidence that DNA translocation in HB27 is an energy-dependent process. DNA uptake studies with genomic DNA of Bacteria, Archaea and Eukarya revealed that Thermus thermophilus HB27 takes up DNA from members of all three domains of life. We propose that the extraordinary broad substrate specificity of the highly efficient Thermus thermophilus HB27 DNA uptake system may contribute significantly to thermoadaptation of Thermus thermophilus HB27 and to interdomain DNA transfer in hot environments. PMID:16939619

  14. Substrate-Specific Development of Thermophilic Bacterial Consortia by Using Chemically Pretreated Switchgrass.

    PubMed

    Eichorst, Stephanie A; Joshua, Chijioke; Sathitsuksanoh, Noppadon; Singh, Seema; Simmons, Blake A; Singer, Steven W

    2014-12-01

    Microbial communities that deconstruct plant biomass have broad relevance in biofuel production and global carbon cycling. Biomass pretreatments reduce plant biomass recalcitrance for increased efficiency of enzymatic hydrolysis. We exploited these chemical pretreatments to study how thermophilic bacterial consortia adapt to deconstruct switchgrass (SG) biomass of various compositions. Microbial communities were adapted to untreated, ammonium fiber expansion (AFEX)-pretreated, and ionic-liquid (IL)-pretreated SG under aerobic, thermophilic conditions using green waste compost as the inoculum to study biomass deconstruction by microbial consortia. After microbial cultivation, gravimetric analysis of the residual biomass demonstrated that both AFEX and IL pretreatment enhanced the deconstruction of the SG biomass approximately 2-fold. Two-dimensional nuclear magnetic resonance (2D-NMR) experiments and acetyl bromide-reactive-lignin analysis indicated that polysaccharide hydrolysis was the dominant process occurring during microbial biomass deconstruction, and lignin remaining in the residual biomass was largely unmodified. Small-subunit (SSU) rRNA gene amplicon libraries revealed that although the dominant taxa across these chemical pretreatments were consistently represented by members of the Firmicutes, the Bacteroidetes, and Deinococcus-Thermus, the abundance of selected operational taxonomic units (OTUs) varied, suggesting adaptations to the different substrates. Combining the observations of differences in the community structure and the chemical and physical structure of the biomass, we hypothesize specific roles for individual community members in biomass deconstruction. PMID:25261509

  15. X-ray Crystallographic Structure of Thermophilic Rhodopsin: IMPLICATIONS FOR HIGH THERMAL STABILITY AND OPTOGENETIC FUNCTION.

    PubMed

    Tsukamoto, Takashi; Mizutani, Kenji; Hasegawa, Taisuke; Takahashi, Megumi; Honda, Naoya; Hashimoto, Naoki; Shimono, Kazumi; Yamashita, Keitaro; Yamamoto, Masaki; Miyauchi, Seiji; Takagi, Shin; Hayashi, Shigehiko; Murata, Takeshi; Sudo, Yuki

    2016-06-01

    Thermophilic rhodopsin (TR) is a photoreceptor protein with an extremely high thermal stability and the first characterized light-driven electrogenic proton pump derived from the extreme thermophile Thermus thermophilus JL-18. In this study, we confirmed its high thermal stability compared with other microbial rhodopsins and also report the potential availability of TR for optogenetics as a light-induced neural silencer. The x-ray crystal structure of TR revealed that its overall structure is quite similar to that of xanthorhodopsin, including the presence of a putative binding site for a carotenoid antenna; but several distinct structural characteristics of TR, including a decreased surface charge and a larger number of hydrophobic residues and aromatic-aromatic interactions, were also clarified. Based on the crystal structure, the structural changes of TR upon thermal stimulation were investigated by molecular dynamics simulations. The simulations revealed the presence of a thermally induced structural substate in which an increase of hydrophobic interactions in the extracellular domain, the movement of extracellular domains, the formation of a hydrogen bond, and the tilting of transmembrane helices were observed. From the computational and mutational analysis, we propose that an extracellular LPGG motif between helices F and G plays an important role in the thermal stability, acting as a "thermal sensor." These findings will be valuable for understanding retinal proteins with regard to high protein stability and high optogenetic performance. PMID:27129243

  16. Nicotinamidase from the thermophilic archaeon Acidilobus saccharovorans: structural and functional characteristics.

    PubMed

    Stekhanova, T N; Bezsudnova, E Y; Mardanov, A V; Osipov, E M; Ravin, N V; Skryabin, K G; Popov, V O

    2014-01-01

    Nicotinamidase is involved in the maintenance of NAD+ homeostasis and in the NAD+ salvage pathway of most prokaryotes, and it is considered as a possible drug target. The gene (ASAC_0847) encoding a hypothetical nicotinamidase has been found in the genome of the thermophilic archaeon Acidilobus saccharovorans. The product of this gene, NA_As0847, has been expressed in Escherichia coli, isolated, and characterized as a Fe(2+)-containing nicotinamidase (k(cat)/K(m) = 427 mM(-1)·sec(-1))/pyrazinamidase (k(cat)/K(m) = 331 mM(-1)·sec(-1)). NA_As0847 is a homodimer with molecular mass 46.4 kDa. The enzyme has high thermostability (T(1/2) (60°C) = 180 min, T(1/2) (80°C) = 35 min) and thermophilicity (T(opt) = 90°C, E(a) = 30.2 ± 1.0 kJ/mol) and broad pH interval of activity, with the optimum at pH 7.5. Special features of NA_As0847 are the presence of Fe2+ instead of Zn2+ in the active site of the enzyme and inhibition of the enzyme activity by Zn2+ at micromolar concentrations. Analysis of the amino acid sequence revealed a new motif of the metal-binding site (DXHXXXDXXEXXXWXXH) for homological archaeal nicotinamidases. PMID:24512664

  17. Mesophilic and thermophilic anaerobic laboratory-scale digestion of Nannochloropsis microalga residues.

    PubMed

    Kinnunen, H V; Koskinen, P E P; Rintala, J

    2014-03-01

    This paper studies methane production using a marine microalga, Nannochloropsis sp. residue from biodiesel production. Residue cake from Nannochloropsis, oils wet-extracted, had a methane potential of 482LCH4kg(-1) volatile solids (VS) in batch assays. However, when dry-extracted, the methane potential of residue cake was only 194LCH4kg(-1) VS. In semi-continuous reactor trials with dry-extracted residue cake, a thermophilic reactor produced 48% higher methane yield (220LCH4kg(-1)VS) than a mesophilic reactor (149LCH4kg(-1)VS). The thermophilic reactor was apparently inhibited due to ammonia with organic loading rate (OLR) of 2kgVSm(-3)d(-1) (hydraulic retention time (HRT) 46d), whereas the mesophilic reactor performed with OLR of 3kgVSm(-3)d(-1) (HRT 30d). Algal salt content did not inhibit digestion. Additional methane (18-33% of primary digester yield) was produced during 100d post-digestion. PMID:24462882

  18. Decreasing ammonia inhibition in thermophilic methanogenic bioreactors using carbon fiber textiles.

    PubMed

    Sasaki, Kengo; Morita, Masahiko; Hirano, Shin-ichi; Ohmura, Naoya; Igarashi, Yasuo

    2011-05-01

    Ammonia accumulation is one of the main causes of the loss of methane production observed during fermentation. We investigated the effect of addition of carbon fiber textiles (CFT) to thermophilic methanogenic bioreactors with respect to ammonia tolerance during the process of degradation of artificial garbage slurry, by comparing the performance of the reactors containing CFT with the performance of reactors without CFT. Under total ammonia-N concentrations of 3,000 mg L(-1), the reactors containing CFT were found to mediate stable removal of organic compounds and methane production. Under these conditions, high levels of methanogenic archaea were retained at the CFT, as determined by 16S rRNA gene analysis for methanogenic archaea. In addition, Methanobacterium sp. was found to be dominant in the suspended fraction, and Methanosarcina sp. was dominant in the retained fraction of the reactors with CFT. However, the reactors without CFT had lower rates of removal of organic compounds and production of methane under total ammonia-N concentrations of 1,500 mg L(-1). Under this ammonia concentration, a significant accumulation of acetate was observed in the reactors without CFT (130.0 mM), relative to the reactors with CFT (4.2 mM). Only Methanobacterium sp. was identified in the reactors without CFT. These results suggest that CFT enables stable proliferation of aceticlastic methanogens by preventing ammonia inhibition. This improves the process of stable garbage degradation and production of methane in thermophilic bioreactors that include high levels of ammonia. PMID:21468711

  19. The formation of illite from nontronite by mesophilic and thermophilic bacterial reaction

    USGS Publications Warehouse

    Jaisi, D.P.; Eberl, D.D.; Dong, H.; Kim, J.

    2011-01-01

    The formation of illite through the smectite-to-illite (S-I) reaction is considered to be one of the most important mineral reactions occurring during diagenesis. In biologically catalyzed systems, however, this transformation has been suggested to be rapid and to bypass the high temperature and long time requirements. To understand the factors that promote the S-I reaction, the present study focused on the effects of pH, temperature, solution chemistry, and aging on the S-I reaction in microbially mediated systems. Fe(III)-reduction experiments were performed in both growth and non-growth media with two types of bacteria: mesophilic (Shewanella putrefaciens CN32) and thermophilic (Thermus scotoductus SA-01). Reductive dissolution of NAu-2 was observed and the formation of illite in treatment with thermophilic SA-01 was indicated by X-ray diffraction (XRD) and high-resolution transmission electron microscopy (HRTEM). A basic pH (8.4) and high temperature (65??C) were the most favorable conditions forthe formation of illite. A long incubation time was also found to enhance the formation of illite. K-nontronite (non-permanent fixation of K) was also detected and differentiated from the discrete illite in the XRD profiles. These results collectively suggested that the formation of illite associated with the biologically catalyzed smectite-to-illite reaction pathway may bypass the prolonged time and high temperature required for the S-I reaction in the absence of microbial activity.

  20. Development and application of a simulation model for the thermophilic oxic process for treating swine waste.

    PubMed

    Jeon, Kyoungho; Nakano, Kazunori; Nishimura, Osamu

    2011-01-01

    The thermophilic oxic process (TOP) is a composting process that enables simultaneous complete decomposition and evaporation of organic waste under high temperature conditions supported by well-balanced calorific value control. To develop the simulation model for TOP, three-dimensional relationships among decomposition rate constant, temperature (20-70 °C) and moisture content (30-70%) were determined for swine waste and cooking oil based on the oxygen consumption rate during a thermophilic oxic decomposition reaction. The decomposition rate of swine waste and cooking oil under various moisture contents was described by the Arrhenius equation. The optimal temperature and moisture content were 60 °C and 60% for swine waste and 60 °C and 50% for cooking oil, respectively. The simulation model for TOP was constructed on the basis of the carbon, heat, and moisture balance. The validation of the simulation model was examined by comparing the measured temperature in the TOP reactor to that estimated by the simulation. The simulation model was proven by comparing experimental and calculated values. The relationship between the injection calorific value and the process mechanism of TOP was interpreted by the simulation model. On the basis of their relationship during TOP, the appropriate process conditions were discussed. PMID:20934200

  1. Isolation and characterization of an H/sub 2/-oxidizing thermophilic methanogen

    SciTech Connect

    Ferguson, T.J.; Mah, R.A.

    1983-01-01

    A thermophilic methanogen was isolated from enrichment cultures originally inoculated with sludge from an anaerobic kelp digester (55 degrees C). This isolate exhibited a temperature optimum of 55 to 60 degrees C and a maximum near 70 degrees C. Growth occurred throughout the pH range of 5.5 to 9.0, with optimal growth near pH 7.2. Altough 4% salt was present in the isolation medium, salt was not required for optimal growth. The thermophile utilized formate or H/sub 2/CO/sub 2/ but not acetate, methanol, or methylamines for growth and methanogenesis. Growth in complex medium was very rapid, and a minimum doubling time of 1.8 hours was recorded in media supplemented with rumen fluid. Growth in defined media required the addition of acetate and an unknown factor(s) from digester supernatant, rumen fluid, or Trypticase. Cells in liquid culture were oval to coccoid, 0.7 to 1.8 ..mu.. meters in diameter, often occurring in pairs. The cells were easily lysed upon exposure to oxygen or 0.08 mg of sodium dodecyl sulfate per ml. The isolate was sensitive to tetracycline and chloramphenicol but not penicillin G or cycloserine. The DNA base composition was 59.69 mol% guanine plus cytosine. (Refs. 34).

  2. Antimicrobial Protein Candidates from the Thermophilic Geobacillus sp. Strain ZGt-1: Production, Proteomics, and Bioinformatics Analysis.

    PubMed

    Alkhalili, Rawana N; Bernfur, Katja; Dishisha, Tarek; Mamo, Gashaw; Schelin, Jenny; Canbäck, Björn; Emanuelsson, Cecilia; Hatti-Kaul, Rajni

    2016-01-01

    A thermophilic bacterial strain, Geobacillus sp. ZGt-1, isolated from Zara hot spring in Jordan, was capable of inhibiting the growth of the thermophilic G. stearothermophilus and the mesophilic Bacillus subtilis and Salmonella typhimurium on a solid cultivation medium. Antibacterial activity was not observed when ZGt-1 was cultivated in a liquid medium; however, immobilization of the cells in agar beads that were subjected to sequential batch cultivation in the liquid medium at 60 °C showed increasing antibacterial activity up to 14 cycles. The antibacterial activity was lost on protease treatment of the culture supernatant. Concentration of the protein fraction by ammonium sulphate precipitation followed by denaturing polyacrylamide gel electrophoresis separation and analysis of the gel for antibacterial activity against G. stearothermophilus showed a distinct inhibition zone in 15-20 kDa range, suggesting that the active molecule(s) are resistant to denaturation by SDS. Mass spectrometric analysis of the protein bands around the active region resulted in identification of 22 proteins with molecular weight in the range of interest, three of which were new and are here proposed as potential antimicrobial protein candidates by in silico analysis of their amino acid sequences. Mass spectrometric analysis also indicated the presence of partial sequences of antimicrobial enzymes, amidase, and dd-carboxypeptidase. PMID:27548162

  3. Modelling of the acid base properties of two thermophilic bacteria at different growth times

    NASA Astrophysics Data System (ADS)

    Heinrich, Hannah T. M.; Bremer, Phil J.; McQuillan, A. James; Daughney, Christopher J.

    2008-09-01

    Acid-base titrations and electrophoretic mobility measurements were conducted on the thermophilic bacteria Anoxybacillus flavithermus and Geobacillus stearothermophilus at two different growth times corresponding to exponential and stationary/death phase. The data showed significant differences between the two investigated growth times for both bacterial species. In stationary/death phase samples, cells were disrupted and their buffering capacity was lower than that of exponential phase cells. For G. stearothermophilus the electrophoretic mobility profiles changed dramatically. Chemical equilibrium models were developed to simultaneously describe the data from the titrations and the electrophoretic mobility measurements. A simple approach was developed to determine confidence intervals for the overall variance between the model and the experimental data, in order to identify statistically significant changes in model fit and thereby select the simplest model that was able to adequately describe each data set. Exponential phase cells of the investigated thermophiles had a higher total site concentration than the average found for mesophilic bacteria (based on a previously published generalised model for the acid-base behaviour of mesophiles), whereas the opposite was true for cells in stationary/death phase. The results of this study indicate that growth phase is an important parameter that can affect ion binding by bacteria, that growth phase should be considered when developing or employing chemical models for bacteria-bearing systems.

  4. Cellulose degradation and ethanol production by thermophilic bacteria using mineral growth medium

    SciTech Connect

    Ahn, Hyung-Jun; Lynd, L.R.

    1996-12-31

    Growth of thermophilic cellulase-utilizing bacteria in a vitimin-free growth medium is reported for both a previously described strain, Clostridium thermoclelum 31549, and now isolates HJA1 and HJA2. Formation of fermentation products with and without vitamins was similar for strains HJA1 and HJA2 as well as for the enrichment cultures from which these stains were derived. Strain HJA2 was maintained in continous culture on a vitamin-free mineral medium with Avicel as the carbon source for over a week. At a 38 h residence time, Avicel conversion was higher (81%) at pH 6.42 than at pH 6.97 (73%) or at 6.01 (58%). Ethanol and acetate were produced in significant amounts by strain JHA2 at all pH values tested (6.97, 6.42, 6.01). Lactic acid was the primary fermentation product at pH 6.97, but was not a significant product at both the lower values. Efforts to grow thermophilic, cellulose-utilizing bacteria at pH < 6.0 were unsuccessful for described strains, new isolates, and enrichment cultures. 4 refs., 4 tabs.

  5. LIGHT WATER MODERATED NEUTRONIC REACTOR

    DOEpatents

    Christy, R.F.; Weinberg, A.M.

    1957-09-17

    A uranium fuel reactor designed to utilize light water as a moderator is described. The reactor core is in a tank at the bottom of a substantially cylindrical cross-section pit, the core being supported by an apertured grid member and comprised of hexagonal tubes each containing a pluralily of fuel rods held in a geometrical arrangement between end caps of the tubes. The end caps are apertured to permit passage of the coolant water through the tubes and the fuel elements are aluminum clad to prevent corrosion. The tubes are hexagonally arranged in the center of the tank providing an amulus between the core and tank wall which is filled with water to serve as a reflector. In use, the entire pit and tank are filled with water in which is circulated during operation by coming in at the bottom of the tank, passing upwardly through the grid member and fuel tubes and carried off near the top of the pit, thereby picking up the heat generated by the fuel elements during the fission thereof. With this particular design the light water coolant can also be used as the moderator when the uranium is enriched by fissionable isotope to an abundance of U/sup 235/ between 0.78% and 2%.

  6. Moderate hyperhomocysteinemia and immune activation.

    PubMed

    Schroecksnadel, K; Frick, B; Wirleitner, B; Winkler, C; Schennach, H; Fuchs, D

    2004-02-01

    Moderate hyperhomocysteinemia is associated with an increased risk of atherosclerosis, thrombosis and neurodegenerative diseases. Homocysteine accumulation in the blood can be due to many underlying causes, which may interact with each other, e.g. genetic disposition and B-vitamin status. The role of the sulfur-containing amino acid homocysteine in the pathogenesis of diseases remains unclear, even if many studies suggest a causal relationship between homocysteine-mediated processes like oxidative stress, NO-inactivation and endothelial deficiency and atherogenesis. Proposed mechanisms of action of homocysteine are discussed, and the question is addressed, whether effects that are attributed to homocysteine, are not rather the consequence of folate and vitamin B12-deficiency. Deficiency of these B-vitamins in parallel with moderate hyperhomocysteinemia is often found in patients with enhanced activation of the cellular immune system, like Alzheimer's disease, rheumatoid arthritis and also vascular diseases. In patients with these diseases an association between homocysteine metabolism, oxidative stress and immune activation exists. On the one hand proliferation of immunocompetent cells having an enhanced demand for B-vitamins leads to the accumulation of homocysteine. On the other hand macrophages stimulated by TH1-type cytokine interferon-gamma form reactive oxygen species (ROS), which oxidize antioxidants, lipoproteins and oxidation-sensitive B-vitamins. Thereby Th1-type immune response could contribute importantly to the development of hyperhomocysteinemia, and may also be a major determinant of disease progression. PMID:14965213

  7. Mesophilic and thermophilic anaerobic biodegradability of water hyacinth pre-treated at 80 {sup o}C

    SciTech Connect

    Ferrer, Ivet; Campos, Elena; Flotats, Xavier

    2010-10-15

    Water hyacinth (Eichornia crassipes) is a fast growing aquatic plant which causes environmental problems in continental water bodies. Harvesting and handling this plant becomes an issue, and focus has been put on the research of treatment alternatives. Amongst others, energy production through biomethanation has been proposed. The aim of this study was to assess the anaerobic biodegradability of water hyacinth under mesophilic and thermophilic conditions. The effect of a thermal sludge pre-treatment at 80 {sup o}C was also evaluated. To this end, anaerobic biodegradability tests were carried out at 35 {sup o}C and 55 {sup o}C, with raw and pre-treated water hyacinth. According to the results, the thermal pre-treatment enhanced the solubilisation of water hyacinth (i.e. increase in the soluble to total chemical oxygen demand (COD)) from 4% to 12% after 30 min. However, no significant effect was observed on the methane yields (150-190 L CH{sub 4}/kg volatile solids). Initial methane production rates for thermophilic treatments were two fold those of mesophilic ones (6-6.5 L vs. 3-3.5 L CH{sub 4}/kg COD.day). Thus, higher methane production rates might be expected from thermophilic reactors working at short retention times. The study of longer low temperature pre-treatments or pre-treatments at elevated temperatures coupled to thermophilic reactors should be considered in the future.

  8. Effect of recycling thermophilic sludge on the activated-sludge process. Report for October 1984-June 1989

    SciTech Connect

    Prakasam, T.B.S.; Soszynski, S.; Zenz, D.R.; Lue-Hing, C.; Blyth, L.

    1990-01-01

    A full-scale investigation was undertaken at Chicago's Hanover Park Water Reclamation Plant (WRP) to study whether the net sludge production from the WRP could be reduced by implementing a scheme developed by W. Torpey et al. (1984). In this process, sludge is withdrawn from a thermophilic digester operated in series with a mesophilic digester and partially recycled into the aeration tanks of the activated sludge system. The Hanover Park WRP, which has a design flow capacity of 45,420 million cu m per day (12 mgd), was split into a control section and an experimental section. The two sections were operated similarly except the thermophilic drawoff was recycled into the aeration tanks of the experimental section from a digester system consisting of mesophilic and thermophilic digesters operated in series. A reduction in net sludge production of about 10 percent was achieved at 40 percent recycle of thermophilic sludge into the aeration tanks at the Hanover Park WRP. This was a much smaller reduction than obtained by Torpey et al. Sludge dewatering properties were not improved. The recycling produced no adverse effects on quality of the treated wastewater.

  9. Draft Genome Sequence of a Thermophilic Cyanobacterium from the Family Oscillatoriales (Strain MTP1) from the Chalk River, Colorado

    PubMed Central

    Grogger, Melanie; Mraz, Megan; Veverka, Donald

    2016-01-01

    The draft genome (57.7% GC, 7,647,882 bp) of the novel thermophilic cyanobacterium MTP1 was determined by metagenomics of an enrichment culture. The genome shows that it is in the family Oscillatoriales and encodes multiple heavy metal resistances as well as the capacity to make exopolysaccharides. PMID:26893415

  10. Draft Genome Sequence of a Thermophilic Cyanobacterium from the Family Oscillatoriales (Strain MTP1) from the Chalk River, Colorado.

    PubMed

    Hallenbeck, Patrick C; Grogger, Melanie; Mraz, Megan; Veverka, Donald

    2016-01-01

    The draft genome (57.7% GC, 7,647,882 bp) of the novel thermophilic cyanobacterium MTP1 was determined by metagenomics of an enrichment culture. The genome shows that it is in the family Oscillatoriales and encodes multiple heavy metal resistances as well as the capacity to make exopolysaccharides. PMID:26893415

  11. Cold moderators for pulsed neutron sources

    SciTech Connect

    Carpenter, J.M.

    1990-01-01

    This paper reviews cold moderators in pulsed neutron sources and provides details of the performance of different cold moderator materials and configurations. Analytical forms are presented which describe wavelength spectra and emission time distributions. Several types of cooling arrangements used in pulsed source moderators are described. Choices of materials are surveyed. The author examines some of the radiation damage effects in cold moderators, including the phenomenon of burping'' in irradiated cold solid methane. 9 refs., 15 figs., 4 tabs.

  12. Low to moderate temperature nanolaminate heater

    DOEpatents

    Eckels, J. Del; Nunes, Peter J.; Simpson, Randall L.; Hau-Riege, Stefan; Walton, Chris; Carter, J. Chance; Reynolds, John G.

    2011-01-11

    A low to moderate temperature heat source comprising a high temperature energy source modified to output low to moderate temperatures wherein the high temperature energy source modified to output low to moderate temperatures is positioned between two thin pieces to form a close contact sheath. In one embodiment the high temperature energy source modified to output low to moderate temperatures is a nanolaminate multilayer foil of reactive materials that produces a heating level of less than 200.degree. C.

  13. Complete genome of the cellulolytic thermophile Acidothermus cellulolyticus 11B provides insights into its ecophysiological and evolutionary adaptations

    SciTech Connect

    Xie, Gary; Detter, John C; Bruce, David C; Challacombe, Jean F; Brettin, Thomas S; Necsulea, Anamaria; Daubin, Vincent; Medigue, Claudine; Adney, William S; Xu, Xin C; Lapidus, Alla; Pujic, Pierre; Berry, Alison M; Barabote, Ravi D; Leu, David; Normand, Phillipe

    2009-01-01

    We present here the complete 2.4 MB genome of the actinobacterial thermophile, Acidothermus cellulolyticus 11B, that surprisingly reveals thermophilic amino acid usage in only the cytosolic subproteome rather than its whole proteome. Thermophilic amino acid usage in the partial proteome implies a recent, ongoing evolution of the A. cellulolyticus genome since its divergence about 200-250 million years ago from its closest phylogenetic neighbor Frankia, a mesophilic plant symbiont. Differential amino acid usage in the predicted subproteomes of A. cellulolyticus likely reflects a stepwise evolutionary process of modern thermophiles in general. An unusual occurrence of higher G+C in the non-coding DNA than in the transcribed genome reinforces a late evolution from a higher G+C common ancestor. Comparative analyses of the A. cellulolyticus genome with those of Frankia and other closely-related actinobacteria revealed that A. cellulolyticus genes exhibit reciprocal purine preferences at the first and third codon positions, perhaps reflecting a subtle preference for the dinucleotide AG in its mRNAs, a possible adaptation to a thermophilic environment. Other interesting features in the genome of this cellulolytic, hot-springs dwelling prokaryote reveal streamlining for adaptation to its specialized ecological niche. These include a low occurrence of pseudo genes or mobile genetic elements, a flagellar gene complement previously unknown in this organism, and presence of laterally-acquired genomic islands of likely ecophysiological value. New glycoside hydrolases relevant for lignocellulosic biomass deconstruction were identified in the genome, indicating a diverse biomass-degrading enzyme repertoire several-fold greater than previously characterized, and significantly elevating the industrial value of this organism.

  14. Complete genome of the cellulolytic thermophile Acidothermus cellulolyticus 11B provides insights into its ecophysiological and evolutionary adaptations

    SciTech Connect

    Xie, Gary; Detter, Chris; Bruce, David; Challacome, Jean F; Brettin, Thomas S; Barabote, Ravi D; Leu, David; Normand, Philippe; Necsula, Anamaria; Daubin, Vincent; Medigue, Claudine; Xu, Xin C; Lapidus, Alla; Pujic, Pierre; Richardson, Paul; Berry, Alison M

    2008-01-01

    We present here the complete 2.4 MB genome of the actinobacterial thermophile, Acidothermus cellulolyticus lIB, that surprisingly reveals thermophilic amino acid usage in only the cytosolic subproteome rather than its whole proteome. Thermophilic amino acid usage in the partial proteome implies a recent, ongoing evolution of the A. cellulolyticus genome since its divergence about 200-250 million years ago from its closest phylogenetic neighbor Frankia, a mesophilic plant symbiont. Differential amino acid usage in the predicted subproteomes of A. cellulolyticus likely reflects a stepwise evolutionary process of modern thermophiles in general. An unusual occurrence of higher G+C in the non-coding DNA than in the transcribed genome reinforces a late evolution from a higher G+C common ancestor. Comparative analyses of the A. cellulolyticus genome with those of Frankia and other closely-related actinobacteria revealed that A. cellulolyticus genes exhibit reciprocal purine preferences at the first and third codon positions, perhaps reflecting a subtle preference for the dinucleotide AG in its mRNAs, a possible adaptation to a thermophilic environment. Other interesting features in the genome of this cellulolytic, hot-springs dwelling prokaryote reveal streamlining for adaptation to its specialized ecological niche. These include a low occurrence of pseudogenes or mobile genetic elements, a flagellar gene complement previously unknown in this organism, and presence of laterally-acquired genomic islands of likely ecophysiological value. New glycoside hydrolases relevant for lignocellulosic biomass deconstruction were identified in the genome, indicating a diverse biomass-degrading enzyme repertoire several-fold greater than previously characterized, and significantly elevating the industrial value of this organism.

  15. High rate mesophilic, thermophilic, and temperature phased anaerobic digestion of waste activated sludge: a pilot scale study.

    PubMed

    Bolzonella, David; Cavinato, Cristina; Fatone, Francesco; Pavan, Paolo; Cecchi, Franco

    2012-06-01

    The paper reports the findings of a two-year pilot scale experimental trial for the mesophilic (35°C), thermophilic (55°C) and temperature phased (65+55°C) anaerobic digestion of waste activated sludge. During the mesophilic and thermophilic runs, the reactor operated at an organic loading rate of 2.2 kgVS/m(3)d and a hydraulic retention time of 20 days. In the temperature phased run, the first reactor operated at an organic loading rate of 15 kgVS/m(3)d and a hydraulic retention time of 2 days while the second reactor operated at an organic loading rate of 2.2 kgVS/m(3)d and a hydraulic retention time of 18 days (20 days for the whole temperature phased system). The performance of the reactor improved with increases in temperature. The COD removal increased from 35% in mesophilic conditions, to 45% in thermophilic conditions, and 55% in the two stage temperature phased system. As a consequence, the specific biogas production increased from 0.33 to 0.45 and to 0.49 m(3)/kgVS(fed) at 35, 55, and 65+55°C, respectively. The extreme thermophilic reactor working at 65°C showed a high hydrolytic capability and a specific yield of 0.33 g COD (soluble) per gVS(fed). The effluent of the extreme thermophilic reactor showed an average concentration of soluble COD and volatile fatty acids of 20 and 9 g/l, respectively. Acetic and propionic acids were the main compounds found in the acids mixture. Because of the improved digestion efficiency, organic nitrogen and phosphorus were solubilised in the bulk. Their concentration, however, did not increase as expected because of the formation of salts of hydroxyapatite and struvite inside the reactor. PMID:22305642

  16. Deep-Subterranean Microbial Habitats in the Hishikari Epithermal Gold Mine: Active Thermophilic Microbial Communities and Endolithic Ancient Microbial Relicts.

    NASA Astrophysics Data System (ADS)

    Hirayama, H.; Takai, K.; Inagaki, F.; Horikoshi, K.

    2001-12-01

    Deep subterranean microbial community structures in an epithermal gold-silver deposit, Hishikari gold mine, southern part of Kyusyu Japan, were evaluated through the combined use of enrichment culture methods and culture-independent molecular surveys. The geologic setting of the Hishikari deposit is composed of three lithologies; basement oceanic sediments of the Cretaceous Shimanto Supergroup, Quaternary andesites, and auriferous quartz vein. We studied the drilled core rock of these, and the geothermal hot waters from the basement aquifers collected by means of the dewatering system located at the deepest level in the mining sites. Culture-independent molecular phylogenetic analyses of PCR-amplified ribosomal DNA (rDNA) recovered from drilled cores suggested that the deep-sea oceanic microbial communities were present as ancient indigenous relicts confined in the Shimanto basement. On the other hand, genetic signals of active thermophilic microbial communities, mainly consisting of thermophilic hydrogen-oxidizer within Aquificales, thermophilic methanotroph within g-Proteobacteria and yet-uncultivated bacterium OPB37 within b-Proteobacteria, were detected with these of oceanic relicts from the subterranean geothermal hot aquifers (temp. 70-100ºC). Successful cultivation and FISH analyses strongly supported that these thermophilic lithotrophic microorganisms could be exactly active and they grew using geochemically produced hydrogen and methane gasses as nutrients. Based on these results, the deep-subsurface biosphere occurring in the Hishikari epithermal gold mine was delineated as endolithic ancient microbial relicts and modern habitats raising active lithotrophic thermophiles associated with the geological and geochemical features of the epithermal gold deposit.

  17. MODERATOR ELEMENTS FOR UNIFORM POWER NUCLEAR REACTOR

    DOEpatents

    Balent, R.

    1963-03-12

    This patent describes a method of obtaining a flatter flux and more uniform power generation across the core of a nuclear reactor. The method comprises using moderator elements having differing moderating strength. The elements have an increasing amount of the better moderating material as a function of radial and/or axial distance from the reactor core center. (AEC)

  18. Effect of the liquid upflow velocity on thermophilic sulphate reduction in acidifying granular sludge reactors.

    PubMed

    Lens, P N; Korthout, D; van Lier, J B; Hulshoff Pol, L W; Lettinga, G

    2001-02-01

    The effect of the superficial liquid upflow velocity on the acidifying and sulfate reducing capacity of thermophilic (55 degrees C; pH 6.0) granular sludge bed reactors treating partly acidified wastewater was investigated. A comparison was made between a UASB and an EGSB reactor, operated at an upflow velocity of 1 m.h-1 and 6.8 m.h-1, respectively. Both reactors were inoculated with a mixture of mesophilic sulphidogenic, thermophilic sulphidogenic and thermophilic methanogenic sludge (ratio 2:1:1). They were fed a synthetic wastewater containing starch, sucrose, lactate, propionate and acetate and a low sulphate concentration (COD/SO4(2-) ratio of 10). At the end of the experiment, the sulphate level of the influent was slightly increased to a COD/SO4(2-) ratio of 8. The reactors were operated at a hydraulic retention time of about 5 h and the imposed volumetric organic loading rates (OLR) ranged from 4.9 to 40.0 g COD l-1d-1. When imposing an OLR of 40.0 g COD l-1d-1, the acidification efficiency dropped to 80% and the sulphate reduction efficiency decreased to 50% in the UASB reactor. In the EGSB reactor, the sulphate reduction efficiency dropped to 30% directly following the OLR increase to 40 g COD l-1d-1, but recovered rapidly to 100% (at an OLR of 35 g COD l-1d-1) until the end of the experiment. In the UASB reactor, there was a net acetate and propionate production. At the higher organic loading rates, propionate was converted to n-butyrate and n-valerate. These back reactions did not occur in the EGSB reactor, in which an active methanogenic population developed, leading to a net acetate removal (up to 50%) and a high gas loading rate (up to 8.5 l l-1d-1). In both reactors, the effluent sulphide concentration was always below 200 mg l-1, of which about 90% was present as undissociated H2S (under the given conditions--pH 5.8-6.1 and 55 degrees C). The biogas (including CH4 and CO2) production rates in the UASB were very low, i.e. < 31 biogas l-1 reactor d

  19. Geobacillus zalihae sp. nov., a thermophilic lipolytic bacterium isolated from palm oil mill effluent in Malaysia

    PubMed Central

    Rahman, Raja Noor Zaliha Raja Abd; Leow, Thean Chor; Salleh, Abu Bakar; Basri, Mahiran

    2007-01-01

    Background Thermophilic Bacillus strains of phylogenetic Bacillus rRNA group 5 were described as a new genus Geobacillus. Their geographical distribution included oilfields, hay compost, hydrothermal vent or soils. The members from the genus Geobacillus have a growth temperatures ranging from 35 to 78°C and contained iso-branched saturated fatty acids (iso-15:0, iso-16:0 and iso-17:0) as the major fatty acids. The members of Geobacillus have similarity in their 16S rRNA gene sequences (96.5–99.2%). Thermophiles harboring intrinsically stable enzymes are suitable for industrial applications. The quest for intrinsically thermostable lipases from thermophiles is a prominent task due to the laborious processes via genetic modification. Results Twenty-nine putative lipase producers were screened and isolated from palm oil mill effluent in Malaysia. Of these, isolate T1T was chosen for further study as relatively higher lipase activity was detected quantitatively. The crude T1 lipase showed high optimum temperature of 70°C and was also stable up to 60°C without significant loss of crude enzyme activity. Strain T1T was a Gram-positive, rod-shaped, endospore forming bacterium. On the basic of 16S rDNA analysis, strain T1T was shown to belong to the Bacillus rRNA group 5 related to Geobacillus thermoleovorans (DSM 5366T) and Geobacillus kaustophilus (DSM 7263T). Chemotaxonomic data of cellular fatty acids supported the affiliation of strain T1T to the genus Geobacillus. The results of physiological and biochemical tests, DNA/DNA hybridization, RiboPrint analysis, the length of lipase gene and protein pattern allowed genotypic and phenotypic differentiation of strain T1T from its validly published closest phylogenetic neighbors. Strain T1T therefore represents a novel species, for which the name Geobacillus zalihae sp. nov. is proposed, with the type strain T1T (=DSM 18318T; NBRC 101842T). Conclusion Strain T1T was able to secrete extracellular thermostable lipase into

  20. Complete Genome Sequence of the Aerobic CO-Oxidizing Thermophile Thermomicrobium roseum

    PubMed Central

    Wu, Dongying; Raymond, Jason; Wu, Martin; Chatterji, Sourav; Ren, Qinghu; Graham, Joel E.; Bryant, Donald A.; Robb, Frank; Colman, Albert; Tallon, Luke J.; Badger, Jonathan H.; Madupu, Ramana; Ward, Naomi L.; Eisen, Jonathan A.

    2009-01-01

    In order to enrich the phylogenetic diversity represented in the available sequenced bacterial genomes and as part of an “Assembling the Tree of Life” project, we determined the genome sequence of Thermomicrobium roseum DSM 5159. T. roseum DSM 5159 is a red-pigmented, rod-shaped, Gram-negative extreme thermophile isolated from a hot spring that possesses both an atypical cell wall composition and an unusual cell membrane that is composed entirely of long-chain 1,2-diols. Its genome is composed of two circular DNA elements, one of 2,006,217 bp (referred to as the chromosome) and one of 919,596 bp (referred to as the megaplasmid). Strikingly, though few standard housekeeping genes are found on the megaplasmid, it does encode a complete system for chemotaxis including both chemosensory components and an entire flagellar apparatus. This is the first known example of a complete flagellar system being encoded on a plasmid and suggests a straightforward means for lateral transfer of flagellum-based motility. Phylogenomic analyses support the recent rRNA-based analyses that led to T. roseum being removed from the phylum Thermomicrobia and assigned to the phylum Chloroflexi. Because T. roseum is a deep-branching member of this phylum, analysis of its genome provides insights into the evolution of the Chloroflexi. In addition, even though this species is not photosynthetic, analysis of the genome provides some insight into the origins of photosynthesis in the Chloroflexi. Metabolic pathway reconstructions and experimental studies revealed new aspects of the biology of this species. For example, we present evidence that T. roseum oxidizes CO aerobically, making it the first thermophile known to do so. In addition, we propose that glycosylation of its carotenoids plays a crucial role in the adaptation of the cell membrane to this bacterium's thermophilic lifestyle. Analyses of published metagenomic sequences from two hot springs similar to the one from which this strain

  1. Identification and Characterization of a New 7-Aminocephalosporanic Acid Deacetylase from Thermophilic Bacterium Alicyclobacillus tengchongensis

    PubMed Central

    Ding, Jun-Mei; Yu, Ting-Ting; Han, Nan-Yu; Yu, Jia-Lin; Li, Jun-Jun; Yang, Yun-Juan; Tang, Xiang-Hua; Xu, Bo; Zhou, Jun-Pei

    2015-01-01

    ABSTRACT Deacetylation of 7-aminocephalosporanic acid (7-ACA) at position C-3 provides valuable starting material for producing semisynthetic β-lactam antibiotics. However, few enzymes have been characterized in this process before now. Comparative analysis of the genome of the thermophilic bacterium Alicyclobacillus tengchongensis revealed a hypothetical protein (EstD1) with typical esterase features. The EstD1 protein was functionally cloned, expressed, and purified from Escherichia coli BL21(DE3). It indeed displayed esterase activity, with optimal activity at around 65°C and pH 8.5, with a preference for esters with short-chain acyl esters (C2 to C4). Sequence alignment revealed that EstD1 is an SGNH hydrolase with the putative catalytic triad Ser15, Asp191, and His194, which belongs to carbohydrate esterase family 12. EstD1 can hydrolyze acetate at the C-3 position of 7-aminocephalosporanic acid (7-ACA) to form deacetyl-7-ACA, which is an important starting material for producing semisynthetic β-lactam antibiotics. EstD1 retained more than 50% of its initial activity when incubated at pH values ranging from 4 to 11 at 65°C for 1 h. To the best of our knowledge, this enzyme is a new SGNH hydrolase identified from thermophiles that is able to hydrolyze 7-ACA. IMPORTANCE Deacetyl cephalosporins are highly valuable building blocks for the industrial production of various kinds of semisynthetic β-lactam antibiotics. These compounds are derived mainly from 7-ACA, which is obtained by chemical or enzymatic processes from cephalosporin C. Enzymatic transformation of 7-ACA is the main method because of the adverse effects chemical deacylation brought to the environment. SGNH hydrolases are widely distributed in plants. However, the tools for identifying and characterizing SGNH hydrolases from bacteria, especially from thermophiles, are rather limited. Here, our work demonstrates that EstD1 belongs to the SGNH family and can hydrolyze acetate at the C-3 position of

  2. Protein Dynamics and Stability: The Distribution of Atomic Fluctuations in Thermophilic and Mesophilic Dihydrofolate Reductase Derived Using Elastic Incoherent Neutron Scattering

    SciTech Connect

    Meinhold, Lars; Clement, David; Tehei, M; Daniel, R. M.; Finney, J.L.; Smith, Jeremy C

    2008-11-01

    The temperature dependence of the dynamics of mesophilic and thermophilic dihydrofolate reductase is examined using elastic incoherent neutron scattering. It is demonstrated that the distribution of atomic displacement amplitudes can be derived from the elastic scattering data by assuming a (Weibull) functional form that resembles distributions seen in molecular dynamics simulations. The thermophilic enzyme has a significantly broader distribution than its mesophilic counterpart. Furthermore, although the rate of increase with temperature of the atomic mean-square displacements extracted from the dynamic structure factor is found to be comparable for both enzymes, the amplitudes are found to be slightly larger for the thermophilic enzyme. Therefore, these results imply that the thermophilic enzyme is the more flexible of the two.

  3. Effect of surfactant on hydrolysis products accumulation and short-chain fatty acids (SCFA) production during mesophilic and thermophilic fermentation of waste activated sludge: kinetic studies.

    PubMed

    Zhang, Peng; Chen, Yinguang; Zhou, Qi

    2010-09-01

    In the presence of surfactant sodium dodecylbenzene sulfonate (SDBS) the hydrolysis products accumulation and the short-chain fatty acids (SCFA) production during waste activated sludge fermentation under mesophilic and thermophilic conditions was compared with that at room temperature. In order to understand the mechanism of significant amounts of mesophilic and thermophilic hydrolysis products and SCFA observed in the presence of surfactant, the kinetic models at different SDBS dosages were developed. It was found that SDBS increased the mesophilic and thermophilic hydrolysis rate significantly, and the maximum specific utilization of hydrolysis products increased at low SDBS and decreased at high one. However, the observed maximum specific utilization of SCFA decreased seriously with SDBS increase. In the presence of SDBS the decay rate of acidogenic bacteria not only was lower than that in the absence of SDBS but decreased with the increase of SDBS under either mesophilic or thermophilic conditions. PMID:20409704

  4. Production of Sporotrichum thermophile xylanase by solid state fermentation utilizing deoiled Jatropha curcas seed cake and its application in xylooligosachharide synthesis.

    PubMed

    Sadaf, Ayesha; Khare, S K

    2014-02-01

    De-oiled Jatropha curcas seed cake, a plentiful by-product of biodiesel industry was used as substrate for the production of a useful xylanase from Sporotrichum thermophile in solid state fermentation. Under the optimized conditions, 1025U xylanase/g (deoiled seed cake) was produced. The xylanase exhibited half life of 4h at 45°C and 71.44min at 50°C respectively. It was stable in a broad pH range of 7.0-11.0. Km and Vmax were 12.54mg/ml and 454.5U/ml/min respectively. S. thermophile xylanase is an endoxylanase free of exoxylanase activity, hence advantageous for xylan hydrolysis to produce xylooligosachharides. Hydrolysis of oat spelt xylan by S. thermophile xylanase yielded 73% xylotetraose, 15.4% xylotriose and 10% xylobiose. The S. thermophile endoxylanase thus seem potentially useful in the food industries. PMID:24362246

  5. Draft Genome Sequences of Seven Thermophilic Spore-Forming Bacteria Isolated from Foods That Produce Highly Heat-Resistant Spores, Comprising Geobacillus spp., Caldibacillus debilis, and Anoxybacillus flavithermus

    PubMed Central

    Berendsen, Erwin M.; Wells-Bennik, Marjon H. J.; Krawczyk, Antonina O.; de Jong, Anne; van Heel, Auke; Holsappel, Siger; Eijlander, Robyn T.

    2016-01-01

    Here, we report the draft genomes of five strains of Geobacillus spp., one Caldibacillus debilis strain, and one draft genome of Anoxybacillus flavithermus, all thermophilic spore-forming Gram-positive bacteria. PMID:27151781

  6. Removal of Sulfur Compounds from Coal by the Thermophilic Organism Sulfolobus acidocaldarius

    PubMed Central

    Kargi, Fikret; Robinson, James M.

    1982-01-01

    The thermophilic, reduced-sulfur, iron-oxidizing bacterium Sulfolobus acidocaldarius was used for the removal of sulfur compounds from coal. The inclusion of complex nutrients such as yeast extract and peptone, and chemical oxidizing agents, 0.01 M FeCl3 into leaching medium, reduced the rate and the extent of sulfur removal from coal. The rate of sulfur removal by S. acidocaldarius was strongly dependent on the sulfur content of the coal and on the total external surface area of coal particles. Approximately 96% of inorganic sulfur was removed from a 5% slurry of coal which had an initial total sulfur content of 4% and an inorganic (pyritic S and sulfate) sulfur content of 2.1%. This resulted in removal of 50% of initial total sulfur present in coal. PMID:16346112

  7. Application of thermophilic enzymes and water jet system to cassava pulp.

    PubMed

    Chaikaew, Siriporn; Maeno, Yuka; Visessanguan, Wonnop; Ogura, Kota; Sugino, Gaku; Lee, Seung-Hwan; Ishikawa, Kazuhiko

    2012-12-01

    Co-production of fermentable sugars and nanofibrillated cellulose from cassava pulp was achieved by the combination of thermophilic enzymes (endoglucanase, β-glucosidase, and α-amylase) and a new atomization system (Star Burst System; SBS), which employs opposing water jets. The SBS represents a key technology for providing cellulose nanofibers and improving the enzymatic saccharification of cassava pulp. Depending on the enzymes used, the production of glucose from cassava pulp treated with the SBS was 1.2- to 2.5-fold higher than that from pulp not treated with the SBS. Nanofibrillated cellulose with the gel-like property in suspension was produced (yield was over 90%) by α-amylase treatment, which completely released trapped starch granules from the fibrous cell wall structure of cassava pulp pretreated with the SBS. The SBS provides an environmentally low-impact pretreatment system for processing biomass material into value-added products. PMID:23073093

  8. Malbranchea cinnamomea: A thermophilic fungal source of catalytically efficient lignocellulolytic glycosyl hydrolases and metal dependent enzymes.

    PubMed

    Mahajan, Chhavi; Basotra, Neha; Singh, Surender; Di Falco, Marcos; Tsang, Adrian; Chadha, B S

    2016-01-01

    This study reports thermophilic fungus Malbranchea cinnamomea as an important source of lignocellulolytic enzymes. The secretome analysis using LC-MS/MS orbitrap showed that fungus produced a spectrum of glycosyl hydrolases (cellulase/hemicellulase), polysaccharide lyases (PL) and carbohydrate esterases (CE) in addition to cellobiose dehydrogenase (CDH) indicating the presence of functional classical and oxidative cellulolytic mechanisms. The protein fractions in the secretome resolved by ion exchange chromatography were analyzed for ability to hydrolyze alkali treated carrot grass (ATCG) in the presence of Mn(2+)/Cu(2+). This strategy in tandem with peptide mass fingerprinting led to identification of metal dependent protein hydrolases with no apparent hydrolytic activity, however, showed 5.7 folds higher saccharification in presence of Mn(2+). Furthermore, adding different protein fractions to commercial cellulase (Novozymes: Cellic CTec2) resulted in enhanced hydrolysis of ATCG ranging between 1.57 and 3.43 folds indicating the enzymes from M. cinnamomea as catalytically efficient. PMID:26476165

  9. Isolation and characterization of a novel thermophilic Bacillus strain degrading long-chain n-alkanes.

    PubMed

    Wang, Lei; Tang, Yun; Wang, Shuo; Liu, Ru-Lin; Liu, Mu-Zhi; Zhang, Yan; Liang, Feng-Lai; Feng, Lu

    2006-08-01

    A thermophilic Bacillus strain NG80-2 growing within the temperature range of 45-73 degrees C (optimum at 65 degrees C) was isolated from a deep subterranean oil-reservoir in northern China. The strain was able to utilize crude oil and liquid paraffin as the sole carbon sources for growth, and the growth with crude oil was accompanied by the production of an unknown emulsifying agent. Further examination showed that NG80-2 degraded and utilized only long-chain (C15-C36) n-alkanes, but not short-chain (C8-C14) n-alkanes and those longer than C40. Based on phenotypic and phylogenic analyses, NG80-2 was identified as Geobacillus thermodenitrificans. The strain NG80-2 may be potentially used for oily-waste treatment at elevated temperature, a condition which greatly accelerates the biodegradation rate, and for microbial enhancing oil recovery process. PMID:16604274

  10. Amylase production in solid state fermentation by the thermophilic fungus Thermomyces lanuginosus.

    PubMed

    Kunamneni, Adinarayana; Permaul, Kugen; Singh, Suren

    2005-08-01

    The production of extracellular amylase by the thermophilic fungus Thermomyces lanuginosus was studied in solid state fermentation (SSF). Solid substrates such as wheat bran, molasses bran, rice bran, maize meal, millet cereal, wheat flakes, barley bran, crushed maize, corncobs and crushed wheat were studied for enzyme production. Growth on wheat bran gave the highest amylase activity. The maximum enzyme activity obtained was 534 U/g of wheat bran under optimum conditions of an incubation period of 120 h, an incubation temperature of 50 degrees C, an initial moisture content of 90%, a pH of 6.0, an inoculum level of 10% (v/w), a salt solution concentration of 1.5:10 (v/w) and a ratio of substrate weight to flask volume of 1:100 with soluble starch (1% w/w) and peptone (1% w/w) as supplements. PMID:16198259

  11. Comparison of non-agitated and agitated batch, thermophilic anaerobic digestion of sugarbeet tailings.

    PubMed

    Tian, Zhuoli; Chauliac, Diane; Pullammanappallil, Pratap

    2013-02-01

    Sugar beet tailings were anaerobically digested at non-agitated and agitated conditions in identical thermophilic batch reactors. The average methane yield in the agitated digester was only 74% of that in the non-agitated digester. Ninety percent of the ultimate methane yield was produced in approximately 5 days in the non-agitated digester whereas it took 12 days in agitated digester. Even upon using an active inoculum from non-agitated digester the methane rate and yield was low in the agitated digester. On the other hand when the poorly performing inoculum from the agitated digester was transferred to the non-agitated digester, its activity was immediately enhanced. The non-agitated digester harbored a diverse microbial community with phylotypes Methanoculleus and Methanosarcina being dominant methanogens. Methanosaeta was the only methanogen detected in the agitated digester. It also contained a hydrogen-producing bacterial phylotype Petrotoga in high proportion which was not detected in the other digester. PMID:23262019

  12. Performance optimization and validation of ADM1 simulations under anaerobic thermophilic conditions.

    PubMed

    Atallah, Nabil M; El-Fadel, Mutasem; Ghanimeh, Sophia; Saikaly, Pascal; Abou-Najm, Majdi

    2014-12-01

    In this study, two experimental sets of data each involving two thermophilic anaerobic digesters treating food waste, were simulated using the Anaerobic Digestion Model No. 1 (ADM1). A sensitivity analysis was conducted, using both data sets of one digester, for parameter optimization based on five measured performance indicators: methane generation, pH, acetate, total COD, ammonia, and an equally weighted combination of the five indicators. The simulation results revealed that while optimization with respect to methane alone, a commonly adopted approach, succeeded in simulating methane experimental results, it predicted other intermediary outputs less accurately. On the other hand, the multi-objective optimization has the advantage of providing better results than methane optimization despite not capturing the intermediary output. The results from the parameter optimization were validated upon their independent application on the data sets of the second digester. PMID:25463805

  13. Bioleaching of spent hydrotreating catalyst by acidophilic thermophile Acidianus brierleyi: Leaching mechanism and effect of decoking.

    PubMed

    Bharadwaj, Abhilasha; Ting, Yen-Peng

    2013-02-01

    Bioleaching of spent hydrotreating catalyst by thermophillic archae Acidianus brierleyi was investigated. The spent catalyst (containing Al, Fe, Ni and Mo as major elements) was characterized, and the effect of pretreatment (decoking) on two-step and spent medium leaching was examined at 1% w/v pulp density. Decoking resulted in removal of carbonaceous deposits and volatile impurities, and affected the solubility of metal compounds through oxidization of the metal sulfides. Nearly 100% extraction was achieved using spent medium leaching for Fe, Ni and Mo, and 67% for Al. Bioleaching reduced nickel concentration in the leachate below the regulated levels for safe waste disposal. Chemical (i.e. abiotic) leaching using equimolar concentration of sulfuric acid produced by the bacteria during two-step process achieved a lower leaching efficiency (by up to 30%). Results indicated that A. brierleyi successfully leached heavy metals from spent catalyst. PMID:23334026

  14. Hospital hydrotherapy pools treated with ultra violet light: bad bacteriological quality and presence of thermophilic Naegleria.

    PubMed Central

    De Jonckheere, J. F.

    1982-01-01

    The microbiological quality of eight halogenated and two u.v.-treated hydrotherapy pools in hospitals was investigated. The microbiological quality of halogenated hydrotherapy pools was comparable to halogenated public swimming pools, although in some Pseudomonas aeruginosa and faecal pollution indicators were more frequent due to bad management. On the other hand u.v.-treated hydrotherapy pools had very bad microbiological quality. Apart from faecal pollution indicators, P. aeruginosa was present in very high numbers. Halogenated hydrotherapy pools were not highly contaminated with amoebae, and Naegleria spp. were never detected. On the other hand u.v.-treated pools contained very high numbers of thermophilic Naegleria. The Naegleria isolated were identified as N. lovaniensis, a species commonly found in association with N. fowleri. Isoenzyme analysis showed a different type of N. lovaniensis was present in each of two u.v.-treated pools. Images Plate 1 PMID:7061835

  15. Genetics of thermophilic bacteria. Final progress report, May 1, 1984--April 30, 1991

    SciTech Connect

    Welker, N.E.

    1991-12-31

    Organisms adapted to high temperature have evolved a variety of unique solutions to the biochemical problems imposed by this environment. Adaptation is commonly used to describe the biochemical properties of organisms which have become adapted to their environment (genetic adaptation). It can also mean the direct response-at the cellular level-of an organism to changes in temperature (physiological adaptation). Thermophilic bacilli (strains of Bacillus stearothermophilus) can exhibit a variety of biochemical adaptations in response to changes in temperature. These include changes in the composition and stability of the membrane, metabolic potential, the transport of amino acids, regulatory mechanisms, ribose methylation of tRNA, protein thermostability, and nutritional requirements. The objectives of the research were to develop efficient and reliable genetic systems to analyze and manipulate B. Stearothermophilus, and to use these systems initiate a biochemical, molecular, and genetic investigations of genes that are required for growth at high temperature.

  16. Glucose-sensing proteins from mesophilic and thermophilic bacteria as new tools in diabetes monitoring

    NASA Astrophysics Data System (ADS)

    D'Auria, S.; Rossi, Mose; Lakowicz, Joseph R.

    2001-05-01

    We developed a new method of glucose sensing using inactive forms of glucose oxidase from Aspergillus niger and glucose dehydrogenase from the thermophilic microorganism Thermoplasma acidophilum. Glucose oxidase was rendered inactive by removal of the FAD cofactor. The resulting apo- glucose oxidase still binds glucose as observed from a decrease in its intrinsic tryptophan fluorescence. 8- Anilino-1-naphthalene sulfonic acid (ANS) was found to bind spontaneously to apo-glucose oxidase as seen from an enhancement of the ANS fluorescence. The steady state intensity of the bound ANS decreased 25% upon binding of glucose, and the mean lifetime of the bound ANS decreased about 40%. These spectral changes occurred with a midpoint from 10 to 20 mM glucose, which is comparable to the Ko of holo-glucose oxidase. These results suggest that apo- glucose oxidase can be used as a reversible non-consuming sensor for glucose.

  17. Production and characterization of thermostable alpha-amylase by thermophilic Geobacillus stearothermophilus.

    PubMed

    Al-Qodah, Zakaria

    2006-01-01

    Studies on the alpha-amylase-producing thermophilic bacterium isolated and identified from a hot spring in Jordan and designated as Geobacillus stearothermophilus JT2 were carried out. The optimum conditions for growth and enzyme production were pH 7 and 55 degrees C. The study of the kinetics of cellular growth indicated a mu(max) of 0.22/h, a K(s) of 1.2 g/L, a tau(d) of 3.15 h and a Y(x/s) of 0.43 g cell/g starch. In addition, the activation energy for growth and death were estimated and found to be 30.5 and 210 J/mol, respectively. The effect of different carbon and nitrogen sources on the cellular growth was tested. PMID:16927263

  18. Some Biochemical Properties of an Acido-Thermophilic Archae-Bacterium Sulfolobus Acidocaldarius

    NASA Astrophysics Data System (ADS)

    Oshima, Tairo; Ohba, Masayuki; Wagaki, Takayoshi

    1984-12-01

    To elucidate the phylogenic status of archaebacteria, some basic cellular components of an acido-thermophilic archaebacterium,Sulfolobus acidocaldarius, were studied. Poly(A) containing RNA was present in the cells, and performed the role of mRNA in a cell-free extract of reticulocyte or the archaebacteria. Poly(A) containing RNA was also found in other archaebacterial cells. The absence of cap structure was suggested in these RNAs. The cell-free protein synthesis using the archaebacterial extract was inhibited by anisomycin, a specific inhibitor for eukaryotic ribosomes. Two unique membrane-bound ATPases were detected. Based on resistance to H+-ATPase inhibitors, these enzymes seemed not to be F0F1-ATPase.

  19. North Western Spain hot springs are a source of lipolytic enzyme-producing thermophilic microorganisms.

    PubMed

    Deive, Francisco J; Alvarez, María S; Sanromán, M Angeles; Longo, Maria A

    2013-02-01

    Several hot springs in Galicia (North Western Spain) have been investigated as potential sources of lipolytic enzyme-producing thermophilic microorganisms. After isolating 12 esterase producing strains, 9 of them were assured to be true lipase producers, and consequently grown in submerged cultures, obtaining high extracellular activities by two of them. Furthermore, a preliminary partial characterization of the crude lipase, obtained by ultrafiltration of the cell-free culture supernatant, was carried out at several pH and temperature values. It is outstanding that several enzymes turned out to be multiextremozymes, since they had their optimum temperature and pH at typical values from thermoalkalophiles. The thermal stability in aqueous solution of the crude enzymes was also assayed, and the influence of some potential enzyme stabilizing compounds was tested. Finally, the viability of the selected microorganisms has been demonstrated at bioreactor scale. PMID:22763779

  20. Identification and characterization of thermophilic Synechococcus spp. isolates from Asian geothermal springs.

    PubMed

    Jing, Hongmei; Liu, Hongbin; Pointing, Stephen B

    2007-04-01

    Two thermophilic cyanobacterial strains, Ts and Bs, collected from Asian geothermal springs were identified morphologically and phylogenetically as Synechococcus in the order Chroococcales and were isolated into axenic cultures. In addition to the high similarities between their full 16S rRNA gene sequences, both strains also shared similar pigment profiles and fatty acid compositions but with varied ratios. Strain Ts had elevated levels of photoprotective pigments such as carotenoid and scytonemin even after prolonged culture under identical laboratory conditions, whereas strain Bs produced more chlorophyll a per unit cell volume, perhaps resulting from UV adaptation in the natural habitats. In addition, strain Ts had more content than strain Bs in terms of the total fatty acids and the proportion of unsaturated fatty acids. Neither isolate was able to fix nitrogen, and they had zero susceptibility to ampicillin and streptomycin. PMID:17612602

  1. Effect of oxygen on the microbial activities of thermophilic anaerobic biomass.

    PubMed

    Pedizzi, C; Regueiro, L; Rodriguez-Verde, I; Lema, J M; Carballa, M

    2016-07-01

    Low oxygen levels (μgO2L(-1)) in anaerobic reactors are quite common and no relevant consequences are expected. On the contrary, higher concentrations could affect the process. This work aimed to study the influence of oxygen (4.3 and 8.8mgO2L(-1), respectively) on the different microbial activities (hydrolytic, acidogenic and methanogenic) of thermophilic anaerobic biomass and on the methanogenic community structure. Batch tests in presence of oxygen were conducted using specific substrates for each biological activity and a blank (with minimum oxygen) was included. No effect of oxygen was observed on the hydrolytic and acidogenic activities. In contrast, the methane production rate decreased by 40% in all oxygenated batches and the development of active archaeal community was slower in presence of 8.8mgO2L(-1). However, despite this sensitivity of methanogens to oxygen at saturation levels, the inhibition was reversible. PMID:27020398

  2. Cellulolytic Activity of Thermophilic Bacilli Isolated from Tattapani Hot Spring Sediment in North West Himalayas.

    PubMed

    Priya, Indu; Dhar, M K; Bajaj, B K; Koul, Sanjana; Vakhlu, Jyoti

    2016-06-01

    Eight thermophilic bacterial strains were isolated from Tattapani Hot spring and screened for various hydrolytic enzymes including cellulases. The isolated bacterial strains were identified as Geobacillus thermodenitrificans IP_WH1(KP842609), Bacillus licheniformis IP_WH2(KP842610), B. aerius IP_WH3(KP842611), B. licheniformis IP_WH4(KP842612), B. licheniformis IP_60Y(KP842613), G. thermodenitrificans IP_60A1(KP842614), Geobacillus sp. IP_60A2(KP842615) and Geobacillus sp. IP_80TP(KP842616) after 16S ribotying. Out of the eight isolates Geobacillus sp. IP_80TP grew best at 80 °C whereas rest of the isolates showed optimal growth at 60 °C. G. thermodenitrificans IP_WH1 produced a thermotolerant cellulase with maximum activity at 60 °C. PMID:27570317

  3. Glycerol acts as alternative electron sink during syngas fermentation by thermophilic anaerobe Moorella thermoacetica.

    PubMed

    Kimura, Zen-ichiro; Kita, Akihisa; Iwasaki, Yuki; Nakashimada, Yutaka; Hoshino, Tamotsu; Murakami, Katsuji

    2016-03-01

    Moorella thermoacetica is an anaerobic thermophilic acetogen that is capable of fermenting sugars, H(2)/CO(2) and syngas (H(2)/CO). For this reason, this bacterium is potentially useful for biotechnology applications, particularly the production of biofuel from CO(2). A soil isolate of M. thermoacetica, strain Y72, produces both ethanol and acetate from H(2)/CO(2); however, the maximum concentrations of these two products are too low to enable commercialization of the syngas fermentation process. In the present study, glycerol was identified as a novel electron sink among the fermentation products of strain Y72. Notably, a 1.5-fold increase in the production of ethanol (1.4 mM) was observed in cultures supplemented with glycerol during syngas fermentation. This discovery is expected to aid in the development of novel methods that allow for the regulation of metabolic pathways to direct and increase the production of desirable fermentative compounds. PMID:26452417

  4. The performance of a thermophilic microbial fuel cell fed with synthesis gas.

    PubMed

    Hussain, A; Mehta, P; Raghavan, V; Wang, H; Guiot, S R; Tartakovsky, B

    2012-08-10

    This study demonstrated electricity generation in a thermophilic microbial fuel cell (MFC) operated on synthesis gas (syngas) as the sole electron donor. At 50°C, a volumetric power output of 30-35 mWL(R)(-1) and a syngas conversion efficiency of 87-98% was achieved. The observed pathway of syngas conversion to electricity primarily consisted of a two-step process, where the carbon monoxide and hydrogen were first converted to acetate, which was then consumed by the anodophilic bacteria to produce electricity. A denaturing gradient gel electrophoresis (DGGE) analysis of the 16S rDNA revealed the presence of Geobacter species, Acetobacter, methanogens and several uncultured bacteria and archaea in the anodic chamber. PMID:22759536

  5. Development of a Quantitative PCR Assay for Thermophilic Spore-Forming Geobacillus stearothermophilus in Canned Food.

    PubMed

    Nakano, Miyo

    2015-01-01

    The thermophilic spore forming bacteria Geobacillus stearothermophilus is recognized as a major cause of spoilage in canned food. A quantitative real-time PCR assay was developed to specifically detect and quantify the species G. stearothermophilus in samples from canned food. The selected primer pairs amplified a 163-bp fragment of the 16S rRNA gene in a specific PCR assay with a detection limit of 12.5 fg of pure culture DNA, corresponding to DNA extracted from approximately 0.7 CFU/mL of G. stearothermophilus. Analysis showed that the bacterial species G. stearothermophilus was not detected in any canned food sample. Our approach presented here will be useful for tracking or quantifying species G. stearotethermophilus in canned food and ingredients. PMID:26412704

  6. Biodegradation of Aliphatic-Aromatic Copolyesters by Thermomonospora fusca and Other Thermophilic Compost Isolates

    PubMed Central

    Kleeberg, Ilona; Hetz, Claudia; Kroppenstedt, Reiner Michael; Müller, Rolf-Joachim; Deckwer, Wolf-Dieter

    1998-01-01

    Random aliphatic-aromatic copolyesters synthesized from 1,4-butanediol, adipic acid, and terephthalic acid (BTA) have excellent thermal and mechanical properties and are biodegradable by mixed cultures (e.g., in compost). Over 20 BTA-degrading strains were isolated by using compost as a microbial source. Among these microorganisms, thermophilic actinomycetes obviously play an outstanding role and appear to dominate the initial degradation step. Two actinomycete strains exhibited about 20-fold higher BTA degradation rates than usually observed in a common compost test. These isolates were identified as Thermomonospora fusca strains. They appeared to be particularly suitable for establishment of rapid degradation tests and were used in comparative studies on the biodegradation of various polyesters. PMID:9572944

  7. Complete genome sequence of thermophilic Bacillus smithii type strain DSM 4216(T).

    PubMed

    Bosma, Elleke F; Koehorst, Jasper J; van Hijum, Sacha A F T; Renckens, Bernadet; Vriesendorp, Bastienne; van de Weijer, Antonius H P; Schaap, Peter J; de Vos, Willem M; van der Oost, John; van Kranenburg, Richard

    2016-01-01

    Bacillus smithii is a facultatively anaerobic, thermophilic bacterium able to use a variety of sugars that can be derived from lignocellulosic feedstocks. Being genetically accessible, it is a potential new host for biotechnological production of green chemicals from renewable resources. We determined the complete genomic sequence of the B. smithii type strain DSM 4216(T), which consists of a 3,368,778 bp chromosome (GenBank accession number CP012024.1) and a 12,514 bp plasmid (GenBank accession number CP012025.1), together encoding 3880 genes. Genome annotation via RAST was complemented by a protein domain analysis. Some unique features of B. smithii central metabolism in comparison to related organisms included the lack of a standard acetate production pathway with no apparent pyruvate formate lyase, phosphotransacetylase, and acetate kinase genes, while acetate was the second fermentation product. PMID:27559429

  8. Isolation and identification of thermophilic and mesophylic proteolytic bacteria from shrimp paste "Terasi"

    NASA Astrophysics Data System (ADS)

    Murwani, R.; Supriyadi, Subagio, Trianto, A.; Ambariyanto

    2015-12-01

    Terasi is a traditional product generally made of fermented shrimp. There were many studies regarding lactic acid bacteria of terasi but none regarding proteolitic bacteria. This study was conducted to isolate and identify the thermophilic and mesophylic proteolytic bacteria from terasi. In addition, the effect of different salt concentrations on the growth of the isolated proteolytic bacteria with the greatest proteolytic activity was also studied. Terasi samples were obtained from the Northern coast region of Java island i.e. Jepara, Demak and Batang. The study obtained 34 proteolytic isolates. Four isolates were identified as Sulfidobacillus, three isolates as Vibrio / Alkaligenes / Aeromonas, two isolates as Pseudomonas, 21 isolates as Bacillus, three isolates as Kurthia/ Caryophanon and one isolates as Amphibacillus. The growth of proteolytic bacteria was affected by salt concentration. The largest growth was found at 0 ppm salt concentrations and growth was declined as salt concentration increased. Maximum growth at each salt concentration tested was found at 8 hours incubation.

  9. Influence of EPS isolated from thermophilic sulphate-reducing bacteria on carbon steel corrosion.

    PubMed

    Dong, Ze Hua; Liu, Tao; Liu, Hong Fang

    2011-05-01

    Extracellular polymeric substances (EPS) were isolated by centrifugation of thermophilic sulphate-reducing bacteria (SRB) grown in API-RP38 culture medium. The protein and polysaccharide fractions were quantified and the highest concentrations were extracted from a 14-day old culture. The effect of EPS on carbon steel corrosion was investigated by electrochemical techniques. At 30°C, a small amount of EPS in 3% NaCl solution inhibited corrosion, whilst excessive amounts of EPS facilitated corrosion. In addition, the inhibition efficiency of EPS decreased with temperature due to thermal desorption of the EPS. The results suggest that adsorbed EPS layers could be beneficial to anti-corrosion by hindering the reduction of oxygen. However, the accumulation of an EPS film could stimulate the anodic dissolution of the underlying steel by chelation of Fe2+ ions. PMID:21604218

  10. Genome sequence of the moderately thermophilic, amino-acid-degrading and sulfur-reducing bacterium Thermovirga lienii type strain (Cas60314T)

    SciTech Connect

    Goker, Markus; Saunders, Elizabeth H; Lapidus, Alla L.; Nolan, Matt; Lucas, Susan; Hammon, Nancy; Deshpande, Shweta; Cheng, Jan-Fang; Han, Cliff; Tapia, Roxanne; Goodwin, Lynne A.; Pitluck, Sam; Liolios, Konstantinos; Mavromatis, K; Pagani, Ioanna; Ivanova, N; Mikhailova, Natalia; Pati, Amrita; Chen, Amy; Palaniappan, Krishna; Land, Miriam L; Chang, Yun-Juan; Jeffries, Cynthia; Brambilla, Evelyne-Marie; Rohde, Manfred; Spring, Stefan; Detter, J. Chris; Woyke, Tanja; Bristow, James; Eisen, Jonathan; Markowitz, Victor; Hugenholtz, Philip; Kyrpides, Nikos C; Klenk, Hans-Peter

    2012-01-01

    Thermovirga lienii Dahle and Birkeland 2006 is a member to the genomically so far uncharacterized genus Thermovirga in the phylum 'Synergistetes'. Members of the only recently (2007) proposed phylum 'Synergistetes' are of interest because of their isolated phylogenetic position and their diverse habitats, e.g. from man to oil well. The genome of T. lienii Cas60314T is only the 5th genome sequence (3rd completed) from this phylum to be published. Here we describe the features of this organism, together with the complete genome sequence and annotation. The 1,999,646 bp long genome (including one plasmid) with its 1,914 protein-coding and 59 RNA genes is a part of the Genomic Encyclopedia of Bacteria and Archaea project.

  11. [Cobalt(III)-EDTA]- Reduction by Thermophilic Methanogen Methanothermobacter Thermautotrophicus

    SciTech Connect

    Singh, Rajesh; Dong, Hailiang; Liu, Deng; Marts, Amy R.; Tierney, David L.; Almquist, Catherine B.

    2015-06-30

    Cobalt is a metal contaminant at high temperature radioactive waste disposal sites. In previous studies have largely focused on mesophilic microorganisms to remediate cobalt, despite the presence of thermophilic microorganisms at such sites. In this study,Methanothermobacter thermautotrophicus, a thermophilic methanogen, was used to reduce Co(III) in the form of [Co(III)–EDTA]-. Bioreduction experiments were conducted in a growth medium with H2/CO2 as a growth substrate at initial Co(III) concentrations of 1, 2, 4, 7, and 10 mM. At low Co(III) concentrations (< 4 mM), a complete reduction was observed within a week. Wet chemistry, X-ray absorption near-edge structure (XANES) and electron paramagnetic resonance (EPR) analyses were all consistent in revealing the reduction kinetics. But, at higher concentrations (7 and 10 mM) the reduction extents only reached 69.8% and 48.5%, respectively, likely due to the toxic effect of Co(III) to the methanogen cells as evidenced by a decrease in total cellular protein at these Co(III) concentrations. Methanogenesis was inhibited by Co(III) bioreduction, possibly due to impaired cell growth and electron diversion from CO2 to Co(III). Overall, our results demonstrated the ability of M. thermautotrophicus to reduce Co(III) to Co(II) and its potential application for remediating 60Co contaminant at high temperature subsurface radioactive waste disposal sites.

  12. Structure and Stability of the Dimeric Triosephosphate Isomerase from the Thermophilic Archaeon Thermoplasma acidophilum

    PubMed Central

    Park, Sang Ho; Kim, Hyoun Sook; Park, Mi Seul; Moon, Sojin; Song, Mi Kyung; Park, Han Su; Hahn, Hyunggu; Kim, Soon-Jong; Bae, Euiyoung; Kim, Hyun-Jung; Han, Byung Woo

    2015-01-01

    Thermoplasma acidophilum is a thermophilic archaeon that uses both non-phosphorylative Entner-Doudoroff (ED) pathway and Embden-Meyerhof-Parnas (EMP) pathway for glucose degradation. While triosephosphate isomerase (TPI), a well-known glycolytic enzyme, is not involved in the ED pathway in T. acidophilum, it has been considered to play an important role in the EMP pathway. Here, we report crystal structures of apo- and glycerol-3-phosphate-bound TPI from T. acidophilum (TaTPI). TaTPI adopts the canonical TIM-barrel fold with eight α-helices and parallel eight β-strands. Although TaTPI shares ~30% sequence identity to other TPIs from thermophilic species that adopt tetrameric conformation for enzymatic activity in their harsh physiological environments, TaTPI exists as a dimer in solution. We confirmed the dimeric conformation of TaTPI by analytical ultracentrifugation and size-exclusion chromatography. Helix 5 as well as helix 4 of thermostable tetrameric TPIs have been known to play crucial roles in oligomerization, forming a hydrophobic interface. However, TaTPI contains unique charged-amino acid residues in the helix 5 and adopts dimer conformation. TaTPI exhibits the apparent Td value of 74.6°C and maintains its overall structure with some changes in the secondary structure contents at extremely acidic conditions (pH 1–2). Based on our structural and biophysical analyses of TaTPI, more compact structure of the protomer with reduced length of loops and certain patches on the surface could account for the robust nature of Thermoplasma acidophilum TPI. PMID:26709515

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

    PubMed

    Nguyen, Peter Q; Silberg, Jonathan J

    2010-07-01

    Many proteins can be split into fragments that exhibit enhanced function upon fusion to interacting proteins. While this strategy has been widely used to create protein-fragment complementation assays (PCAs) for discovering protein-protein interactions within mesophilic organisms, similar assays have not yet been developed for studying natural and engineered protein complexes at the temperatures where thermophilic microbes grow. We describe the development of a selection for protein-protein interactions within Thermus thermophilus that is based upon growth complementation by fragments of Thermotoga neapolitana adenylate kinase (AK(Tn)). Complementation studies with an engineered thermophile (PQN1) that is not viable above 75 degrees C because its adk gene has been replaced by a Geobacillus stearothermophilus ortholog revealed that growth could be restored at 78 degrees C by a vector that coexpresses polypeptides corresponding to residues 1-79 and 80-220 of AK(Tn). In contrast, PQN1 growth was not complemented by AK(Tn) fragments harboring a C156A mutation within the zinc-binding tetracysteine motif unless these fragments were fused to Thermotoga maritima chemotaxis proteins that heterodimerize (CheA and CheY) or homodimerize (CheX). This enhanced complementation is interpreted as arising from chemotaxis protein-protein interactions, since AK(Tn)-C156A fragments having only one polypeptide fused to a chemotaxis protein did not complement PQN1 to the same extent. This selection increases the maximum temperature where a PCA can be used to engineer thermostable protein complexes and to map protein-protein interactions. PMID:20418388

  14. Biochemical characterization of a thermostable HNH endonuclease from deep-sea thermophilic bacteriophage GVE2.

    PubMed

    Zhang, Likui; Huang, Yanchao; Xu, Dandan; Yang, Lixiang; Qian, Kaicheng; Chang, Guozhu; Gong, Yong; Zhou, Xiaojian; Ma, Kesen

    2016-09-01

    His-Asn-His (HNH) proteins are a very common family of small nucleic acid-binding proteins that are generally associated with endonuclease activity and are found in all kingdoms of life. Although HNH endonucleases from mesophiles have been widely investigated, the biochemical functions of HNH endonucleases from thermophilic bacteriophages remain unknown. Here, we characterized the biochemical properties of a thermostable HNH endonuclease from deep-sea thermophilic bacteriophage GVE2. The recombinant GVE2 HNH endonuclease exhibited non-specific cleavage activity at high temperature. The optimal temperature of the GVE2 HNH endonuclease for cleaving DNA was 60-65 °C, and the enzyme retained its DNA cleavage activity even after heating at 100 °C for 30 min, suggesting the enzyme is a thermostable endonuclease. The GVE2 HNH endonuclease cleaved DNA over a wide pH spectrum, ranging from 5.5 to 9.0, and the optimal pH for the enzyme activity was 8.0-9.0. Furthermore, the GVE2 HNH endonuclease activity was dependent on a divalent metal ion. While the enzyme is inactive in the presence of Cu(2+), the GVE2 HNH endonuclease displayed cleavage activity of varied efficiency with Mn(2+), Mg(2+), Ca(2+), Fe(2+), Co(2+), Zn(2+), and Ni(2+). The GVE2 HNH endonuclease activity was inhibited by NaCl. This study provides the basis for determining the role of this endonuclease in life cycle of the bacteriophage GVE2 and suggests the potential application of the enzyme in molecular biology and biotechnology. PMID:27131500

  15. Bacterial community structure in treated sewage sludge with mesophilic and thermophilic anaerobic digestion.

    PubMed

    Stiborova, Hana; Wolfram, Jan; Demnerova, Katerina; Macek, Tomas; Uhlik, Ondrej

    2015-11-01

    Stabilized sewage sludge is applied to agricultural fields and farmland due to its high organic matter content. The aim of this study was to investigate the effects of two types of sludge stabilization, mesophilic anaerobic digestion (MAD) and thermophilic anaerobic digestion (TAD), on bacterial communities in sludge, including the presence of pathogenic microorganisms. Bacterial community structure and phylogenetic diversity were analyzed in four sewage sludge samples from the Czech Republic. Analysis of 16S ribosomal RNA (rRNA) genes showed that investigated sludge samples harbor diverse bacterial populations with only a few taxa present across all samples. Bacterial diversity was higher in sludge samples after MAD versus TAD treatment, and communities in MAD-treated sludge shared the highest genetic similarities. In all samples, the bacterial community was dominated by reads affiliated with Proteobacteria. The sludge after TAD treatment had considerably higher number of reads of thermotolerant/thermophilic taxa, such as the phyla Deinococcus-Thermus and Thermotogae or the genus Coprothermobacter. Only one operational taxonomic unit (OTU), which clustered with Rhodanobacter, was detected in all communities at a relative abundance >1 %. All of the communities were screened for the presence of 16S rRNA gene sequences of pathogenic bacteria using a database of 122 pathogenic species and ≥98 % identity threshold. The abundance of such sequences ranged between 0.23 and 1.57 % of the total community, with lower numbers present after the TAD treatment, indicating its higher hygienization efficiency. Sequences clustering with nontuberculous mycobacteria were present in all samples. Other detected sequences of pathogenic bacteria included Streptomyces somaliensis, Acinetobacter calcoaceticus, Alcaligenes faecalis, Gordonia spp., Legionella anisa, Bordetella bronchiseptica, Enterobacter aerogenes, Brucella melitensis, and Staphylococcus aureus. PMID:25921720

  16. The Structure of a Thermophilic Kinase Shapes Fitness upon Random Circular Permutation.

    PubMed

    Jones, Alicia M; Mehta, Manan M; Thomas, Emily E; Atkinson, Joshua T; Segall-Shapiro, Thomas H; Liu, Shirley; Silberg, Jonathan J

    2016-05-20

    Proteins can be engineered for synthetic biology through circular permutation, a sequence rearrangement in which native protein termini become linked and new termini are created elsewhere through backbone fission. However, it remains challenging to anticipate a protein's functional tolerance to circular permutation. Here, we describe new transposons for creating libraries of randomly circularly permuted proteins that minimize peptide additions at their termini, and we use transposase mutagenesis to study the tolerance of a thermophilic adenylate kinase (AK) to circular permutation. We find that libraries expressing permuted AKs with either short or long peptides amended to their N-terminus yield distinct sets of active variants and present evidence that this trend arises because permuted protein expression varies across libraries. Mapping all sites that tolerate backbone cleavage onto AK structure reveals that the largest contiguous regions of sequence that lack cleavage sites are proximal to the phosphotransfer site. A comparison of our results with a range of structure-derived parameters further showed that retention of function correlates to the strongest extent with the distance to the phosphotransfer site, amino acid variability in an AK family sequence alignment, and residue-level deviations in superimposed AK structures. Our work illustrates how permuted protein libraries can be created with minimal peptide additions using transposase mutagenesis, and it reveals a challenge of maintaining consistent expression across permuted variants in a library that minimizes peptide additions. Furthermore, these findings provide a basis for interpreting responses of thermophilic phosphotransferases to circular permutation by calibrating how different structure-derived parameters relate to retention of function in a cellular selection. PMID:26976658

  17. Roles of Thermophiles and Fungi in Bitumen Degradation in Mostly Cold Oil Sands Outcrops.

    PubMed

    Wong, Man-Ling; An, Dongshan; Caffrey, Sean M; Soh, Jung; Dong, Xiaoli; Sensen, Christoph W; Oldenburg, Thomas B P; Larter, Steve R; Voordouw, Gerrit

    2015-10-01

    Oil sands are surface exposed in river valley outcrops in northeastern Alberta, where flat slabs (tablets) of weathered, bitumen-saturated sandstone can be retrieved from outcrop cliffs or from riverbeds. Although the average yearly surface temperature of this region is low (0.7°C), we found that the temperatures of the exposed surfaces of outcrop cliffs reached 55 to 60°C on sunny summer days, with daily maxima being 27 to 31°C. Analysis of the cooccurrence of taxa derived from pyrosequencing of 16S/18S rRNA genes indicated that an aerobic microbial network of fungi and hydrocarbon-, methane-, or acetate-oxidizing heterotrophic bacteria was present in all cliff tablets. Metagenomic analyses indicated an elevated presence of fungal cytochrome P450 monooxygenases in these samples. This network was distinct from the heterotrophic community found in riverbeds, which included fewer fungi. A subset of cliff tablets had a network of anaerobic and/or thermophilic taxa, including methanogens, Firmicutes, and Thermotogae, in the center. Long-term aerobic incubation of outcrop samples at 55°C gave a thermophilic microbial community. Analysis of residual bitumen with a Fourier transform ion cyclotron resonance mass spectrometer indicated that aerobic degradation proceeded at 55°C but not at 4°C. Little anaerobic degradation was observed. These results indicate that bitumen degradation on outcrop surfaces is a largely aerobic process with a minor anaerobic contribution and is catalyzed by a consortium of bacteria and fungi. Bitumen degradation is stimulated by periodic high temperatures on outcrop cliffs, which cause significant decreases in bitumen viscosity. PMID:26209669

  18. Response surface optimization of substrates for thermophilic anaerobic codigestion of sewage sludge and food waste.

    PubMed

    Kim, Hyun-Woo; Shin, Hang-Sik; Han, Sun-Kee; Oh, Sae-Eun

    2007-03-01

    This study investigated the effects of food waste constituents on thermophilic (55 degrees C) anaerobic codigestion of sewage sludge and food waste by using statistical techniques based on biochemical methane potential tests. Various combinations of grain, vegetable, and meat as cosubstrate were tested, and then the data of methane potential (MP), methane production rate (MPR), and first-order kinetic constant of hydrolysis (kH) were collected for further analyses. Response surface methodology by the Box-Behnken design can verify the effects and their interactions of three variables on responses efficiently. MP was mainly affected by grain, whereas MPR and kH were affected by both vegetable and meat. Estimated polynomial regression models can properly explain the variability of experimental data with a high-adjusted R2 of 0.727, 0.836, and 0.915, respectively. By applying a series of optimization techniques, it was possible to find the proper criteria of cosubstrate. The optimal cosubstrate region was suggested based on overlay contours of overall mean responses. With the desirability contour plots, it was found that optimal conditions of cosubstrate for the maximum MPR (56.6 mL of CH4/g of chemical oxygen demand [COD]/day) were 0.71 g of COD/L of grain, 0.18 g of COD/L of vegetable, and 0.38 g of COD/L of meat by the simultaneous consideration of MP, MPR, and kH. Within the range of each factor examined, the corresponding optimal ratio of sewage sludge to cosubstrate was 71:29 as the COD basis. Elaborate discussions could yield practical operational strategies for the enhanced thermophilic anaerobic codigestion of sewage sludge and food waste. PMID:17385597

  19. Effects of selected thermophilic microorganisms on crude oils at elevated temperatures and pressures. Final report

    SciTech Connect

    Premuzic, E.T.; Lin, M.S.

    1995-07-01

    During the past several years, a considerable amount of work has been carried out showing that microbially enhanced oil recovery (MEOR) is promising and the resulting biotechnology may be deliverable. At the Brookhaven National Laboratory (BNL), systematic studies have been conducted which dealt with the effects of thermophilic and thermoadapted bacteria on the chemical and physical properties of selected types of crude oils at elevated temperatures and pressures. Particular attention was paid to heavy crude oils from Venezuela, California, Alabama, Arkansas, Wyoming, Alaska, and other oil producing areas. Current studies indicate that during the biotreatment several chemical and physical properties of crude oils are affected. The oils are (1) emulsified; (2) acidified; (3) there is a qualitative and quantitative change in light and heavy fractions of the crudes; (4) there are chemical changes in fractions containing sulfur compounds; (5) there is an apparent reduction in the concentration of trace metals; (6) the qualitative and quantitative changes appear to be microbial species dependent; and (7) there is a distinction between {open_quotes}biodegraded{close_quotes} and {open_quotes}biotreated{close_quotes} oils. Preliminary results indicate the introduced microorganisms may become the dominant species in the bioconversion of oils. These studies also indicate the biochemical interactions between crude oils and microorganisms follow distinct trends, characterized by a group of chemical markers. Core-flooding experiments have shown significant additional crude oil recoveries are achievable with thermophilic microorganisms at elevated temperatures similar to those found in oil reservoirs. In addition, the biochemical treatment of crude oils has technological applications in downstream processing of crude oils such as in upgrading of low grade oils and the production of hydrocarbon based detergents.

  20. A thermophilic and acid stable family-10 xylanase from the acidophilic fungus Bispora sp. MEY-1.

    PubMed

    Luo, Huiying; Li, Jiang; Yang, Jun; Wang, Hui; Yang, Yuhui; Huang, Huoqing; Shi, Pengjun; Yuan, Tiezheng; Fan, Yunliu; Yao, Bin

    2009-09-01

    A complete gene, xyl10C, encoding a thermophilic endo-1,4-beta-xylanase (XYL10C), was cloned from the acidophilic fungus Bispora sp. MEY-1 and expressed in Pichia pastoris. XYL10C shares highest nucleotide and amino acid sequence identities of 57.3 and 49.7%, respectively, with a putative xylanase from Aspergillus fumigatus Af293 of glycoside hydrolase family 10. A high expression level in P. pastoris (73,400 U ml(-1)) was achieved in a 3.7-l fermenter. The purified recombinant XYL10C was thermophilic, exhibiting maximum activity at 85 degrees C, which is higher than that reported from any fungal xylanase. The enzyme was also highly thermostable, exhibiting approximately 100% of the initial activity after incubation at 80 degrees C for 60 min and >87% of activity at 90 degrees C for 10 min. The half lives of XYL10C at 80 and 85 degrees C were approximately 45 and 3 h, respectively. It had two activity peaks at pH 3.0 and 4.5-5.0 (maximum), respectively, and was very acid stable, retaining more than 80% activity after incubation at pH 1.5-6.0 for 1 h. The enzyme was resistant to Co(2+), Mn(2+), Cr(3+) and Ag(+). The specific activity of XYL10C for oat spelt xylan was 18,831 U mg(-1). It also had wide substrate specificity and produced simple products (65.1% xylose, 25.0% xylobiose and 9.9% xylan polymer) from oat spelt xylan. PMID:19655217

  1. Translational control of small heat shock genes in mesophilic and thermophilic cyanobacteria by RNA thermometers

    PubMed Central

    Cimdins, Annika; Klinkert, Birgit; Aschke-Sonnenborn, Ursula; Kaiser, Friederike M; Kortmann, Jens; Narberhaus, Franz

    2014-01-01

    Cyanobacteria constitute a heterogeneous phylum of oxygen-producing, photosynthetic prokaryotes. They are susceptible to various stress conditions like heat, salt, or light stress, all inducing the cyanobacterial heat shock response (HSR). Cyanobacterial small heat shock proteins (sHsps) are known to preserve thylakoid membrane integrity under stress conditions, thereby protecting the photosynthesis machinery. In Synechocystis sp PCC 6803, synthesis of the sHsp Hsp17 is regulated by an RNA thermometer (RNAT) in the 5′-untranslated region (5′-UTR) of the hsp17 mRNA. RNATs are direct temperature sensors that control expression of many bacterial heat shock and virulence genes. They hinder translation at low temperatures by base pairing, thus blocking ribosome access to the mRNA.   To explore the temperature range in which RNATs act, we studied various RNAT candidates upstream of sHsp genes from mesophilic and thermophilic cyanobacteria. The mesophilic cyanobacteria Anabaena variabilis and Nostoc sp chromosomally encode two sHsps each. Reporter gene studies suggested RNAT-mediated post-transcriptional regulation of shsp expression in both organisms. Detailed structural analysis of the two A. variabilis candidates revealed two novel RNAT types. The first, avashort, regulates translation primarily by masking of the AUG translational start codon. The second, featuring an extended initial hairpin, thus named avalong, presumably makes use of complex tertiary interaction. The 5′-UTR of the small heat shock gene hspA in the thermophile Thermosynechococcus elongatus is predicted to adopt an extended secondary structure. Structure probing revealed that the ribosome binding site was blocked at temperatures below 55 °C. The results of this study demonstrate that cyanobacteria commonly use RNATs to control expression of their small heat shock genes. PMID:24755616

  2. Translational control of small heat shock genes in mesophilic and thermophilic cyanobacteria by RNA thermometers.

    PubMed

    Cimdins, Annika; Klinkert, Birgit; Aschke-Sonnenborn, Ursula; Kaiser, Friederike M; Kortmann, Jens; Narberhaus, Franz

    2014-01-01

    Cyanobacteria constitute a heterogeneous phylum of oxygen-producing, photosynthetic prokaryotes. They are susceptible to various stress conditions like heat, salt, or light stress, all inducing the cyanobacterial heat shock response (HSR). Cyanobacterial small heat shock proteins (sHsps) are known to preserve thylakoid membrane integrity under stress conditions, thereby protecting the photosynthesis machinery. In Synechocystis sp PCC 6803, synthesis of the sHsp Hsp17 is regulated by an RNA thermometer (RNAT) in the 5'-untranslated region (5'-UTR) of the hsp17 mRNA. RNATs are direct temperature sensors that control expression of many bacterial heat shock and virulence genes. They hinder translation at low temperatures by base pairing, thus blocking ribosome access to the mRNA.   To explore the temperature range in which RNATs act, we studied various RNAT candidates upstream of sHsp genes from mesophilic and thermophilic cyanobacteria. The mesophilic cyanobacteria Anabaena variabilis and Nostoc sp chromosomally encode two sHsps each. Reporter gene studies suggested RNAT-mediated post-transcriptional regulation of shsp expression in both organisms. Detailed structural analysis of the two A. variabilis candidates revealed two novel RNAT types. The first, avashort, regulates translation primarily by masking of the AUG translational start codon. The second, featuring an extended initial hairpin, thus named avalong, presumably makes use of complex tertiary interaction. The 5'-UTR of the small heat shock gene hspA in the thermophile Thermosynechococcus elongatus is predicted to adopt an extended secondary structure. Structure probing revealed that the ribosome binding site was blocked at temperatures below 55 °C. The results of this study demonstrate that cyanobacteria commonly use RNATs to control expression of their small heat shock genes. PMID:24755616

  3. Detection of Putatively Thermophilic Anaerobic Methanotrophs in Diffuse Hydrothermal Vent Fluids

    PubMed Central

    Huber, Julie A.; Chernyh, Nikolay A.; Bonch-Osmolovskaya, Elizaveta A.; Lebedinsky, Alexander V.

    2013-01-01

    The anaerobic oxidation of methane (AOM) is carried out by a globally distributed group of uncultivated Euryarchaeota, the anaerobic methanotrophic arachaea (ANME). In this work, we used G+C analysis of 16S rRNA genes to identify a putatively thermophilic ANME group and applied newly designed primers to study its distribution in low-temperature diffuse vent fluids from deep-sea hydrothermal vents. We found that the G+C content of the 16S rRNA genes (PGC) is significantly higher in the ANME-1GBa group than in other ANME groups. Based on the positive correlation between the PGC and optimal growth temperatures (Topt) of archaea, we hypothesize that the ANME-1GBa group is adapted to thrive at high temperatures. We designed specific 16S rRNA gene-targeted primers for the ANME-1 cluster to detect all phylogenetic groups within this cluster, including the deeply branching ANME-1GBa group. The primers were successfully tested both in silico and in experiments with sediment samples where ANME-1 phylotypes had previously been detected. The primers were further used to screen for the ANME-1 microorganisms in diffuse vent fluid samples from deep-sea hydrothermal vents in the Pacific Ocean, and sequences belonging to the ANME-1 cluster were detected in four individual vents. Phylotypes belonging to the ANME-1GBa group dominated in clone libraries from three of these vents. Our findings provide evidence of existence of a putatively extremely thermophilic group of methanotrophic archaea that occur in geographically and geologically distinct marine hydrothermal habitats. PMID:23183981

  4. Roles of Thermophiles and Fungi in Bitumen Degradation in Mostly Cold Oil Sands Outcrops

    PubMed Central

    Wong, Man-Ling; An, Dongshan; Caffrey, Sean M.; Soh, Jung; Dong, Xiaoli; Sensen, Christoph W.; Oldenburg, Thomas B. P.; Larter, Steve R.

    2015-01-01

    Oil sands are surface exposed in river valley outcrops in northeastern Alberta, where flat slabs (tablets) of weathered, bitumen-saturated sandstone can be retrieved from outcrop cliffs or from riverbeds. Although the average yearly surface temperature of this region is low (0.7°C), we found that the temperatures of the exposed surfaces of outcrop cliffs reached 55 to 60°C on sunny summer days, with daily maxima being 27 to 31°C. Analysis of the cooccurrence of taxa derived from pyrosequencing of 16S/18S rRNA genes indicated that an aerobic microbial network of fungi and hydrocarbon-, methane-, or acetate-oxidizing heterotrophic bacteria was present in all cliff tablets. Metagenomic analyses indicated an elevated presence of fungal cytochrome P450 monooxygenases in these samples. This network was distinct from the heterotrophic community found in riverbeds, which included fewer fungi. A subset of cliff tablets had a network of anaerobic and/or thermophilic taxa, including methanogens, Firmicutes, and Thermotogae, in the center. Long-term aerobic incubation of outcrop samples at 55°C gave a thermophilic microbial community. Analysis of residual bitumen with a Fourier transform ion cyclotron resonance mass spectrometer indicated that aerobic degradation proceeded at 55°C but not at 4°C. Little anaerobic degradation was observed. These results indicate that bitumen degradation on outcrop surfaces is a largely aerobic process with a minor anaerobic contribution and is catalyzed by a consortium of bacteria and fungi. Bitumen degradation is stimulated by periodic high temperatures on outcrop cliffs, which cause significant decreases in bitumen viscosity. PMID:26209669

  5. Moderately luminous Type II supernovae

    NASA Astrophysics Data System (ADS)

    Inserra, C.; Pastorello, A.; Turatto, M.; Pumo, M. L.; Benetti, S.; Cappellaro, E.; Botticella, M. T.; Bufano, F.; Elias-Rosa, N.; Harutyunyan, A.; Taubenberger, S.; Valenti, S.; Zampieri, L.

    2013-07-01

    Context. Core-collapse Supernovae (CC-SNe) descend from progenitors more massive than about 8 M⊙. Because of the young age of the progenitors, the ejecta may eventually interact with the circumstellar medium (CSM) via highly energetic processes detectable in the radio, X-ray, ultraviolet (UV) and, sometimes, in the optical domains. Aims: In this paper we present ultraviolet, optical and near infrared observations of five Type II SNe, namely SNe 2009dd, 2007pk, 2010aj, 1995ad, and 1996W. Together with few other SNe they form a group of moderately luminous Type II events. We investigate the photometric similarities and differences among these bright objects. We also attempt to characterise them by analysing the spectral evolutions, in order to find some traces of CSM-ejecta interaction. Methods: We collected photometry and spectroscopy with several telescopes in order to construct well-sampled light curves and spectral evolutions from the photospheric to the nebular phases. Both photometry and spectroscopy indicate a degree of heterogeneity in this sample. Modelling the data of SNe 2009dd, 2010aj and 1995ad allows us to constrain the explosion parameters and the properties of the progenitor stars. Results: The light curves have luminous peak magnitudes (-16.95 < MB < -18.70). The ejected masses of 56Ni for three SNe span a wide range of values (2.8 × 10-2 M⊙ < M(56Ni)< 1.4 × 10-1 M⊙), while for a fourth (SN 2010aj) we could determine a stringent upper limit (7 × 10-3 M⊙). Clues of interaction, such as the presence of high velocity (HV) features of the Balmer lines, are visible in the photospheric spectra of SNe 2009dd and 1996W. For SN 2007pk we observe a spectral transition from a Type IIn to a standard Type II SN. Modelling the observations of SNe 2009dd, 2010aj and 1995ad with radiation hydrodynamics codes, we infer kinetic plus thermal energies of about 0.2-0.5 foe, initial radii of 2-5 × 1013 cm and ejected masses of ~5.0-9.5 M⊙. Conclusions: These

  6. USGS MODERATE RESOLUTION LAND IMAGING

    NASA Astrophysics Data System (ADS)

    Dwyer, J. L.; Willems, J. S.

    2009-12-01

    For the past 37 years, the Landsat series of satellites has provided continuous data of the Earth’s land masses, coastal boundaries, and coral reefs creating an unprecedented comprehensive record of landscape dynamics. Landsat 5 and 7 continue to capture hundreds of images of the Earth’s surface each day. In mid-December 2008, the USGS made the entire Landsat archive available to everyone, anywhere, at anytime via the Internet at no cost to the user. The opening of the Landsat archive, the longest record of the terrestrial environment, is a revolution that will affect the future of moderate resolution Earth observations, enabling scientists to address research questions and develop operational applications that were previously cost prohibitive. In addition, the time-series data richness of the archive allows for the development of essential climate variables used to monitor the causes and consequences of lands cover change as a function of climate variability and anthropogenic influences. Landsat is unique as a single source of systematic, global land observations in terms of the number of spectral bands, global collection capacity, image quality, and the proven fidelity of its calibrated sensors. Through the Land Remote Sensing Policy Act of 1992 and the Presidential Decision Direct/NSTC-3 (1994), as amended on October 16, 2000, the U.S Geological Survey (USGS) is charged to ensure the continuity of Landsat data. To accomplish this, the USGS, in partnership with the National Aeronautics and Space Administration (NASA), is currently preparing for the launch of the Landsat Data Continuity Mission (LDCM) in December 2012, the eighth satellite in the Landsat Program. The LDCM will ensure the continuation of the Landsat record and will consist of significant improvements in radiometric response and additional spectral bands, from which high quality data products will be generated and accessible to users at no cost.

  7. Anoxybacillus kamchatkensis sp. nov., a novel thermophilic facultative aerobic bacterium with a broad pH optimum from the Geyser valley, Kamchatka.

    PubMed

    Kevbrin, Vadim V; Zengler, Karsten; Lysenko, Anatolii M; Wiegel, Juergen

    2005-10-01

    A facultative aerobic, moderately thermophilic, spore forming bacterium, strain JW/VK-KG4 was isolated from an enrichment culture obtained from the Geyser valley, a geo-thermally heated environment located in the Kamchatka peninsula (Far East region of Russia). The cells were rod shaped, motile, peritrichous flagellated stained Gram positive and had a Gram positive type cell wall. Aerobically, the strain utilized a range of carbohydrates including glucose, fructose, trehalose, proteinuous substrates, and pectin as well. Anaerobically, only carbohydrates are utilized. When growing on carbohydrates, the strain required yeast extract and vitamin B(12). Anaerobically, glucose was fermented to lactate as main product and acetate, formate, ethanol as minor products. Aerobically, even in well-aerated cultures (agitated at 500 rpm), glucose oxidation was incomplete and lactate and acetate were found in culture supernatants as by-products. Optimal growth of the isolate was observed at pH(25 C) 6.8-8.5 and 60 degrees C. The doubling times on glucose at optimal growth conditions were 34 min (aerobically) and 40 min (anaerobically). The G+C content was 42.3 mol% as determined by T(m) assay. Sequence analysis of the 16S rRNA gene indicated an affiliation of strain JW/VK-KG4 with Anoxybacillus species. Based on its morphology, physiology, phylogenetic relationship and its low DNA-DNA homology with validly published species of Anoxybacillus, it is proposed that strain JW/VK-KG4 represents a new species in the genus Anoxybacillus as A. kamchatkensis sp. nov. The type strain for the novel species is JW/VK-KG4(T) (=DSM 14988, =ATCC BAA-549). The GenBank accession number for the 16S rDNA sequence is AF510985. PMID:16142505

  8. Unique plasmids generated via pUC replicon mutagenesis in an error-prone thermophile derived from Geobacillus kaustophilus HTA426.

    PubMed

    Kobayashi, Jyumpei; Tanabiki, Misaki; Doi, Shohei; Kondo, Akihiko; Ohshiro, Takashi; Suzuki, Hirokazu

    2015-11-01

    The plasmid pGKE75-catA138T, which comprises pUC18 and the catA138T gene encoding thermostable chloramphenicol acetyltransferase with an A138T amino acid replacement (CATA138T), serves as an Escherichia coli-Geobacillus kaustophilus shuttle plasmid that confers moderate chloramphenicol resistance on G. kaustophilus HTA426. The present study examined the thermoadaptation-directed mutagenesis of pGKE75-catA138T in an error-prone thermophile, generating the mutant plasmid pGKE75(αβ)-catA138T responsible for substantial chloramphenicol resistance at 65°C. pGKE75(αβ)-catA138T contained no mutation in the catA138T gene but had two mutations in the pUC replicon, even though the replicon has no apparent role in G. kaustophilus. Biochemical characterization suggested that the efficient chloramphenicol resistance conferred by pGKE75(αβ)-catA138T is attributable to increases in intracellular CATA138T and acetyl-coenzyme A following a decrease in incomplete forms of pGKE75(αβ)-catA138T. The decrease in incomplete plasmids may be due to optimization of plasmid replication by RNA species transcribed from the mutant pUC replicon, which were actually produced in G. kaustophilus. It is noteworthy that G. kaustophilus was transformed with pGKE75(αβ)-catA138T using chloramphenicol selection at 60°C. In addition, a pUC18 derivative with the two mutations propagated in E. coli at a high copy number independently of the culture temperature and high plasmid stability. Since these properties have not been observed in known plasmids, the outcomes extend the genetic toolboxes for G. kaustophilus and E. coli. PMID:26319877

  9. Unique Plasmids Generated via pUC Replicon Mutagenesis in an Error-Prone Thermophile Derived from Geobacillus kaustophilus HTA426

    PubMed Central

    Kobayashi, Jyumpei; Tanabiki, Misaki; Doi, Shohei; Kondo, Akihiko; Ohshiro, Takashi

    2015-01-01

    The plasmid pGKE75-catA138T, which comprises pUC18 and the catA138T gene encoding thermostable chloramphenicol acetyltransferase with an A138T amino acid replacement (CATA138T), serves as an Escherichia coli-Geobacillus kaustophilus shuttle plasmid that confers moderate chloramphenicol resistance on G. kaustophilus HTA426. The present study examined the thermoadaptation-directed mutagenesis of pGKE75-catA138T in an error-prone thermophile, generating the mutant plasmid pGKE75αβ-catA138T responsible for substantial chloramphenicol resistance at 65°C. pGKE75αβ-catA138T contained no mutation in the catA138T gene but had two mutations in the pUC replicon, even though the replicon has no apparent role in G. kaustophilus. Biochemical characterization suggested that the efficient chloramphenicol resistance conferred by pGKE75αβ-catA138T is attributable to increases in intracellular CATA138T and acetyl-coenzyme A following a decrease in incomplete forms of pGKE75αβ-catA138T. The decrease in incomplete plasmids may be due to optimization of plasmid replication by RNA species transcribed from the mutant pUC replicon, which were actually produced in G. kaustophilus. It is noteworthy that G. kaustophilus was transformed with pGKE75αβ-catA138T using chloramphenicol selection at 60°C. In addition, a pUC18 derivative with the two mutations propagated in E. coli at a high copy number independently of the culture temperature and high plasmid stability. Since these properties have not been observed in known plasmids, the outcomes extend the genetic toolboxes for G. kaustophilus and E. coli. PMID:26319877

  10. Patient Moderator Interaction in Online Health Communities

    PubMed Central

    Huh, Jina; McDonald, David W.; Hartzler, Andrea; Pratt, Wanda

    2013-01-01

    An increasing number of people visit online health communities to share experiences and seek health information. Although studies have enumerated reasons for patients’ visits to online communities for health information from peers, we know little about how patients gain health information from the moderators in these communities. We qualitatively analyze 480 patient and moderator posts from six communities to understand how moderators fulfill patients’ information needs. Our findings show that patients use the community as an integral part of their health management practices. Based on our results, we suggest enhancements to moderated online health communities for their unique role to support patient care. PMID:24551364

  11. The effect of shock loading on the performance of a thermophilic anaerobic contact reactor at constant organic loading rate

    PubMed Central

    2014-01-01

    The influences of organic loading disturbances on the process performance of a thermophilic anaerobic contact reactor treating potato-processing wastewater were investigated. For this purpose, while the reactor was operated at steady state conditions with organic loading rate of 5.5 kg COD/m3 · day, an instant acetate concentration increase (1 g/L) was introduced to the reactor. During the shock loading test of acetate, it was observed that the overall process performance was adversely affected by all the shock loading, however, the system reached steady state conditions less than 24 hours of operation indicating that thermophilic anaerobic contact reactor is resistant to shock loading and be capable of returning its normal conditions within a short time period. PMID:24872886

  12. [Pilot study of thermal treatment/thermophilic anaerobic digestion process treating waste activated sludge of high solid content].

    PubMed

    Wu, Jing; Wang, Guang-qi; Cao, Zhi-ping; Li, Zhong-hua; Hu, Yu-ying; Wang, Kai-jun; Zu, Jian-e

    2014-09-01

    A pilot-scale experiment about the process of "thermal pretreatment at 70°C/thermophilic anaerobic digestion" of waste activated sludge of high solid content (8% -9% ) was conducted. The process employed thermal treatment of 3 days to accelerate the hydrolysis and thermophilic digestion to enhance anaerobic reaction. Thus it was good at organic removal and stabilization. When the solid retention time (SRT) was longer than 20 days, the VSS removal rate was greater than 42. 22% and it was linearly correlated to the SRT of the aerobic digestion with the R2 of 0. 915 3. It was suggested that SRT of anaerobic digestion was 25 days in practice. VSS removal rate and biogas production rate of the pilot experiment were similar to those of the run-well traditional full-scale sludge anaerobic digestion plants (solid content 3% -5% ) and the plant of high solid content using German technique. PMID:25518666

  13. Glutaraldehyde cross-linking of immobilized thermophilic esterase on hydrophobic macroporous resin for application in poly(ε-caprolactone) synthesis.

    PubMed

    Wang, Min; Shi, Hui; Wu, Di; Han, Haobo; Zhang, Jianxu; Xing, Zhen; Wang, Shuang; Li, Quanshun

    2014-01-01

    The immobilized thermophilic esterase from Archaeoglobus fulgidus was successfully constructed through the glutaraldehyde-mediated covalent coupling after its physical adsorption on a hydrophobic macroporous resin, Sepabeads EC-OD. Through 0.05% glutaraldehyde treatment, the prevention of enzyme leaching and the maintenance of catalytic activity could be simultaneously realized. Using the enzymatic ring-opening polymerization of ε-caprolactone as a model, effects of organic solvents and reaction temperature on the monomer conversion and product molecular weight were systematically investigated. After the optimization of reaction conditions, products were obtained with 100% monomer conversion and Mn values lower than 1010 g/mol. Furthermore, the cross‑linked immobilized thermophilic esterase exhibited an excellent operational stability, with monomer conversion values exceeding 90% over the course of 12 batch reactions, still more than 80% after 16 batch reactions. PMID:25006789

  14. Efficient production of 2,3-butanediol from corn stover hydrolysate by using a thermophilic Bacillus licheniformis strain.

    PubMed

    Li, Lixiang; Li, Kun; Wang, Kai; Chen, Chao; Gao, Chao; Ma, Cuiqing; Xu, Ping

    2014-10-01

    In this study, a thermophilic Bacillus licheniformis strain X10 was newly isolated for 2,3-butanediol (2,3-BD) production from lignocellulosic hydrolysate. Strain X10 could utilize glucose and xylose simultaneously without carbon catabolite repression. In addition, strain X10 possesses high tolerance to fermentation inhibitors including furfural, vanillin, formic acid, and acetic acid. In a fed-batch fermentation, 74.0g/L of 2,3-BD was obtained from corn stover hydrolysate, with a productivity of 2.1g/Lh and a yield of 94.6%. Thus, this thermophilic B. licheniformis strain is a candidate for the development of efficient industrial production of 2,3-BD from corn stover hydrolysate. PMID:25151068

  15. Hydrogen Limitation and Syntrophic Growth among Natural Assemblages of Thermophilic Methanogens at Deep-sea Hydrothermal Vents

    PubMed Central

    Topçuoğlu, Begüm D.; Stewart, Lucy C.; Morrison, Hilary G.; Butterfield, David A.; Huber, Julie A.; Holden, James F.

    2016-01-01

    Thermophilic methanogens are common autotrophs at hydrothermal vents, but their growth constraints and dependence on H2 syntrophy in situ are poorly understood. Between 2012 and 2015, methanogens and H2-producing heterotrophs were detected by growth at 80°C and 55°C at most diffuse (7–40°C) hydrothermal vent sites at Axial Seamount. Microcosm incubations of diffuse hydrothermal fluids at 80°C and 55°C demonstrated that growth of thermophilic and hyperthermophilic methanogens is primarily limited by H2 availability. Amendment of microcosms with NH4+ generally had no effect on CH4 production. However, annual variations in abundance and CH4 production were observed in relation to the eruption cycle of the seamount. Microcosm incubations of hydrothermal fluids at 80°C and 55°C supplemented with tryptone and no added H2 showed CH4 production indicating the capacity in situ for methanogenic H2 syntrophy. 16S rRNA genes were found in 80°C microcosms from H2-producing archaea and H2-consuming methanogens, but not for any bacteria. In 55°C microcosms, sequences were found from H2-producing bacteria and H2-consuming methanogens and sulfate-reducing bacteria. A co-culture of representative organisms showed that Thermococcus paralvinellae supported the syntrophic growth of Methanocaldococcus bathoardescens at 82°C and Methanothermococcus sp. strain BW11 at 60°C. The results demonstrate that modeling of subseafloor methanogenesis should focus primarily on H2 availability and temperature, and that thermophilic H2 syntrophy can support methanogenesis within natural microbial assemblages and may be an important energy source for thermophilic autotrophs in marine geothermal environments. PMID:27547206

  16. Superoxide Dismutase Structures, Stability, Mechanism and Insights into the Human Disease Amyotrophic Lateral Sclerosis from Eukaryotic Thermophile Alvinella pompejana

    PubMed Central

    Shin, David S.; DiDonato, Michael; Barondeau, David P.; Hura, Greg L.; Hitomi, Chiharu; Berglund, J. Andrew; Getzoff, Elizabeth D.; Cary, S. Craig; Tainer, John A.

    2009-01-01

    Summary Prokaroytic thermophiles supply stable human protein homologs for structural biology; yet, eukaryotic thermophiles would provide more similar macromolecules plus those missing in microbes. Alvinella pompejana is a deep-sea hydrothermal-vent worm that has been found in temperatures averaging as high as 68 °C, with spikes up to 84 °C. Here, we used Cu, Zn superoxide dismutase (SOD) to test if this eukaryotic thermophile can provide insights into macromolecular mechanisms and stability, by supplying better stable mammalian homologs for structural biology and other biophysical characterizations than those from prokaryotic thermophiles. Identification, cloning, characterization, X-ray scattering (SAXS) and crystal structure determinations show that Alvinella pompejana SOD (ApSOD) is super-stable, homologous, and informative. SAXS solution analyses identify the human-like ApSOD dimer. The crystal structure shows the active site at 0.99 Å resolution, plus anchoring interaction motifs in loops and termini accounting for enhanced stability of ApSOD versus human SOD. Such stabilizing features may reduce movements that promote inappropriate intermolecular interactions, such as amyloid-like filaments found in SOD mutants causing the neurodegenerative disease familial amyotrophic lateral sclerosis or Lou Gehrig’s disease. ApSOD further provides a long-sought SOD product complex at 1.35 Å resolution, suggesting a unified inner sphere mechanism for catalysis involving metal ion movement. Notably, this proposed mechanism resolves apparent paradoxes regarding electron transfer. These results extend knowledge of SOD stability and catalysis, and suggest that the eukaryote A. pompejana provides macromolecules highly similar to those from humans, but with enhanced stability more suitable for scientific and medical applications. PMID:19063897

  17. Hydrogen Limitation and Syntrophic Growth among Natural Assemblages of Thermophilic Methanogens at Deep-sea Hydrothermal Vents.

    PubMed

    Topçuoğlu, Begüm D; Stewart, Lucy C; Morrison, Hilary G; Butterfield, David A; Huber, Julie A; Holden, James F

    2016-01-01

    Thermophilic methanogens are common autotrophs at hydrothermal vents, but their growth constraints and dependence on H2 syntrophy in situ are poorly understood. Between 2012 and 2015, methanogens and H2-producing heterotrophs were detected by growth at 80°C and 55°C at most diffuse (7-40°C) hydrothermal vent sites at Axial Seamount. Microcosm incubations of diffuse hydrothermal fluids at 80°C and 55°C demonstrated that growth of thermophilic and hyperthermophilic methanogens is primarily limited by H2 availability. Amendment of microcosms with NH4 (+) generally had no effect on CH4 production. However, annual variations in abundance and CH4 production were observed in relation to the eruption cycle of the seamount. Microcosm incubations of hydrothermal fluids at 80°C and 55°C supplemented with tryptone and no added H2 showed CH4 production indicating the capacity in situ for methanogenic H2 syntrophy. 16S rRNA genes were found in 80°C microcosms from H2-producing archaea and H2-consuming methanogens, but not for any bacteria. In 55°C microcosms, sequences were found from H2-producing bacteria and H2-consuming methanogens and sulfate-reducing bacteria. A co-culture of representative organisms showed that Thermococcus paralvinellae supported the syntrophic growth of Methanocaldococcus bathoardescens at 82°C and Methanothermococcus sp. strain BW11 at 60°C. The results demonstrate that modeling of subseafloor methanogenesis should focus primarily on H2 availability and temperature, and that thermophilic H2 syntrophy can support methanogenesis within natural microbial assemblages and may be an important energy source for thermophilic autotrophs in marine geothermal environments. PMID:27547206

  18. Evaluation of thermophilic anaerobic digestion processes for full-scale Class A biosolids disinfection at Hyperion Treatment Plant.

    PubMed

    Iranpour, R; Cox, H H J

    2007-05-01

    This paper describes 5 phases of full-scale testing at the City of Los Angeles Hyperion Treatment Plant (HTP) for producing Class A biosolids (U.S. EPA Part 503 Biosolids Rule) by thermophilic anaerobic digestion. Phases I and II were tests with a two-stage continuous-batch process in a thermophilic battery of six digesters and a designated post-digestion train that was isolated from mesophilic operations. These tests demonstrated that digester outflow biosolids met the Class A limits for fecal coliforms and Salmonella sp. However, fecal coliform densities sharply increased during post-digestion. The recurrence was possibly related to a combination of a large drop of the biosolids temperature after the dewatering centrifuges and contamination of thermophilically digested biosolids from mesophilic operations. Phase III was conducted after insulation and electrical heat-tracing of the post-digestion train to maintain a biosolids temperature throughout post-digestion at about the same level as in the digester outflow. Biosolids monitoring at the last points of plant control (silos at Truck Loading Facility and farm for land application) indicated that fecal coliform recurrence was prevented. After completing the conversion of HTP to thermophilic operation, certification tests of Phases IV and V demonstrated Class A compliance of a two-stage continuous-batch process under Alternatives 1 and 3 of the Part 503 Biosolids Rule, respectively. HTP received the permit for Class A (indeed exceptional quality) biosolids land application in Kern County, California, in December 2002 under Alternative 3. Since 2003, HTP has consistently complied with the federal and local standards for Class A biosolids, indicating that Class A limits can be met under conditions less stringent than defined by the Alternative 1 time-temperature requirement for batch treatment. PMID:17054113

  19. Comprehensive microbial analysis of combined mesophilic anaerobic-thermophilic aerobic process treating high-strength food wastewater.

    PubMed

    Jang, Hyun Min; Ha, Jeong Hyub; Park, Jong Moon; Kim, Mi-Sun; Sommer, Sven G

    2015-04-15

    A combined mesophilic anaerobic-thermophilic aerobic process was used to treat high-strength food wastewater in this study. During the experimental period, most of solid residue from the mesophilic anaerobic reactor (R1) was separated by centrifugation and introduced into the thermophilic aerobic reactor (R2) for further digestion. Then, thermophilic aerobically-digested sludge was reintroduced into R1 to enhance reactor performance. The combined process was operated with two different Runs: Run I with hydraulic retention time (HRT) = 40 d (corresponding OLR = 3.5 kg COD/m(3) d) and Run II with HRT = 20 d (corresponding OLR = 7 kg COD/m(3)). For a comparison, a single-stage mesophilic anaerobic reactor (R3) was operated concurrently with same OLRs and HRTs as the combined process. During the overall digestion, all reactors showed high stability without pH control. The combined process demonstrated significantly higher organic matter removal efficiencies (over 90%) of TS, VS and COD and methane production than did R3. Quantitative real-time PCR (qPCR) results indicated that higher populations of both bacteria and archaea were maintained in R1 than in R3. Pyrosequencing analysis revealed relatively high abundance of phylum Actinobacteria in both R1 and R2, and a predominance of phyla Synergistetes and Firmicutes in R3 during Run II. Furthermore, R1 and R2 shared genera (Prevotella, Aminobacterium, Geobacillus and Unclassified Actinobacteria), which suggests synergy between mesophilic anaerobic digestion and thermophilic aerobic digestion. For archaea, in R1 methanogenic archaea shifted from genus Methanosaeta to Methanosarcina, whereas genera Methanosaeta, Methanobacterium and Methanoculleus were predominant in R3. The results demonstrated dynamics of key microbial populations that were highly consistent with an enhanced reactor performance of the combined process. PMID:25689817

  20. Biochemical characterization of a thermophilic β-mannanase from Talaromyces leycettanus JCM12802 with high specific activity.

    PubMed

    Wang, Caihong; Luo, Huiying; Niu, Canfang; Shi, Pengjun; Huang, Huoqing; Meng, Kun; Bai, Yingguo; Wang, Kun; Hua, Huifang; Yao, Bin

    2015-02-01

    Thermophilic β-mannanases are of increasing importance for wide industrial applications. In the current study, gene cloning, functional expression in Pichia pastoris, and characterization of a thermophilic β-mannanase (Man5A) from thermophilic Talaromyces leycettanus JCM12802 are reported. Deduced Man5A exhibits the highest identity with a putative β-mannanase from Talaromyces stipitatus ATCC10500 (70.3 %) and is composed of an N-terminal signal peptide, a fungal-type carbohydrate-binding module (CBM) of family 1, and a catalytic domain of glycosyl hydrolase (GH) family 5 at the C-terminus. Two recombinant proteins with different glycosylation levels, termed Man5A1 (72 kDa) and Man5A2 (60 kDa), were identified after purification. Both enzymes were thermophilic, exhibiting optimal activity at 85-90 °C, and were highly stable at 70 °C. Man5A1 and Man5A2 had a pH optimum of 4.5 and 4.0, respectively, and were highly stable over the broad pH range of 3.0-10.0. Most metal ions and sodium dodecyl sulfate (SDS) had no effect on the enzymatic activities. Man5A1 and Man5A2 exhibited high specific activity (2,160 and 1,800 U/mg, respectively) when using locust bean gum as the substrate. The CBM1 and two key residues D191 and R286 were found to affect Man5A thermostability. Man5A displays a classical four-site-binding mode, hydrolyzing mannooligosaccharides into smaller units, galactomannan into mannose and mannobiose, and glucomanman into mannose, mannobiose, and mannopentaose, respectively. All these properties make Man5A a good candidate for extensive applications in the bioconversion, pulp bleaching, textile, food, and feed industries. PMID:25104029

  1. The chemostat study of metabolic distribution in extreme-thermophilic (70°C) mixed culture fermentation.

    PubMed

    Zhang, Fang; Chen, Yun; Dai, Kun; Zeng, Raymond J

    2014-12-01

    The effects of pH, hydrogen partial pressure (PH2), and influent glucose concentration on the metabolic distribution in chemostat were investigated in this work in extreme-thermophilic mixed culture fermentation (MCF) process. The results showed that acetate, ethanol, and hydrogen were the main metabolites. A shift of ethanol to acetate and hydrogen was observed as pH increasing from 4.0 to 7.0 or PH2 decreasing from 0.64 to 0.05 atm. The maximum hydrogen yield was 3.16 ± 0.16 mol/mol glucose at PH2 0.05 atm. Lactate was only accumulated at low pH or high influent glucose concentration, while others such as butyrate and formate were rather low. Thermodynamic analysis illustrated that a mixture of acetate, ethanol, and/or lactate was essential for hydrogen production in extreme-thermophilic MCF. The hydrogen-producing rate was also calculated, and the maximum value was 2.2 ± 0.1 L/(L-reactor/day) at PH2 0.05 atm. Except hydrogen, other metabolites, such as liquid fatty acids and biofuels, could also be the producing targets in extreme-thermophilic MCF. PMID:25341404

  2. Effect of limited air exposure and comparative performance between thermophilic and mesophilic solid-state anaerobic digestion of switchgrass.

    PubMed

    Sheets, Johnathon P; Ge, Xumeng; Li, Yebo

    2015-03-01

    Switchgrass is an attractive feedstock for biogas production via anaerobic digestion (AD). Many studies have used switchgrass for liquid anaerobic digestion (L-AD), but few have used switchgrass for solid-state anaerobic digestion (SS-AD). Limited air exposure to the reactor headspace has been adopted in commercial scale anaerobic digesters for different applications. However, little research has examined the effect of limited air exposure on biogas production during SS-AD. In this study, the effects of air exposure and total solids (TS) content on SS-AD performance were evaluated under mesophilic (36±1°C) and thermophilic (55±0.3°C) conditions. Limited air exposure did not significantly influence the methane yield during SS-AD. Thermophilic SS-AD had greater methane yields (102-145LCH4kg(-1)VSadded) than mesophilic SS-AD (88-113LCH4kg(-1)VSadded). Both mesophilic SS-AD (73-136GJ) and thermophilic SS-AD (2-95GJ) produced positive net energy based on a theoretical 'garage-type' SS-AD digester operating in a temperate climate. PMID:25618499

  3. Biogas Upgrading via Hydrogenotrophic Methanogenesis in Two-Stage Continuous Stirred Tank Reactors at Mesophilic and Thermophilic Conditions.

    PubMed

    Bassani, Ilaria; Kougias, Panagiotis G; Treu, Laura; Angelidaki, Irini

    2015-10-20

    This study proposes an innovative setup composed by two stage reactors to achieve biogas upgrading coupling the CO2 in the biogas with external H2 and subsequent conversion into CH4 by hydrogenotrophic methanogenesis. In this configuration, the biogas produced in the first reactor was transferred to the second one, where H2 was injected. This configuration was tested at both mesophilic and thermophilic conditions. After H2 addition, the produced biogas was upgraded to average CH4 content of 89% in the mesophilic reactor and 85% in the thermophilic. At thermophilic conditions, a higher efficiency of CH4 production and CO2 conversion was recorded. The consequent increase of pH did not inhibit the process indicating adaptation of microorganisms to higher pH levels. The effects of H2 on the microbial community were studied using high-throughput Illumina random sequences and full-length 16S rRNA genes extracted from the total sequences. The relative abundance of archaeal community markedly increased upon H2 addition with Methanoculleus as dominant genus. The increase of hydrogenotrophic methanogens and syntrophic Desulfovibrio and the decrease of aceticlastic methanogens indicate a H2-mediated shift toward the hydrogenotrophic pathway enhancing biogas upgrading. Moreover, Thermoanaerobacteraceae were likely involved in syntrophic acetate oxidation with hydrogenotrophic methanogens in absence of aceticlastic methanogenesis. PMID:26390125

  4. Anaerobic co-digestion of steam-treated Quercus serrata chips and sewage sludge under mesophilic and thermophilic conditions.

    PubMed

    Wang, Feng; Hidaka, Taira; Sakurai, Kensuke; Tsumori, Jun

    2014-08-01

    The biodegradation of Quercus serrata chips was evaluated by anaerobic digestion under various steam explosion conditions. In continuous experiments, untreated chips (W₀) and chips steam-treated at less than 1.0 MPa (W₁) and 2.0 MPa (W₄) were co-digested with sewage sludge (S₁ and S₂) taken from two different wastewater treatment plants. The apparent methane yield of W₁ and W₄ co-digested with S₁ (thermophilic) was 261 dm(3)/kgVS (volatile solids) and 248 dm(3)/kgVS, respectively. The apparent methane yield of W₄ co-digested with S₂ was 258 dm(3)/kgVS (mesophilic) and 271 dm(3)/kgVS (thermophilic). Methane production was inhibited by W₀ due to components released during hydrolysis. The methane conversion ratio of pretreated chips obtained in batch experiments varied from 40.5% to 53.8% (mesophilic) and from 49.0% to 63.7% (thermophilic). The methane conversion ratio increased with decreasing acid-soluble lignin content in the chips. PMID:24926605

  5. Nitrogen removal characteristics analyzed with gas and microbial community in thermophilic aerobic digestion for piggery waste treatment.

    PubMed

    Lee, J W; Lee, H W; Kim, S W; Lee, S Y; Park, Y K; Han, J H; Choi, S I; Yi, Y S; Yun, Z

    2004-01-01

    In order to characterize the nitrogen conversion characteristics in a thermophilic aerobic digestion (TAD) system, a laboratory study has been conducted with the analysis of effluent gas and microbial community in the sludge samples. The lab TAD system was operated with HRT of 3 days and 60 degrees C. Based on the nitrogen mass balance, it has been found that about 2/3 of the daily load of nitrogen was converted to the gaseous form of nitrogen whereas cellular transformation and unmetabolized nitrogen accounted for about 1/3. Among the gaseous nitrogen transformation, significant amount of influent nitrogen had been converted to N2 gas (29% of influent N) and N2O (9% of influent N). Ammonia conversion was only 28% of influent N. The detection of N2O gas is a clear indication of the biological nitrogen reduction process in the thermophilic aerobic digester. No conclusive evidence for the existence of aerobic deammonification has been found. The microbial community analysis showed that thermophilic bacteria such as Bacillus thermocloacae, Bacillus sp. and Clostridial groups dominated in this TAD reactor. The diverse microbial community in TAD sludge may play an important role in removing both strong organics and nitrogen from piggery waste. PMID:15137444

  6. Reducing waste contamination from animal-processing plants by anaerobic thermophilic fermentation and by flesh fly digestion.

    PubMed

    Marchaim, U; Gelman, A; Braverman, Y

    2003-01-01

    There is currently no market in Israel for the large amounts of waste from fish- and poultry-processing plants. Therefore, this waste is incinerated, as part of the measures to prevent the spread of pathogens. Anaerobic methanogenic thermophilic fermentation (AMTF) of wastes from the cattle-slaughtering industry was examined previously, as an effective system to treat pathogenic bacteria, and in this article, we discuss a combined method of digestion by thermophilic anaerobic bacteria and by flesh flies, as a means of waste treatment. The AMTF process was applied to the wastes on a laboratory scale, and digestion by rearing of flesh fly (Phaenicia sericata) and housefly (Musca domestica) larvae on the untreated raw material was done on a small scale and showed remarkable weight conversion to larvae. The yield from degradation of poultry waste by flesh fly was 22.47% (SD = 3.89) and that from fish waste degradation was 35.34% (SD = 12.42), which is significantly higher than that from rearing houseflies on a regular rearing medium. Bacterial contents before and after thermophilic anaerobic digestion, as well as the changes in the chemical composition of the components during the rearing of larvae, were also examined. PMID:12794287

  7. From mesophilic to thermophilic digestion: the transitions of anaerobic bacterial, archaeal, and fungal community structures in sludge and manure samples.

    PubMed

    Sun, Weimin; Yu, Guangwei; Louie, Tiffany; Liu, Tong; Zhu, Chengsheng; Xue, Gang; Gao, Pin

    2015-12-01

    The shift of microbial communities during a transition from mesophilic anaerobic digestion (MAD) to thermophilic anaerobic digestion (TAD) was characterized in two treatments. One treatment was inoculated with sludge and the other was inoculated with manure. In this study, methane was produced both in MAD and TAD, but TAD has slightly more methane produced than MAD. A broad phylogenetic spectrum of bacterial, archaeal, and fungal taxa at thermophilic conditions was detected. Coprothermobacter, Bacillus, Haloplasma, Clostridiisalibacter, Methanobacterium, Methanothermobacter, Saccharomycetales, Candida, Alternaria, Cladosporium, and Penicillium were found almost exclusively in TAD, suggesting their adaptation to thermophilic conditions and ecological roles in digesting the organic compounds. The characterization of the lesser-known fungal community revealed that fungi probably constituted an important portion of the overall community within TAD and contributed to this process by degrading complex organic compounds. The shift of the microbial communities between MAD and TAD implied that temperature drastically affected the microbial diversity in anaerobic digestion. In addition, the difference in microbial communities between sludge and manure indicated that different source of inoculum also affected the microbial diversity and community. PMID:26245681

  8. Mesophilic and thermophilic anaerobic digestion of the liquid fraction of pressed biowaste for high energy yields recovery.

    PubMed

    Micolucci, Federico; Gottardo, Marco; Cavinato, Cristina; Pavan, Paolo; Bolzonella, David

    2016-02-01

    Deep separate collection of the organic fraction of municipal solid waste generates streams with relatively low content of inert material and high biodegradability. This material can be conveniently treated to recovery both energy and material by means of simplified technologies like screw-press and extruder: in this study, the liquid fraction generated from pressed biowaste from kerbside and door-to-door collection was anaerobically digested in both mesophilic and thermophilic conditions while for the solid fraction composting is suggested. Continuous operation results obtained both in mesophilic and thermophilic conditions indicated that the anaerobic digestion of pressed biowaste was viable at all operating conditions tested, with the greatest specific gas production of 0.92m(3)/kgVSfed at an organic loading rate of 4.7kgVS/m(3)d in thermophilic conditions. Based on calculations the authors found that the expected energy recovery is highly positive. The contents of heavy metals and pathogens of fed substrate and effluent digestates were analyzed, and results showed low levels (below End-of-Waste 2014 criteria limits) for both the parameters thus indicating the good quality of digestate and its possible use for agronomic purposes. Therefore, both energy and material were effectively recovered. PMID:26427935

  9. Thermophilic sludge digestion improves energy balance and nutrient recovery potential in full-scale municipal wastewater treatment plants.

    PubMed

    De Vrieze, Jo; Smet, Davey; Klok, Jacob; Colsen, Joop; Angenent, Largus T; Vlaeminck, Siegfried E

    2016-10-01

    The conventional treatment of municipal wastewater by means of activated sludge is typically energy demanding. Here, the potential benefits of: (1) the optimization of mesophilic digestion; and (2) transitioning to thermophilic sludge digestion in three wastewater treatment plants (Tilburg-Noord, Land van Cuijk and Bath) in the Netherlands is evaluated, including a full-scale trial validation in Bath. In Tilburg-Noord, thermophilic sludge digestion covered the energy requirements of the plant (102%), whereas 111% of sludge operational treatment costs could be covered in Bath. Thermophilic sludge digestion also resulted in a strong increase in nutrient release. The potential for nutrient recovery was evaluated via: (1) stripping/absorption of ammonium; (2) autotrophic removal of ammonium via partial nitritation/anammox; and (3) struvite precipitation. This research shows that optimization of sludge digestion may lead to a strong increase in energy recovery, sludge treatment costs reduction, and the potential for advanced nutrient management in full-scale sewage treatment plants. PMID:27423372

  10. Best Practices in Online Conference Moderation

    ERIC Educational Resources Information Center

    De Schutter, Adrienne; Fahrni, Patricia; Rudolph, Jim

    2004-01-01

    Facilitation by a moderator is crucial to a purposeful and productive conference. The moderator keeps the session focused, and ensures that all participants receive feedback regarding their contributions to the discussion. These functions are particularly important in the otherwise impersonal context of online discussion. The current report…

  11. Pharmacodynamic Considerations for Moderate and Deep Sedation

    PubMed Central

    Becker, Daniel E.

    2012-01-01

    Moderate and deep sedation can be provided using various classes of drugs, each having unique mechanisms of action. While drugs within a given classification share similar mechanisms and effects, certain classes demonstrate superior efficacy but added concern regarding safety. This continuing education article will highlight essential principles of pharmacodynamics and apply these to drugs commonly used to produce moderate and deep sedation. PMID:22428972

  12. Assessing Moderator Variables: Two Computer Simulation Studies.

    ERIC Educational Resources Information Center

    Mason, Craig A.; And Others

    1996-01-01

    A strategy is proposed for conceptualizing moderating relationships based on their type (strictly correlational and classically correlational) and form, whether continuous, noncontinuous, logistic, or quantum. Results of computer simulations comparing three statistical approaches for assessing moderator variables are presented, and advantages of…

  13. Integrating Mediators and Moderators in Research Design

    ERIC Educational Resources Information Center

    MacKinnon, David P.

    2011-01-01

    The purpose of this article is to describe mediating variables and moderating variables and provide reasons for integrating them in outcome studies. Separate sections describe examples of moderating and mediating variables and the simplest statistical model for investigating each variable. The strengths and limitations of incorporating mediating…

  14. High rate mesophilic, thermophilic, and temperature phased anaerobic digestion of waste activated sludge: A pilot scale study

    SciTech Connect

    Bolzonella, David; Cavinato, Cristina; Fatone, Francesco; Pavan, Paolo; Cecchi, Franco

    2012-06-15

    Highlights: Black-Right-Pointing-Pointer High temperatures were tested in single and two-stage anaerobic digestion of waste activated sludge. Black-Right-Pointing-Pointer The increased temperature demonstrated the possibility of improving typical yields of the conventional mesophilic process. Black-Right-Pointing-Pointer The temperature phased anaerobic digestion process (65 + 55 Degree-Sign C) showed the best performances with yields of 0.49 m{sup 3}/kgVS{sub fed}. Black-Right-Pointing-Pointer Ammonia and phosphate released from solids destruction determined the precipitation of struvite in the reactor. - Abstract: The paper reports the findings of a two-year pilot scale experimental trial for the mesophilic (35 Degree-Sign C), thermophilic (55 Degree-Sign C) and temperature phased (65 + 55 Degree-Sign C) anaerobic digestion of waste activated sludge. During the mesophilic and thermophilic runs, the reactor operated at an organic loading rate of 2.2 kgVS/m{sup 3}d and a hydraulic retention time of 20 days. In the temperature phased run, the first reactor operated at an organic loading rate of 15 kgVS/m{sup 3}d and a hydraulic retention time of 2 days while the second reactor operated at an organic loading rate of 2.2 kgVS/m{sup 3}d and a hydraulic retention time of 18 days (20 days for the whole temperature phased system). The performance of the reactor improved with increases in temperature. The COD removal increased from 35% in mesophilic conditions, to 45% in thermophilic conditions, and 55% in the two stage temperature phased system. As a consequence, the specific biogas production increased from 0.33 to 0.45 and to 0.49 m{sup 3}/kgVS{sub fed} at 35, 55, and 65 + 55 Degree-Sign C, respectively. The extreme thermophilic reactor working at 65 Degree-Sign C showed a high hydrolytic capability and a specific yield of 0.33 gCOD (soluble) per gVS{sub fed}. The effluent of the extreme thermophilic reactor showed an average concentration of soluble COD and volatile

  15. Insight into Glycoside Hydrolases for Debranched Xylan Degradation from Extremely Thermophilic Bacterium Caldicellulosiruptor lactoaceticus

    PubMed Central

    Jia, Xiaojing; Mi, Shuofu; Wang, Jinzhi; Qiao, Weibo; Peng, Xiaowei; Han, Yejun

    2014-01-01

    Caldicellulosiruptor lactoaceticus 6A, an anaerobic and extremely thermophilic bacterium, uses natural xylan as carbon source. The encoded genes of C. lactoaceticus 6A for glycoside hydrolase (GH) provide a platform for xylan degradation. The GH family 10 xylanase (Xyn10A) and GH67 α-glucuronidase (Agu67A) from C. lactoaceticus 6A were heterologously expressed, purified and characterized. Both Xyn10A and Agu67A are predicted as intracellular enzymes as no signal peptides identified. Xyn10A and Agu67A had molecular weight of 47.0 kDa and 80.0 kDa respectively as determined by SDS-PAGE, while both appeared as homodimer when analyzed by gel filtration. Xyn10A displayed the highest activity at 80°C and pH 6.5, as 75°C and pH 6.5 for Agu67A. Xyn10A had good stability at 75°C, 80°C, and pH 4.5–8.5, respectively, and was sensitive to various metal ions and reagents. Xyn10A possessed hydrolytic activity towards xylo-oligosaccharides (XOs) and beechwood xylan. At optimum conditions, the specific activity of Xyn10A was 44.6 IU/mg with beechwood xylan as substrate, and liberated branched XOs, xylobiose, and xylose. Agu67A was active on branched XOs with methyl-glucuronic acids (MeGlcA) sub-chains, and primarily generated XOs equivalents and MeGlcA. The specific activity of Agu67A was 1.3 IU/mg with aldobiouronic acid as substrate. The synergistic action of Xyn10A and Agu67A was observed with MeGlcA branched XOs and xylan as substrates, both backbone and branched chain of substrates were degraded, and liberated xylose, xylobiose, and MeGlcA. The synergism of Xyn10A and Agu67A provided not only a thermophilic method for natural xylan degradation, but also insight into the mechanisms for xylan utilization of C. lactoaceticus. PMID:25184498

  16. Thermophilic nitrate-reducing microorganisms prevent sulfate reduction in cold marine sediments incubated at high temperature

    NASA Astrophysics Data System (ADS)

    Nepomnyashchaya, Yana; Rezende, Julia; Hubert, Casey

    2014-05-01

    Hydrogen sulphide produced during metabolism of sulphate-reducing microorganisms (SRM) is toxic, corrosive and causes detrimental oil reservoir souring. During secondary oil recovery, injecting oil reservoirs with seawater that is rich in sulphate and that also cools high temperature formations provides favourable growth conditions for SRM. Nitrate addition can prevent metabolism of SRM by stimulating nitrate-reducing microorganisms (NRM). The investigations of thermophilic NRM are needed to develop mechanisms to control the metabolism of SRM in high temperature oil field ecosystems. We therefore established a model system consisting of enrichment cultures of cold surface marine sediments from the Baltic Sea (Aarhus Bay) that were incubated at 60°C. Enrichments contained 25 mM nitrate and 40 mM sulphate as potential electron acceptors, and a mixture of the organic substrates acetate, lactate, propionate, butyrate (5 mM each) and yeast extract (0.01%) as potential carbon sources and electron donors. Slurries were incubated at 60°C both with and without initial pasteurization at 80°C for 2 hours. In the enrichments containing both nitrate and sulphate, the concentration of nitrate decreased indicating metabolic activity of NRM. After a four-hour lag phase the rate of nitrate reduction increased and the concentration of nitrate dropped to zero after 10 hours of incubation. The concentration of nitrite increased as the reduction of nitrate progressed and reached 16.3 mM after 12 hours, before being consumed and falling to 4.4 mM after 19-day of incubation. No evidence for sulphate reduction was observed in these cultures during the 19-day incubation period. In contrast, the concentration of sulphate decreased up to 50% after one week incubation in controls containing only sulphate but no nitrate. Similar sulfate reduction rates were seen in the pasteurized controls suggesting the presence of heat resistant SRM, whereas nitrate reduction rates were lower in the

  17. Champagne Pool (New Zealand) Thermophiles Yield Insights into the Evolution of Microbial Arsenic Resistance

    NASA Astrophysics Data System (ADS)

    Hug, K.; Krikowa, F.; Morgan, X.; Maher, W. A.; Stott, M. B.; Moreau, J. W.

    2011-12-01

    Arsenic is a highly toxic metalloid typically enriched in geothermal waters due to aqueous weathering of arsenic-bearing minerals. Investigation of enzymatic pathways by which thermophilic microorganisms cope with toxic arsenic levels may yield insights into the evolution of arsenic resistance mechanisms on the early Earth. At Wai-O-Tapu in the Taupo Volcanic Zone on the North Island of New Zealand, hot springs with temperatures of 30-90°C and elemental sulfur concentrations (expressed as equivalent sulfate) from 340 to 850 mg/l establish a range of environmental conditions. Total arsenic concentrations varied from 0.083 mg/l to 56 mg/l. Arsenic speciation analysis elucidated various biogeochemical arsenic transformations occurring within different springs. For example, in the Alum Cliff spring oxidizing conditions (Eh = 225 mV) were expected to stabilize dissolved arsenate (AsO43-). However, HPLC-ICPMS analyses yielded dissolved arsenate and arsenite (AsO33-) concentrations of 0.25 mg/l versus 43.3 mg/l, respectively, and point towards microbial arsenate reduction as the likely mechanism for arsenic redox transformation. 16S rRNA gene cloning of Alum Cliff DNA showed a predominantly archaeal population with the dominant clone "AC1_A1" most closely related (99% sequence similarity, NCBI BLAST°) to the uncultured Sulfolobus clone "ChP_97P" found in Champagne Pool (Childs et al., 2008). The closest isolated relative to AC1_A1 is Sulfolobus tokodaii str. TW with a sequence similarity of 94%. Arsenic speciation measurements from the Alum Cliff spring suggest that clone AC1_A1 features the arsenate reduction resistance mechanism, and we hypothesize therefore that an arsC (homolog or analog) provides this functionality. The organic arsenic species monomethylarsonic acid (MMA) and dimethylarsinic acid (DMA), detected via HPLC-ICPMS at concentrations ranging from 1 μg/l to 12 μg/l in various springs, may also implicate microbial methyl-group transfers as an active

  18. Solid-methane moderator systems at KENS

    SciTech Connect

    Furusaka, M.

    1997-09-01

    An overview of the target-moderator-reflector assembly (TMRA) systems at the Neutron Science Laboratory (KENS) at the High Energy Accelerator Research Organization (KEK) is described together with the historical changes. Because of the optimized design of TMRA, the neutron-generation efficiency is very high. The characteristics of the cold moderator system, such as the absolute intensity, spectrum and radiation damage to it, are also described. There are a number of new neutron-scattering instruments which are being viewed at the moderator. A brief description of the proposed TMRA for our future project is also given. (auth)

  19. JACKETED FUEL ELEMENTS FOR GRAPHITE MODERATED REACTORS

    DOEpatents

    Szilard, L.; Wigner, E.P.; Creutz, E.C.

    1959-05-12

    Fuel elements for a heterogeneous, fluid cooled, graphite moderated reactor are described. The fuel elements are comprised of a body of natural uranium hermetically sealed in a jacket of corrosion resistant material. The jacket, which may be aluminum or some other material which is non-fissionable and of a type having a low neutron capture cross-section, acts as a barrier between the fissioning isotope and the coolant or moderator or both. The jacket minimizes the tendency of the moderator and coolant to become radioactive and/or contaminated by fission fragments from the fissioning isotope.

  20. Comparison of the microbial communities in solid-state anaerobic digestion (SS-AD) reactors operated at mesophilic and thermophilic temperatures.

    PubMed

    Li, Yueh-Fen; Nelson, Michael C; Chen, Po-Hsu; Graf, Joerg; Li, Yebo; Yu, Zhongtang

    2015-01-01

    The microbiomes involved in liquid anaerobic digestion process have been investigated extensively, but the microbiomes underpinning solid-state anaerobic digestion (SS-AD) are poorly understood. In this study, microbiome composition and temporal succession in batch SS-AD reactors, operated at mesophilic or thermophilic temperatures, were investigated using Illumina sequencing of 16S rRNA gene amplicons. A greater microbial richness and evenness were found in the mesophilic than in the thermophilic SS-AD reactors. Firmicutes accounted for 60 and 82 % of the total Bacteria in the mesophilic and in the thermophilic SS-AD reactors, respectively. The genus Methanothermobacter dominated the Archaea in the thermophilic SS-AD reactors, while Methanoculleus predominated in the mesophilic SS-AD reactors. Interestingly, the data suggest syntrophic acetate oxidation coupled with hydrogenotrophic methanogenesis as an important pathway for biogas production during the thermophilic SS-AD. Canonical correspondence analysis (CCA) showed that temperature was the most influential factor in shaping the microbiomes in the SS-AD reactors. Thermotogae showed strong positive correlation with operation temperature, while Fibrobacteres, Lentisphaerae, Spirochaetes, and Tenericutes were positively correlated with daily biogas yield. This study provided new insight into the microbiome that drives SS-AD process, and the findings may help advance understanding of the microbiome in SS-AD reactors and the design and operation of SS-AD systems. PMID:25194839

  1. Disinhibition of the ammonium nitrogen in autothermal thermophilic aerobic digestion for sewage sludge by chemical precipitation.

    PubMed

    Yuan, Haiping; Xu, Changwen; Zhu, Nanwen

    2014-10-01

    Magnesium ammonium phosphate (MAP) precipitation was introduced to remove ammonium nitrogen (NH4(+)-N) in autothermal thermophilic aerobic digestion (ATAD) in this study by addition of MgCl2 · 6H2O and NaH2PO4 · 2H2O. The results showed that the lowest NH4(+)-N concentration was found in the D2 digester after 2nd day dosing treatment and 38.12% of VS removal efficiency was obtained after 15 days ATAD treatment. Sludge stabilization was achieved in the D2 digester 6 days earlier than the non-dosing digester when 8.7 g/L MgCl2 · 6H2O and 6.7 g/L NaH2PO4 · 2H2O were added into the digester. Furthermore, the highest VS removal efficiency of 40.03% was observed after 21 days digestion in D2 digesters. Therefore, MAP precipitation was an effective method for the ammonium nitrogen disinhibition when 8.7 g/L MgCl2 · 6H2O and 6.7 g/L NaH2PO4 · 2H2O were added into on the 2nd day after the digester startup. PMID:25108267

  2. Formation of magnetite and iron-rich carbonates by thermophilic iron-reducing bacteria

    SciTech Connect

    Zhang, C.; Liu, S.; Roh, Y.; Cole, D.; Phelps, T.; Vali, H.; Kirschvink, J.L.; Onsttot, T.; McKay, D.

    1997-06-01

    Laboratory experiments were performed to study the formation of iron minerals by a thermophilic (45 to 75 C) fermentative iron-reducing bacterial culture (TOR39) obtained from the deep subsurface. Using amorphous Fe(III) oxyhydroxide as an electron acceptor and glucose as an electron donor, TOR39 produced magnetite and iron-rich carbonates at conditions consistent, on a thermodynamic basis, with Eh ({minus}200 mV to {minus}415 mV) and pH (6.2 to 7.7) values determined for these experiments. Analyses of the precipitating solid phases by X-ray diffraction showed that the starting amorphous Fe(III) oxyhydroxide was nearly completely converted to magnetite and Fe-rich carbonate after 20 days of incubation. Increasing bicarbonate concentration in the chemical milieu resulted in increased proportions of siderite relative to magnetite and the addition of MgCl{sub 2} caused the formation of magnesium-rich carbonate in addition to siderite. The results suggest that the TOR39 bacterial culture may have the capacity to form magnetite and iron-rich carbonates in a variety of geochemical conditions. These results may have significant implications for studying the past biogenic activities in the Martian meteorite ALH84001.

  3. Encapsulated in silica: genome, proteome and physiology of the thermophilic bacterium Anoxybacillus flavithermus

    SciTech Connect

    Saw, Jimmy H; Mountain, Bruce W; Feng, Lu; Omelchenko, Marina V; Saito, Jennifer A; Stott, Matthew B; Li, Dan; Zhao, Guang; Wu, Junli; Galperin, Michael Y; Dunfield, Peter F; Wang, Lei; Alam, Maqsudul

    2008-01-01

    Gram-positive bacteria of the genus Anoxybacillus have been found in diverse thermophilic habitats, such as geothermal hot springs and manure, and in processed foods such as gelatin and milk powder. Anoxybacillus flavithermus is a facultatively anaerobic bacterium found in super-saturated silica solutions and in opaline silica sinter. The ability of A. flavithermus to grow in super-saturated silica solutions makes it an ideal subject to study the processes of sinter formation, which might be similar to the biomineralization processes that occurred at the dawn of life. We report here the complete genome sequence of A. flavithermus strain WK1, isolated from the waste water drain at the Wairakei geothermal power station in New Zealand. It consists of a single chromosome of 2,846,746 base pairs and is predicted to encode 2,863 proteins. In silico genome analysis identified several enzymes that could be involved in silica adaptation and biofilm formation, and their predicted functions were experimentally validated in vitro. Proteomic analysis confirmed the regulation of biofilm-related proteins and crucial enzymes for the synthesis of long-chain polyamines as constituents of silica nanospheres. Microbial fossils preserved in silica and silica sinters are excellent objects for studying ancient life, a new paleobiological frontier. An integrated analysis of the A. flavithermus genome and proteome provides the first glimpse of metabolic adaptation during silicification and sinter formation. Comparative genome analysis suggests an extensive gene loss in the Anoxybacillus/Geobacillus branch after its divergence from other bacilli.

  4. Mass and Energy Balances of Dry Thermophilic Anaerobic Digestion Treating Swine Manure Mixed with Rice Straw.

    PubMed

    Zhou, Sheng; Zhang, Jining; Zou, Guoyan; Riya, Shohei; Hosomi, Masaaki

    2015-01-01

    To evaluate the feasibility of swine manure treatment by a proposed Dry Thermophilic Anaerobic Digestion (DT-AD) system, we evaluated the methane yield of swine manure treated using a DT-AD method with rice straw under different C/N ratios and solid retention time (SRT) and calculated the mass and energy balances when the DT-AD system is used for swine manure treatment from a model farm with 1000 pigs and the digested residue is used for forage rice production. A traditional swine manure treatment Oxidation Ditch system was used as the study control. The results suggest that methane yield using the proposed DT-AD system increased with a higher C/N ratio and shorter SRT. Correspondently, for the DT-AD system running with SRT of 80 days, the net energy yields for all treatments were negative, due to low biogas production and high heat loss of digestion tank. However, the biogas yield increased when the SRT was shortened to 40 days, and the generated energy was greater than consumed energy when C/N ratio was 20 : 1 and 30 : 1. The results suggest that with the correct optimization of C/N ratio and SRT, the proposed DT-AD system, followed by using digestate for forage rice production, can attain energy self-sufficiency. PMID:26609436

  5. Characterization of a novel thermostable N-acylhomoserine lactonase from the thermophilic bacterium Thermaerobacter marianensis.

    PubMed

    Morohoshi, Tomohiro; Tominaga, Yoshiaki; Someya, Nobutaka; Ikeda, Tsukasa

    2015-07-01

    Thermaerobacter marianensis is an extremely thermophilic bacterium, which was isolated from the Mariana Trench, with an optimal growth temperature of approximately 75 °C. N-Acylhomoserine lactone (AHL) is a quorum-sensing signal molecule used by many gram-negative bacteria. Here, we report the identification of an AHL-degrading gene homolog (designated aiiT) in the genome of T. marianensis JCM 10246. AiiT has 59.7%, 21.2%, and 11.2% identity to AhlS from Solibacillus silvestris, AiiA from Bacillus cereus, and AidC from Chryseobacterium sp., respectively. Homologs of aiiT were also found in Thermaerobacter nagasakiensis, T. composti, and T. subterraneus. A purified AiiT-maltose binding fusion showed high AHL-degrading activity against N-hexanoyl-L-homoserine lactone, N-octanoyl-L-homoserine lactone, and N-decanoyl-L-homoserine lactone at temperatures ranging from 40 to 80 °C. HPLC analysis revealed that AiiT functions as an AHL-lactonase that catalyzes AHL ring opening by hydrolyzing lactones. AiiT displayed maximal activity at high temperatures (60-80 °C) and showed higher thermostability than other AHL lactonases. PMID:25529553

  6. Complete genome sequences of Geobacillus sp. WCH70, a thermophilic strain isolated from wood compost.

    PubMed

    Brumm, Phillip J; Land, Miriam L; Mead, David A

    2016-01-01

    Geobacillus sp. WCH70 was one of several thermophilic organisms isolated from hot composts in the Middleton, WI area. Comparison of 16 S rRNA sequences showed the strain may be a new species, and is most closely related to G. galactosidasius and G. toebii. The genome was sequenced, assembled, and annotated by the DOE Joint Genome Institute and deposited at the NCBI in December 2009 (CP001638). The genome of Geobacillus species WCH70 consists of one circular chromosome of 3,893,306 bp with an average G + C content of 43 %, and two circular plasmids of 33,899 and 10,287 bp with an average G + C content of 40 %. Among sequenced organisms, Geobacillus sp. WCH70 shares highest Average Nucleotide Identity (86 %) with G. thermoglucosidasius strains, as well as similar genome organization. Geobacillus sp. WCH70 appears to be a highly adaptable organism, with an exceptionally high 125 annotated transposons in the genome. The organism also possesses four predicted restriction-modification systems not found in other Geobacillus species. PMID:27123157

  7. celB, a gene coding for a bifunctional cellulase from the extreme thermophile "Caldocellum saccharolyticum".

    PubMed Central

    Saul, D J; Williams, L C; Grayling, R A; Chamley, L W; Love, D R; Bergquist, P L

    1990-01-01

    "Caldocellum saccharolyticum" is an obligatory anaerobic thermophilic bacterium. A gene from this organism, designated celB, has been cloned in Escherichia coli as part of a bacteriophage lambda gene library. This gene produces a thermostable cellulase that shows both endoglucanase and exoglucanase activities on test substrates and is able to degrade crystalline cellulose to glucose. The sequence of celB has homology with both exo- and endoglucanases described by others. It appears to have a central domain without enzymatic activity which is joined to the enzymatic domains by runs of amino acids rich in proline and threonine (PT boxes). Deletion analysis shows that the exoglucanase activity is located in the amino-terminal domain of the enzyme and that endoglucanase activity is located in the carboxy-terminal domain. There are internal transcriptional and translational start sites within the gene. The intact gene has been cloned into a temperature-inducible expression vector, pJLA602, and overexpressed in E. coli. Polyacrylamide gel electrophoresis showed that celB produced a protein with a molecular weight of 118,000 to 120,000. A number of smaller proteins with activity against carboxymethyl cellulose and 4-methyl umbelliferyl-beta-D-cellobioside were also produced. These are believed to be the result of alternative translational start sites and/or proteolytic degradation products of the translated gene product. Images PMID:2126700

  8. Toxicity of Select Organic Acids to the Slightly Thermophilic Acidophile Acidithiobaccillus Caldus

    SciTech Connect

    John E Aston; William A Apel; Brady D Lee; Brent M Peyton

    2009-02-01

    Acidithiobacillus caldus is a thermophilic acidophile found in commercial biomining, acid mine drainage systems, and natural environments. Previous work has characterized A. caldus as a chemolithotrophic autotroph capable of utilizing reduced sulfur compounds under aerobic conditions. Organic acids are especially toxic to chemolithotrophs in low-pH environments, where they diffuse more readily into the cell and deprotonate within the cytoplasm. In the present study, the toxic effects of oxaloacetate, pyruvate, 2-ketoglutarate, acetate, malate, succinate, and fumarate on A. caldus strain BC13 were examined under batch conditions. All tested organic acids exhibited some inhibitory effect. Oxaloacetate was observed to inhibit growth completely at a concentration of 250 µM, whereas other organic acids were completely inhibitory at concentrations of between 1,000 and 5,000 µM. In these experiments, the measured concentrations of organic acids decreased with time, indicating uptake or assimilation by the cells. Phospholipid fatty acid analyses indicated an effect of organic acids on the cellular envelope. Notable differences included an increase in cyclic fatty acids in the presence of organic acids, indicating possible instability of the cellular envelope. This was supported by field emission scanning-electron micrographs showing blebbing and sluffing in cells grown in the presence of organic acids.

  9. Perchlorate and chlorate reduction by the Crenarchaeon Aeropyrum pernix and two thermophilic Firmicutes.

    PubMed

    Liebensteiner, Martin G; Pinkse, Martijn W H; Nijsse, Bart; Verhaert, Peter D E M; Tsesmetzis, Nicolas; Stams, Alfons J M; Lomans, Bart P

    2015-12-01

    This study reports the ability of one hyperthermophilic and two thermophilic microorganisms to grow anaerobically by the reduction of chlorate and perchlorate. Physiological, genomic and proteome analyses suggest that the Crenarchaeon Aeropyrum pernix reduces perchlorate with a periplasmic enzyme related to nitrate reductases, but that it lacks a functional chlorite-disproportionating enzyme (Cld) to complete the pathway. Aeropyrum pernix, previously described as a strictly aerobic microorganism, seems to rely on the chemical reactivity of reduced sulfur compounds with chlorite, a mechanism previously reported for perchlorate-reducing Archaeoglobus fulgidus. The chemical oxidation of thiosulfate (in excessive amounts present in the medium) and the reduction of chlorite result in the release of sulfate and chloride, which are the products of a biotic-abiotic perchlorate reduction pathway in Ae. pernix. The apparent absence of Cld in two other perchlorate-reducing microorganisms, Carboxydothermus hydrogenoformans and Moorella glycerini strain NMP, and their dependence on sulfide for perchlorate reduction is consistent with the observations made on Ar. fulgidus. Our findings suggest that microbial perchlorate reduction at high temperature differs notably from the physiology of perchlorate- and chlorate-reducing mesophiles and that it is characterized by the lack of a chlorite dismutase and is enabled by a combination of biotic and abiotic reactions. PMID:26332065

  10. Carbon isotope fractionation by thermophilic phototrophic sulfur bacteria: evidence for autotrophic growth in natural populations

    NASA Technical Reports Server (NTRS)

    Madigan, M. T.; Takigiku, R.; Lee, R. G.; Gest, H.; Hayes, J. M.

    1989-01-01

    Purple phototrophic bacteria of the genus Chromatium can grow as either photoautotrophs or photoheterotrophs. To determine the growth mode of the thermophilic Chromatium species, Chromatium tepidum, under in situ conditions, we have examined the carbon isotope fractionation patterns in laboratory cultures of this organism and in mats of C. tepidum which develop in sulfide thermal springs in Yellowstone National Park. Isotopic analysis (13C/12C) of total carbon, carotenoid pigments, and bacteriochlorophyll from photoautotrophically grown cultures of C. tepidum yielded 13C fractionation factors near -20%. Cells of C. tepidum grown on excess acetate, wherein synthesis of the Calvin cycle enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase ribulose bisphosphate carboxylase) was greatly repressed, were isotopically heavier, fractionation factors of ca. -7% being observed. Fractionation factors determined by isotopic analyses of cells and pigment fractions of natural populations of C. tepidum growing in three different sulfide thermal springs in Yellowstone National Park were approximately -20%, indicating that this purple sulfur bacterium grows as a photoautotroph in nature.

  11. Cloning, expression and characterization of a novel thermophilic polygalacturonase from Caldicellulosiruptor bescii DSM 6725.

    PubMed

    Chen, Yanyan; Sun, Dejun; Zhou, Yulai; Liu, Liping; Han, Weiwei; Zheng, Baisong; Wang, Zhi; Zhang, Zuoming

    2014-01-01

    We cloned the gene ACM61449 from anaerobic, thermophilic Caldicellulosiruptor bescii, and expressed it in Escherichia coli origami (DE3). After purification through thermal treatment and Ni-NTA agarose column extraction, we characterized the properties of the recombinant protein (CbPelA). The optimal temperature and pH of the protein were 72 °C and 5.2, respectively. CbPelA demonstrated high thermal-stability, with a half-life of 14 h at 70 °C. CbPelA also showed very high activity for polygalacturonic acid (PGA), and released monogalacturonic acid as its sole product. The Vmax and Km of CbPelA were 384.6 U·mg⁻¹ and 0.31 mg·mL⁻¹, respectively. CbPelA was also able to hydrolyze methylated pectin (48% and 10% relative activity on 20%-34% and 85% methylated pectin, respectively). The high thermo-activity and methylated pectin hydrolization activity of CbPelA suggest that it has potential applications in the food and textile industry. PMID:24705464

  12. Complete genome sequences of Geobacillus sp. WCH70, a thermophilic strain isolated from wood compost

    DOE PAGESBeta

    Brumm, Phillip; Land, Miriam L.; Mead, David

    2016-04-27

    Geobacillus sp. WCH70 was one of several thermophilic organisms isolated from hot composts in the Middleton, WI area. Comparison of 16 S rRNA sequences showed the strain may be a new species, and is most closely related to G. galactosidasius and G. toebii. The genome was sequenced, assembled, and annotated by the DOE Joint Genome Institute and deposited at the NCBI in December 2009 (CP001638). The genome of Geobacillus species WCH70 consists of one circular chromosome of 3,893,306 bp with an average G + C content of 43 %, and two circular plasmids of 33,899 and 10,287 bp with anmore » average G + C content of 40 %. Among sequenced organisms, Geobacillus sp. WCH70 shares highest Average Nucleotide Identity (86 %) with G. thermoglucosidasius strains, as well as similar genome organization. Geobacillus sp. WCH70 appears to be a highly adaptable organism, with an exceptionally high 125 annotated transposons in the genome. The organism also possesses four predicted restriction-modification systems not found in other Geobacillus species.« less

  13. Total solid content drives hydrogen production through microbial selection during thermophilic fermentation.

    PubMed

    Motte, Jean-Charles; Trably, Eric; Hamelin, Jérôme; Escudié, Renaud; Bonnafous, Anaïs; Steyer, Jean-Philippe; Bernet, Nicolas; Delgenès, Jean-Philippe; Dumas, Claire

    2014-08-01

    In this study, the effect of total solid content (TS) on thermophilic hydrogen production from wheat straw was investigated. Six TS contents ranging from wet to dry conditions (10-34%TS) were tested in batch tests. A decrease of H₂ yields was observed and three statistical groups were distinguished according to the TS content: wet conditions (10% and 14%TS) with 15.3 ± 1.6 NmlH₂ gTS(-1), intermediate conditions (19%TS) with 6.4 ± 1.0 NmlH₂ gTS(-1) and dry conditions (25-34%TS) with 3.4 ± 0.8 NmlH₂ gTS(-1). Such a decrease in biohydrogen yields was related to a metabolic shift with an accumulation of lactic acid under dry conditions. Concomitantly, a microbial population shift was observed with a dominance of species related to the class Clostridia under wet conditions, and a co-dominance of members of Bacilli, Clostridia classes and Bacteroidetes phylum under dry conditions. PMID:24951274

  14. In Vitro Antibiofilm Activity of an Exopolysaccharide from the Marine Thermophilic Bacillus licheniformis T14.

    PubMed

    Spanò, Antonio; Laganà, Pasqualina; Visalli, Giuseppa; Maugeri, Teresa L; Gugliandolo, Concetta

    2016-05-01

    The development of antibiofilm strategies is of major interest in contrasting bacterial pathogenic biofilms. A novel fructose and fucose rich exopolysaccharide (EPS1-T14) produced by the recently described thermophilic Bacillus licheniformis T14, isolated from a shallow hydrothermal vent of Panarea Island (Eolian Island, Italy), was evaluated for its effects on biofilm formation by multiresistant clinical strains of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus. The antibiofilm activity of EPS1-T14 was assessed by microtiter plate assays and visualized by confocal laser scanning microscopic images. EPS1-T14, with molecular weight of 1000 kDa, reduced biofilm formation on abiotic surfaces without affecting bacterial vitality. The novel EPS1-T14 is a water-soluble, noncytotoxic exopolymer able to prevent biofilm formation and its use may represent a promising therapeutic strategy for combating bacterial biofilm-associated infections. EPS1-T14 as antiadhesive biomolecule could be useful for novel prospective in medical and nonmedical applications. PMID:26750122

  15. A cambialistic superoxide dismutase in the thermophilic photosynthetic bacterium Chloroflexus aurantiacus.

    PubMed

    Lancaster, Vanessa L; LoBrutto, Russell; Selvaraj, Fabiyola M; Blankenship, Robert E

    2004-06-01

    Superoxide dismutase from the thermophilic anoxygenic photosynthetic bacterium Chloroflexus aurantiacus was cloned, purified, and characterized. This protein is in the manganese- and iron-containing family of superoxide dismutases and is able to use both manganese and iron catalytically. This appears to be the only soluble superoxide dismutase in C. aurantiacus. Iron and manganese cofactors were identified by using electron paramagnetic resonance spectroscopy and were quantified by atomic absorption spectroscopy. By metal enrichment of growth media and by performing metal fidelity studies, the enzyme was found to be most efficient with manganese incorporated, yet up to 30% of the activity was retained with iron. Assimilation of iron or manganese ions into superoxide dismutase was also found to be affected by the growth conditions. This enzyme was also found to be remarkably thermostable and was resistant to H2O2 at concentrations up to 80 mM. Reactive oxygen defense mechanisms have not been previously characterized in the organisms belonging to the phylum Chloroflexi. These systems are of interest in C. aurantiacus since this bacterium lives in a hyperoxic environment and is subject to high UV radiation fluxes. PMID:15150226

  16. Effect of temperature on bacterial species diversity in thermophilic solid-waste composting.

    PubMed Central

    Strom, P F

    1985-01-01

    Continuously thermophilic composting was examined with a 4.5-liter reactor placed in an incubator maintained at representative temperatures. Feed was a mixture of dried table scraps and shredded newspaper wetted to 55% moisture. One run at 49 degrees C (run A) employed a 1:4 feed-to-compost ratio, while the other runs used a 10:1 ratio and were incubated at 50, 55, 60, or 65 degrees C. Due to self-heating, internal temperatures of the composting mass were 0 to 7 degrees C hotter than the incubator. Two full-scale composting plants (at Altoona, Pa., and Leicester, England) were also examined. Plate counts per gram (dry weight) on Trypticase soy broth (BBL Microbiology Systems) with 2% agar ranged from 0.7 X 10(9) to 5.3 X 10(9) for laboratory composting and 0.02 X 10(9) to 7.4 X 10(9) for field composting. Fifteen taxa were isolated, including 10 of genus Bacillus, which dominated all samples except that from run A. Species diversity decreased markedly in laboratory composting at 60 degrees C and above, but was similar for the three runs incubated at 49, 50, and 55 degrees C. The maximum desirable composting temperature based on species diversity is thus 60 degrees C, the same as that previously recommended based on measures of the rate of decomposition. PMID:4083885

  17. Methylotrophy in the thermophilic Bacillus methanolicus, basic insights and application for commodity production from methanol.

    PubMed

    Müller, Jonas E N; Heggeset, Tonje M B; Wendisch, Volker F; Vorholt, Julia A; Brautaset, Trygve

    2015-01-01

    Using methanol as an alternative non-food feedstock for biotechnological production offers several advantages in line with a methanol-based bioeconomy. The Gram-positive, facultative methylotrophic and thermophilic bacterium Bacillus methanolicus is one of the few described microbial candidates with a potential for the conversion of methanol to value-added products. Its capabilities of producing and secreting the commercially important amino acids L-glutamate and L-lysine to high concentrations at 50 °C have been demonstrated and make B. methanolicus a promising target to develop cell factories for industrial-scale production processes. B. methanolicus uses the ribulose monophosphate cycle for methanol assimilation and represents the first example of plasmid-dependent methylotrophy. Recent genome sequencing of two physiologically different wild-type B. methanolicus strains, MGA3 and PB1, accompanied with transcriptome and proteome analyses has generated fundamental new insight into the metabolism of the species. In addition, multiple key enzymes representing methylotrophic and biosynthetic pathways have been biochemically characterized. All this, together with establishment of improved tools for gene expression, has opened opportunities for systems-level metabolic engineering of B. methanolicus. Here, we summarize the current status of its metabolism and biochemistry, available genetic tools, and its potential use in respect to overproduction of amino acids. PMID:25431011

  18. Effects of transient temperature increases on odor production from thermophilic anaerobic digestion.

    PubMed

    Iranpour, R; Alatriste-Mondragon, F; Cox, H H J; Haug, R T

    2005-01-01

    The City of Los Angeles, Bureau of Sanitation, has implemented thermophilic anaerobic sludge digestion at the Hyperion and Terminal Island Treatment Plants (HTP and TITP). A two-stage continuous-batch process was established at HTP, while a single-stage sequencing batch process was established at TITP. This was to evaluate compliance with the Class A pathogen reduction requirements of U.S. EPA 40 CFR Part 503. A rapid increase of the digester temperature at TITP from 57.5 to 65.5 degrees C caused an increase of the volatile fatty acid to alkalinity ratio, a decline in digester performance, and an elevated production of methyl mercaptan and hydrogen sulfide. A rapid increase of the digester temperature at HTP from 54 to 58 degrees C caused an elevated production of methyl mercaptan, but the effect on the volatile fatty acid to alkalinity ratio and digester performance was insignificant. It is likely that these effects observed at TITP and HTP were transient responses to rapid changes in temperature. PMID:16180433

  19. Thermophilic two-stage dry anaerobic digestion of model garbage with ammonia stripping.

    PubMed

    Yabu, Hironori; Sakai, Chikako; Fujiwara, Tomoko; Nishio, Naomichi; Nakashimada, Yutaka

    2011-03-01

    To avoid the inhibition of methane production by ammonia that occurs during the degradation of garbage, anaerobic digestion with prior ammonia production and subsequent stripping was investigated. In the ammonia production phase, the maximum ammonia concentration was approximately 2800 mg N/kg of total wet sludge in the range of 4 days of sludge retention time, indicating that only 43% of total nitrogen in the model garbage was converted to ammonia. The model garbage from which ammonia was produced and stripped was subjected to semi-continuous thermophilic dry anaerobic digestion over 180 days. The gas yield was in the range of 0.68 to 0.75 Nm(3)/kg volatile solid, and it decreased with the decrease of the sludge retention time. The ammonia-nitrogen concentration in the sludge was kept below 3000 mg N/kg total wet sludge. Microbial community structure analysis revealed that the phylum Firmicutes dominated in the ammonia production, but the community structure changed at different sludge retention times. In dry anaerobic digestion, the dominant bacteria shifted from the phylum Thermotogae to Firmicutes. The dominant archaeon was the genus Methanothermobacter, but the ratio of Methanosarcina increased during the process of dry anaerobic digestion. PMID:21094085

  20. Heat stable alkaline phosphatase from thermophiles. Final report, March-October 1993

    SciTech Connect

    Combie, J.D.; Runnion, K.N.; Williamson, M.L.

    1994-07-01

    Alkaline phosphatase has been the most widely used enzyme for colorimetric immunoassays. The current potential for this enzyme lies in biosensors, fieldable assay kits, biotechnology applications, degradation of certain nerve agents and pesticides and detoxification of heavy metal waste streams. While the commercial source of this enzyme is predominantly from mammalian tissues, expanded commercial application is restricted by the enzyme's instability at elevated temperatures. Although alkaline phosphatases are ubiquitous in nature, two isolates out of 44 alkaline phosphatase producing isolates occurring in habitats at 50 deg C and above have been isolated possessing extremely stable enzymes. One enzyme retained 98% of original activity following boiling for 1 hr. The secretion of the enzyme by the organism is an added benefit promoting efficient and economical production capability. Procedures for the screening, isolation, and optimal growth and fermentation of organisms acquired from geothermal sources located in Yellowstone National Park, WY are described. Purification was most effectively achieved using size exclusion chromatography where 101% of the activity and 33% of the crude mother liquor protein were recovered. Although the presence of manganese in the assay buffer was observed to significantly elevate the enzyme's catalytic activity, a precipitate incompatibility with calcium chloride, a requirement for high temperature stability, prohibits its use. Bacteria, Fermentation, Alkaline phosphatase, Biosensors, Biotechnology, Heat stable enzymes, Biochemistry, Bioremediation, Thermophilic microorganisms.