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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.

  2. 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

  3. 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.

  4. 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

    2014-08-28

    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.

  5. 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

  6. 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.

  7. Structural and Biochemical Studies of a Moderately Thermophilic Exonuclease I from Methylocaldum szegediense

    PubMed Central

    Fei, Li; Tian, SiSi; Moysey, Ruth; Misca, Mihaela; Barker, John J.; Smith, Myron A.; McEwan, Paul A.; Pilka, Ewa S.; Crawley, Lauren; Evans, Tom; Sun, Dapeng

    2015-01-01

    A novel exonuclease, designated as MszExo I, was cloned from Methylocaldum szegediense, a moderately thermophilic methanotroph. It specifically digests single-stranded DNA in the 3ʹ to 5ʹ direction. The protein is composed of 479 amino acids, and it shares 47% sequence identity with E. coli Exo I. The crystal structure of MszExo I was determined to a resolution of 2.2 Å and it aligns well with that of E. coli Exo I. Comparative studies revealed that MszExo I and E. coli Exo I have similar metal ion binding affinity and similar activity at mesophilic temperatures (25–47°C). However, the optimum working temperature of MszExo I is 10°C higher, and the melting temperature is more than 4°C higher as evaluated by both thermal inactivation assays and DSC measurements. More importantly, two thermal transitions during unfolding of MszExo I were monitored by DSC while only one transition was found in E. coli Exo I. Further analyses showed that magnesium ions not only confer structural stability, but also affect the unfolding of MszExo I. MszExo I is the first reported enzyme in the DNA repair systems of moderately thermophilic bacteria, which are predicted to have more efficient DNA repair systems than mesophilic ones. PMID:25658953

  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. Differential bioleaching of copper by mesophilic and moderately thermophilic acidophilic consortium enriched from same copper mine water sample.

    PubMed

    Marhual, N P; Pradhan, N; Kar, R N; Sukla, L B; Mishra, B K

    2008-11-01

    Three acidophilic enrichment consortium were developed from mine water sample of copper mine site at Khetri, India were compared for their copper leaching efficiency. Out of these one was mesophilic (35 degrees C) and two were moderately thermophilic (50 degrees C). Consortia were named as mesophilic acidophilic chemolithotrophic consortia (MACC), thermophilic acidophilic chemolithotrophic consortia (TACC), and Sulfobacillus acidophilic consortia (SAC). Copper extraction ability of both the thermophilic consortia (77-78% extraction) was almost double to that of mesophilic consortia (40% extraction) at 10% pulp density after 55 days. Both the thermophilic consortia were equally effective in leaching of other metals like Ni, Co, Zn, Mn. After 55 days, the percentage of extractions of copper by TACC was 76, 74, 67, 48 and 45 at 5%, 10%, 15%, 20% and 30% pulp density, respectively. Total number of bacteria was maximum at 5% pulp density which decreases with increase in pulp density. Sulfobacillus-like bacteria were seen in the Sulfobacillus enrichment cultures. Moderately thermophilic consortia proved to be better in leaching performance than the mesophilic counterpart.

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

    PubMed Central

    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

  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. Isolation and characterization of a moderately thermophilic anaerobic alkaliphile, Clostridium paradoxum sp. nov.

    SciTech Connect

    Youhong Li; Wiegel, J.; Mandelco, L.

    1993-07-01

    Alkaliphilic, moderately thermophilic anaerobic bacteria able to grow above pH 10.5 and 55{degrees}C were isolated from various sewage plants in the United States. The strains were motile with two to six peritrichous flagella and formed round to slightly oval terminal spores in terminally distended and slightly enlarged cells. Sporulated cells remained motile. The pH range for growth was between 7.0 and 11.1, with an optimum of around 10.1. At pH 10.1 the temperature range for growth was between 30 and 63{degrees}C, with an optimum of 56{degrees}C. The shortest observed doubling time (glucose) was around 16 min at 56{degrees}C and pH 10.1. No dissimilatory sulfate reduction was detected. The organism utilized glucose, fructose, sucrose, maltose, and pyruvate but required yeast extract or tryptone for growth. Optimal NaCl concentrations for growth were between 50 and 200 mM. The guanine-plus-cytosine content was 30.0 {+-} 0.10 mol%. On the basis of unique properties and 16S rRNA analysis, the strains are placed in a new species, Clostridium paradoxum, referring to the unusual retainment of motility by sporulated cells. Strain JW-YL-7 (DSM 7308) is designated as the type strain.

  15. Proteomic analysis of membrane proteins from a radioresistant and moderate thermophilic bacterium Deinococcus geothermalis.

    PubMed

    Tian, Bing; Wang, Hu; Ma, Xiaoqiong; Hu, Yaping; Sun, Zongtao; Shen, Shaochuan; Wang, Fei; Hua, Yuejin

    2010-10-01

    Deinococcus geothermalis is a radioresistant and moderate thermophilic bacterium. Little was known about the membrane or membrane associated proteins of this bacterium. This study established the membrane subproteome profile of D. geothermalis, using 1-D PAGE and LC-MS/MS analysis following Triton X-114 detergent extraction. A total of 552 proteins from the membrane preparations were identified from two independent trials. In the total identified proteins, 117 were membrane subproteomic proteins, and 89 of them were described for the first time in D. geothermalis including fimbrial pilin (Dgeo_2038), cytochrome bd ubiquinol oxidase (Dgeo_2705) and multi-sensor (Dgeo_2096). The major membrane subproteomic proteins were distributed into 18 functional groups including nutrient transport and metabolism, energy production and conversion, cell wall/membrane biogenesis and a poorly characterized subclass. The identifications of Deinococcus-specific proteins, such as cell surface receptor IPT/TIG (Dgeo_1119) and four hypothetical proteins, demonstrated the special protein composition and functions in the cell membrane of Deinococcus. The results provide a basis for quantitative proteomic analysis, which will facilitate the understanding of the adaptation of this organism to different environmental stresses and the development of strategies for bioremediation of environmental waste.

  16. 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.

  17. Clostridium tepidiprofundi sp. nov., a moderately thermophilic bacterium from a deep-sea hydrothermal vent.

    PubMed

    Slobodkina, G B; Kolganova, T V; Tourova, T P; Kostrikina, N A; Jeanthon, C; Bonch-Osmolovskaya, E A; Slobodkin, A I

    2008-04-01

    A moderately thermophilic, anaerobic bacterium (strain SG 508T) was isolated from a hydrothermal vent chimney located at 1 degrees N on the East Pacific Rise at a depth of 2650 m. Cells of strain SG 508T were straight to slightly curved rods, 0.4-0.6 microm in diameter and 2.0-3.0 microm in length. Spore formation was observed only below pH 5.5. The temperature range for growth was 22-60 degrees C, with optimum growth at 50 degrees C. The pH range for growth was 4.0-8.5, with optimum growth at pH 6.0-6.8. Growth of strain SG 508T was observed at NaCl concentrations ranging from 1.0 to 6.0 % (w/v), with optimum growth at 2.5 % (w/v). Substrates utilized by strain SG 508T included casein, peptone, tryptone, yeast extract, beef extract, starch, maltose and glucose. The products of glucose fermentation were ethanol, acetate, H2, formate and CO2. Strain SG 508T was able to reduce elemental sulfur to hydrogen sulfide. The DNA G+C content of strain SG 508T was 30.9 mol%. 16S rRNA gene sequence analysis revealed that the isolated organism belonged to cluster I of the genus Clostridium. On the basis of its physiological properties and data from phylogenetic analyses, strain SG 508T is considered to represent a novel species of the genus Clostridium, for which the name Clostridium tepidiprofundi sp. nov. is proposed. The type strain is SG 508T (=DSM 19306T =VKM B-2459T).

  18. 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.

  19. 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

  20. 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

  1. 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

  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. 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

  4. Cloning, expression, crystallization and preliminary X-ray characterization of cytochrome c{sub 552} from a moderate thermophilic bacterium, Hydrogenophilus thermoluteolus

    SciTech Connect

    Ichiki, Shin-ichi; Nakamura, Shota; Ohkubo, Tadayasu; Kobayashi, Yuji; Hasegawa, Jun; Uchiyama, Susumu; Nishihara, Hirofumi; Mizuta, Keiko; Sambongi, Yoshihiro

    2005-04-01

    Cytochrome c{sub 552} of a moderate thermophile, H. thermoluteolus, was overexpressed in E. coli and crystallized for X-ray diffraction study. The amino-acid sequence of cytochrome c{sub 552} (PH c{sub 552}) from a moderately thermophilic bacterium, Hydrogenophilus thermoluteolus, was more than 50% identical to that of cytochrome c from an extreme thermophile, Hydrogenobacter thermophilus (HT c{sub 552}), and from a mesophile, Pseudomonas aeruginosa (PA c{sub 551}). The PH c{sub 552} gene was overexpressed as a correctly processed holoprotein in the Escherichia coli periplasm. The overexpressed PH c{sub 552} has been crystallized by vapour diffusion from polyethylene glycol 4000 pH 6.5. The crystals belong to space group C222{sub 1}, with unit-cell parameters a = 48.98, b = 57.99, c = 56.20 Å. The crystals diffract X-rays to around 2.1 Å resolution.

  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. Ferroplasma cupricumulans sp. nov., a novel moderately thermophilic, acidophilic archaeon isolated from an industrial-scale chalcocite bioleach heap.

    PubMed

    Hawkes, Rebecca B; Franzmann, Peter D; O'Hara, Graham; Plumb, Jason J

    2006-12-01

    A new species of Archaea was isolated from an industrial mineral sulphide bioleach heap. Strain BH2, a non-motile pleomorphic coccus, was capable of chemomixotrophic growth on ferrous sulphate and yeast extract. Growth was not supported in the absence of yeast extract. Phylogenetic analysis based on the 16S rRNA gene showed that strain BH2 was most closely related to the species Ferroplasma acidiphilum; however, it showed only 95% sequence similarity with this species. Strain BH2 had a temperature optimum of 53.6 degrees C and a temperature range for growth between 22 and 63 degrees C. Thus, it is the first moderately thermophilic member of the genus Ferroplasma. The optimum pH for the growth of the strain occurred between pH 1.0 and 1.2 and the lowest pH at which growth was observed was 0.4. Based on 16S rRNA gene sequence analysis and other physiological characteristics, strain BH2 constitutes a new species within the genus Ferroplasma. The name Ferroplasma cupricumulans is proposed for the new species and strain BH2 (DSM 16651) is proposed as the type strain.

  7. Biological conversion of synthesis gas. [Chlorobium thiosulfatophilum, chlorobium phaeobacteroides, and Rhodospirillum rubrum

    SciTech Connect

    Not Available

    1992-01-06

    The anaerobic bacterium Rhodospirillum rubrum has been chosen for catalysis of the biological water gas shift reaction. Two bacteria, Chlorobium thiosulfatophilum and Chlorobium phaeobacteroides, are being evaluated as candidates for H{sub 2}S conversion to elemental sulfur. Since these latter two organisms both grow and convert H{sub 2}S in batch culture using standard basal medium, the choice of a suitable bacterium must be made in consideration of specific growth and uptake rates. Produced elemental sulfur stability against further oxidation to sulfate, and minimal use of H{sub 2} as a producing agent must also be considered. The effects of temperature on the performance of R. rubrum were evaluated. It was found that the cell concentration was highest at temperatures of 25 and 30{degree}C, and that the specific uptake rate was highest at temperatures of 30, 32 and 34{degree}C. No growth was observed at 37{degree}C. Also, temperature did not affect the yield of H{sub 2} from CO. Thus, R. rubrum may be used for biological rates gas shift at any temperature between 30 and 34{degree}C, although growth is maximized at lower temperatures. Preliminary studies with C. thiosulfatophilum showed rapid utilization of H{sub 2}S from the gas and liquid phases with subsequent production of elemental sulfur. Elemental sulfur production interfered with cell concentrations measurements, although a technique has been developed to rectify this problem.

  8. Sorption of metals by Chlorobium spp.

    PubMed

    Garcia-Gil, J; Borrego, C

    1997-12-01

    The capacity of two species of green phototrophic sulfur bacteria, Chlorobium limicola and C. phaeobacteroides, to sorb several metal ions (Mn2+, Fe2+, Ni2+, Cu2+, Zn2+, Cd2+ and Pb2+) has been tested in laboratory batch cultures at increasing concentrations up to 2,000 mumol/l. Except for nickel--which was not sorbed to bacterial cells--the rest of metals tested were bound in a fast and passive process, which was mathematically described by means of Freundlich isotherms models. The sorption capacity of the two species studied were found to be dependent on the metal involved, whereas no differences were observed in the sorption intensity, suggesting that in all cases the sorption process proceeds in a similar way. Further, the comparison of the sorption intensity values as well as the metal recovery index (Ri), for both species, revealed that C. phaeobacteroides was more efficient that C. limicola to attach metal ions. The ecological significance of this ability in the water column of some stratified lakes, where coinciding maxima of ferrous iron and green photosynthetic sulfur bacteria are frequently found, is discussed.

  9. Amplification of ribulose biphosphate carboxylase/oxygenase large subunit (RuBisCO LSU) gene fragments from Thiobacillus ferrooxidans and a moderate thermophile using polymerase chain reaction.

    PubMed

    Holden, P J; Brown, R W

    1993-07-01

    Southern blot analysis of DNA from an iron-oxidising moderate thermophile NMW-6 and from Thiobacillus ferrooxidans strain TFI-35 demonstrated sequences homologous to the RuBisCO LSU gene of Synechococcus. DNA fragments (457 bp) encoding part of the RuBisCO LSU gene (amino acids 73-200) were amplified from the genomic DNA of Thiobacillus ferrooxidans and the moderate thermophile NMW-6 using the polymerase chain reaction (PCR) technique (Saiki et al. (1985) Science 233, 1350-1354). A comparison with the LSU sequences from T. ferrooxidans, Alcaligenes eutrophus, Chromatium vinosum, Synechococcus and Spinacea oleracea, which all have RuBisCOs with a hexadecameric structure, showed that the RuBisCO LSU gene sequence from NMW-6 appeared to be most closely related to that of the hydrogen bacterium A. eutrophus which showed 71.9% homology at the amino acid level. Despite its physiological similarity, T. ferrooxidans showed only 64.1% homology to the amino acid sequence from NMW-6 and had the lowest DNA homology (60.9%) of the hexadecameric type RuBisCOs. In the region sequenced, T. ferrooxidans and the RuBisCOs of the phototrophs C. vinosum, Synechococcus and S. oleracea, had 17 residues that were completely conserved which were substituted in both NMW-6 and A. eutrophus, 11 of these being identical substitutions. Comparison of the nucleotide and derived amino acid sequences of the RuBisCO LSU fragment from T. ferrooxidans with other RuBisCO sequences indicated a closer relationship to the hexadecameric type LSU genes of photosynthetic origin than to that of A. eutrophus. The T. ferrooxidans amino acid sequence showed 93.8%, 78.9% and 77.3% homology, respectively, to the C. vinosum, Synechococcus and S. oleracea (spinach) sequences but only 56.2% to A. eutrophus. The DNA sequence from Rhodospirillum rubrum, which has the atypical large subunit dimer RuBisCO structure with no small subunit, showed 39.2% and 42.7% homology, respectively, with the sequences of NMW-6 and T

  10. Methylomarinovum caldicuralii gen. nov., sp. nov., a moderately thermophilic methanotroph isolated from a shallow submarine hydrothermal system, and proposal of the family Methylothermaceae fam. nov.

    PubMed

    Hirayama, Hisako; Abe, Mariko; Miyazaki, Masayuki; Nunoura, Takuro; Furushima, Yasuo; Yamamoto, Hiroyuki; Takai, Ken

    2014-03-01

    A novel methane-oxidizing bacterium, strain IT-9(T), was isolated from a shallow submarine hydrothermal system occurring in a coral reef in Japan. Strain IT-9(T) was a Gram-negative, aerobic, motile, coccoid or oval-shaped bacterium with the distinctive intracytoplasmic membrane arrangement of a type I methanotroph. Strain IT-9(T) was a moderately thermophilic, obligate methanotroph that grew on methane and methanol at 30-55 °C (optimum 45-50 °C). The strain possessed the particulate methane monooxygenase (pMMO). The ribulose monophosphate pathway was operative for carbon assimilation. NaCl was required for growth within a concentration range of 1-5 % (optimum 3 %). The hao gene encoding hydroxylamine oxidoreductase (HAO) involved in nitrification was detected by a PCR experiment. The major phospholipid fatty acids were C16 : 0 and C18 : 1ω7c. The major isoprenoid quinone was Q-8. The DNA G+C content was 66.0 mol%. The 16S rRNA gene sequence of strain IT-9(T) was only moderately related to the sequences of members of the closest genera Methylohalobius (94.1 % similarity) and Methylothermus (91.7-91.9 % similarity); however, those sequences formed a deeply branching monophyletic group within the order Methylococcales. Phylogenies based on 16S rRNA gene sequences, deduced partial PmoA sequences and deduced partial Hao sequences and physiological and chemotaxonomic characteristics revealed that strain IT-9(T) represents a novel species of a new genus, for which the name Methylomarinovum caldicuralii gen. nov., sp. nov. is proposed. The type strain of Methylomarinovum caldicuralii is IT-9(T) ( = JCM 13666(T) = DSM 19749(T)). In addition, we propose a new family, Methylothermaceae fam. nov., in the order Methylococcales, to accommodate the genera Methylothermus, Methylohalobius and Methylomarinovum. The genera Methylothermus and Methylohalobius have been recognized as being distinct from other genera in the methane-oxidizing order

  11. Methylomarinovum caldicuralii gen. nov., sp. nov., a moderately thermophilic methanotroph isolated from a shallow submarine hydrothermal system, and proposal of the family Methylothermaceae fam. nov.

    PubMed

    Hirayama, Hisako; Abe, Mariko; Miyazaki, Masayuki; Nunoura, Takuro; Furushima, Yasuo; Yamamoto, Hiroyuki; Takai, Ken

    2014-03-01

    A novel methane-oxidizing bacterium, strain IT-9(T), was isolated from a shallow submarine hydrothermal system occurring in a coral reef in Japan. Strain IT-9(T) was a Gram-negative, aerobic, motile, coccoid or oval-shaped bacterium with the distinctive intracytoplasmic membrane arrangement of a type I methanotroph. Strain IT-9(T) was a moderately thermophilic, obligate methanotroph that grew on methane and methanol at 30-55 °C (optimum 45-50 °C). The strain possessed the particulate methane monooxygenase (pMMO). The ribulose monophosphate pathway was operative for carbon assimilation. NaCl was required for growth within a concentration range of 1-5 % (optimum 3 %). The hao gene encoding hydroxylamine oxidoreductase (HAO) involved in nitrification was detected by a PCR experiment. The major phospholipid fatty acids were C16 : 0 and C18 : 1ω7c. The major isoprenoid quinone was Q-8. The DNA G+C content was 66.0 mol%. The 16S rRNA gene sequence of strain IT-9(T) was only moderately related to the sequences of members of the closest genera Methylohalobius (94.1 % similarity) and Methylothermus (91.7-91.9 % similarity); however, those sequences formed a deeply branching monophyletic group within the order Methylococcales. Phylogenies based on 16S rRNA gene sequences, deduced partial PmoA sequences and deduced partial Hao sequences and physiological and chemotaxonomic characteristics revealed that strain IT-9(T) represents a novel species of a new genus, for which the name Methylomarinovum caldicuralii gen. nov., sp. nov. is proposed. The type strain of Methylomarinovum caldicuralii is IT-9(T) ( = JCM 13666(T) = DSM 19749(T)). In addition, we propose a new family, Methylothermaceae fam. nov., in the order Methylococcales, to accommodate the genera Methylothermus, Methylohalobius and Methylomarinovum. The genera Methylothermus and Methylohalobius have been recognized as being distinct from other genera in the methane-oxidizing order

  12. Thermophilic microorganisms in biomining.

    PubMed

    Donati, Edgardo Rubén; Castro, Camila; Urbieta, María Sofía

    2016-11-01

    Biomining is an applied biotechnology for mineral processing and metal extraction from ores and concentrates. This alternative technology for recovering metals involves the hydrometallurgical processes known as bioleaching and biooxidation where the metal is directly solubilized or released from the matrix for further solubilization, respectively. Several commercial applications of biomining can be found around the world to recover mainly copper and gold but also other metals; most of them are operating at temperatures below 40-50 °C using mesophilic and moderate thermophilic microorganisms. Although biomining offers an economically viable and cleaner option, its share of the world´s production of metals has not grown as much as it was expected, mainly considering that due to environmental restrictions in many countries smelting and roasting technologies are being eliminated. The slow rate of biomining processes is for sure the main reason of their poor implementation. In this scenario the use of thermophiles could be advantageous because higher operational temperature would increase the rate of the process and in addition it would eliminate the energy input for cooling the system (bioleaching reactions are exothermic causing a serious temperature increase in bioreactors and inside heaps that adversely affects most of the mesophilic microorganisms) and it would decrease the passivation of mineral surfaces. In the last few years many thermophilic bacteria and archaea have been isolated, characterized, and even used for extracting metals. This paper reviews the current status of biomining using thermophiles, describes the main characteristics of thermophilic biominers and discusses the future for this biotechnology.

  13. Thermophilic microorganisms in biomining.

    PubMed

    Donati, Edgardo Rubén; Castro, Camila; Urbieta, María Sofía

    2016-11-01

    Biomining is an applied biotechnology for mineral processing and metal extraction from ores and concentrates. This alternative technology for recovering metals involves the hydrometallurgical processes known as bioleaching and biooxidation where the metal is directly solubilized or released from the matrix for further solubilization, respectively. Several commercial applications of biomining can be found around the world to recover mainly copper and gold but also other metals; most of them are operating at temperatures below 40-50 °C using mesophilic and moderate thermophilic microorganisms. Although biomining offers an economically viable and cleaner option, its share of the world´s production of metals has not grown as much as it was expected, mainly considering that due to environmental restrictions in many countries smelting and roasting technologies are being eliminated. The slow rate of biomining processes is for sure the main reason of their poor implementation. In this scenario the use of thermophiles could be advantageous because higher operational temperature would increase the rate of the process and in addition it would eliminate the energy input for cooling the system (bioleaching reactions are exothermic causing a serious temperature increase in bioreactors and inside heaps that adversely affects most of the mesophilic microorganisms) and it would decrease the passivation of mineral surfaces. In the last few years many thermophilic bacteria and archaea have been isolated, characterized, and even used for extracting metals. This paper reviews the current status of biomining using thermophiles, describes the main characteristics of thermophilic biominers and discusses the future for this biotechnology. PMID:27628339

  14. Chlorobium aminolipid: a new membrane lipid from green sulfur bacteria

    SciTech Connect

    Olson, J.M.; Shaw, E.K.; Gaffney, J.S.; Scandella, C.J.

    1983-01-01

    A preliminary chemical characterization of the aminolipid from Chlorobium limicola f. thiosulfatophilum shows that this lipid contains no phosphorus, glycerol, sugar, ornithine or lysine. The lipid has an R/sub f/-value of 0.03 in chloroform-methanol-ammonia (13:5:1 v/v) on silica gel TLC plates. In chloroform-methanol-water (65:25:4) the R/sub f/- value is 0.16. The absorption spectrum indicates bands at 225 and 231 nm, and the fluorescence (maximum emission at 340 nm) shows excitation maxima at 232, 288, and approx. 325 nm. Acid hydrolysis of the lipid yields fluorescent substances A (ninhydrin positive) and B (negative) in addition to myristic acid (C 14:0). The fluorescence emission and excitation spectra of substance A are very similar to those of the intact lipid. Although these excitation spectra resemble the absorption spectra of 7,8-dihydropterins (bands at approx. 230, approx. 280, and 320-330 nm at pH 7), the weight of evidence appears to be against substance A being a pterin. Nevertheless the UV absorption and fluorescence spectra do indicate that the amino moiety (substance A) is an aromatic heterocyclic component.

  15. 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.

  16. 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

  17. 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

  18. 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

  19. 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

  20. 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

  1. Tepidisphaera mucosa gen. nov., sp. nov., a moderately thermophilic member of the class Phycisphaerae in the phylum Planctomycetes, and proposal of a new family, Tepidisphaeraceae fam. nov., and a new order, Tepidisphaerales ord. nov.

    PubMed

    Kovaleva, O L; Merkel, A Yu; Novikov, A A; Baslerov, R V; Toshchakov, S V; Bonch-Osmolovskaya, E A

    2015-02-01

    Three strains of facultatively aerobic, moderately thermophilic bacteria were isolated from terrestrial hot springs in Baikal Lake region and Kamchatka (Russia). Cells of the new isolates were cocci reproducing by binary fission. The temperature range for growth was between 20 and 56 °C and the pH range for growth from pH 4.5 to 8.5, with optimal growth at 47-50 °C and pH 7.0-7.5. The organisms were chemoheterotrophs preferring sugars and polysaccharides as growth substrates. 16S rRNA gene sequences of strains 2842, 2813 and 2918Kr were nearly identical (99.7-100 % similarity) and indicated that the strains belonged to the phylum Planctomycetes. The phylogenetically closest cultivated relatives were Algisphaera agarilytica 06SJR6-2(T) and Phycisphaera mikurensis FYK2301M01(T) with 16S rRNA gene sequence similarity values of 82.4 and 80.3 %, respectively. The novel strains differed from them by higher growth temperature, sensitivity to NaCl concentration above 3.0 % and by their cellular fatty acids profile. On the basis of phylogenetic and physiological data, strains 2842(T), 2813 and 2918Kr represent a novel genus and species for which we propose the name Tepidisphaera mucosa sp. nov. The type strain is 2842(T) ( = VKM B-2832(T) = JCM 19875(T)). We also propose that Tepidisphaera gen. nov. is the type genus of a novel family, Tepidisphaeraceae fam. nov. and a novel order, Tepidisphaerales ord. nov. PMID:25404483

  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. 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.

  5. Exciton dynamics in the chlorosomal antennae of the green bacteria Chloroflexus aurantiacus and Chlorobium tepidum.

    PubMed Central

    Prokhorenko, V I; Steensgaard, D B; Holzwarth, A R

    2000-01-01

    The energy transfer processes in isolated chlorosomes from green bacteria Chlorobium tepidum and Chloroflexus aurantiacus have been studied at low temperatures (1.27 K) by two-pulse photon echo and one-color transient absorption techniques with approximately 100 fs resolution. The decay of the coherence in both types of chlorosomes is characterized by four different dephasing times stretching from approximately 100 fs up to 300 ps. The fastest component reflects dephasing that is due to interaction of bacteriochlorophylls with the phonon bath, whereas the other components correspond to dephasing due to different energy transfer processes such as distribution of excitation along the rod-like aggregates, energy exchange between different rods in the chlorosome, and energy transfer to the base plate. As a basis for the interpretation of the excitation dephasing and energy transfer pathways, a superlattice-like structural model is proposed based on recent experimental data and computer modeling of the Bchl c aggregates (1994. Photosynth. Res. 41:225-233.) This model predicts a fine structure of the Q(y) absorption band that is fully supported by the present photon echo data. PMID:11023914

  6. Two genes encoding new carotenoid-modifying enzymes in the green sulfur bacterium Chlorobium tepidum.

    PubMed

    Maresca, Julia A; Bryant, Donald A

    2006-09-01

    The green sulfur bacterium Chlorobium tepidum produces chlorobactene as its primary carotenoid. Small amounts of chlorobactene are hydroxylated by the enzyme CrtC and then glucosylated and acylated to produce chlorobactene glucoside laurate. The genes encoding the enzymes responsible for these modifications of chlorobactene, CT1987, and CT0967, have been identified by comparative genomics, and these genes were insertionally inactivated in C. tepidum to verify their predicted function. The gene encoding chlorobactene glucosyltransferase (CT1987) has been named cruC, and the gene encoding chlorobactene lauroyltransferase (CT0967) has been named cruD. Homologs of these genes are found in the genomes of all sequenced green sulfur bacteria and filamentous anoxygenic phototrophs as well as in the genomes of several nonphotosynthetic bacteria that produce similarly modified carotenoids. The other bacteria in which these genes are found are not closely related to green sulfur bacteria or to one another. This suggests that the ability to synthesize modified carotenoids has been a frequently transferred trait.

  7. Thermophilic Life Inside the Permafrost

    NASA Astrophysics Data System (ADS)

    Mironov, V. A.; Rivkina, E. M.; Vishnivetskaya, T. A.; Shcherbakova, V. A.; Gilichinsky, D. A.

    2010-04-01

    Anaerobic thermophiles and hyperthermophiles detected in permanently frozen volcanic sediments and freshly fallen volcanic ash. Volcanoes of Kamchatka peninsula (Russia) and Antarctic island Deception.

  8. 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.

  9. 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.

  10. 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.

  11. 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

  12. 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

    2011-10-02

    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.

  13. Diversity and biosynthesis of compatible solutes in hyper/thermophiles.

    PubMed

    Empadinhas, Nuno; da Costa, Milton S

    2006-09-01

    The accumulation of compatible solutes, either by uptake from the medium or by de novo synthesis, is a general response of microorganisms to osmotic stress. The diversity of compatible solutes is large but falls into a few major chemical categories, such as carbohydrates or their derivatives and amino acids or their derivatives. This review deals with compatible solutes found in thermophilic or hyperthermophilic bacteria and archaea that have not been commonly identified in microorganisms growing at low and moderate temperatures. The response to NaCl stress of Thermus thermophilus is an example of how a thermophilic bacterium responds to osmotic stress by compatible solute accumulation. Emphasis is made on the pathways leading to the synthesis of mannosylglycerate and glucosylglycerate that have been recently elucidated in several hyper/thermophilic microorganisms. The role of compatible solutes in the thermoprotection of these fascinating microorganisms is also discussed.

  14. Continuous Thermophilic Composting12

    PubMed Central

    Schulze, K. L.

    1962-01-01

    Under complete mixing conditions, aerobic decomposition of mixed organic waste materials has been maintained continuously in the thermophilic phase in a 55-gal rotating drum. Temperatures ranged between 53 and 70 C. Raw material was added daily or every second day in amounts up to 18 lb per 100 lb of decomposing material. The weight of material removed ranged between 42 and 60% of the raw material added. Factors influencing the operation of the composting unit were studied in detail. Images FIG. 2 PMID:13909559

  15. 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

  16. Genetic manipulation of carotenoid biosynthesis in the green sulfur bacterium Chlorobium tepidum.

    PubMed

    Frigaard, Niels-Ulrik; Maresca, Julia A; Yunker, Colleen E; Jones, A Daniel; Bryant, Donald A

    2004-08-01

    The green sulfur bacterium Chlorobium tepidum is a strict anaerobe and an obligate photoautotroph. On the basis of sequence similarity with known enzymes or sequence motifs, nine open reading frames encoding putative enzymes of carotenoid biosynthesis were identified in the genome sequence of C. tepidum, and all nine genes were inactivated. Analysis of the carotenoid composition in the resulting mutants allowed the genes encoding the following six enzymes to be identified: phytoene synthase (crtB/CT1386), phytoene desaturase (crtP/CT0807), zeta-carotene desaturase (crtQ/CT1414), gamma-carotene desaturase (crtU/CT0323), carotenoid 1',2'-hydratase (crtC/CT0301), and carotenoid cis-trans isomerase (crtH/CT0649). Three mutants (CT0180, CT1357, and CT1416 mutants) did not exhibit a discernible phenotype. The carotenoid biosynthetic pathway in C. tepidum is similar to that in cyanobacteria and plants by converting phytoene into lycopene using two plant-like desaturases (CrtP and CrtQ) and a plant-like cis-trans isomerase (CrtH) and thus differs from the pathway known in all other bacteria. In contrast to the situation in cyanobacteria and plants, the construction of a crtB mutant completely lacking carotenoids demonstrates that carotenoids are not essential for photosynthetic growth of green sulfur bacteria. However, the bacteriochlorophyll a contents of mutants lacking colored carotenoids (crtB, crtP, and crtQ mutants) were decreased from that of the wild type, and these mutants exhibited a significant growth rate defect under all light intensities tested. Therefore, colored carotenoids may have both structural and photoprotection roles in green sulfur bacteria. The ability to manipulate the carotenoid composition so dramatically in C. tepidum offers excellent possibilities for studying the roles of carotenoids in the light-harvesting chlorosome antenna and iron-sulfur-type (photosystem I-like) reaction center. The phylogeny of carotenogenic enzymes in green sulfur

  17. 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.

  18. 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.

  19. 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

  20. The three-dimensional structure of CsmA: a small antenna protein from the green sulfur bacterium Chlorobium tepidum.

    PubMed

    Pedersen, Marie Østergaard; Underhaug, Jarl; Dittmer, Jens; Miller, Mette; Nielsen, Niels Chr

    2008-08-20

    The structure of the chlorosome baseplate protein CsmA from Chlorobium tepidum in a 1:1 chloroform:methanol solution was determined using liquid-state NMR spectroscopy. The data reveal that the 59-residue protein is predominantly alpha-helical with a long helical domain extending from residues V6 to L36, containing a putative bacteriochlorophyll a binding domain, and a short helix in the C-terminal part extending from residues M41 to G49. These elements are compatible with a model of CsmA having the long N-terminal alpha-helical stretch immersed into the lipid monolayer confining the chlorosome and the short C-terminal helix protruding outwards, thus available for interaction with the Fenna-Matthews-Olson antenna protein.

  1. 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/).

  2. 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.

  3. [Thermophilic prokaryotes from deep subterranean habitats].

    PubMed

    Slobodkin, A I; Slobodkina, G B

    2014-01-01

    The deep continental biosphere consists of geologically isolated ecosystems differing in their physicochemical, geological, and trophic parameters. Most of the deep ecosystems exist at elevated temperatures (50-120 degrees C), which favor the development of thermophilic microorganisms. In many cases, indigenous nature of subsurface microorganisms is questionable due to problems of collecting representative and non-contaminated samples. In spite of the numerous studies on the deep biosphere microbial communities, the number of cultivated thermophiles isolated from subsurface environments not associated with petroleum deposits does not exceed 30 species. More than half of the thermophilic species isolated from deep subsurface belong to the Firmicutes. Majority of the underground thermophiles are subsurface strict or facultative anaerobes, with capacity for sulfate and iron reduction are notably widespread. Most thermophilic subsurface microorganisms are organotrophs, although chemolithoautotrophic thermophiles also have been reported. This review deals with the phylogenetic diversity and physiological properties of the cultivated thermophilic prokaryotes isolated from various deep subterranean habitats.

  4. Pump-probe anisotropies of Fenna-Matthews-Olson protein trimers from Chlorobium tepidum: a diagnostic for exciton localization?

    PubMed Central

    Savikhin, S; Buck, D R; Struve, W S

    1997-01-01

    Exciton calculations on symmetric and asymmetric Fenna-Matthews-Olson (FMO) trimers, combined with absorption difference anisotropy measurements on FMO trimers from the green bacterium Chlorobium tepidum, suggest that real samples exhibit sufficient diagonal energy disorder so that their laser-excited exciton states are noticeably localized. Our observed anisotropies are clearly inconsistent with 21-pigment exciton simulations based on a threefold-symmetric FMO protein. They are more consistent with a 7-pigment model that assumes that the laser-prepared states are localized within a subunit of the trimer. Differential diagonal energy shifts of 50 cm(-1) between symmetry-related pigments in different subunits are large enough to cause sharp localization in the stationary states; these shifts are commensurate with the approximately 95 cm(-1) inhomogeneous linewidth of the lowest exciton levels. Experimental anisotropies (and by implication steady-state linear and circular dichroism) likely arise from statistical averaging over states with widely contrasting values of these observables, in consequence of their sensitivity to diagonal energy disorder. PMID:9336204

  5. Ethanol production by engineered thermophiles.

    PubMed

    Olson, Daniel G; Sparling, Richard; Lynd, Lee R

    2015-06-01

    We compare a number of different strategies that have been pursued to engineer thermophilic microorganisms for increased ethanol production. Ethanol production from pyruvate can proceed via one of four pathways, which are named by the key pyruvate dissimilating enzyme: pyruvate decarboxylase (PDC), pyruvate dehydrogenase (PDH), pyruvate formate lyase (PFL), and pyruvate ferredoxin oxidoreductase (PFOR). For each of these pathways except PFL, we see examples where ethanol production has been engineered with a yield of >90% of the theoretical maximum. In each of these cases, this engineering was achieved mainly by modulating expression of native genes. We have not found an example where a thermophilic ethanol production pathway has been transferred to a non-ethanol-producing organism to produce ethanol at high yield. A key reason for the lack of transferability of ethanol production pathways is the current lack of understanding of the enzymes involved. PMID:25745810

  6. Thermophilic anaerobic oxidation of methane by marine microbial consortia

    PubMed Central

    Holler, Thomas; Widdel, Friedrich; Knittel, Katrin; Amann, Rudolf; Kellermann, Matthias Y; Hinrichs, Kai-Uwe; Teske, Andreas; Boetius, Antje; Wegener, Gunter

    2011-01-01

    The anaerobic oxidation of methane (AOM) with sulfate controls the emission of the greenhouse gas methane from the ocean floor. AOM is performed by microbial consortia of archaea (ANME) associated with partners related to sulfate-reducing bacteria. In vitro enrichments of AOM were so far only successful at temperatures ⩽25 °C; however, energy gain for growth by AOM with sulfate is in principle also possible at higher temperatures. Sequences of 16S rRNA genes and core lipids characteristic for ANME as well as hints of in situ AOM activity were indeed reported for geothermally heated marine environments, yet no direct evidence for thermophilic growth of marine ANME consortia was obtained to date. To study possible thermophilic AOM, we investigated hydrothermally influenced sediment from the Guaymas Basin. In vitro incubations showed activity of sulfate-dependent methane oxidation between 5 and 70 °C with an apparent optimum between 45 and 60 °C. AOM was absent at temperatures ⩾75 °C. Long-term enrichment of AOM was fastest at 50 °C, yielding a 13-fold increase of methane-dependent sulfate reduction within 250 days, equivalent to an apparent doubling time of 68 days. The enrichments were dominated by novel ANME-1 consortia, mostly associated with bacterial partners of the deltaproteobacterial HotSeep-1 cluster, a deeply branching phylogenetic group previously found in a butane-amended 60 °C-enrichment culture of Guaymas sediments. The closest relatives (Desulfurella spp.; Hippea maritima) are moderately thermophilic sulfur reducers. Results indicate that AOM and ANME archaea could be of biogeochemical relevance not only in cold to moderate but also in hot marine habitats. PMID:21697963

  7. Electron transfer kinetics in purified reaction centers from the green sulfur bacterium Chlorobium tepidum studied by multiple-flash excitation.

    PubMed

    Kusumoto, N; Sétif, P; Brettel, K; Seo, D; Sakurai, H

    1999-09-14

    Reaction center preparations from the green sulfur bacterium Chlorobium tepidum, which contain monoheme cytochrome c, were studied by flash-absorption spectroscopy in the near-UV, visible, and near-infrared regions. The decay kinetics of the photooxidized primary donor P840(+), together with the amount of photooxidized cytochrome c, were analyzed along a series of four flashes spaced by 1 ms: 95% of the P840(+) was reduced by cytochrome c with a t(1/2) of approximately 65 micros after the first flash, 80% with a t(1/2) of approximately 100 micros after the second flash, and 23% with a t(1/2) of approximately 100 micros after the third flash; after the fourth flash, almost no cytochrome c oxidation occurred. The observed rates, the establishment of redox equilibrium after each flash, and the total amount of photooxidizable cytochrome c are consistent with the presence of two equivalent cytochrome c molecules per photooxidizable P840. The data are well fitted assuming a standard free energy change DeltaG degrees of -53 meV for electron transfer from one cytochrome c to P840(+), DeltaG degrees being independent of the oxidation state of the other cytochrome c. These observations support a model with two monoheme cytochromes c which are symmetrically arranged around the reaction center core. From the ratio of menaquinone-7 to the bacteriochlorophyll pigment absorbing at 663 nm, it was estimated that our preparations contain 0.6-1.2 menaquinone-7 molecules per reaction center. However, no transient signal due to menaquinone could be observed between 360 and 450 nm in the time window from 10 ns to 4 micros. No recombination reaction between the primary partners P840(+) and A(0)(-) could be detected under normal conditions. Such a recombination was observed (t(1/2) approximately 19 ns) under highly reducing conditions or after accumulation of three electrons on the acceptor side during a series of flashes, showing that the secondary acceptors can stabilize three electrons

  8. 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.

  9. 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.

  10. Thermophilic biological nitrogen removal in industrial wastewater treatment.

    PubMed

    Lopez-Vazquez, C M; Kubare, M; Saroj, D P; Chikamba, C; Schwarz, J; Daims, H; Brdjanovic, D

    2014-01-01

    Nitrification is an integral part of biological nitrogen removal processes and usually the limiting step in wastewater treatment systems. Since nitrification is often considered not feasible at temperatures higher than 40 °C, warm industrial effluents (with operating temperatures higher than 40 °C) need to be cooled down prior to biological treatment, which increases the energy and operating costs of the plants for cooling purposes. This study describes the occurrence of thermophilic biological nitrogen removal activity (nitritation, nitratation, and denitrification) at a temperature as high as 50 °C in an activated sludge wastewater treatment plant treating wastewater from an oil refinery. Using a modified two-step nitrification-two-step denitrification mathematical model extended with the incorporation of double Arrhenius equations, the nitrification (nitrititation and nitratation) and denitrification activities were described including the cease in biomass activity at 55 °C. Fluorescence in situ hybridization (FISH) analyses revealed that Nitrosomonas halotolerant and obligatehalophilic and Nitrosomonas oligotropha (known ammonia-oxidizing organisms) and Nitrospira sublineage II (nitrite-oxidizing organism (NOB)) were observed using the FISH probes applied in this study. In particular, this is the first time that Nitrospira sublineage II, a moderatedly thermophilic NOB, is observed in an engineered full-scale (industrial) wastewater treatment system at temperatures as high as 50 °C. These observations suggest that thermophilic biological nitrogen removal can be attained in wastewater treatment systems, which may further contribute to the optimization of the biological nitrogen removal processes in wastewater treatment systems that treat warm wastewater streams.

  11. Thermophilic fungi isolated from a heated aquatic habitat

    SciTech Connect

    Ellis, D.H.

    1980-09-01

    The cooling pond for the Hazelwood Power Station, Morwell, Victoria, Australia, was studied for the presence of thermophilic fungi. A total of five species was recorded: Chaetomium thermophile, Aspergillus fumigatus, Humicola grisea, Humicola insolens, and Thermomyces lanuginosus. (ACR)

  12. 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.

  13. 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.

  14. 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

  15. 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

  16. 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

  17. 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.

  18. 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.

  19. 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.

  20. Menaquinone-7 in the reaction center complex of the green sulfur bacterium Chlorobium vibrioforme functions as the electron acceptor A1.

    PubMed

    Kjaer, B; Frigaard, N U; Yang, F; Zybailov, B; Miller, M; Golbeck, J H; Scheller, H V

    1998-03-10

    Photosynthetically active reaction center complexes were prepared from the green sulfur bacterium Chlorobium vibrioforme NCIMB 8327, and the content of quinones was determined by extraction and high-performance liquid chromatography. The analysis showed a stoichiometry of 1.7 molecules of menaquinone-7/reaction center. No other quinones were detected in the isolated reaction centers, whereas membrane preparations also contained chlorobiumquinone. The possible involvement of quinones in electron transport was investigated by electron paramagnetic resonance (EPR) spectroscopy. A highly anisotropic radical was detected by Q-band EPR spectroscopy in both membranes and isolated reaction centers following dark reduction with sodium dithionite and photoaccumulation at 205 K. At 34 GHz, the EPR spectrum is characterized by a g tensor with gxx = 2.0063, gyy = 2.0052, gzz = 2.0020 and delta B of 0.7 mT, consistent with its identification as a quinone. This spectrum is highly similar in terms of g values and line widths to photoaccumulated A1- in photosystem I of Synechococcus sp. PCC 7002. The results indicate that menaquinone-7 in the green sulfur bacterial reaction center is analogous to phylloquinone in photosystem I.

  1. 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

  2. Solubilization of insoluble phosphates by thermophilic fungi.

    PubMed

    Singh, C P; Mishra, M M; Yadav, K S

    1980-01-01

    The solubilization of tricalcium phosphate and rock phosphate and assimilation of solubilized P by thermophilic fungi isolated from compost were studied. The solubilization of tricalcium phosphate was greater than that of rock phosphate on inoculation with fungi in liquid medium, but growth of most of the fungi was greater in rock phosphate. Torula thermophila solubilized tricalcium phosphate maximally. There was solubilization of rock phosphate in semi-solid lignocellulose medium by Aspergillus fumigatus.

  3. Characterization of a recently purified thermophilic DNase from a novel thermophilic fungus.

    PubMed

    Landry, Kyle S; Levin, Robert E

    2014-08-01

    A newly isolated thermophilic fungus was found to produce a partially inducible extracellular DNase. This manuscript focuses on the characterization of this novel thermophilic DNase in terms of optimal enzyme conditions, molecular weight, and certain kinetic properties. The DNase was found to be inactivated by the presence of EDTA demonstrating its dependence on metal cofactors for activity. Maximum activity occurred at pH 6.0 with no activity at pH 2.0 or 10.0. The optimal temperature for the purified DNase was 65 °C. The thermophilic DNase was found to be an exonuclease with an estimated molecular weight of 56 kDa.

  4. 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.

  5. Thermophilic methane production and oxidation in compost.

    PubMed

    Jäckel, Udo; Thummes, Kathrin; Kämpfer, Peter

    2005-04-01

    Methane cycling within compost heaps has not yet been investigated in detail. We show that thermophilic methane oxidation occurred after a lag phase of up to one day in 4-week old, 8-week old and mature (>10-week old) compost material. The potential rate of methane oxidation was between 2.6 and 4.1 micromol CH4(gdw)(-1)h(-1). Profiles of methane concentrations within heaps of different ages indicated that 46-98% of the methane produced was oxidised by methanotrophic bacteria. The population size of thermophilic methanotrophs was estimated at 10(9) cells (gdw)(-1), based on methane oxidation rates. A methanotroph (strain KTM-1) was isolated from the highest positive step of a serial dilution series. This strain belonged to the genus Methylocaldum, which contains thermotolerant and thermophilic methanotrophs. The closest relative organism on the basis of 16S rRNA gene sequence identity was M. szegediense (>99%), a species originally isolated from hot springs. The temperature optimum (45-55 degrees C) for methane oxidation within the compost material was identical to that of strain KTM-1, suggesting that this strain was well adapted to the conditions in the compost material. The temperatures measured in the upper layer (0-40 cm) of the compost heaps were also in this range, so we assume that these organisms are capable of effectively reducing the potential methane emissions from compost.

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

    SciTech Connect

    Stoner, D.L.; Miller, K.S.; Fife, D.J.; Larsen, E.D.; Tolle, C.R.; Johnson, J.A.

    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 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.

  8. 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.

  9. 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

  10. Thermophilic anaerobic digestion: the best option for waste treatment.

    PubMed

    Suryawanshi, P C; Chaudhari, A B; Kothari, R M

    2010-03-01

    After introducing thermophilic anaerobic digestion (AD), characteristics of thermophilic methanogens are provided. Accordingly, (a) site of occurrence, (b) morphological characteristics (shape and motility), (c) biochemical characteristics (Gram character and % G+C profile), (d) nutritional characteristics (NaCl requirement and substrate specificity), and (e) growth characteristics (pH and temperature) of thermophilic methanogens are described. Some studies of the thermophilic AD are cited with their operational management problems. Subsequently, strategies to maximize net energy production are given, including mode of heating the bioreactors, role of agitation to promote AD performance and mode/intensity of mixing. Finally, advantages as well as drawbacks of AD under thermophilic conditions are given, concluding with its applications. PMID:20148754

  11. Sequencing mesophilic and thermophilic anaerobic digesters. Final report

    SciTech Connect

    Not Available

    1982-12-01

    This project employed two laboratory bench scale, complete-mix anaerobic sludge digesters arranged in a series configuration. The first digester was operated at 35/sup 0/C (mesophilic) and the second at 50/sup 0/C (thermophilic). A portion of the thermophilic sludge was recycled through an aeration basin. As a comparison to the mesophilic-thermophilic sequencing, a mesophilic-mesophilic digester sequence, without sludge recycle to the aeration basin, was operated in parallel to the test units and loaded at an equivalent rate. Conclusions of this study are as follows: in establishing a thermophilic anaerobic digester, a slow-start procedure, in which the temperature is increased at a rate of 0.6/sup 0/C per day with loading, appears to produce a more stable thermophilic digester in a shorter period of time than a quick-start procedure, in which the temperature is increased rapidly with no loading. Even after a year, the slow-start thermophilic digester proved to be unstable once sequencing began. A greater volatile solids, COD, BOD, and grease reduction with a higher gas production was achieved using a mesophilic-mesophilic sequence, probably, in part, due to the instability (volatile acids in the effluent) of the thermophilic digester in the mesophilic-thermophilic sequence. A greater total kjeldahl N (TKN) and total coliform destruction was achieved in the thermophilic digester, however, poor dewatering characteristics, as indicated by the capillary suction time (CST), and an obnoxious odor were also evident. Other than an increase in effluent suspended solids, the recycle of thermophilic sludge to an aeration basin produced no discernable effect. 9 figs., 3 tabs.

  12. 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.

  13. 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

  14. Thermophile bacteria in permafrost: model for astrobiology

    NASA Astrophysics Data System (ADS)

    Gilichinsky, D.; Rivkina, E.; Shcherbakova, V.; Laurinavichius, K.; Kholodov, A.; Abramov, A.

    2003-04-01

    According the NASA point of view, one way to have liquid water on Mars at shallow depths would be through subglacial volcanism. Such volcano-ice interactions could be going on beneath the polar caps of Mars today, or even within the adjacent permafrost around the margins of the ice caps. This is why one of the Earth's models, close to extraterrestrial environment, represented by active volcanoes in permafrost areas and the main question is - does such econishes as volcanoes and associated environment contain recently microbial communities? The first step of this study was carried out on volcano Stromboli (Italy), using the marine water samples extracted from the borehole near the island marine coast, surrounding the volcano. According the temperatures (45^oC), this thermal water has the hydraulic connection with volcano. Microscopy analyses of studied water shown the presence of different morphological types of microorganisms: small mobile roads, coccoid and sarcina-like organisms and long fixed roads, as well as rest forms (spores and cysts). To separate this community on marine and volcano microorganisms, the common mineral media with added CO_2, acetate or glucose-peptone as a source of carbon were used for culturing, and Fe3+, S^o, SO_42- were added as a electron acceptors. We attempt to isolate thermophilic anaerobic microorganisms of different metabolic groups - methanogens, acetogens, iron-, sulfur- and sulfate-reducers, and to test each group of microorganisms on the presence of halophilic forms. After 24 hours of incubation at temperatures varied 55 to 85^o, the grow relatively the control media was observed at CO_2+H_2 and glucose-peptone media. Microscopy study of preparations showed small coccus of irregular shape that was unable to reduce S^o or SO_42-. During the subsequent re-seeding were obtained the enrichment cultures of themophilic bacteria, genetically closed to genera Thermococcus: heterotrophic, growing up to 95^oC with the growth optimum at

  15. 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.

  16. Hydrogenase activity in the thermophile mastigocladus laminosus

    SciTech Connect

    Benemann, J.R.; Miyamoto, K.; Hallenbeck, P.C.; Murry, M.A.

    1982-06-30

    Hydrogenase activity in the thermophilic cyanobacterium, Mastigocladus laminosus was studied both in vivo and in vitro. In vivo hydrogen consumption required oxygen but not light, was about ten-fold higher than in mesophilic cyanobacteria, and was relatively insensitive to carbon monoxide. H/sub 2/-supported acetylene reduction in reductant-limited cultures was a light-dependent, but O/sub 2/-independent reaction. In vitro hydrogen evolution was unaffected by carbon monoxide, and this activity could be partially purified using a procedure developed for Anabaena cylindrica.

  17. 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.

  18. Coal desulfurization by leaching involving acidophilic and thermophilic microorganisms

    SciTech Connect

    Murr, L.E.; Mehta, A.P.

    1982-03-01

    It was shown that thermophilic microorganisms can increase the rate and volume of pyrite leaching from pulverized, high-sulfur coal. This occurs not only because of the elevated temperature of operation possible, but also the apparent accelerated catalytic activity of thermophilic microorganisms as compared to autotropic microbes such as T. ferrooxidans. The ability of thermophilic microogranisms to successfully leach pyrite in coal as demonstrated in the study points toward an even greater potential for the develpment of a successful and economically viable large-scale desulfurization process involving biochemical leaching. (JMT)

  19. 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

  20. 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

  1. 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

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

    PubMed

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

    2014-01-01

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

  3. 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.

  4. 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

  5. Decomposition of intact chicken feathers by a thermophile in combination with an acidulocomposting garbage-treatment process.

    PubMed

    Shigeri, Yasushi; Matsui, Tatsunobu; Watanabe, Kunihiko

    2009-11-01

    In order to develop a practical method for the decomposition of intact chicken feathers, a moderate thermophile strain, Meiothermus ruber H328, having strong keratinolytic activity, was used in a bio-type garbage-treatment machine working with an acidulocomposting process. The addition of strain H328 cells (15 g) combined with acidulocomposting in the garbage machine resulted in 70% degradation of intact chicken feathers (30 g) within 14 d. This degradation efficiency is comparable to a previous result employing the strain as a single bacterium in flask culture, and it indicates that strain H328 can promote intact feather degradation activity in a garbage machine currently on the market.

  6. Decomposition of intact chicken feathers by a thermophile in combination with an acidulocomposting garbage-treatment process.

    PubMed

    Shigeri, Yasushi; Matsui, Tatsunobu; Watanabe, Kunihiko

    2009-11-01

    In order to develop a practical method for the decomposition of intact chicken feathers, a moderate thermophile strain, Meiothermus ruber H328, having strong keratinolytic activity, was used in a bio-type garbage-treatment machine working with an acidulocomposting process. The addition of strain H328 cells (15 g) combined with acidulocomposting in the garbage machine resulted in 70% degradation of intact chicken feathers (30 g) within 14 d. This degradation efficiency is comparable to a previous result employing the strain as a single bacterium in flask culture, and it indicates that strain H328 can promote intact feather degradation activity in a garbage machine currently on the market. PMID:19897897

  7. Thermophilic growth and enzymatic thermostability are polyphyletic traits within Chaetomiaceae.

    PubMed

    van den Brink, Joost; Facun, Kryss; de Vries, Michel; Stielow, J Benjamin

    2015-12-01

    Thermophilic fungi have the potential to produce industrial-relevant thermostable enzymes, in particular for the degradation of plant biomass. Sordariales is one of the few fungal orders containing several thermophilic taxa, of which many have been associated with the production of thermostable enzymes. The evolutionary affiliation of Sordariales fungi, especially between thermophiles and non-thermophilic relatives, is however poorly understood. Phylogenetic analysis within the current study was based on sequence data, derived from a traditional Sanger and highly multiplexed targeted next generation sequencing approach of 45 isolates. The inferred phylogeny and detailed growth analysis rendered the trait 'thermophily' as polyphyletic within Chaetomiaceae (Sordariales, Sordariomycetes), and characteristic to: Myceliophthora spp., Thielavia terrestris, Chaetomium thermophilum, and Mycothermus thermophilus. Compared to mesophiles, the isolates within thermophilic taxa produced enzyme mixtures with the highest thermostability of known cellulase activities. Temperature profiles of the enzyme activities correlated strongly with the optimal growth temperatures of the isolates but not with their phylogenetic relationships. This strong correlation between growth and enzyme characteristics indicated that detailed analysis of growth does give predictive information on enzyme physiology. The variation in growth and enzyme characteristics reveals these fungi as an excellent platform to better understand fungal thermophily and enzyme thermostability.

  8. 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.

  9. Microbial influenced corrosion by thermophilic bacteria

    NASA Astrophysics Data System (ADS)

    Lata, Suman; Sharma, Chhaya; Singh, Ajay

    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.

  10. Mathematical modelling of autothermal thermophilic aerobic digesters.

    PubMed

    Gomez, J; de Gracia, M; Ayesa, E; Garcia-Heras, J L

    2007-03-01

    This paper presents a new mathematical model for Autothermal Thermophilic Aerobic Digesters. The reactor has been modelled as two completely mixed volumes to separately predict the behaviour of the liquid and gaseous phases as well as the interrelation between them. The model includes biochemical transformations based on the standard Activated Sludge Models of IWA, as well as physico-chemical transformations associated with the chemical equilibria and the mass transfer between the liquid and the gaseous phases similar to those proposed in the ADM1 of IWA. An energy balance has also been included in the model in order to predict the temperature of the system. This thermal balance takes into account all those biochemical and physico-chemical transformations that entail the most relevant heat interchanges. Reactor performance has been explored by simulation in two different scenarios: in the first where it acts as the initial stage in a Dual system, and in the second where it acts as a single-stage treatment. Each scenario enabled the identification of the relevance of the different parameters. PMID:17258787

  11. Habitat, applications and genomics of the aerobic, thermophilic genus Geobacillus.

    PubMed

    McMullan, G; Christie, J M; Rahman, T J; Banat, I M; Ternan, N G; Marchant, R

    2004-04-01

    Thermophilic bacteria belonging to Bacillus genetic group 5 have been reclassified as being members of Geobacillus gen. nov., with G. stearothermophilus as the type strain. Geobacillus species, literally meaning earth or soil Bacillus, are widely distributed and readily isolated from natural and man-made thermophilic biotopes. Work within our group has however shown that an abundance of genetically distinct Geobacillus isolates can be obtained from temperate Irish soils. As with many thermophiles there is considerable interest in potential industrial application of these bacteria and their gene products. This review describes two novel applications for Geobacillus isolates, firstly in the metabolism of the herbicide glyphosate and secondly in the metabolism of quorum-sensing signal molecules from Gram-negative bacteria. Finally the current state of the art is described for Bacillus genomics, with details given of three independent genome-sequencing projects of Geobacillus isolates.

  12. 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

  13. Inhibitory Effects of Ferrihydrite on a Thermophilic Methanogenic Community

    PubMed Central

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

    2014-01-01

    The addition of ferrihydrite to methanogenic microbial communities obtained from a thermophilic anaerobic digester suppressed methanogenesis in a dose-dependent manner. The amount of reducing equivalents consumed by the reduction of iron was significantly smaller than that expected from the decrease in the production of CH4, which suggested that competition between iron-reducing microorganisms and methanogens was not the most significant cause for the suppression of methanogenesis. Microbial community analyses revealed that the presence of ferrihydrite markedly affected the bacterial composition, but not the archaeal composition. These results indicate that the presence of ferrihydrite directly and indirectly suppresses thermophilic methanogenesis. PMID:24859310

  14. 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.

  15. [Optimization of culture condition of a moderately thermophilic acidophilic Ferroplasma thermophilum].

    PubMed

    Zhou, Hongbo; Peng, Juanhua; Zhang, Ruiyong; Chen, Chen; Zhang, Rubing; Gu, Guohua; Qiu, Guanzhou

    2008-06-01

    The extreme microorganisms Ferroplasma spp., play an important role in bioleaching of sulphide ores at low pH value and temperatures around 50 degrees C. Without cell wall, Ferroplasma spp. is sensitive to pulp density, shearing force and heavy metal ions. Thus it is difficult to obtain their high cell density cultures, which limits the large-scale industrial application. In this paper, the optimum culture conditions of Ferroplasma thermophilum were studied by shaking culture. The results showed that the optimum culture conditions are as follows: 50 degrees C, initial pH 0.5, 50 mL working volume in 250 mL shaking-flask, inorganic nitrogen source (NH4)2SO4. The optimum combination of FeSO4.7H2O, yeast extract and peptone was determined by orthogonal experiments, including FeSO4.7H2O 40 g/L, yeast extract 0.3 g/L, peptone 0.2 g/L. Under the optimum culture conditions, the cell density was up to 6.3 x 10(7) cell/mL, and the oxidation of 40 g/L ferrous sulfate heptahydrate was finished in less than 72 hours. The results might provide information for scale-up of archaeon culture as well as its industrial application.

  16. 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

  17. Genome sequence of the moderately thermophilic halophile Flexistipes sinusarabici strain (MAS10).

    PubMed

    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

    2011-10-15

    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.

  18. 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.

  19. 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.

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

    PubMed Central

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

    2013-01-01

    Anaerobaculum mobile Menes and Muxí 2002 is one of three described species of the genus Anaerobaculum, family Synergistaceae, phylum Synergistetes. This anaerobic and motile bacterium ferments a range of carbohydrates and mono- and dicarboxylic acids with acetate, hydrogen 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 sequenced genome. Here we describe the features of this bacterium, together with the complete 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. PMID:23961311

  1. 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

  2. Complete genome sequence of the moderate thermophile Anaerobaculum mobile type strain (NGA(T)).

    PubMed

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

    2013-04-15

    Anaerobaculum mobile Menes and Muxí 2002 is one of three described species of the genus Anaerobaculum, family Synergistaceae, phylum Synergistetes. This anaerobic and motile bacterium ferments a range of carbohydrates and mono- and dicarboxylic acids with acetate, hydrogen and CO2 as end products. A. mobile NGA(T) is the first member of the genus Anaerobaculum and the sixth member of the phylum Synergistetes with a completely sequenced genome. Here we describe the features of this bacterium, together with the complete 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 G enomic E ncyclopedia of Bacteria and Archaea project.

  3. Georgenia sediminis sp. nov., a moderately thermophilic actinobacterium isolated from sediment.

    PubMed

    You, Zhi-Qing; Li, Jie; Qin, Sheng; Tian, Xin-Peng; Wang, Fa-Zuo; Zhang, Si

    2013-11-01

    A Gram-stain-positive actinobacterium, designated strain SCSIO 15020(T), was isolated from sediment of the South China Sea, and characterized by using a polyphasic approach. The temperature range for growth was 24-60 °C, with optimal growth occurring at 50 °C. The pH range for growth was 6-10 (optimum pH 8-9). The NaCl concentration range for growth was 0-5 % (w/v). The peptidoglycan type was A4α. Polar lipids contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside and an unknown polar lipid. The major menaquinone was MK-8(H4); MK-7(H4) was present as a minor component. The major fatty acids (>5 %) were anteiso-C15 : 0, iso-C15 : 0 and iso-C16 : 0. The DNA G+C content of strain SCSIO 15020(T) was 73.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain SCSIO 15020(T) belonged to the genus Georgenia, with the closest neighbours being Georgenia muralis 1A-C(T) (96.3 % similarity), Georgenia thermotolerans TT02-04(T) (95.7 %) and Georgenia ruanii YIM 004(T) (95.6 %). Based on evidence from the present polyphasic study, strain SCSIO 15020(T) is considered to represent a novel species of the genus Georgenia, for which the name Georgenia sediminis sp. nov. is proposed. The type strain is SCSIO 15020(T) ( = DSM 25884(T) = NBRC 108941(T)).

  4. (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.

  5. [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.

  6. [Characteristics of thermophilic ASBR treating thermal-hydrolyzed sewage sludge].

    PubMed

    Ouyang, Er-Ming; Wang, Wei

    2010-10-01

    The characteristics of thermophilic anaerobic sequencing batch reactor (ASBR) treating thermal-hydrolyzed sewage sludge were investigated. The results showed that increasing the OLR (COD) of the thermophilic ASBR treating thermal-hydrolyzed sewage sludge from 7.762 kg/(m3 x d) to 13.106 kg/(m3 x d) lead to the "acidification" of the reactor with the accumulation of the volatile fatty acid (VFA) and the drop of the pH and gas production. This acidification is recoverability. The OLR(COD) of the thermophilic ASBR treating thermal-hydrolyzed sewage sludge can reach to 10 kg/(m3 x d) after the system was recovered. Under the OLR(COD) of 2.523, 4.196, 7.762 and 10.091 kg/(m3 x d), the methane yield (CH4/COD(input)) of thermophilic ASBR were 250, 247, 219 and 187 mL respectively. There are significant linear relationship among OLR and MPR, the methane yield. With the increase of the OLR, the methane production rate increased while the methane yield reduced.

  7. 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

  8. 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.

  9. 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

  10. Electrochemical Performance and Microbial Characterization of Thermophilic Microbial Fuel Cells

    NASA Astrophysics Data System (ADS)

    Wrighton, K. C.; Agbo, P.; Brodie, E. L.; Weber, K. A.; Desantis, T. Z.; Anderson, G. L.; Coates, J. D.

    2007-12-01

    Significant research effort is currently focused on microbial fuel cells (MFC) as a source of renewable energy. To date, most of these efforts have concentrated on MFCs operating at mesophilic temperatures. However, many previous studies have reported on the superiority of thermophilic conditions in anaerobic digestion and demonstrated a net gain in energy yield, in terms of methane, relative to the increased energy requirements of operation. Because of this, our recent studies on MFCs focused on investigating the operation and microbiology associated with thermophilic MFCs operating at 55°C. Over 100-day operation, these MFCs were highly stable and achieved a maximum power density of 24mW/m2 and a columbic efficiency of 89 percent with acetate as the sole electron donor. In order to characterize the microbial community involved in thermophilic electricity generation, DNA and RNA were isolated from the electrode and PhyloChip analyses performed. Exploring the changes in the microbial community over time in electricity producing MFC revealed an increase in relative abundance of populations belonging to the Firmicutes, Chloroflexi, and alpha Proteobacteria by at least one order of magnitude. In contrast, these populations decreased in the open circuit and no electron donor amended controls. In order to better characterize the active microbial populations, we enriched and isolated a novel organism, strain JR, from samples collected from an operating MFC. Based on 16S rRNA sequence analysis strain JR was a member of the family Peptococcaceae, within the Phylum Firmicutes, clustering with Thermincola ferriacetica (98 percent similarity). Phenotypic characterization revealed that strain JR was capable of thermophilic dissimilatory reduction of insoluble electron acceptors such as amorphous Fe(III); as well as reduction of the model quinone 2,6-anthraquinone disulfonate. Thermincola strain JR was also capable of producing current by coupling acetate oxidation to anodic

  11. [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.

  12. 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

  13. Integration between chemical oxidation and membrane thermophilic biological process.

    PubMed

    Bertanza, G; Collivignarelli, M C; Crotti, B M; Pedrazzani, R

    2010-01-01

    Full scale applications of activated sludge thermophilic aerobic process for treatment of liquid wastes are rare. This experimental work was carried out at a facility, where a thermophilic reactor (1,000 m(3) volume) is operated. In order to improve the global performance of the plant, it was decided to upgrade it, by means of two membrane filtration units (ultrafiltration -UF-, in place of the final sedimentation, and nanofiltration -NF-). Subsequently, the integration with chemical oxidation (O(3) and H(2)O(2)/UV processes) was taken into consideration. Studied solutions dealt with oxidation of both the NF effluents (permeate and concentrate). Based on experimental results and economic evaluation, an algorithm was proposed for defining limits of convenience of this process.

  14. 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.

  15. Use of bacteriophage for the selective isolation of thermophilic actinomycetes from composted eucalyptus bark.

    PubMed

    Kurtböke, D I; Murphy, N E; Sivasithamparam, K

    1993-01-01

    A method was developed to reduce the numbers of thermophilic bacteria on isolation plates, which in turn facilitated the detection and isolation of thermophilic actinomycetes. The method involves exposing the test material to bacteriophage suspensions prior to inoculation on isolation plates. This method was applied to composted eucalyptus bark samples, which were then inoculated on R8 and 1/2 TSA + 0.2% casein hydrolysate agar plates. The phage susceptibility of thermophilic bacteria provided a selective means of reducing their numbers on isolation plates and hence increased the numbers of Thermomonospora, Saccharopolyspora rectivirgula, and thermophilic Streptomyces spp. on these media in comparison with the numbers recorded from control plates.

  16. 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

  17. Deferribacter thermophilus gen. nov., sp. nov., a novel thermophilic manganese- and iron-reducing bacterium isolated from a petroleum reservoir.

    PubMed

    Greene, A C; Patel, B K; Sheehy, A J

    1997-04-01

    A thermophilic anaerobic bacterium, designated strain BMAT (T = type strain), was isolated from the production water of Beatrice oil field in the North Sea (United Kingdom). The cells were straight to bent rods (1 to 5 by 0.3 to 0.5 microns) which stained gram negative. Strain BMAT obtained energy from the reduction of manganese (IV), iron(III), and nitrate in the presence of yeast extract, peptone, Casamino Acids, tryptone, hydrogen, malate, acetate, citrate, pyruvate, lactate, succinate, and valerate. The isolate grew optimally at 60 degrees C (temperature range for growth, 50 to 65 degrees C) and in the presence of 2% (wt/vol) NaCl (NaCl range for growth, 0 to 5% [wt/vol]). The DNA base composition was 34 mol% G + C. Phylogenetic analyses of the 16S rRNA gene indicated that strain BMAT is a member of the domain Bacteria. The closest known bacterium is the moderate thermophile Flexistipes sinusarabici (similarity value, 88%). Strain BMAT possesses phenotypic and phylogenetic traits that do not allow its classification as a member of any previously described genus; therefore, we propose that this isolate should be described as a member of a novel species of a new genus, Deferribacter thermophilus gen. nov., sp. nov.

  18. Thermophilic biofilter for SO2 removal: performance and microbial characteristics.

    PubMed

    Zhang, Jingying; Li, Lin; Liu, Junxin

    2015-03-01

    A bench-scale thermophilic biofilter was applied to remove SO2 at 60°C in the present study. The SO2 concentration in the inlet stream ranged from 100mg/m(3) to 200mg/m(3). An average SO2 removal efficiency of 93.10% was achieved after developing acclimated organisms that can degrade SO2. The thermophilic biofilter effectively reduced SO2, with a maximum elimination capacity of 50.67g/m(3)/h at a loading rate of 51.44g/m(3)/h. Removal efficiency of the thermophilic biofilter was largely influenced by the water containing rate of the packing materials. The SO2 transfer in the biofilter included adsorption by the packing materials, dissolution in liquid, and microbial degradation. The main product of SO2 degradation was SO4(2-). The temporal shifts in the bacterial community that formed in the biofilter were determined through polymerase chain reaction-denaturing gradient gel electrophoresis and DNA sequence analysis. These shifts revealed a correlation between biofilter performance and bacterial community structure.

  19. 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

  20. 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.

  1. 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).

  2. 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)

  3. 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...

  4. 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

  5. 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

  6. Complete genome sequence of the thermophilic Acidobacteria, Pyrinomonas methylaliphatogenes type strain K22(T).

    PubMed

    Lee, Kevin C Y; Morgan, Xochitl C; Power, Jean F; Dunfield, Peter F; Huttenhower, Curtis; Stott, Matthew B

    2015-01-01

    Strain K22(T) is the type species of the recently- described genus Pyrinomonas, in subdivision 4 of the phylum Acidobacteria (Int J Syst Evol Micr. 2014; 64(1):220-7). It was isolated from geothermally-heated soil from Mt. Ngauruhoe, New Zealand, using low-nutrient medium. P. methylaliphatogenes K22(T) has a chemoheterotrophic metabolism; it can hydrolyze a limited range of simple carbohydrates and polypeptides. Its cell membrane is dominated by iso-branching fatty acids, and up to 40 % of its lipid content is membrane-spanning and ether lipids. It is obligately aerobic, thermophilic, moderately acidophilic, and non-spore-forming. The 3,788,560 bp genome of P. methylaliphatogenes K22(T) has a G + C content of 59.36 % and contains 3,189 protein-encoding and 55 non-coding RNA genes. Genomic analysis was consistent with nutritional requirements; in particular, the identified transporter classes reflect the oligotrophic nature of this strain.

  7. Purification and characterization of thermostable α-amylase from thermophilic Anoxybacillus flavithermus.

    PubMed

    Agüloğlu Fincan, S; Enez, B; Özdemir, S; Matpan Bekler, F

    2014-02-15

    This study reports on the purification and characterization of thermostable α-amylase (α-1-4 D-glucan glucanohydrolase EC 3.2.1.1) from a newly isolated Anoxybacillus flavithermus. A. flavithermus was used, which was isolated from hot water springs of Ömer, Afyonkarahisar, Turkey. The gram-positive, spore-forming, motile, moderately thermophilic bacteria were found to be a strain of A. flavithermus analysed by 16S rRNA comparison. The optimal conditions for bacterial growth were determined to be at 20 thh, 55 °C and pH 6.0. Maximum α-amylase activity was obtained at 55 °C at pH 7.0 after 24h of incubation. Thermostable α-amylase from A. flavithermus was purified by 70% (NH4)2SO4 and ion-exchange chromatography (5.2-fold; 65.8% yield). The molecular weight of α-amylase was 60 kDa, as estimated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The α-amylase hydrolyzed soluble starch at 55 °C with Km: 0.005 mM and Vmax: 3.5 μmol min(-1).

  8. 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.

  9. 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.

  10. 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

  11. 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.

  12. Occurrence of Isocitrate Lyase in a Thermophilic Bacillus Species

    PubMed Central

    Daron, Harlow H.

    1967-01-01

    A thermophilic, sporeforming bacterium has been isolated from soil on a medium containing acetate as a carbon source. This organism is similar to Bacillus stearothermophilus in most respects but differs in its inability to hydrolyze starch. Isocitrate lyase is present in cell-free extracts of organisms grown in a medium with acetate as a carbon source. The specific activity was 400 times lower in extracts of organisms utilizing glucose as a carbon source. With crude extracts, enzyme activity was strongly stimulated by Mg++, but cysteine and ethylenediaminetetraacetate had little effect. It appeared to be more heat-stable than the pure isocitrate lyase from Pseudomonas indigofera. Images PMID:6020570

  13. 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

  14. 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

  15. 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

  16. 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

  17. 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.

  18. Isolation and Characterization of an H2-Oxidizing Thermophilic Methanogen

    PubMed Central

    Ferguson, Thomas J.; Mah, Robert A.

    1983-01-01

    A thermophilic methanogen was isolated from enrichment cultures originally inoculated with sludge from an anaerobic kelp digester (55°C). This isolate exhibited a temperature optimum of 55 to 60°C and a maximum near 70°C. Growth occurred throughout the pH range of 5.5 to 9.0, with optimal growth near pH 7.2. Although 4% salt was present in the isolation medium, salt was not required for optimal growth. The thermophile utilized formate or H2-CO2 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 h 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 μm 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. Images PMID:16346171

  19. Efficient Plant Biomass Degradation by Thermophilic Fungus Myceliophthora heterothallica

    PubMed Central

    van den Brink, Joost; van Muiswinkel, Gonny C. J.; Theelen, Bart; Hinz, Sandra W. A.

    2013-01-01

    Rapid and efficient enzymatic degradation of plant biomass into fermentable sugars is a major challenge for the sustainable production of biochemicals and biofuels. Enzymes that are more thermostable (up to 70°C) use shorter reaction times for the complete saccharification of plant polysaccharides compared to hydrolytic enzymes of mesophilic fungi such as Trichoderma and Aspergillus species. The genus Myceliophthora contains four thermophilic fungi producing industrially relevant thermostable enzymes. Within this genus, isolates belonging to M. heterothallica were recently separated from the well-described species M. thermophila. We evaluate here the potential of M. heterothallica isolates to produce efficient enzyme mixtures for biomass degradation. Compared to the other thermophilic Myceliophthora species, isolates belonging to M. heterothallica and M. thermophila grew faster on pretreated spruce, wheat straw, and giant reed. According to their protein profiles and in vitro assays after growth on wheat straw, (hemi-)cellulolytic activities differed strongly between M. thermophila and M. heterothallica isolates. Compared to M. thermophila, M. heterothallica isolates were better in releasing sugars from mildly pretreated wheat straw (with 5% HCl) with a high content of xylan. The high levels of residual xylobiose revealed that enzyme mixtures of Myceliophthora species lack sufficient β-xylosidase activity. Sexual crossing of two M. heterothallica showed that progenies had a large genetic and physiological diversity. In the future, this will allow further improvement of the plant biomass-degrading enzyme mixtures of M. heterothallica. PMID:23241981

  20. 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

  1. 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

  2. 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

  3. 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.

  4. Biomass production and energy source of thermophiles in a Japanese alkaline geothermal pool.

    PubMed

    Kimura, Hiroyuki; Mori, Kousuke; Nashimoto, Hiroaki; Hattori, Shohei; Yamada, Keita; Koba, Keisuke; Yoshida, Naohiro; Kato, Kenji

    2010-02-01

    Microbial biomass production has been measured to investigate the contribution of planktonic bacteria to fluxations in dissolved organic matter in marine and freshwater environments, but little is known about biomass production of thermophiles inhabiting geothermal and hydrothermal regions. The biomass production of thermophiles inhabiting an 85 degrees C geothermal pool was measured by in situ cultivation using diffusion chambers. The thermophiles' growth rates ranged from 0.43 to 0.82 day(-1), similar to those of planktonic bacteria in marine and freshwater habitats. Biomass production was estimated based on cellular carbon content measured directly from the thermophiles inhabiting the geothermal pool, which ranged from 5.0 to 6.1 microg C l(-1) h(-1). This production was 2-75 times higher than that of planktonic bacteria in other habitats, because the cellular carbon content of the thermophiles was much higher. Quantitative PCR and phylogenetic analysis targeting 16S rRNA genes revealed that thermophilic H2-oxidizing bacteria closely related to Calderobacterium and Geothermobacterium were dominant in the geothermal pool. Chemical analysis showed the presence of H2 in gases bubbling from the bottom of the geothermal pool. These results strongly suggested that H2 plays an important role as a primary energy source of thermophiles in the geothermal pool.

  5. Interface matters: the stiffness route to stability of a thermophilic tetrameric malate dehydrogenase.

    PubMed

    Kalimeri, Maria; Girard, Eric; Madern, Dominique; Sterpone, Fabio

    2014-01-01

    In this work we investigate by computational means the behavior of two orthologous bacterial proteins, a mesophilic and a thermophilic tetrameric malate dehydrogenase (MalDH), at different temperatures. Namely, we quantify how protein mechanical rigidity at different length- and time-scales correlates to protein thermophilicity as commonly believed. In particular by using a clustering analysis strategy to explore the conformational space of the folded proteins, we show that at ambient conditions and at the molecular length-scale the thermophilic variant is indeed more rigid that the mesophilic one. This rigidification is the result of more efficient inter-domain interactions, the strength of which is further quantified via ad hoc free energy calculations. When considered isolated, the thermophilic domain is indeed more flexible than the respective mesophilic one. Upon oligomerization, the induced stiffening of the thermophilic protein propagates from the interface to the active site where the loop, controlling the access to the catalytic pocket, anchors down via an extended network of ion-pairs. On the contrary in the mesophilic tetramer the loop is highly mobile. Simulations at high temperature, could not re-activate the mobility of the loop in the thermophile. This finding opens questions on the similarities of the binding processes for these two homologues at their optimal working temperature and suggests for the thermophilic variant a possible cooperative role of cofactor/substrate.

  6. Interface Matters: The Stiffness Route to Stability of a Thermophilic Tetrameric Malate Dehydrogenase

    PubMed Central

    Kalimeri, Maria; Girard, Eric; Madern, Dominique; Sterpone, Fabio

    2014-01-01

    In this work we investigate by computational means the behavior of two orthologous bacterial proteins, a mesophilic and a thermophilic tetrameric malate dehydrogenase (MalDH), at different temperatures. Namely, we quantify how protein mechanical rigidity at different length- and time-scales correlates to protein thermophilicity as commonly believed. In particular by using a clustering analysis strategy to explore the conformational space of the folded proteins, we show that at ambient conditions and at the molecular length-scale the thermophilic variant is indeed more rigid that the mesophilic one. This rigidification is the result of more efficient inter-domain interactions, the strength of which is further quantified via ad hoc free energy calculations. When considered isolated, the thermophilic domain is indeed more flexible than the respective mesophilic one. Upon oligomerization, the induced stiffening of the thermophilic protein propagates from the interface to the active site where the loop, controlling the access to the catalytic pocket, anchors down via an extended network of ion-pairs. On the contrary in the mesophilic tetramer the loop is highly mobile. Simulations at high temperature, could not re-activate the mobility of the loop in the thermophile. This finding opens questions on the similarities of the binding processes for these two homologues at their optimal working temperature and suggests for the thermophilic variant a possible cooperative role of cofactor/substrate. PMID:25437494

  7. Moderate views of abortion.

    PubMed

    Sumner, L W

    1997-01-01

    This essay offers a moderate view of abortion that imposes a time limit for unrestricted abortion and specific indications for later abortions. The introduction notes that the discussion will provide a defense for this policy based on a moral analysis but that other options for moderates, especially options provided by freestanding views (the defense of which does not rest on any prior commitment about the morality of abortion), will also be considered. The next section considers the moral status of the fetus grounded in a criterion of moral standing that stipulates the necessary characteristics to achieve moral standing. This discussion concludes that a fetus acquires moral standing only when it becomes sentient. Section 3 moves the argument from ethics to politics to prove that a moderate policy must place no limitations on abortion before the time the fetus becomes sentient because before that time the fetus has no interest for the state to protect. The final section notes that some pro-choice advocates may be happier with the moderate policy proposed than with its controversial defense based on the moral status of the fetus and that another defense of a moderate policy could be based on a finding that the ethical issue can not be decided and that no view about abortion ethics is more reasonable than any other. The essay concludes that the ethical debate is ultimately unavoidable. PMID:12348328

  8. 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)

  9. Hydrogen moderator performance calculations

    SciTech Connect

    Picton, D. J.; Beynon, T. D.; Broome, T. A.

    1997-09-01

    A comparison was made between MCNP calculations and experimental measurements of the neutron spectrum from the liquid hydrogen moderator on ISIS. The calculations were performed for varying ortho/para concentrations, and demonstrated a best fit for 100% para-hydrogen. The agreement between the measured and calculated results was good below 2Å (i.e. for energies above 20 meV) but significant deviations were seen for longer wavelengths. A second study used the MCNP code for a detailed comparison of the time distributions and neutron spectra from poisoned liquid hydrogen and liquid methane moderators. The results indicate that the replacement of a liquid methane moderator with liquid hydrogen, in order to eliminate radiation damage effects, is an option which can be seriously considered. (auth)

  10. Growth of thermophilic and hyperthermophilic Fe(III)-reducing microorganisms on a ferruginous smectite as the sole electron acceptor.

    PubMed

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

    2008-01-01

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

  11. 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.

  12. 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

  13. 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

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 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

  19. [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.

  20. Vibrational entropy differences between mesophile and thermophile proteins and their use in protein engineering

    PubMed Central

    Frappier, Vincent; Najmanovich, Rafael

    2015-01-01

    We recently introduced ENCoM, an elastic network atomic contact model, as the first coarse-grained normal mode analysis method that accounts for the nature of amino acids and can predict the effect of mutations on thermostability based on changes vibrational entropy. In this proof-of-concept article, we use pairs of mesophile and thermophile homolog proteins with identical structures to determine if a measure of vibrational entropy based on normal mode analysis can discriminate thermophile from mesophile proteins. We observe that in around 60% of cases, thermophile proteins are more rigid at equivalent temperatures than their mesophile counterpart and this difference can guide the design of proteins to increase their thermostability through series of mutations. We observe that mutations separating thermophile proteins from their mesophile orthologs contribute independently to a decrease in vibrational entropy and discuss the application and implications of this methodology to protein engineering. PMID:25367089

  1. Types and distribution of obligate thermophilic bacteria in man-made and natural thermal gradients.

    PubMed

    Ramaley, R F; Bitzinger, K

    1975-07-01

    The types and distribution of obligate thermophilic bacteria were found to be similar in a thermal gradient resulting from man-made thermal pollution and the thermal gradients of two natural hot springs located in Colorado.

  2. Vibrational entropy differences between mesophile and thermophile proteins and their use in protein engineering.

    PubMed

    Frappier, Vincent; Najmanovich, Rafael

    2015-04-01

    We recently introduced ENCoM, an elastic network atomic contact model, as the first coarse-grained normal mode analysis method that accounts for the nature of amino acids and can predict the effect of mutations on thermostability based on changes vibrational entropy. In this proof-of-concept article, we use pairs of mesophile and thermophile homolog proteins with identical structures to determine if a measure of vibrational entropy based on normal mode analysis can discriminate thermophile from mesophile proteins. We observe that in around 60% of cases, thermophile proteins are more rigid at equivalent temperatures than their mesophile counterpart and this difference can guide the design of proteins to increase their thermostability through series of mutations. We observe that mutations separating thermophile proteins from their mesophile orthologs contribute independently to a decrease in vibrational entropy and discuss the application and implications of this methodology to protein engineering.

  3. 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.

  4. Characterization of a thermophilic bacteriophage of Geobacillus kaustophilus.

    PubMed

    Marks, Timothy J; Hamilton, Paul T

    2014-10-01

    GBK2 is a bacteriophage, isolated from a backyard compost pile, that infects the thermophile Geobacillus kaustophilus. GBK2 has a circularly permuted genome of 39,078 bp with a G+C content of 43 %. Annotation of the genome reveals 62 putative open reading frames (ORFs), 25 of which (40.3 %) show homology to known proteins and 37 of which (59.7 %) are proteins with unknown functions. Twelve of the identified ORFs had the greatest homology to genes from the phage SPP1, a phage that infects the mesophile Bacillus subtilis. The overall genomic arrangement of GBK2 is similar to that of SPP1, with the majority of GBK2 SPP1-like genes coding for proteins involved in DNA replication and metabolism.

  5. 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.

  6. Thermophilic Mixed Culture of Bacteria Utilizing Methanol for Growth1

    PubMed Central

    Snedecor, Bradley; Cooney, Charles L.

    1974-01-01

    A thermophilic mixed population of bacteria, capable of utilizing methanol as its sole carbon-energy source at temperatures up to 65 C, was selected by enrichment and studied. A maximal cellular yield of 0.42 g per g of methanol was observed at 50 to 56 C. The maximal specific growth rate of the mixed population in continuous culture at 56 C was greater than 0.32 per h. The amino acid profile of the mixed culture indicated that a high quality protein was produced and the protein content was 71%. The properties of this culture and its ability to grow at elevated temperatures are discussed in terms of single-cell protein production and the treatment of industrial waste. Images PMID:16349996

  7. The hemicellulases from the ethanologenic thermophile: Themoanaerobacter ethanolius

    SciTech Connect

    Wiegel, J.

    1991-05-01

    Previously, we had only obtained extremely low xylanase activity in cultures of {ital Thermoanaerobacter ethanolicus} strain JW200, despite demonstrated xylan hydrolysis. We were not able to increase the enzyme activity or concentrate it in solution. Therefore, we decided to isolate new strains of thermophilic anaerobes with higher xylanase activity as a future source for cloning xylanases into {ital T. ethanolicus}. We now have organisms exhibiting 100-fold higher xylanase activity than JW200, but still cannot isolate or concentrate the enzyme activity except at very low yields. We have concentrated and partially purified a xylanase from strain N.D. using preparative matrix-free isoelectric focusing. We have also purified to homogeneity and partially characterized a xylosidase from {ital T. ethanolicus}. We have detected and measured arabinosidase and acetyl esterase activity in {ital T.ethanolicus}, {ital Clostridium thermohydrosulfuricum} and strain N.D. 7 refs., 2 tabs. (MHB)

  8. [The thermophilic streptomycetes flora in milk powders and condensed milk products (author's transl)].

    PubMed

    Falkowski, J

    1978-08-01

    247 specimens of powdered milk and 165 of condensed milk were tested for their contamination with thermophile Streptomycetes. Colonies of these contaminants were isolated from all specimens of powdered milk and from 73 samples of condensed sweeted milk. The isolated strains corresponded with the following species of thermophilic Streptomycetes: Thermoactinomyces vulgaris, Tsiklinsky 1899, Thermoactinomyces vulgaris, Tsiklinsky 1899 "giant colonies", Micromonospora sp. (Agre et al., 1).

  9. 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

  10. 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

  11. 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

  12. 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

  13. 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.

  14. Efficiency of autothermal thermophilic aerobic digestion and thermophilic anaerobic digestion of municipal wastewater sludge in removing Salmonella spp. and indicator bacteria.

    PubMed

    Zábranská, J; Dohányos, M; Jenícek, P; Růziciková, H; Vránová, A

    2003-01-01

    The study is focused on the comparison of autothermal thermophilic aerobic digestion, thermophilic and mesophilic anaerobic digestion, based on long-term monitoring of all processes in full-scale wastewater treatment plants, with an emphasis on the efficiency in destroying pathogens. The hygienisation effect was evaluated as a removal of counts of indicator bacteria, thermotolerant coliforms and enterococci as CFU/g total sludge solids and a frequency of a positive Salmonella spp. detection. Both thermophilic technologies of municipal wastewater sludge stabilisation had the capability of producing sludge A biosolids suitable for agricultural land application when all operational parameters (mainly temperature, mixing and retention time) were stable and maintained at an appropriate level.

  15. 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).

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

    NASA Astrophysics Data System (ADS)

    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.

  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. Caloramator boliviensis sp. nov., a thermophilic, ethanol-producing bacterium isolated from a hot spring.

    PubMed

    Crespo, Carla; Pozzo, Tania; Karlsson, Eva Nordberg; Alvarez, Maria Teresa; Mattiasson, Bo

    2012-07-01

    A novel moderately thermophilic, anaerobic, ethanol-producing bacterial strain, 45B(T), was isolated from a mixed sediment water sample collected from a hot spring at Potosi, Bolivia. The cells were straight to slightly curved rods approximately 2.5 µm long and 0.5 µm wide. The strain was Gram-stain-variable, spore-forming and monotrichously flagellated. Growth of the strain was observed at 45-65 °C and pH 5.5-8.0, with optima of 60 °C and pH 6.5. The substrates utilized by strain 45B(T) were xylose, cellobiose, glucose, arabinose, sucrose, lactose, maltose, fructose, galactose, mannose, glycerol, xylan, carboxymethylcellulose and yeast extract. The main fermentation product from xylose and cellobiose was ethanol (0.70 and 0.45 g ethanol per gram of consumed sugar, respectively). Acetate, lactate, propionate, carbon dioxide and hydrogen were also produced in minor quantities. 1,3-Propanediol was produced when glycerol-containing medium was supplemented with yeast extract. The major cellular fatty acids were anteiso-C(15:0), C(16:0), iso-C(16:0), C(15:1), iso-C(14:0), C(13:0) and C(14:0). The polar lipids diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, an aminoglycolipid and 15 other unidentified lipids were predominant. The DNA G+C content of strain 45B(T) was 32.6 mol%. Phylogenetic analysis based on 16S rRNA gene sequence similarity revealed that strain 45B(T) is located within the Gram-type positive Bacillus-Clostridium branch of the phylogenetic tree. On the basis of morphological and physiological properties and phylogenetic analysis, strain 45B(T) represents a novel species, for which the name Caloramator boliviensis sp. nov. is proposed; the type strain is 45B(T) (=DSM 22065(T)=CCUG 57396(T)).

  19. Conjugative plasmid transfer from Escherichia coli is a versatile approach for genetic transformation of thermophilic Bacillus and Geobacillus species.

    PubMed

    Tominaga, Yurie; Ohshiro, Takashi; Suzuki, Hirokazu

    2016-05-01

    We previously demonstrated efficient transformation of the thermophile Geobacillus kaustophilus HTA426 using conjugative plasmid transfer from Escherichia coli BR408. To evaluate the versatility of this approach to thermophile transformation, this study examined genetic transformation of various thermophilic Bacillus and Geobacillus spp. using conjugative plasmid transfer from E. coli strains. E. coli BR408 successfully transferred the E. coli-Geobacillus shuttle plasmid pUCG18T to 16 of 18 thermophiles with transformation efficiencies between 4.1 × 10(-7) and 3.8 × 10(-2)/recipient. Other E. coli strains that are different from E. coli BR408 in intracellular DNA methylation also generated transformants from 9 to 15 of the 18 thermophiles, including one that E. coli BR408 could not transform, although the transformation efficiencies of these strains were generally lower than those of E. coli BR408. The conjugation was performed by simple incubation of an E. coli donor and a thermophile recipient without optimization of experimental conditions. Moreover, thermophile transformants were distinguished from abundant E. coli donor only by high temperature incubation. These observations suggest that conjugative plasmid transfer, particularly using E. coli BR408, is a facile and versatile approach for plasmid introduction into thermophilic Bacillus and Geobacillus spp., and potentially a variety of other thermophiles.

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

    PubMed

    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

  1. 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

  2. Phthalic acid esters found in municipal organic waste: enhanced anaerobic degradation under hyper-thermophilic conditions.

    PubMed

    Hartmann, H; Ahring, B K

    2003-01-01

    Contamination of the organic fraction of municipal solid waste (OFMSW) with xenobiotic compounds and their fate during anaerobic digestion was investigated. The phthalic acid ester di-(2-ethylhexyl)phthalate (DEHP) was identified as the main contaminant in OFMSW in concentrations more than half of the threshold value for the use as fertilizer on agricultural soil in Denmark. Analysis of DEHP in samples before and after large-scale anaerobic digesters revealed higher concentrations of DEHP per kg dry matter in the effluent than in the influent. The concentration of DEHP and DBP (dibutylphthalate) in OFMSW was monitored in the influent and effluent of anaerobic thermophilic (55 degrees C) and hyper-thermophilic (68 degrees C) laboratory-scale reactor systems. In the thermophilic reactors with a hydraulic retention time (HRT) of 15 days 38-70% of DBP was removed, but no consistent removal of DEHP was observed. However, after treatment of the effluent from the thermophilic reactor in a hyper-thermophilic digester (HRT: 5 days) 34-53% of the DEHP content was removed and the DBP removal was increased to further 62-74%. Removal rates (k(h)) of DEHP and DBP were found to be 0.11-0.32 d(-1) and 0.41-0.79 d(-1), which is much higher than in previous investigations. It can be concluded that the higher removal rates are due to the higher temperature and higher initial concentrations per kg dry matter. These results suggest that the limiting factor for DEHP degradation is the bioavailability, which is enhanced at higher temperature and higher degradation of solid organic matter, to which the highly hydrophobic DEHP is adsorbed. The investigated reactor configuration with a thermophilic and a hyper-thermophilic treatment is, therefore, a good option for combining high rate degradation of organic matter with high biogas yields and efficient reduction of the phthalic acid ester contamination.

  3. 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

  4. Phenylobacterium lituiforme sp. nov., a moderately thermophilic bacterium from a subsurface aquifer, and emended description of the genus Phenylobacterium.

    PubMed

    Kanso, Sungwan; Patel, Bharat K C

    2004-11-01

    A facultative anaerobic bacterium, strain FaiI3(T), was isolated from samples collected from the free-flowing waters of a bore well (Fairlea Bore, registration number 3768) which taps into the Australian Great Artesian Basin subsurface thermal aquifer. Strain FaiI3(T) developed yellow to pale-yellow colonies (0.5-1.5 mm) after 48 h. The non-spore forming rods (0.5x1-3 microm) were slightly curved, occurred singly and as pairs and were motile with a single polar flagellum. Cells tended to form clumps in liquid medium and rosettes were commonly observed. The cells stained Gram-negative and electron micrographs of thin sections revealed a multi-layered complex Gram-negative cell wall structure. Strain FaiI3(T) grew optimally at 40-41 degrees C, with growth observed at 45 degrees C but not at 50 degrees C. The pH growth range was between pH 6 and 9 and optimal growth occurred between pH 6 and 6.5. Strain FaiI3(T) grew best with yeast extract as the sole carbon and energy source. Peptone, yeast extract, acetate, xylose, sucrose, glucose, glycerol, succinate, butyrate, lactate, fumarate, citrate, L-phenylalanine, cellobiose and gelatin supported growth but maltose, fructose, glycine, ethanol, benzoate and oxalate did not. Tyrosine was produced from L-phenylalanine. Strain FaiI3(T) was catalase-positive and oxidase-negative and did not hydrolyse starch. Growth was inhibited by neomycin, tetracycline, streptomycin, chloramphenicol, ampicillin, vancomycin and spectinomycin. The G+C content was determined to be 66.5+/-0.5 mol%. On the basis of the 16S rRNA gene sequence analysis, strain FaiI3(T) was assigned as a novel species of the genus Phenylobacterium, Phenylobacterium lituiforme sp. nov. in the order Caulobacterales, subclass alpha-Proteobacteria, class Proteobacteria. The type strain is FaiI3(T) (=ATCC BAA-294(T)=DSM 14363(T)).

  5. 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.

  6. Desulfotomaculum ferrireducens sp. nov., a moderately thermophilic sulfate-reducing and dissimilatory Fe(III)-reducing bacterium isolated from compost.

    PubMed

    Yang, Guiqin; Guo, Junhui; Zhuang, Li; Yuan, Yong; Zhou, Shungui

    2016-08-01

    A novel dissimilatory Fe(III)-reducing bacterium, designated strain GSS09T, was isolated from a compost sample by using a solid medium containing acetate and ferrihydrite as electron donor and electron acceptor, respectively. Cells of strain GSS09T were anaerobic, Gram-stain-positive, motile, endospore-forming and rod-shaped. Growth occurred at 30-55 °C (optimum 50 °C), at pH 6.5-9.0 (optimum pH 7.5) and in the presence of 0-3 % (w/v) NaCl (optimum 1 %). Both sulfur compounds such as sulfate, sulfite and thiosulfate and Fe(III) oxides such as ferrihydrite could be utilized as electron acceptors. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain GSS09T was related closely to Desulfotomaculum hydrothermale Lam5T (94.5 % sequence similarity). The major fatty acids were C16 : 0 and C16 : 1ω7c/C16 : 1ω6c. The G+C content of the genomic DNA was 49.1 mol%. On the basis of phylogenetic analysis, phenotypic characterization and physiological tests, strain GSS09T is considered to represent a novel species of the genus Desulfotomaculum, for which the name Desulfotomaculum ferrireducens sp. nov. is proposed. The type strain is GSS09T (=KCTC 15523T=MCCC 1K01254T). PMID:27153808

  7. Thermophilic biofiltration of H2S and isolation of a thermophilic and heterotrophic H2S-degrading bacterium, Bacillus sp. TSO3.

    PubMed

    Ryu, Hee-Wook; Yoo, Sun-Kyung; Choi, Jung Min; Cho, Kyung-Suk; Cha, Daniel K

    2009-08-30

    Thermophilic biofiltration of H(2)S-containing gas was studied at 60 degrees C using polyurethane (PU) cubes and as a packing material and compost as a source of thermophilic microorganisms. The performance of biofilter was enhanced by pH control and addition of yeast extract (YE). With YE supplement and pH control, H(2)S removal efficiency remained above 95% up to an inlet concentration of 950 ppmv at a space velocity (SV) of 50h(-1) (residence time=1.2 min). H(2)S removal efficiency strongly correlated with the inverse of H(2)S inlet concentrations and gas flow rates. Thermophilic, sulfur-oxidizing bacteria, TSO3, were isolated from the biofilter and identified as Bacillus sp., which had high similarity value (99%) with Bacillus thermoleovorans. The isolate TSO3 was able to degrade H(2)S without a lag period at 60 degrees C in liquid cultures as well as in the biofilter. High H(2)S removal efficiencies were sustained with a periodic addition of YE. This study demonstrated that an application of thermophilic microorganism for a treatment of hot gases may be an economically attractive option since expensive pre-cooling of gases to accommodate mesophilic processes is not required.

  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. Reduction of hexavalent chromium by the thermophilic methanogen Methanothermobacter thermautotrophicus

    DOE PAGES

    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

  10. 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

  11. 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

  12. 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

  13. 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.

  14. Ribosomes, Polyribosomes, and Deoxyribonucleic Acid from Thermophilic, Mesophilic, and Psychrophilic Clostridia

    PubMed Central

    Irwin, Carol C.; Akagi, James M.; Himes, Richard H.

    1973-01-01

    Analysis of deoxyribonucleic acid (DNA) from four species of Clostridium, including two thermophiles, a mesophile, and a psychrophile, revealed no obvious relationship between growth temperature and DNA base composition. The melting temperatures (Tm) of the DNA from the four species varied no more among the thermophilic, mesophilic, and psychrophilic species than among many related mesophilic species. Characterization of ribosomes from the clostridia by means of optical rotatory dispersion yielded similar spectra in common with other unrelated organisms. Only small differences were noted in the base composition of ribosomal ribonucleic acid (RNA) and in the amino acid composition of ribosomal proteins, including half-cystine content, as determined by cysteic acid analysis, and accessible sulfhydryl groups, as determined by titration with dithiobis (2-nitrobenzoic acid). Except for the two thermophiles, the ribosomal protein electrophoretic patterns were dissimilar. No unusual thermal stability was manifested in the Tm values of thermophile ribosomal RNA. However, thermophile ribosome Tm values (69 C) were higher than were mesophile and psychrophile Tm values (64 C). Ribosomes from the four clostridial species were also examined in regard to the effect of heat on their functional integrity, measured by their activity in poly U-directed 14C-phenylaline incorporation, and their gross physical integrity, measured by sucrose gradient analysis. The Td, 5 values (temperature which produces 50% inactivation after 5 min) was found to be 70 and 72 C for the two thermophiles C. tartarivorum and C. thermosaccharolyticum, respectively; 57 C for a mesophile, C. pasteurianum; and 53 C for a psychrophile, Clostridium sp. strain 69. At 55 C, little effect was seen on the thermophile ribosomes, but the mesophile ribosomes lost 90% of their activity in 1 hr, and psychrophile ribosomes lost 100% of their activity within 10 min. According to sucrose gradient profiles, heating at 55 C

  15. 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

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

    DOE PAGES

    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

  17. 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.

  18. 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

  19. Synonymous codon usage is subject to selection in thermophilic bacteria

    PubMed Central

    Lynn, David J.; Singer, Gregory A. C.; Hickey, Donal A.

    2002-01-01

    The patterns of synonymous codon usage, both within and among genomes, have been extensively studied over the past two decades. Despite the accumulating evidence that natural selection can shape codon usage, it has not been possible to link a particular pattern of codon usage to a specific external selective force. Here, we have analyzed the patterns of synonymous codon usage in 40 completely sequenced prokaryotic genomes. By combining the genes from several genomes (more than 80 000 genes in all) into a single dataset for this analysis, we were able to investigate variations in codon usage, both within and between genomes. The results show that synonymous codon usage is affected by two major factors: (i) the overall G+C content of the genome and (ii) growth at high temperature. This study focused on the relationship between synonymous codon usage and the ability to grow at high temperature. We have been able to eliminate both phylogenetic history and lateral gene transfer as possible explanations for the characteristic pattern of codon usage among the thermophiles. Thus, these results demonstrate a clear link between a particular pattern of codon usage and an external selective force. PMID:12364606

  20. CO Metabolism in the Thermophilic Acetogen Thermoanaerobacter kivui

    PubMed Central

    Weghoff, Marie Charlotte

    2016-01-01

    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. 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.

  2. Involvement of thermophilic archaea in the biocorrosion of oil pipelines.

    PubMed

    Davidova, Irene A; Duncan, Kathleen E; Perez-Ibarra, B Monica; Suflita, Joseph M

    2012-07-01

    Two thermophilic archaea, strain PK and strain MG, were isolated from a culture enriched at 80°C from the inner surface material of a hot oil pipeline. Strain PK could ferment complex organic nitrogen sources (e.g. yeast extract, peptone, tryptone) and was able to reduce elemental sulfur (S°), Fe(3+) and Mn(4+) . Phylogenetic analysis revealed that the organism belonged to the order Thermococcales. Incubations of this strain with elemental iron (Fe°) resulted in the abiotic formation of ferrous iron and the accumulation of volatile fatty acids during yeast extract fermentation. The other isolate, strain MG, was a H(2) :CO(2) -utilizing methanogen, phylogenetically affiliated with the genus Methanothermobacter family. Co-cultures of the strains grew as aggregates that produced CH(4) without exogenous H(2) amendment. The co-culture produced the same suite but greater concentrations of fatty acids from yeast extract than did strain PK alone. Thus, the physiological characteristics of organisms both alone and in combination could conceivably contribute to pipeline corrosion. The Thermococcus strain PK could reduce elemental sulfur to sulfide, produce fatty acids and reduce ferric iron. The hydrogenotrophic methanogen strain MG enhanced fatty acid production by fermentative organisms but could not couple the dissolution Fe° with the consumption of water-derived H(2) like other methanogens.

  3. 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

  4. Cellulose- and xylan-degrading thermophilic anaerobic bacteria from biocompost.

    PubMed

    Sizova, M V; Izquierdo, J A; Panikov, N S; Lynd, L R

    2011-04-01

    Nine thermophilic cellulolytic clostridial isolates and four other noncellulolytic bacterial isolates were isolated from self-heated biocompost via preliminary enrichment culture on microcrystalline cellulose. All cellulolytic isolates grew vigorously on cellulose, with the formation of either ethanol and acetate or acetate and formate as principal fermentation products as well as lactate and glycerol as minor products. In addition, two out of nine cellulolytic strains were able to utilize xylan and pretreated wood with roughly the same efficiency as for cellulose. The major products of xylan fermentation were acetate and formate, with minor contributions of lactate and ethanol. Phylogenetic analyses of 16S rRNA and glycosyl hydrolase family 48 (GH48) gene sequences revealed that two xylan-utilizing isolates were related to a Clostridium clariflavum strain and represent a distinct novel branch within the GH48 family. Both isolates possessed high cellulase and xylanase activity induced independently by either cellulose or xylan. Enzymatic activity decayed after growth cessation, with more-rapid disappearance of cellulase activity than of xylanase activity. A mixture of xylan and cellulose was utilized simultaneously, with a significant synergistic effect observed as a reduction of lag phase in cellulose degradation.

  5. 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

  6. 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

  7. Isobutanol production at elevated temperatures in thermophilic Geobacillus thermoglucosidasius.

    PubMed

    Lin, Paul P; Rabe, Kersten S; Takasumi, Jennifer L; Kadisch, Marvin; Arnold, Frances H; Liao, James C

    2014-07-01

    The potential advantages of biological production of chemicals or fuels from biomass at high temperatures include reduced enzyme loading for cellulose degradation, decreased chance of contamination, and lower product separation cost. In general, high temperature production of compounds that are not native to the thermophilic hosts is limited by enzyme stability and the lack of suitable expression systems. Further complications can arise when the pathway includes a volatile intermediate. Here we report the engineering of Geobacillus thermoglucosidasius to produce isobutanol at 50°C. We prospected various enzymes in the isobutanol synthesis pathway and characterized their thermostabilities. We also constructed an expression system based on the lactate dehydrogenase promoter from Geobacillus thermodenitrificans. With the best enzyme combination and the expression system, 3.3g/l of isobutanol was produced from glucose and 0.6g/l of isobutanol from cellobiose in G. thermoglucosidasius within 48h at 50°C. This is the first demonstration of isobutanol production in recombinant bacteria at an elevated temperature.

  8. Characterization of thermophilic consortia from two souring oil reservoirs.

    PubMed

    Mueller, R F; Nielsen, P H

    1996-09-01

    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.

  9. Effect of cobalt on the anaerobic thermophilic conversion of methanol.

    PubMed

    Paulo, Paula L; Jiang, Bo; Cysneiros, Denise; Stams, Alfons J M; Lettinga, Gatze

    2004-02-20

    The importance of cobalt on the anaerobic conversion of methanol under thermophilic conditions was studied in three parallel lab-scale UASB-reactors and in cobalt-limited enriched cultures. Reactors R1, R2, and R3 were fed with methanol in a bicarbonate-buffered medium, supplied with iron and macronutrients: in R1 all metals were supplied (control), R2 was cobalt deprived, and in R3 all metals were deprived. In the 136 days of continuous experiment, a drop in performance was observed over the last 30 days. Particularly in R3, both methanol removal and methane formation dropped by 7.1% and 13.7%, respectively, compared to the control reactor, R1. When the medium was cobalt-deprived, acetate was not produced and, as a consequence, the enriched consortium lost its capacity to degrade acetate, indicating that the acetotrophic microorganisms were washed out. The addition of 0.5 microM of cobalt to a cobalt-deprived enrichment culture led to acetate accumulation. The results obtained in this study indicate that the mixed consortium requires a proper amount of cobalt, and its addition to a concentration of 0.1 microM leads to the highest methanol conversion rate, with methane as the sole end product from methanol.

  10. Anaerobic digestion of municipal solid wastes: dry thermophilic performance.

    PubMed

    Forster-Carneiro, T; Pérez, M; Romero, L I

    2008-11-01

    The purpose of this study was to analyze the performance of two laboratory-scale reactors (5.0L) treating organic fraction of municipal solid waste (OFMSW): source sorted OFMSW (SS_OFMSW) obtained from a university restaurant and mechanically selected municipal fraction (MS_OFMSW) obtained from a Municipal Treatment Plant placed in Cadiz-Spain. Discontinuous reactors operated at thermophilic (55 degrees C) and dry (20% total solid) conditions. Different decomposition patterns were observed: (1) the SS_OFMSW exhibited the classical waste decomposition pattern with a fast start up phase beginning within 0-5 days and 20-30 and a subsequent stabilization phase. The VS removal was 45% with a cumulative biogas of 120L in approx. 60 days; (2) the MS_OFMSW showed a methanogenic pattern throughout the whole experimental period (60 days) and this gave higher levels of organic biodegradation (56%VSr) and biogas production (82L). Both processes were completed and a high level of cumulative methane production was achieved in less than 60 days, proximally 25-30L. PMID:18434139

  11. Modeling temperature variations in a pilot plant thermophilic anaerobic digester.

    PubMed

    Valle-Guadarrama, Salvador; Espinosa-Solares, Teodoro; López-Cruz, Irineo L; Domaschko, Max

    2011-05-01

    A model that predicts temperature changes in a pilot plant thermophilic anaerobic digester was developed based on fundamental thermodynamic laws. The methodology utilized two simulation strategies. In the first, model equations were solved through a searching routine based on a minimal square optimization criterion, from which the overall heat transfer coefficient values, for both biodigester and heat exchanger, were determined. In the second, the simulation was performed with variable values of these overall coefficients. The prediction with both strategies allowed reproducing experimental data within 5% of the temperature span permitted in the equipment by the system control, which validated the model. The temperature variation was affected by the heterogeneity of the feeding and extraction processes, by the heterogeneity of the digestate recirculation through the heating system and by the lack of a perfect mixing inside the biodigester tank. The use of variable overall heat transfer coefficients improved the temperature change prediction and reduced the effect of a non-ideal performance of the pilot plant modeled.

  12. 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

  13. Illumination enhances methane production from thermophilic anaerobic digestion.

    PubMed

    Tada, C; Tsukahara, K; Sawayama, S

    2006-07-01

    Incandescent lamp illumination enhanced methane production from a thermophilic anaerobic digestion reactor (55 degrees C) supplied with glucose. After 10 days of operation, the volume of methane produced from light reactors was approximately 2.5 times higher than that from dark reactors. A comparison of the carbon balance between light and dark conditions showed that methane produced from hydrogen and carbon dioxide in the light reactors was higher than that from the dark reactors. When hydrogen or acetate was fed into the reactors, methane production with added hydrogen was faster and higher under light conditions than under dark conditions. The use of blue light-emitting diodes also enhanced methane production over that under dark conditions. The 16S rRNA gene copy numbers for Methanothermobacter spp. in the light reactor and in the dark reactor were at the same level. The copy number for Methanosarcina spp. in the light reactors was approximately double than that in the dark reactors. These results suggest that blue light enhances the methanogenic activity of hydrogenotrophic methanogens.

  14. Identification of thermophilic bacteria in solid-waste composting.

    PubMed

    Strom, P F

    1985-10-01

    The thermophilic microbiota of solid-waste composting, with major emphasis on Bacillus spp., was examined with Trypticase soy broth (BBL Microbiology Systems) with 2% agar as the initial plating medium. Five 4.5-liter laboratory units at 49 to 69 degrees C were fed a mixture of dried table scraps and shredded newspaper. The composting plants treating refuse at Altoona, Pa., and refuse-sludge at Leicester, England, were also sampled. Of 652 randomly picked colonies, 87% were identified as Bacillus spp. Other isolates included two genera of unidentified nonsporeforming bacteria (one of gram-negative small rods and the other of gram-variable coccobacilli), the actinomycetes Streptomyces spp. and Thermoactinomyces sp., and the fungus Aspergillus fumigatus. Among the Bacillus isolates, the following, in order of decreasing frequency, were observed: B. circulans complex, B. stearothermophilus, B. coagulans types A and B, B. licheniformis, B. brevis, B. sphaericus, Bacillus spp. types i and ii, and B. subtilis. About 15% of the Bacillus isolates could be assigned to species only by allowing for greater variability in one or more characteristics than has been reported by other authors for their strains. In particular, growth at higher temperatures than previously reported was found for strains of several species. A small number of Bacillus isolates (less than 2%) could not be assigned to any recognized species. PMID:4083886

  15. Use of thermophilic bacteria for bioremediation of petroleum contaminants

    SciTech Connect

    Al-Maghrabi, I.M.A.; Bin Aqil, A.O.; Chaalal, O.; Islam, M.R.

    1999-01-01

    Several strains of thermophilic bacteria were isolated from the environment of the United Arab Emirates. These bacteria show extraordinary resistance to heat and have their maximum growth rate around 60--80 C. This article investigates the potential of using these facultative bacteria for both in situ and ex situ bioremediation of petroleum contaminants. In a series of batch experiments, bacterial growth was observed using a computer image analyzer following a recently developed technique. These experiments showed clearly that the growth rate is enhanced in the presence of crude oil. This is coupled with a rapid degradation of the crude oil. These bacteria were found to be ideal for breaking down long-chain organic molecules at a temperature of 40 C, which is the typical ambient temperature of the Persian Gulf region. The same strains of bacteria are also capable of surviving in the presence of the saline environment that can prevail in both sea water and reservoir connate water. This observation prompted further investigation into the applicability of the bacteria in microbial enhanced oil recovery. In the United Arab Emirates, the reservoirs are typically at a temperature of around 85 C. Finally, the performance of the bacteria is tested in a newly developed bioreactor that uses continuous aeration through a transverse slotted pipe. This reactor also uses mixing without damaging the filamentous bacteria. In this process, the mechanisms of bioremediation are identified.

  16. 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.

  17. Complete Genome Sequence of Anoxybacillus flavithermus TNO-09.006, a Thermophilic Sporeformer Associated with a Dairy-Processing Environment.

    PubMed

    Caspers, Martien P M; Boekhorst, Jos; Abee, Tjakko; Siezen, Roland J; Kort, Remco

    2013-01-01

    Spores of thermophilic spore-forming bacteria are a common cause of contamination in dairy products. We isolated the thermophilic strain Anoxybacillus flavithermus TNO-09.006 from a milk-processing plant, and we report the complete genome of this isolate consisting of a single chromosome of 2.65 Mb. PMID:23469333

  18. Complete genome sequence of Geobacillus thermoglucosidans TNO-09.020, a thermophilic sporeformer associated with a dairy-processing environment.

    PubMed

    Zhao, Yu; Caspers, Martien P; Abee, Tjakko; Siezen, Roland J; Kort, Remco

    2012-08-01

    Thermophilic spore-forming bacteria are a common cause of contamination in dairy products. We isolated the thermophilic strain Geobacillus thermoglucosidans TNO-09.020 from a milk processing plant and report the complete genome of a dairy plant isolate consisting of a single chromosome of 3.75 Mb. PMID:22815439

  19. 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

  20. 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.

  1. 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.

  2. Immunological detection of lipopolysaccharide antigens of thermophilic campylobacters captured on polymyxin-coated polyester cloth.

    PubMed

    Gomi, K; Brooks, B; Blais, B; Garcia, M; Yamazaki, H

    1996-05-01

    Cholate-extracted lipopolysaccharide (LPS) antigens from thermophilic campylobacters were captured on polymyxin-coated polyester cloth. The captured antigens were detected by sequential reactions with rabbit anti-Campylobacter antibody, anti-rabbit IgG peroxidase conjugate and chromogenic peroxidase substrate. A polyclonal rabbit antibody elicited against a single Campylobacter-jejuni strain detected the reference strains of the twenty most frequently isolated thermophilic campylobacters in the Lior serotyping scheme. Moreover, LPS antigens of six C. Jejuni Penner serotypes fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and examined by immunoblotting were recognized by four antisera prepared against homologous and heterologous Penner serotypes. The results suggest the potential application of polymyxin-cloth enzyme immunoassay for rapid detection of thermophilic campylobacters where monoclonal antibodies can be raised to possible common LPS epitopes.

  3. The influence of cell surface properties of thermophilic streptococci on attachment to stainless steel.

    PubMed

    Flint, S H; Brooks, J D; Bremer, P J

    1997-10-01

    The quality of milk products is threatened by the formation of biofilms of thermophilic streptococci on the internal surfaces of plate heat exchangers used in milk processing. Although attachment to stainless steel surfaces is one of the first stages in the development of a biofilm, the mechanisms involved in attachment have not been reported. The cell surface properties of 12 strains of thermophilic streptococci were examined to determine their importance in attachment to stainless steel surfaces. Hydrophobicity, extracellular polysaccharide production and cell surface charge varied between the different strains but could not be related to numbers attaching. Treating the cells with sodium metaperiodate, lysozyme or trichloroacetic acid to disrupt cell surface polysaccharide had no effect on attachment. Treatment with trypsin or sodium dodecyl sulphate to remove cell surface proteins resulted in a 100-fold reduction in the number of bacteria attaching. This result suggests that the surface proteins of the thermophilic streptococci are important in their attachment to stainless steel. PMID:9351231

  4. Cultivation and properties of Echinamoeba thermarum n. sp., an extremely thermophilic amoeba thriving in hot springs.

    PubMed

    Baumgartner, Manuela; Yapi, Ahoua; Gröbner-Ferreira, Regina; Stetter, Karl O

    2003-08-01

    Here we describe a new, extremely thermophilic amoeba growing between 33 degrees C and 57 degrees C ( Topt.=50 degrees C). Isolates had been obtained from hot springs at Agnano Terme (Italy), Yellowstone National Park (USA), Kamchatka (Russia), and the Arenal Volcano (Costa Rica). They could be cultured monoxenically on a thermophilic alpha-proteobacterium. The morphology of the amoeba was studied using a microscope situated under a heatable polyacrylate hood. At 50 degrees C, the cells appeared flat with an irregular triangular or elongate shape, sometimes exhibiting fine spine-like subpseudopodia. On average, they were 22 microm long and 11 microm wide and had one nucleus with a central nucleolus. Based on morphology and on SSU rRNA comparisons, the amoeba belonged to the genus Echinamoeba, where it represents a new species. Referring to its extremely thermophilic lifestyle and its hydrothermal habitat, we name it E. thermarum. PMID:12910386

  5. 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

  6. Metagenomics of Thermophiles with a Focus on Discovery of Novel Thermozymes

    PubMed Central

    DeCastro, María-Eugenia; Rodríguez-Belmonte, Esther; González-Siso, María-Isabel

    2016-01-01

    Microbial populations living in environments with temperatures above 50°C (thermophiles) have been widely studied, increasing our knowledge in the composition and function of these ecological communities. Since these populations express a broad number of heat-resistant enzymes (thermozymes), they also represent an important source for novel biocatalysts that can be potentially used in industrial processes. The integrated study of the whole-community DNA from an environment, known as metagenomics, coupled with the development of next generation sequencing (NGS) technologies, has allowed the generation of large amounts of data from thermophiles. In this review, we summarize the main approaches commonly utilized for assessing the taxonomic and functional diversity of thermophiles through metagenomics, including several bioinformatics tools and some metagenome-derived methods to isolate their thermozymes. PMID:27729905

  7. 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.

  8. Thermodynamic assessment of domain-domain interactions and in vitro activities of mesophilic and thermophilic ribosome recycling factors.

    PubMed

    Uchiyama, Susumu; Ohshima, Atsushi; Yoshida, Takuya; Ohkubo, Tadayasu; Kobayashi, Yuji

    2013-07-01

    Ribosome recycling factor (RRF) is essential for bacterial growth. Structural studies revealed that RRF consists of two domains connected by two short polypeptides at the hinge region. Here, we evaluated the intrinsic stabilities (ΔG*s) of the two domains and the free energy of the domain-domain interactions (ΔG(int)) for mesophilic RRF (RRF from Escherichia coli, EcRRF) and thermophilic RRF (RRF from Thermus thermophilus, TtRRF) by using differential scanning calorimetry and circular dichroic spectroscopy. Despite single endothermic peaks, a higher than unity value for the ratio of calorimetric enthalpy to van't Hoff enthalpy of the unfolding indicated the presence of unfolding intermediates for both RRFs. Deconvolution analysis based on statistical thermodynamics employing multiple states of the unfolding process with domain-domain interactions allowed us to determine ΔG*s of each domain and ΔG(int). The results indicated that domain I has a higher unfolding temperature than domain II and that it stabilizes domain II through ΔG(int), giving rise to an apparent single peak of unfolding. Interestingly, the estimated ΔG(int) values of 6.28 kJ/mol for EcRRF and 26.28 kJ/mol for TtRRF reflect the observation that only EcRRF has recycling activity at ambient temperature. Our present study suggests the importance of a moderate ΔG(int) for biological activity of multidomain proteins.

  9. 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).

  10. 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

  11. 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.

  12. Mesophilic and thermophilic biotreatment of BTEX-polluted air in reactors.

    PubMed

    Mohammad, Balsam T; Veiga, María C; Kennes, Christian

    2007-08-15

    This study compares the removal of a mixture of benzene, toluene, ethylbenzene, and all three xylene isomers (BTEX) in mesophilic and thermophilic (50 degrees C) bioreactors. In the mesophilic reactor fungi became dominant after long-term operation, while bacteria dominated in the thermophilic unit. Microbial acclimation was achieved by exposing the biofilters to initial BTEX loads of 2-15 g m(-3) h(-1), at an empty bed residence time of 96 s. After adaptation, the elimination capacities ranged from 3 to 188 g m(-3) h(-1), depending on the inlet load, for the mesophilic biofilter with removal efficiencies reaching 96%. On the other hand, in the thermophilic reactor the average removal efficiency was 83% with a maximum elimination capacity of 218 g m(-3) h(-1). There was a clear positive relationship between temperature gradients as well as CO(2) production and elimination capacities across the biofilters. The gas phase was sampled at different depths along the reactors observing that the percentage pollutant removal in each section was strongly dependant on the load applied. The fate of individual alkylbenzene compounds was checked, showing the unusually high biodegradation rate of benzene at high loads under thermophilic conditions (100%) compared to its very low removal in the mesophilic reactor at such load (<10%). Such difference was less pronounced for the other pollutants. After 210 days of operation, the dry biomass content for the mesophilic and thermophilic reactors were 0.300 and 0.114 g g(-1) (support), respectively, reaching higher removals under thermophilic conditions with a lower biomass accumulation, that is, lower pressure drop.

  13. Coliforms, Enterococci, Thermodurics, Thermophiles, and Psychrophiles in Untreated Farm Pond Waters

    PubMed Central

    Malaney, G. W.; Weiser, H. H.; Turner, R. O.; Van Horn, Marilyn

    1962-01-01

    Untreated waters from ten farm ponds located in central, north central, southeastern, and southwestern Ohio were examined for numbers of coliforms, enterococci, thermodurics, thermophiles, and psychrophiles. The median population densities per 100 ml water for all ponds were: coliforms, 23; enterococci, 3.6; thermodurics, 6,000; thermophiles, 450; psychrophiles, 1,000. The results indicate that these farm pond waters were only lightly polluted and suggest that farm ponds, properly maintained, are a source of raw water of high bacteriological quality, requiring a minimum of treatment to be made suitable for domestic and livestock purposes. PMID:14468809

  14. Quality Matters: Extension of Clusters of Residues with Good Hydrophobic Contacts Stabilize (Hyper)Thermophilic Proteins

    PubMed Central

    2015-01-01

    Identifying determinant(s) of protein thermostability is key for rational and data-driven protein engineering. By analyzing more than 130 pairs of mesophilic/(hyper)thermophilic proteins, we identified the quality (residue-wise energy) of hydrophobic interactions as a key factor for protein thermostability. This distinguishes our study from previous ones that investigated predominantly structural determinants. Considering this key factor, we successfully discriminated between pairs of mesophilic/(hyper)thermophilic proteins (discrimination accuracy: ∼80%) and searched for structural weak spots in E. coli dihydrofolate reductase (classification accuracy: 70%). PMID:24437522

  15. Quality matters: extension of clusters of residues with good hydrophobic contacts stabilize (hyper)thermophilic proteins.

    PubMed

    Rathi, Prakash Chandra; Höffken, Hans Wolfgang; Gohlke, Holger

    2014-02-24

    Identifying determinant(s) of protein thermostability is key for rational and data-driven protein engineering. By analyzing more than 130 pairs of mesophilic/(hyper)thermophilic proteins, we identified the quality (residue-wise energy) of hydrophobic interactions as a key factor for protein thermostability. This distinguishes our study from previous ones that investigated predominantly structural determinants. Considering this key factor, we successfully discriminated between pairs of mesophilic/(hyper)thermophilic proteins (discrimination accuracy: ∼80%) and searched for structural weak spots in E. coli dihydrofolate reductase (classification accuracy: 70%).

  16. 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

  17. 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.

  18. 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

  19. [Characterization of a thermophilic Geobacillus strain DM-2 degrading hydrocarbons].

    PubMed

    Liu, Qing-kun; Wang, Jun; Li, Guo-qiang; Ma, Ting; Liang, Feng-lai; Liu, Ru-lin

    2008-12-01

    A thermophilic Geobacillus strain DM-2 from a deep-subsurface oil reservoir was investigated on its capability of degrading crude oil under various conditions as well as its characters on degrading hydrocarbons in optimal conditions. The results showed that Geobacillus strain DM-2 was able to degrade crude oil under anoxic wide-range conditions with pH ranging from 4.0 to 10.0, high temperature in the range of 45-70 degrees C and saline concentration ranging from 0.2% to 3.0%. Furthermore, the optimal temperature and pH value for utilizing hydrocarbons by the strain were 60 degrees C and 7.0, respectively. Under such optimal conditions, the strain utilized liquid paraffine emulsified by itself as its carbon source for growth; further analysis by gas chromatography (GC) and infrared absorption spectroscopy demonstrated that it was able to degrade n-alkanes (C14-C30), branched-chain alkanes and aromatic hydrocarbons in crude oil and could also utilize long-chain n-alkanes from C16 to C36, among of which the degradation efficiency of C28 was the highest, up to 88.95%. One metabolite of the strain oxidizing alkanes is fatty acid.While utilizing C16 as carbon source for 5 d, only one fatty acid-acetic acid was detected by HPLC and MS as the product, with the amount of 0.312 g/L, which indicated that it degraded n-alkanes with pathway of inferior terminal oxidation,and then followed by a beta-oxidation pathway. Due to its characters of efficient emulsification, high-performance degradation of hydrocarbons and fatty-acid production under high temperature and anoxic condition, the strain DM-2 may be potentially applied to oil-waste treatment and microbial enhanced heavy oil recovery in extreme conditions.

  20. Interaction of gaseous aromatic and aliphatic compounds in thermophilic biofilters.

    PubMed

    Hu, Qing-yuan; Wang, Can

    2015-12-30

    Two thermophilic biofilters were applied in treating a mixture of gaseous aromatic (benzene) and aliphatic compounds (hexane) to evaluate the interaction of the compounds. The performance of the biofilters was investigated in terms of removal efficiencies, elimination capacity, kinetic analysis, interaction indices, and microbial metabolic characteristics. Results showed that the removal performance of benzene was unaffected by the addition of hexane. The removal efficiencies of benzene were maintained at approximately 80% and the biodegradation rate constant was maintained at 120 h(-1). However, the removal efficiencies of hexane decreased significantly from 60% to 20% and the biodegradation rate constant exhibited a distinct decrease from 93.59 h(-1) to 56.32 h(-1). The interaction index of benzene with the addition of hexane was -0.029, which indicated that hexane had little effect on the degradation of benzene. By contrast, the interaction index of hexane by benzene was -0.557, which showed that benzene inhibited the degradation of hexane significantly. Similar conclusions were obtained about the substrate utilization. Moreover, the utilization degree of carbon sources and the microbial metabolic activities in the biofilter treating hexane were significantly improved with the addition of benzene, whereas the addition of hexane had a slight effect on the microbial communities in the biofilter treating benzene. Conclusions could be obtained that when mixtures of benzene and hexane were treated using biofilters, the degradation of benzene, which was more easily degradable, was dominant and unaffected; whereas the degradation of hexane, which was less easily degradable, was inhibited because of the changing of microbes.

  1. Ammoniibacillus agariperforans gen. nov., sp. nov., a thermophilic, agar-degrading bacterium isolated from compost.

    PubMed

    Sakai, Masao; Deguchi, Daigo; Hosoda, Akifumi; Kawauchi, Tomohiro; Ikenaga, Makoto

    2015-02-01

    A thermophilic, agar-degrading bacterium, strain FAB2(T), was isolated from sewage sludge compost. According to phylogenetic analysis based on 16S rRNA gene sequences, strain FAB2(T) belonged to the family Paenibacillaceae within the phylum Firmicutes. However, FAB2(T) was different enough at the genus level from closely related species. The percentages of 16S rRNA gene sequence similarity with related organisms were 90.4 % for Thermobacillus xylanilyticus, 91.8 % for Paenibacillus barengoltzii, 89.4 % for Cohnella lupini, 90.1 % for Fontibacillus aquaticus, and 89.0 % for Saccharibacillus sacchari. Morphological and physiological analyses revealed that the strain was motile, rod-shaped, Gram-stain-positive, aerobic and able to form oval endospores in swollen sporangia. Ammonium was required as a nitrogen source while nitrate, nitrite, urea and glutamate were not utilized. Catalase and oxidase activities were weakly positive and positive, respectively. The bacterium grew in the temperature range of 50-65 °C and in media with pH 7.5 to 9.0. Optimal growth occurred at 60 °C and pH 8.0-8.6. Growth was inhibited at pH≤7.0 and NaCl concentrations ≥2.5 % (w/v). In chemotaxonomic characterization, MK-7 was identified as the dominant menaquinone. Major fatty acids were iso-C16 : 0 and C16 : 0. Dominant polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. Phosphatidylcholine was present in a moderate amount. The diamino acid in the cell wall was meso-diaminopimelic acid. The G+C content of the genomic DNA was 49.5 mol% in a nucleic acid study. On the basis of genetic and phenotypic characteristics, strain FAB2(T) ( = NBRC 109510(T) = KCTC 33130(T)) showed characteristics suitable for classification as the type strain of a novel species of a new genus in the family Paenibacillaceae, for which the name Ammoniibacillus agariperforans gen. nov., sp. nov. is proposed.

  2. Biogenic Mineral Production by a Novel Arsenic-Metabolizing Thermophilic Bacterium from the Alvord Basin, Oregon▿

    PubMed Central

    Ledbetter, Rhesa N.; Connon, Stephanie A.; Neal, Andrew L.; Dohnalkova, Alice; Magnuson, Timothy S.

    2007-01-01

    The Alvord Basin in southeast Oregon contains a variety of hydrothermal features which have never been microbiologically characterized. A sampling of Murky Pot (61°C; pH 7.1) led to the isolation of a novel arsenic-metabolizing organism (YeAs) which produces an arsenic sulfide mineral known as β-realgar, a mineral that has not previously been observed as a product of bacterial arsenic metabolism. YeAs was grown on a freshwater medium and utilized a variety of organic substrates, particularly carbohydrates and organic acids. The temperature range for growth was 37 to 75°C (optimum, 55°C), and the pH range for growth was 6.0 to 8.0 (optimum, pH 7.0 to 7.5). No growth was observed when YeAs was grown under aerobic conditions. The doubling time when the organism was grown with yeast extract and As(V) was 0.71 h. Microscopic examination revealed Gram stain-indeterminate, non-spore-forming, nonmotile, rod-shaped cells, with dimensions ranging from 0.1 to 0.2 μm wide by 3 to 10 μm long. Arsenic sulfide mineralization of cell walls and extracellular arsenic sulfide particulate deposition were observed with electron microscopy and elemental analysis. 16S rRNA gene analysis placed YeAs in the family Clostridiaceae and indicated that the organism is most closely related to the Caloramator and Thermobrachium species. The G+C content was 35%. YeAs showed no detectable respiratory arsenate reductase but did display significant detoxification arsenate reductase activity. The phylogenetic, physiological, and morphological characteristics of YeAs demonstrate that it is an anaerobic, moderately thermophilic, arsenic-reducing bacterium. This organism and its associated metabolism could have major implications in the search for innovative methods for arsenic waste management and in the search for novel biogenic mineral signatures. PMID:17630300

  3. Isolation of butyrate-utilizing bacteria from thermophilic and mesophilic methane-producing ecosystems

    SciTech Connect

    Henson, J.M.

    1983-01-01

    The ability of various ecosystems to convert butyrate to methane was studied in order to isolate the bacteria responsible for the conversion. When thermophilic digester sludge was enriched with butyrate, methane was produced without a lag period. Marine sediments enriched with butyrate required a 2-week incubation period before methanogenesis began. A thermophilic digester was studied in more detail and found by most-probable-number enumeration to have ca. 5 x 10/sup 6/ butyrate-utilizing bactera/ml of sludge. A thermophilic butyrate-utilizing bacterium was isolated in coculture with Methanobacterium thermoautotrophicum and a Methanosarcina sp. This bacterium was a gram-negative, slightly curved rod that occurred singly, was nonmotile, and did not appear to produce spores. The thermophilic digester was infused with butyrate at the rate of 10 ..mu..moles/ml of sludge per day. Biogas production increased by 150%, with the percentage of methane increasing from 58% to 68%. Acetate, propionate, and butyrate did not accumulate. Butyrate-utilizing enrichments from mesophilic ecosystems were used in obtaining cocultures of butyrate-utilizing bacteria. These cocultures served as inocula for attempts to isolate pure cultures of butyrate-utilizing bacteria by use of hydrogenase-containing membrane fragments of Escherichia coli. After a 3-week incubation period, colonies appeared only in inoculated tubes that contained membrane fragments and butyrate.

  4. 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.

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

    PubMed Central

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

    2014-01-01

    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

  6. 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.

  7. 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

    2015-06-11

    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.

  8. 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

  9. Strain and bioprocess improvement of a thermophilic anaerobe for the production of ethanol from wood

    DOE PAGES

    Herring, Christopher D.; Kenealy, William R.; Shaw, A. Joe; Covalla, Sean F.; Olson, Daniel G.; Zhang, Jiayi; Sillers, W. Ryan; Tsakraklides, Vasiliki; Bardsley, John S.; Rogers, Stephen R.; et al

    2016-06-16

    Here, the thermophilic, anaerobic bacterium Thermoanaerobacterium saccharolyticum digests hemicellulose and utilizes the major sugars present in biomass. It was previously engineered to produce ethanol at yields equivalent to yeast. While saccharolytic anaerobes have been long studied as potential biomass-fermenting organisms, development efforts for commercial ethanol production have not been reported.

  10. 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.

  11. Assembly and multiple gene expression of thermophilic enzymes in Escherichia coli for in vitro metabolic engineering.

    PubMed

    Ninh, Pham Huynh; Honda, Kohsuke; Sakai, Takaaki; Okano, Kenji; Ohtake, Hisao

    2015-01-01

    In vitro reconstitution of an artificial metabolic pathway is an emerging approach for the biocatalytic production of industrial chemicals. However, several enzymes have to be separately prepared (and purified) for the construction of an in vitro metabolic pathway, thereby limiting the practical applicability of this approach. In this study, genes encoding the nine thermophilic enzymes involved in a non-ATP-forming chimeric glycolytic pathway were assembled in an artificial operon and co-expressed in a single recombinant Escherichia coli strain. Gene expression levels of the thermophilic enzymes were controlled by their sequential order in the artificial operon. The specific activities of the recombinant enzymes in the cell-free extract of the multiple-gene-expression E. coli were 5.0-1,370 times higher than those in an enzyme cocktail prepared from a mixture of single-gene-expression strains, in each of which a single one of the nine thermophilic enzymes was overproduced. Heat treatment of a crude extract of the multiple-gene-expression cells led to the denaturation of indigenous proteins and one-step preparation of an in vitro synthetic pathway comprising only a limited number of thermotolerant enzymes. Coupling this in vitro pathway with other thermophilic enzymes including the H2 O-forming NADH oxidase or the malate/lactate dehydrogenase facilitated one-pot conversion of glucose to pyruvate or lactate, respectively.

  12. Thermal and spectroscopic characterization of a proton pumping rhodopsin from an extreme thermophile.

    PubMed

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

    2013-07-26

    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.

  13. 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

  14. 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.

  15. 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.

  16. 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

  17. Thermoadaptation trait revealed by the genome sequence of thermophilic Geobacillus kaustophilus

    PubMed Central

    Takami, Hideto; Takaki, Yoshihiro; Chee, Gab-Joo; Nishi, Shinro; Shimamura, Shigeru; Suzuki, Hiroko; Matsui, Satomi; Uchiyama, Ikuo

    2004-01-01

    We present herein the first complete genome sequence of a thermophilic Bacillus-related species, Geobacillus kaustophilus HTA426, which is composed of a 3.54 Mb chromosome and a 47.9 kb plasmid, along with a comparative analysis with five other mesophilic bacillar genomes. Upon orthologous grouping of the six bacillar sequenced genomes, it was found that 1257 common orthologous groups composed of 1308 genes (37%) are shared by all the bacilli, whereas 839 genes (24%) in the G.kaustophilus genome were found to be unique to that species. We were able to find the first prokaryotic sperm protamine P1 homolog, polyamine synthase, polyamine ABC transporter and RNA methylase in the 839 unique genes; these may contribute to thermophily by stabilizing the nucleic acids. Contrasting results were obtained from the principal component analysis (PCA) of the amino acid composition and synonymous codon usage for highlighting the thermophilic signature of the G.kaustophilus genome. Only in the PCA of the amino acid composition were the Bacillus-related species located near, but were distinguishable from, the borderline distinguishing thermophiles from mesophiles on the second principal axis. Further analysis revealed some asymmetric amino acid substitutions between the thermophiles and the mesophiles, which are possibly associated with the thermoadaptation of the organism. PMID:15576355

  18. 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.

  19. 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

  20. 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

  1. 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

  2. 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

  3. Comparison of mesophilic and thermophilic feruloyl esterases: characterization of their substrate specificity for methyl phenylalkanoates.

    PubMed

    Topakas, Evangelos; Christakopoulos, Paul; Faulds, Craig B

    2005-02-23

    The active sites of feruloyl esterases from mesophilic and thermophilic sources were probed using methyl esters of phenylalkanoic acids. Only 13 out of 26 substrates tested were significant substrates for all the enzymes. Lengthening or shortening the aliphatic side chain while maintaining the same aromatic substitutions completely abolished activity for both enzymes, which demonstrates the importance of the correct distance between the aromatic group and the ester bond. Maintaining the phenylpropanoate structure but altering the substitutions of the aromatic ring demonstrated that the type-A esterase from the mesophilic fungus Fusarium oxysporum (FoFaeA) showed a preference for methoxylated substrates, in contrast to the type-B esterase from the same source (FoFaeB) and the thermophilic type-B (StFaeB) and type-C (StFaeC) from Sporotrichum thermophile, which preferred hydroxylated substrates. All four esterases hydrolyzed short chain aliphatic acid (C2-C4) esters of p-nitrophenol, but not the C12 ester of laurate. All the feruloyl esterases were able to release ferulic acid from the plant cell wall material in conjunction with a xylanase, but only the type-A esterase FoFaeA was effective in releasing the 5,5' form of diferulic acid. The thermophilic type-B esterase had a lower catalytic efficiency than its mesophilic counterpart, but released more ferulic acid from plant cell walls.

  4. 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

  5. Thermophilic actinomycetes in cane sugar mills: an aeromicrobiologic and seroepidemiologic study.

    PubMed

    Khan, Z U; Gangwar, M; Gaur, S N; Randhawa, H S

    1995-01-01

    Aerial prevalence of clinically important thermophilic actinomycetes and occurrence of precipitating antibodies against them in sera of 153 exposed workers have been reported. The study was carried out in two cane sugar mills namely, the Upper Doab Sugar Mills and the Ramala Sugar Mills, located in north-west India. In both the sugar mills, T. sacchari was the predominant species, it accounted for 55.1% and 50.3% of the total population of thermophilic actinomycetes, followed by T. vulgaris (19.7% and 23.7%), T. thalpophilus (21.1% and 17.1%), Saccharomonospora viridis (3.4% and 5.0%) and Saccharopolyspora rectivirgula (Faenia rectivirgula) (0.7% and 3.9%), respectively. Precipitating antibodies against thermophilic actinomycetes were demonstrable in 34 (22.2%) workers; T. sacchari alone accounted for 20 of the positive precipitin reactions, followed by S. rectivirgula in 10. The mean absorbance values for IgG antibody activity against T. sacchari as well as S. rectivirgula were found to be elevated significantly in the symptomatic workers than in the asymptomatic workers (p < 0.05) or unexposed controls (p < 0.001). However, the difference in IgG antibody activity was insignificant between precipitin-positive symptomatic workers and precipitin-positive asymptomatic workers. The results indicate that clinically important thermophilic actinomycetes are widely prevalent in cane sugar mills, and T. sacchari and S. rectivirgula are the major species involved in the sensitization of the bagasse workers in India.

  6. Change of PCBs and forms of heavy metals in sewage sludge during thermophilic anaerobic digestion.

    PubMed

    Dąbrowska, Lidia; Rosińska, Agata

    2012-06-01

    Determination of seven congeners of PCBs was carried out for sewage sludge before, during and after thermophilic digestion. The overall content of heavy metals (Zn, Cu, Ni, Cd, Pb, Cr) in sludge before and after digestion was determined. Moreover the concentration of heavy metals in particular chemical fractions of the sludge was analyzed. After the thermophilic digestion total concentration of seven PCBs was reduced by 47%, which suggests that thermophilic digestion affects PCB reduction positively. On the 10th d of the process, concentration of lower chlorinated PCBs increased, whereas those of higher chlorinated PCBs decreased. The thermophilic digestion process showed no accumulation of the studied heavy metals in the mobile fractions (exchangeable and carbonate) of the stabilized sewage sludge, except for nickel. The highest increase in zinc, copper, cadmium, and chromium concentration was observed in the organic-sulfide fraction, whereas the highest increase in lead was found in the residual fraction of the sludge. In case of nickel both fractions of organic-sulfide and exchangeable-carbonate fractions were enriched. PMID:22459422

  7. 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

  8. 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

  9. Evaluation of dairy powder products implicates thermophilic sporeformers as the primary organisms of interest.

    PubMed

    Watterson, M J; Kent, D J; Boor, K J; Wiedmann, M; Martin, N H

    2014-01-01

    Dairy powder products (e.g., sweet whey, nonfat dry milk, acid whey, and whey protein concentrate-80) are of economic interest to the dairy industry. According to the US Dairy Export Council, customers have set strict tolerances (<500 to <1,000/g) for thermophilic and mesophilic spores in dairy powders; therefore, understanding proliferation and survival of sporeforming organisms within dairy powder processing plants is necessary to control and reduce sporeformer counts. Raw, work-in-process, and finished product samples were collected from 4 dairy powder processing facilities in the northeastern United States over a 1-yr period. Two separate spore treatments: (1) 80°C for 12min (to detect sporeformers) and (2) 100°C for 30min (to detect highly heat resistant sporeformers) were applied to samples before microbiological analyses. Raw material, work-in-process, and finished product samples were analyzed for thermophilic, mesophilic, and psychrotolerant sporeformers, with 77.5, 71.0, and 4.6% of samples being positive for those organisms, respectively. Work-in-process and finished product samples were also analyzed for highly heat resistant thermophilic and mesophilic sporeformers, with 63.7 and 42.6% of samples being positive, respectively. Sporeformer prevalence and counts varied considerably by product and plant; sweet whey and nonfat dry milk showed a higher prevalence of thermophilic and mesophilic sporeformers compared with acid whey and whey protein concentrate-80. Unlike previous reports, we found limited evidence for increased spore counts toward the end of processing runs. Our data provide important insight into spore contamination patterns associated with production of different types of dairy powders and support that thermophilic sporeformers are the primary organism of concern in dairy powders.

  10. 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

  11. Aspergillus fumigatus and other thermophilic fungi in nests of wetland birds.

    PubMed

    Korniłłowicz-Kowalska, Teresa; Kitowski, Ignacy

    2013-02-01

    A study was performed on the numbers and species diversity of thermophilic fungi (growing at 45 °C in vitro) in 38 nests of 9 species of wetland birds, taking into account the physicochemical properties of the nests and the bird species. It was found that in nests with the maximum weight (nests of Mute Swan), the number and diversity of thermophilic fungi were significantly greater than in other nests, with lower weight. The diversity of the thermophilic biota was positively correlated with the individual mass of bird and with the level of phosphorus in the nests. The dominant species within the mycobiota under study was Aspergillus fumigatus which inhabited 95% of the nests under study, with average frequency of ca. 650 cfu g(-1) of dry mass of the nest material. In a majority of the nests studied (nests of 7 bird species), the share of A. fumigatus exceeded 50% of the total fungi growing at 45 °C. Significantly higher frequencies of the fungal species were characteristic of the nests of small and medium-sized piscivorous species, compared with the other bird species. The number of A. fumigatus increased with increase in the moisture level of the nests, whereas the frequency of occurrence of that opportunistic pathogen, opposite to the general frequency of thermophilic mycobiota, was negatively correlated with the level of phosphorus in the nest material, and with the body mass and length of the birds. The authors indicate the causes of varied growth of thermophilic fungi in nests of wetland birds and, in particular, present a discussion of the causes of accumulation of A. fumigatus, the related threats to the birds, and its role as a source of transmission in the epidemiological chain of aspergillosis. PMID:23054328

  12. Aspergillus fumigatus and other thermophilic fungi in nests of wetland birds.

    PubMed

    Korniłłowicz-Kowalska, Teresa; Kitowski, Ignacy

    2013-02-01

    A study was performed on the numbers and species diversity of thermophilic fungi (growing at 45 °C in vitro) in 38 nests of 9 species of wetland birds, taking into account the physicochemical properties of the nests and the bird species. It was found that in nests with the maximum weight (nests of Mute Swan), the number and diversity of thermophilic fungi were significantly greater than in other nests, with lower weight. The diversity of the thermophilic biota was positively correlated with the individual mass of bird and with the level of phosphorus in the nests. The dominant species within the mycobiota under study was Aspergillus fumigatus which inhabited 95% of the nests under study, with average frequency of ca. 650 cfu g(-1) of dry mass of the nest material. In a majority of the nests studied (nests of 7 bird species), the share of A. fumigatus exceeded 50% of the total fungi growing at 45 °C. Significantly higher frequencies of the fungal species were characteristic of the nests of small and medium-sized piscivorous species, compared with the other bird species. The number of A. fumigatus increased with increase in the moisture level of the nests, whereas the frequency of occurrence of that opportunistic pathogen, opposite to the general frequency of thermophilic mycobiota, was negatively correlated with the level of phosphorus in the nest material, and with the body mass and length of the birds. The authors indicate the causes of varied growth of thermophilic fungi in nests of wetland birds and, in particular, present a discussion of the causes of accumulation of A. fumigatus, the related threats to the birds, and its role as a source of transmission in the epidemiological chain of aspergillosis.

  13. Identification of thermophilic proteins by incorporating evolutionary and acid dissociation information into Chou's general pseudo amino acid composition.

    PubMed

    Fan, Guo-Liang; Liu, Yan-Ling; Wang, Hui

    2016-10-21

    Thermophilic proteins can thrive stalely at the high temperatures. Identification of thermophilic protein could be helpful to learn the function of protein. Automated prediction of thermophilic protein is an important tool for genome annotation. In this work, a powerful predictor is proposed by combining amino acid composition, evolutionary information, and acid dissociation constant. The overall prediction accuracy of 93.53% was obtained for using the algorithm of support vector machine. In order to check the performance of our method, two low-similarity independent testing datasets are used to test the proposed method. Comparisons with other methods show that the prediction results were better than other existing methods in literature. This indicates that our approach was effective to predict thermophilic proteins. PMID:27396359

  14. Differences in the catalytic mechanisms of mesophilic and thermophilic indole-3-glycerol phosphate synthase enzymes at their adaptive temperatures

    SciTech Connect

    Zaccardi, Margot J.; Mannweiler, Olga; Boehr, David D.

    2012-02-10

    Highlights: Black-Right-Pointing-Pointer Catalytic mechanisms of thermophilic-mesophilic enzymes may differ. Black-Right-Pointing-Pointer Product release is rate-determining for thermophilic IGPS at low temperatures. Black-Right-Pointing-Pointer But at higher temperatures, proton transfer from the general acid is rate-limiting. Black-Right-Pointing-Pointer Rate-determining step is different still for mesophilic IGPS. Black-Right-Pointing-Pointer Both chemical and physical steps of catalysis are important for temperature adaptation. -- Abstract: Thermophilic enzymes tend to be less catalytically-active at lower temperatures relative to their mesophilic counterparts, despite having very similar crystal structures. An often cited hypothesis for this general observation is that thermostable enzymes have evolved a more rigid tertiary structure in order to cope with their more extreme, natural environment, but they are also less flexible at lower temperatures, leading to their lower catalytic activity under mesophilic conditions. An alternative hypothesis, however, is that complementary thermophilic-mesophilic enzyme pairs simply operate through different evolutionary-optimized catalytic mechanisms. In this communication, we present evidence that while the steps of the catalytic mechanisms for mesophilic and thermophilic indole-3-glycerol phosphate synthase (IGPS) enzymes are fundamentally similar, the identity of the rate-determining step changes as a function of temperature. Our findings indicate that while product release is rate-determining at 25 Degree-Sign C for thermophilic IGPS, near its adaptive temperature (75 Degree-Sign C), a proton transfer event, involving a general acid, becomes rate-determining. The rate-determining steps for thermophilic and mesophilic IGPS enzymes are also different at their respective, adaptive temperatures with the mesophilic IGPS-catalyzed reaction being rate-limited before irreversible CO{sub 2} release, and the thermophilic IGPS

  15. 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

  16. 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.

  17. Thermophilic anaerobic digestion of sewage sludge: focus on the influence of the start-up. A review.

    PubMed

    De la Rubia, M A; Riau, V; Raposo, F; Borja, R

    2013-12-01

    The thermophilic anaerobic digestion (TAD) of sewage sludge has often been found to be less stable than mesophilic treatment. In comparison to mesophilic digesters, thermophilic reactors treating sludge are generally characterized by relatively high concentrations of volatile fatty acids (VFA) in the effluent along with poor effluent quality, indicating a lower level of process stability. However, reviewing the literature related to the procedure for obtaining a thermophilic inoculum, it seems that most of the problems associated with the instability and the accumulation of organic intermediates are the result of the manner in which the thermophilic sludge has been obtained. In this paper, the different options available for obtaining an anaerobic digester operating at thermophilic temperature (55°C) have been reviewed. In this light, rapid heating to the target temperature followed by the development of thermophilic microorganisms, which can be determined by VFA dropping to ≤ 500 mg acetic acid L(-1) before increasing the organic loading rate (OLR), has been determined the most suitable means of establishing TAD.

  18. 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

  19. 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)).

  20. Draft genome sequence of Herbinix hemicellulosilytica T3/55 T, a new thermophilic cellulose degrading bacterium isolated from a thermophilic biogas reactor.

    PubMed

    Koeck, Daniela E; Maus, Irena; Wibberg, Daniel; Winkler, Anika; Zverlov, Vladimir V; Liebl, Wolfgang; Pühler, Alfred; Schwarz, Wolfgang H; Schlüter, Andreas

    2015-11-20

    A novel bacterial species was isolated from an industrial-scale biogas plant. The isolate Herbinix hemicellulosilytica T3/55(T) is able to degrade crystalline cellulose. Comparative 16S rRNA gene sequencing demonstrated that the isolate is closely related to environmental samples forming a hitherto unknown sub-cluster within the family Lachnospiraceae. The draft genome sequence of strain T3/55(T) was established and now provides the genetic basis for application of this microorganism in thermophilic degradation of lignocellulosic biomass.

  1. An integrated approach for genome annotation of the eukaryotic thermophile Chaetomium thermophilum

    PubMed Central

    Bock, Thomas; Chen, Wei-Hua; Ori, Alessandro; Malik, Nayab; Silva-Martin, Noella; Huerta-Cepas, Jaime; Powell, Sean T.; Kastritis, Panagiotis L.; Smyshlyaev, Georgy; Vonkova, Ivana; Kirkpatrick, Joanna; Doerks, Tobias; Nesme, Leo; Baßler, Jochen; Kos, Martin; Hurt, Ed; Carlomagno, Teresa; Gavin, Anne-Claude; Barabas, Orsolya; Müller, Christoph W.; van Noort, Vera; Beck, Martin; Bork, Peer

    2014-01-01

    The thermophilic fungus Chaetomium thermophilum holds great promise for structural biology. To increase the efficiency of its biochemical and structural characterization and to explore its thermophilic properties beyond those of individual proteins, we obtained transcriptomics and proteomics data, and integrated them with computational annotation methods and a multitude of biochemical experiments conducted by the structural biology community. We considerably improved the genome annotation of Chaetomium thermophilum and characterized the transcripts and expression of thousands of genes. We furthermore show that the composition and structure of the expressed proteome of Chaetomium thermophilum is similar to its mesophilic relatives. Data were deposited in a publicly available repository and provide a rich source to the structural biology community. PMID:25398899

  2. 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

  3. [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.

  4. 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.

  5. 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

  6. 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.

  7. 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

  8. Structure and flexibility of the thermophilic cold-shock protein of Thermus aquaticus.

    PubMed

    Jin, Bonghwan; Jeong, Ki-Woong; Kim, Yangmee

    2014-08-29

    The thermophilic bacterium Thermus aquaticus is a well-known source of Taq polymerase. Here, we studied the structure and dynamics of the T. aquaticus cold-shock protein (Ta-Csp) to better understand its thermostability using NMR spectroscopy. We found that Ta-Csp has a five-stranded β-barrel structure with five salt bridges which are important for more rigid structure and a higher melting temperature (76 °C) of Ta-Csp compared to mesophilic and psychrophilic Csps. Microsecond to millisecond time scale exchange processes occur only at the β1-β2 surface region of the nucleic acid binding site with an average conformational exchange rate constant of 674 s(-1). The results imply that thermophilic Ta-Csp has a more rigid structure and may not need high structural flexibility to accommodate nucleic acids upon cold shock compared to its mesophile and psychrophile counterparts. PMID:25101648

  9. Crystallization of an alpha-amylase, AmyA, from the thermophilic halophile Halothermothrix orenii.

    PubMed

    Li, Nan; Patel, Bharat K C; Mijts, Benjamin N; Swaminathan, Kunchithapadam

    2002-12-01

    This report is the first crystallographic study of an amylase from an organism that is both thermophilic and halophilic. alpha-Amylase from the thermophilic halophile Halothermothrix orenii (AmyA) is a 515-residue protein. It is stable and significantly active at 338 K in starch solution containing NaCl [up to 25%(w/v)]. Purified recombinant AmyA protein crystallizes in the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 55.126, b = 61.658, c = 147.625 A, using the hanging-drop vapour-diffusion method. The crystal diffracts X-rays to a resolution limit of 1.89 A.

  10. Long-term hydrolytic capacity evaluation of a thermophilic anaerobic digester treating sewage sludge.

    PubMed

    Donoso-Bravo, Andres; Pérez-Elvira, Sara; Wouwer, Alain Vande; Fdz-Polanco, Fernando

    2012-01-01

    This study presents an evaluation of the hydrolytic activity of a continuous thermophilic anaerobic reactor in long-term operation. The hydrolytic coefficient was estimated by fitting a three-reaction model of the anaerobic digestion process with experimental data obtained from a pilot thermophilic digester operated for about 2 years. The model fitting and the cross-validation indicate that this model can represent the behavior of the system in a proper way; moreover, the results show a variation of the hydrolytic capacity of the system throughout the evaluation period. The increase in the hydrolytic coefficient is in agreement with the increase in the organic load applied to the reactor, which shows the capacity of the continuous reactor to select populations according to the input conditions of the system.

  11. 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.

  12. Error-reducing Structure of the Genetic Code Indicates Code Origin in Non-thermophile Organisms

    NASA Astrophysics Data System (ADS)

    Gutfraind, Alexander; Kempf, Achim

    2008-02-01

    During the RNA World, organisms experienced high rates of genetic errors, which implies that there was strong evolutionary pressure to reduce the errors’ phenotypical impact by suitably structuring the still-evolving genetic code. Therefore, the relative rates of the various types of genetic errors should have left characteristic imprints in the structure of the genetic code. Here, we show that, therefore, it is possible to some extent to reconstruct those error rates, as well as the nucleotide frequencies, for the time when the code was fixed. We find evidence indicating that the frequencies of G and C in the genome were not elevated. Since, for thermodynamic reasons, RNA in thermophiles tends to possess elevated G+C content, this result indicates that the fixation of the genetic code occurred in organisms which were either not thermophiles or that the code’s fixation occurred after the rise of DNA.

  13. 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

  14. Combined thermophilic aerobic process and conventional anaerobic digestion: effect on sludge biodegradation and methane production.

    PubMed

    Dumas, C; Perez, S; Paul, E; Lefebvre, X

    2010-04-01

    The efficiency of hyper-thermophilic (65 degrees Celsius) aerobic process coupled with a mesophilic (35 degrees Celsius) digester was evaluated for the activated sludge degradation and was compared to a conventional mesophilic digester. For two Sludge Retention Time (SRT), 21 and 42 days, the Chemical Oxygen Demand (COD) solubilisation and biodegradation processes, the methanisation yield and the aerobic oxidation were investigated during 180 days. The best results were obtained at SRT of 44 days; the COD removal yield was 30% higher with the Mesophilic Anaerobic Digestion/Thermophilic Aerobic Reactor (MAD-TAR) co-treatment. An increase of the sludge intrinsic biodegradability is also observed (20-40%), showing that the unbiodegradable COD in mesophilic conditions becomes bioavailable. However, the methanisation yield was quite similar for both processes at a same SRT. Finally, such a process enables to divide by two the volume of digester with an equivalent efficiency.

  15. 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

  16. Exploring research frontiers in microbiology: recent advances in halophilic and thermophilic extremophiles.

    PubMed

    Averhoff, Beate; Müller, Volker

    2010-01-01

    Extremophilic prokaryotes inhabit ecosystems that are, from a human perspective, extreme, and life in these environments requires far-reaching cellular adaptations. Here, we will describe, for two examples (Thermus thermophilus, Halobacillus halophilus), how thermophilic or halophilic bacteria adapt to their environment; we will describe the molecular basis of sensing and responding to hypersalinity and we will analyze the impact and basis of natural competence for survival in hot environments.

  17. Dry co-digestion of sewage sludge and rice straw under mesophilic and thermophilic anaerobic conditions.

    PubMed

    Chu, Xiangqian; Wu, Guangxue; Wang, Jiaquan; Hu, Zhen-Hu

    2015-12-01

    Dry anaerobic digestion of sewage sludge can recover biogas as energy; however, its low C/N ratio limits it as a single substrate in the anaerobic digestion. Rice straw is an abundant agricultural residue in China, which is rich in carbon and can be used as carbon source. In the present study, the performance of dry co-digestion of sewage sludge and rice straw was investigated under mesophilic (35 °C) and thermophilic (55 °C) conditions. The operational factors impacting dry co-digestion of sewage sludge and rice straw such as C/N ratio, moisture content, and initial pH were explored under mesophilic conditions. The results show that low C/N ratios resulted in a higher biogas production rate, but a lower specific biogas yield; low moisture content of 65 % resulted in the instability of the digestion system and a low specific biogas yield. Initial pH ranging 7.0-9.0 did not affect the performance of the anaerobic digestion. The C/N ratio of 26-29:1, moisture content of 70-80 %, and pH 7.0-9.0 resulted in good performance in the dry mesophilic co-digestion of sewage sludge and rice straw. As compared with mesophilic digestion, thermophilic co-digestion of sewage sludge and rice straw significantly enhanced the degradation efficiency of the substrates and the specific biogas yield (p < 0.05) at the conditions of C/N ratio 26:1, moisture content 80 %, and natural initial pH. Although high concentrations of ammonia-nitrogen (NH4-N, 1500 mg/kg wet weight) were formed during thermophilic digestion, there was no obvious inhibition occurred. The results indicated that rice straw can be used as carbon source for the dry co-digestion of sewage sludge under mesophilic and thermophilic conditions.

  18. 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.

  19. High stability of apo-cytochrome c' from thermophilic Hydrogenophilus thermoluteolus.

    PubMed

    Fujii, Sotaro; Masanari, Misa; Yamanaka, Masaru; Wakai, Satoshi; Sambongi, Yoshihiro

    2014-01-01

    Apo-cytochomes c without heme are usually unstructured. Here we showed that apo-form of thermophilic Hydrogenophilus thermoluteolus cytochrome c' (PHCP) was a monomeric protein with high helix content. Apo-PHCP was thermally stable, possibly due to the hydrophobic residues and ion pairs. PHCP is the first example of a structured apo-cytochrome c', which will expand our view of hemoprotein structure formation.

  20. 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.

  1. Engineering pyruvate decarboxylase-mediated ethanol production in the thermophilic host Geobacillus thermoglucosidasius.

    PubMed

    Van Zyl, L J; Taylor, M P; Eley, K; Tuffin, M; Cowan, D A

    2014-02-01

    This study reports the expression, purification, and kinetic characterization of a pyruvate decarboxylase (PDC) from Gluconobacter oxydans. Kinetic analyses showed the enzyme to have high affinity for pyruvate (120 μM at pH 5), high catalytic efficiency (4.75 × 10(5) M(-1) s(-1) at pH 5), a pHopt of approximately 4.5 and an in vitro temperature optimum at approximately 55 °C. Due to in vitro thermostablity (approximately 40 % enzyme activity retained after 30 min at 65 °C), this PDC was considered to be a suitable candidate for heterologous expression in the thermophile Geobacillus thermoglucosidasius for ethanol production. Initial studies using a variety of methods failed to detect activity at any growth temperature (45-55 °C). However, the application of codon harmonization (i.e., mimicry of the heterogeneous host's transcription and translational rhythm) yielded a protein that was fully functional in the thermophilic strain at 45 °C (as determined by enzyme activity, Western blot, mRNA detection, and ethanol productivity). Here, we describe the first successful expression of PDC in a true thermophile. Yields as high as 0.35 ± 0.04 g/g ethanol per gram of glucose consumed were detected, highly competitive to those reported in ethanologenic thermophilic mutants. Although activities could not be detected at temperatures approaching the growth optimum for the strain, this study highlights the possibility that previously unsuccessful expression of pdcs in Geobacillus spp. may be the result of ineffective transcription/translation coupling.

  2. 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

  3. 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.

  4. 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.

  5. 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

  6. Reducing antibiotic resistance genes, integrons, and pathogens in dairy manure by continuous thermophilic composting.

    PubMed

    Qian, Xun; Sun, Wei; Gu, Jie; Wang, Xiao-Juan; Zhang, Ya-Jun; Duan, Man-Li; Li, Hai-Chao; Zhang, Ran-Ran

    2016-11-01

    This study explored the effects of composting using three temperature regimes, namely, insufficient thermophilic composting (ITC), normal thermophilic composting (NTC), and continuous thermophilic composting (CTC), on antibiotic resistance genes (ARGs), integrons, and human pathogenic bacteria (HPB), as well as the mechanisms involved. The NTC and CTC treatments led to greater decreases in 5/10 ARGs and two integrons than ITC, and the abundances of ARGs (tetC, tetG, and tetQ) and int1 only declined in the NTC and CTC treatments. The abundances of HPB decreased by 82.8%, 76.9%, and 96.9% under ITC, NTC, CTC, respectively. Redundancy analysis showed that both bacterial succession and horizontal gene transfer play important roles in the variation of ARGs, and the changes in different ARGs were due to diverse mechanisms. CTC performed significantly better at reducing ARGs, integrons, and HPB, thus it may be used to manage the public health risks of ARGs in animal manure.

  7. Co-digestion of polylactide and kitchen garbage in hyperthermophilic and thermophilic continuous anaerobic process.

    PubMed

    Wang, Feng; Hidaka, Taira; Tsuno, Hiroshi; Tsubota, Jun

    2012-05-01

    Two series of two-phase anaerobic systems, consisting of a hyperthermophilic (80°C) reactor and a thermophilic (55°C) reactor, fed with a mixture of kitchen garbage (KG) and polylactide (PLA), was compared with a single-phase thermophilic reactor for the overall performance. The result indicated that ammonia addition under hyperthermophilic condition promoted the transformation of PLA particles to lactic acid. The systems with hyperthermophilic treatment had advantages on PLA transformation and methane conversion ratio to the control system. Under the organic loading rate (OLR) of 10.3 g COD/(L day), the PLA transformation ratios of the two-phase systems were 82.0% and 85.2%, respectively, higher than that of the control system (63.5%). The methane conversion ratios of the two-phase systems were 82.9% and 80.8%, respectively, higher than 70.1% of the control system. The microbial community analysis indicated that hyperthermophilic treatment is easily installed to traditional thermophilic anaerobic digestion plants without inoculation of special bacteria.

  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. 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.

  10. 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

  11. Bioenergy feedstock-specific enrichment of microbial populations during high-solids thermophilic deconstruction.

    PubMed

    Reddy, Amitha P; Allgaier, Martin; Singer, Steven W; Hazen, Terry C; Simmons, Blake A; Hugenholtz, Philip; VanderGheynst, Jean S

    2011-09-01

    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.

  12. 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

  13. 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

  14. Strategies for changing temperature from mesophilic to thermophilic conditions in anaerobic CSTR reactors treating sewage sludge.

    PubMed

    Bousková, A; Dohányos, M; Schmidt, J E; Angelidaki, I

    2005-04-01

    Thermophilic anaerobic digestion presents an advantageous way for stabilization of sludge from wastewater treatment plants. Two different strategies for changing operational process temperature from mesophilic (37 degrees C) to thermophilic (55 degrees C) were tested using two continuous flow stirred tank reactors operated at constant organic loading rate of 1.38 g VS/l reactor/day and hydraulic retention time of 20 days. In reactor A, the temperature was increased step-wise: 37 degrees C-->42 degrees C-->47 degrees C-->51 degrees C-->55 degrees C. While in reactor B, the temperature was changed in one-step, from 37 degrees C to the desired temperature of 55 degrees C, The results showed that the overall adaptation of the process for the step-wise temperature increment took 70 days in total and a new change was applied when the process was stabilized as indicated by stable methane production and low volatile fatty acids concentrations. Although the one-step temperature increase caused a severe disturbance in all the process parameters, the system reached a new stable operation after only 30 days indicating that this strategy is the best in changing from mesophilic to thermophilic operation in anaerobic digestion plants.

  15. 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

  16. 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.

  17. 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.

  18. 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.

  19. 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

  20. 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

  1. 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

  2. Characteristics of hydrocarbon hydroxylase genes in a thermophilic aerobic biological system treating oily produced wastewater.

    PubMed

    Liu, Ruyin; Gao, Yingxin; Ji, Yifeng; Zhang, Yu; Yang, Min

    2015-01-01

    Alkane and aromatic hydroxylase genes in a full-scale aerobic system treating oily produced wastewater under thermophilic condition (45-50 °C) in the Jidong oilfield, China, were investigated using clone library and quantitative polymerase chain reaction methods. Rather than the normally encountered integral-membrane non-haem iron monooxygenase (alkB) genes, only CYP153-type P450 hydroxylase genes were detected for the alkane activation, indicating that the terminal oxidation of alkanes might be mainly mediated by the CYP153-type alkane hydroxylases in the thermophilic aerobic process. Most of the obtained CYP153 gene clones showed distant homology with the reference sequences, which might represent novel alkane hydroxylases. For the aromatic activation, the polycyclic aromatic hydrocarbon-ring hydroxylating dioxygenase (PAH-RHD) gene was derived from Gram-negative PAH-degraders belonging to the Burkholderiales order, with a 0.72% relative abundance of PAH-RHD gene to 16S rRNA gene. This was consistent with the result of 16S rRNA gene analysis, indicating that Burkholderiales bacteria might play a key role in the full-scale process of thermophilic hydrocarbon degradation.

  3. 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.

  4. 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

  5. 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

  6. Naphthalene degradation and incorporation of naphthalene-derived carbon into biomass by the thermophile Bacillus thermoleovorans

    SciTech Connect

    Annweiler, E.; Richnow, H.H.; Antranikian, G.; Hebenbrock, S.; Garms, C.; Franke, S.; Francke, W.; Michaelis, W.

    2000-02-01

    The thermophilic aerobic bacterium Bacillus thermoleovorans Hamburg 2 grows at 60 C on naphthalene as the sole source of carbon and energy. In batch cultures, an effective substrate degradation was observed. The carbon balance, including naphthalene, metabolites, biomass, and CO{sub 2}, was determined by the application of [1-{sup 13}C]naphthalene. The incorporation of naphthalene-derived carbon into the bulk biomass as well as into specified biomass fractions such as fatty acids and amino acids was confirmed by coupled gas chromatography-mass spectrometry (GC-MS) and isotope analyses. Metabolites were characterized by GC-MS; the established structures allow tracing the degradation pathway under thermophilic conditions. Apart from typical metabolites of naphthalene degradation known from mesophiles, intermediates such as 2,3-dihydroxynaphthalene, 2-carboxycinnamic acid, and phthalic and benzoic acid were identified for the pathway of this bacterium. These compounds indicate that naphthalene degradation by the thermophilic B. thermoleovorans differs from the known pathways found for mesophilic bacteria.

  7. 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.

  8. 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.

  9. 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

  10. Co-digestion of polylactide and kitchen garbage in hyperthermophilic and thermophilic continuous anaerobic process.

    PubMed

    Wang, Feng; Hidaka, Taira; Tsuno, Hiroshi; Tsubota, Jun

    2012-05-01

    Two series of two-phase anaerobic systems, consisting of a hyperthermophilic (80°C) reactor and a thermophilic (55°C) reactor, fed with a mixture of kitchen garbage (KG) and polylactide (PLA), was compared with a single-phase thermophilic reactor for the overall performance. The result indicated that ammonia addition under hyperthermophilic condition promoted the transformation of PLA particles to lactic acid. The systems with hyperthermophilic treatment had advantages on PLA transformation and methane conversion ratio to the control system. Under the organic loading rate (OLR) of 10.3 g COD/(L day), the PLA transformation ratios of the two-phase systems were 82.0% and 85.2%, respectively, higher than that of the control system (63.5%). The methane conversion ratios of the two-phase systems were 82.9% and 80.8%, respectively, higher than 70.1% of the control system. The microbial community analysis indicated that hyperthermophilic treatment is easily installed to traditional thermophilic anaerobic digestion plants without inoculation of special bacteria. PMID:22409980

  11. [Conversion of acetic acid to methane by thermophiles]. Annual progress report

    SciTech Connect

    Zinder, S.H.

    1994-02-01

    Acetate is the precursor of approximately two-thirds of the methane produced by anaerobic bioreactors and many other methanogenic habitats. Besides their intrinsic interest, thermophilic acetotrophic methanogenic cultures usually grow at least twice as fast as their mesophilic counterparts, making them more amenable to study. In recent years, attention has been mainly focused on the thermophilic acetate utilizing methanogen Methanothrix strain CALS-1. Methanothrix, also called Methanosaeta, is one of only two methanogenic genera known to convert acetate to methane, the other being Methanosarcina. The faster-growing more versatile Methanosarcina has been better studied. However, when one examines anaerobic digestor contents, Methanothrix is often the dominant acetate-utilizing methanogen. As described in previous progress reports, the authors have achieved methanogenesis from acetate in cell-free extracts of Methanothrix strain CALS-1 grown in a pH auxostat. Using these cell extracts, specific activities for methanogenesis from acetate and ATP of 100--300 nmol/min were routinely obtained, levels comparable to the rate in whole cells, which is not usually the case in methanogenic extracts. Recently obtained results are given and discussed for the following: Methanogenesis in crude extracts; Role of the cell membrane in methanogenesis from acetate; Carbon monoxide dehydrogenase; Novel thermophilic cultures converting acetate to methane; and Methanol-utilizing methanogen.

  12. 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

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

    PubMed

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

    2011-11-01

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

  14. 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.

  15. 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.

  16. Thermophilic (55-65 degrees C) and extreme thermophilic (70-80 degrees C) sulfate reduction in methanol and formate-fed UASB reactors.

    PubMed

    Vallero, Marcus V G; Camarero, Eva; Lettinga, Gatze; Lens, Piet N L

    2004-01-01

    The feasibility of thermophilic (55-65 degrees C) and extreme thermophilic (70-80 degrees C) sulfate-reducing processes was investigated in three lab-scale upflow anaerobic sludge bed (UASB) reactors fed with either methanol or formate as the sole substrates and inoculated with mesophilic granular sludge previously not exposed to high temperatures. Full methanol and formate degradation at temperatures up to, respectively, 70 and 75 degrees C, were achieved when operating UASB reactors fed with sulfate rich (COD/SO4(2-)=0.5) synthetic wastewater. Methane-producing archaea (MPA) outcompeted sulfate-reducing bacteria (SRB) in the formate-fed UASB reactor at all temperatures tested (65-75 degrees C). In contrast, SRB outcompeted MPA in methanol-fed UASB reactors at temperatures equal to or exceeding 65 degrees C, whereas strong competition between SRB and MPA was observed in these reactors at 55 degrees C. A short-term (5 days) temperature increase from 55 to 65 degrees C was an effective strategy to suppress methanogenesis in methanol-fed sulfidogenic UASB reactors operated at 55 degrees C. Methanol was found to be a suitable electron donor for sulfate-reducing processes at a maximal temperature of 70 degrees C, with sulfide as the sole mineralization product of methanol degradation at that temperature.

  17. 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

  18. 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

  19. Causal Moderation Analysis Using Propensity Score Methods

    ERIC Educational Resources Information Center

    Dong, Nianbo

    2012-01-01

    This paper is based on previous studies in applying propensity score methods to study multiple treatment variables to examine the causal moderator effect. The propensity score methods will be demonstrated in a case study to examine the causal moderator effect, where the moderators are categorical and continuous variables. Moderation analysis is an…

  20. Evolutionary relationships amongst archaebacteria. A comparative study of 23 S ribosomal RNAs of a sulphur-dependent extreme thermophile, an extreme halophile and a thermophilic methanogen.

    PubMed

    Leffers, H; Kjems, J; Ostergaard, L; Larsen, N; Garrett, R A

    1987-05-01

    The 23 S RNA genes representative of each of the main archaebacterial subkingdoms, Desulfurococcus mobilis an extreme thermophile, Halococcus morrhuae an extreme halophile and Methanobacterium thermoautotrophicum a thermophilic methanogen, were cloned and sequenced. The inferred RNA sequences were aligned with all the available 23 S-like RNAs of other archaebacteria, eubacteria/chloroplasts and the cytoplasm of eukaryotes. Universal secondary structural models containing six major structural domains were refined, and extended, using the sequence comparison approach. Much of the present structure was confirmed but six new helices were added, including one that also exists in the eukaryotic 5.8 S RNA, and extensions were made to several existing helices. The data throw doubt on whether the 5' and 3' ends of the 23 S RNA interact, since no stable helix can form in either the extreme thermophile or the methanogen RNA. A few secondary structural features, specific to the archaebacterial RNAs were identified; two of these were supported by a comparison of the archaebacterial RNA sequences, and experimentally, using chemical and ribonuclease probes. Seven tertiary structural interactions, common to all 23 S-like RNAs, were predicted within unpaired regions of the secondary structural model on the basis of co-variation of nucleotide pairs; two lie in the region of the 23 S RNA corresponding to 5.8 S RNA but they are not conserved in the latter. The flanking sequences of each of the RNAs could base-pair to form long RNA processing stems. They were not conserved in sequence but each exhibited a secondary structural feature that is common to all the archaebacterial stems for both 16 S and 23 S RNAs and constitutes a processing site. Kingdom-specific nucleotides have been identified that are associated with antibiotic binding sites at functional centres in 23 S-like RNAs: in the peptidyl transferase centre (erythromycin-domain V) the archaebacterial RNAs classify with the

  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. 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

  3. 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

  4. 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.

  5. 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.

  6. 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.

  7. Cooperative role of electrical stimulation on microbial metabolism and selection of thermophilic communities for p-fluoronitrobenzene treatment.

    PubMed

    Zhang, Xueqin; Shen, Dongsheng; Feng, Huajun; Wang, Yanfeng; Li, Na; Han, Jingyi; Long, Yuyang

    2015-01-01

    A novel thermophilic bioelectrochemical system (TBES) based on electrical stimulation was established for the enhanced treatment of p-fluoronitrobenzene (p-FNB) wastewater. p-FNB removal rate constant in the TBES was 78.6% higher than that of the mesophilic BES (MBES), the elevation of which owing to high-temperature overtook the rate improvement of 50.8% in the electrocatalytic system (ECS). Additionally, an overwhelming mineralization efficiency of 91.96% ± 5.70% was obtained in the TBES. The superiority of TBES was attributed to the integrated role of electrical stimulation and high-temperature. Electrical stimulation provided an alternative for the microbial growth independent energy requirements, compensating insufficient energy support from p-FNB metabolism under the high-temperature stress. Besides, electrical stimulation facilitated microbial community evolution to form specific thermophilic biocatalysis. The uniquely selected thermophilic microorganisms including Coprothermobacter sp. and other ones cooperated to enhance p-FNB mineralization.

  8. 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.

  9. Complete genome sequence of the strain Defluviitoga tunisiensis L3, isolated from a thermophilic, production-scale biogas plant.

    PubMed

    Maus, Irena; Cibis, Katharina Gabriela; Wibberg, Daniel; Winkler, Anika; Stolze, Yvonne; König, Helmut; Pühler, Alfred; Schlüter, Andreas

    2015-06-10

    An anaerobic, thermophilic bacterium belonging to the phylum Thermotogae was isolated from a rural, thermophilic biogas plant (54°C) producing methane-rich biogas from maize silage, barley, cattle and pig manure. Here we report the first complete genome sequence of the Defluviitoga tunisiensis strain L3, an isolate from the family Thermotogaceae. The strain L3 encodes several genes predicted to be involved in utilization of a large diversity of complex carbohydrates including cellobiose and xylan for the production of acetate, hydrogen (H2) and carbon dioxide (CO2). The genome sequence of D. tunisiensis L3 provides the basis for biotechnological exploitation of genetic determinants playing an important role in thermophilic fermentation processes utilizing renewable primary products.

  10. Thermoanaerobacterium aotearoense sp. nov., a slightly acidophilic, anaerobic thermophile isolated from various hot springs in New Zealand, and emendation of the genus Thermoanaerobacterium

    SciTech Connect

    Shu-Ying Liu; Wiegel, J.; Rainey, F.A.

    1996-04-01

    Six moderately acidophilic, thermophilic bacterial strains with similar properties were isolated from geothermally heated water and sediment samples collected in New Zealand. These Gram stain-negative but Gram type-positive, rod-shaped bacteria formed oval terminal endospores. The cells were peritrichously flagellated and exhibited tumbling motility. At 60{degrees}C the pH range for growth was 3.8 to 6.8, and the optimum pH was 5.2 when the organisms were grown with xylose. At pH 5.2 the temperature range for growth was 35 to 66{degrees}C, and the optimum temperature was 60 to 63{degrees}C. The fermentation products from flucose or xylose were ethanol, acetate, lactate, CO{sub 2}, and H{sub 2}. The DNA G+C content was 34.5 to 35 mol%. On the basis of properties such as formation of elemental sulfur from thiosulfate, growth at acidic pH values at elevated temperatures, and the results of a 16S rRNA sequence comparison performed with previously validly published species belonging to the genus Thermoanaerobacterium, we propose that strain JW/SL-NZ613{sup T} (T = type strain) and five similar strains isolated from samples collected in New Zealand represent a new species, Thermoanaerobacterium aotearoense. Strain JW/SL-NZ613{sup T} (= DSM 10170) is the type strain of this species.

  11. 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.

  12. 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

  13. Anaerobic co-digestion of food waste and chemically enhanced primary-treated sludge under mesophilic and thermophilic conditions.

    PubMed

    Obulisamy, Parthiba Karthikeyan; Chakraborty, Debkumar; Selvam, Ammaiyappan; Wong, Jonathan W C

    2016-12-01

    Anaerobic co-digestion of food waste with primary sewage sludge is beneficial for urban centers, while the optimized conditions reported in the literature are not locally suitable for Hong Kong. Therefore, the present study was aimed to develop an optimized mixing ratio of food waste to chemically enhanced primary-treated sewer sludge (CEPT) for co-digestion using batch tests under mesophilic (37°C) and thermophilic (55°C) conditions. The mixing ratios of 1:1, 1:2, 1:3, 2:1 and 3:1 (v v(-1)) of food waste to CEPT sludge was tested under the following conditions: temperature - 35°C and 55°C; pH - not regulated; agitation - 150 rpm and time - 20 days. The thermophilic incubations led a good hydrolysis rate and 2-12-fold higher enzyme activities than in mesophilic incubations for different mixing ratios. While the acidogenesis were found retarded that leading to 'sour and stuck' digestion for all mixing ratio of food waste to CEPT sludge from thermophilic incubations. The measured zeta potential was most favourable (-5 to -16.8 mV) for methane production under thermophilic incubations; however the CH4 recovery was less than that in mesophilic incubations. The results suggested that the quick hydrolysis and subsequent acid accumulation under thermophilic incubation lead to inhibited methanogenesis at the early stage than in mesophilic systems. It is concluded that buffer addition is therefore required for any mixing ratio of food waste to CEPT sludge for improved CH4 recovery for both mesophilic and thermophilic operations.

  14. A novel process configuration for anaerobic digestion of source-sorted household waste using hyper-thermophilic post-treatment.

    PubMed

    Hartmann, Hinrich; Ahring, Birgitte K

    2005-06-30

    A novel reactor configuration was investigated for anaerobic digestion (AD) of the organic fraction of municipal solid waste (OFMSW). An anaerobic hyper-thermophilic (68 degrees C) reactor R68 was implemented as a post-treatment step for the effluent of a thermophilic reactor R1 (55 degrees C) in order to enhance hydrolysis of recalcitrant organic matter, improve sanitation and ease the stripping of ammonia from the reactor. The efficiency of the combined system was studied in terms of methane yield, volatile solids (VS) reduction, and volatile fatty acid (VFA) production at different hydraulic retention times (HRT). A single-stage thermophilic (55 degrees C) reactor R2 was used as control. VS reduction and biogas yield of the combined system was 78-89% and 640-790 mL/g VS, respectively. While the VS reduction in the combined system was up to 7% higher than in the single-stage treatment, no increase in methane yield was observed. Shifting the HRT of the hyper-thermophilic reactor from 5 days to 3 days resulted in a drop in the methanogenic activity in the hydrolysis reactor to a minimum. Operation of R68 at HRTs of 24-48 h was sufficient to achieve high VS conversion into VFAs. Removal of pathogens was enhanced by the hyper-thermophilic post-treatment. 7% of the ammonia load was removed in the hyper-thermophilic reactor with a flow of headspace gas through the reactor equivalent to four times the biogas flow produced in reactor R1.

  15. Spore-forming thermophilic sulfate-reducing bacteria isolated from North Sea oil field waters

    SciTech Connect

    Rosnes, J.T.; Torsvik, T.; Lien, T. )

    1991-08-01

    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 (C{sub 4} through C{sub 6}) was added as a carbon source and electron donor. Representative strains were isolated and characterized. The isolates grew autotrophically on H{sub 2}-CO{sub 2} 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 78C; the spores were extremely heat resistant and survived 131C 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.

  16. 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

  17. Preservation of microbial communities enriched on lignocellulose under thermophilic and high-solid conditions

    DOE PAGES

    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

  18. 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

  19. 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.

  20. 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

  1. 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

  2. Ecology of Thermophilic Fungi in Mushroom Compost, with Emphasis on Scytalidium thermophilum and Growth Stimulation of Agaricus bisporus Mycelium

    PubMed Central

    Straatsma, Gerben; Samson, Robert A.; Olijnsma, Tineke W.; Op Den Camp, Huub J. M.; Gerrits, Jan P. G.; Van Griensven, Leo J. L. D.

    1994-01-01

    Twenty-two species of thermophilic fungi were isolated from mushroom compost. Scytalidium thermophilum was present in the compost ingredients, fresh straw, horse droppings, and drainage from compost and dominated the fungal biota of compost after preparation. Of 34 species of thermophilic fungi tested, 9 promoted mycelial growth of Agaricus bisporus on sterilized compost: Chaetomium thermophilum, an unidentified Chaetomium sp., Malbranchea sulfurea, Myriococcum thermophilum, S. thermophilum, Stilbella thermophila, Thielavia terrestris, and two unidentified basidiomycetes. These species will be considered for future experiments on inoculation and more controlled preparation of compost. PMID:16349175

  3. 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

  4. Comparative study on the selective chalcopyrite bioleaching of a molybdenite concentrate with mesophilic and thermophilic bacteria.

    PubMed

    Romano, P; Blázquez, M L; Alguacil, F J; Muñoz, J A; Ballester, A; González, F

    2001-03-01

    This study evaluates different bioleaching treatments of a molybdenite concentrate using mesophilic and thermophilic bacterial cultures. Further studies on the chemical leaching and the electrochemical behavior of the MoS(2) concentrate were carried out. Bioleaching tests showed a progressive removal of chalcopyrite from the molybdenite concentrate with an increase in temperature. Chemical leaching tests support the idea of an indirect attack of the concentrate. Electrochemical tests indicate that chalcopyrite dissolution is favored when molybdenite is present. Therefore, this type of bioleaching treatment could be applied to purify molybdenite flotation concentrates by selectively dissolving chalcopyrite. PMID:11257551

  5. Geobacillus icigianus sp. nov., a thermophilic bacterium isolated from a hot spring.

    PubMed

    Bryanskaya, Alla V; Rozanov, Alexey S; Slynko, Nikolay M; Shekhovtsov, Sergey V; Peltek, Sergey E

    2015-03-01

    A Gram-reaction-positive, motile, thermophilic spore-forming strain, G1w1(T), was isolated from a hot spring of the Valley of Geysers, Kamchatka (Russia). Based on data from the present polyphasic taxonomic study, including phylogenetic analysis of 16S rRNA and spo0A gene sequences, the strain is considered to represent a novel species of the genus Geobacillus, for which the name Geobacillus icigianus sp. nov. is proposed. The type strain is G1w1(T) ( = VKM B-2853(T) = DSM 28325(T)).

  6. 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/).

  7. Macroscopic mass and energy balance of a pilot plant anaerobic bioreactor operated under thermophilic conditions.

    PubMed

    Espinosa-Solares, Teodoro; Bombardiere, John; Chatfield, Mark; Domaschko, Max; Easter, Michael; Stafford, David A; Castillo-Angeles, Saul; Castellanos-Hernandez, Nehemias

    2006-01-01

    Intensive poultry production generates over 100,000 t of litter annually in West Virginia and 9 x 10(6) t nationwide. Current available technological alternatives based on thermophilic anaerobic digestion for residuals treatment are diverse. A modification of the typical continuous stirred tank reactor is a promising process being relatively stable and owing to its capability to manage considerable amounts of residuals at low operational cost. A 40-m3 pilot plant digester was used for performance evaluation considering energy input and methane production. Results suggest some changes to the pilot plant configuration are necessary to reduce power consumption although maximizing biodigester performance.

  8. 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/).

  9. Enrichment of Thermophilic Ammonia-Oxidizing Archaea from an Alkaline Hot Spring in the Great Basin, USA

    NASA Astrophysics Data System (ADS)

    Zhang, C.; Huang, Z.; Jiang, H.; Wiegel, J.; Li, W.; Dong, H.

    2010-12-01

    One of the major advances in the nitrogen cycle is the recent discovery of ammonia oxidation by archaea. While culture-independent studies have revealed occurrence of ammonia-oxidizing archaea (AOA) in nearly every surface niche on earth, most of these microorganisms have resisted isolation and so far only a few species have been identified. The Great Basin contains numerous hot springs, which are characterized by moderately high temperature (40-65 degree C) and circumneutral or alkaline pH. Unique thermophilic archaea have been identified based on molecular DNA and lipid biomarkers; some of which may be ammonia oxidizers. This study aims to isolate some of these archaea from a California hot spring that has pH around 9.0 and temperature around 42 degree C. Mat material was collected from the spring and transported on ice to the laboratory. A synthetic medium (SCM-5) was inoculated with the mat material and the culture was incubated under varying temperature (35-65 degree C) and pH (7.0-10.0) conditions using antibiotics to suppress bacterial growth. Growth of the culture was monitored by microscopy, decrease in ammonium and increase in nitrite, and increases in Crenarchaeota and AOA abundances over time. Clone libraries were constructed to compare archaeal community structures before and after the enrichment experiment. Temperature and pH profiles indicated that the culture grew optimally at pH 9.0 and temperature 45 degree C, which are consistent with the geochemical conditions of the natural environment. Phylogenetic analysis showed that the final OTU was distantly related to all known hyperthermophilic archaea. Analysis of the amoA genes showed two OTUs in the final culture; one of them was closely related to Candidatus Nitrososphaera gargensis. However, the enrichment culture always contained bacteria and attempts to separate them from archaea have failed. This highlights the difficulty in bringing AOA into pure culture and suggests that some of the AOA may

  10. 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%.

  11. 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

  12. Quantitative influences of butyrate or propionate on thermophilic production of methane from biomass

    SciTech Connect

    Henson, J.M.; Bordeaux, F.M.; Rivards, C.J.; Smith, P.H.

    1986-02-01

    Sodium butyrate and sodium propionate were continuously infused into separate 4-liter thermophilic digesters. These digesters were operated at 55/sup 0/C, had a retention time of 20 days, and had a pH of 7.8. Infusion rates were started at 10 mM day/sup -1/ and were increased incrementally when new stable external organic acid pool sizes and new stable gas production rates were observed. Stable conditions were obtained in both digesters at an infusion rate of 15 mM day/sup -1/, with methanogenesis elevated over that of control digesters. Calculations based on expected CH/sub 4/ at this infusion rate and measured CH/sub 4/ production in the treated and control digesters, however, showed an approximately 25% inhibition of methanogenesis in both digesters. A digester infused with sodium chloride showed little or no inhibition at this infusion rate, but was totally inhibited when its infusion rate was increased to 20 mM day/sup -1/, and cumulative added NaCl reached 0.38 M. The butyrate and propionate-amended digesters tolerated addition rates of 20 mM day/sup -1/, but both failed when they were increased to 25 mM day/sup -1/. These results indicate that the thermophilic digesters could function stably at higher external pool sizes of butyrate or propionate than routinely observed.

  13. 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

  14. 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

  15. Bacterial community analysis of swine manure treated with autothermal thermophilic aerobic digestion.

    PubMed

    Han, Il; Congeevaram, Shankar; Ki, Dong-Won; Oh, Byoung-Taek; Park, Joonhong

    2011-02-01

    Due to the environmental problems associated with disposal of livestock sludge, many stabilization studies emphasizing on the sludge volume reduction were performed. However, little is known about the microbial risk present in sludge and its stabilized products. This study microbiologically explored the effects of anaerobic lagoon fermentation (ALF) and autothermal thermophilic aerobic digestion (ATAD) on pathogen-related risk of raw swine manure by using culture-independent 16S rDNA cloning and sequencing methods. In raw swine manure, clones closely related to pathogens such as Dialister pneumosintes, Erysipelothrix rhusiopathiae, Succinivibrioan dextrinosolvens, and Schineria sp. were detected. Meanwhile, in the mesophilic ALF-treated swine manure, bacterial community clones closely related to pathogens such as Schineria sp. and Succinivibrio dextrinosolvens were still detected. Interestingly, the ATAD treatment resulted in no detection of clones closely related to pathogens in the stabilized thermophilic bacterial community, with the predominance of novel Clostridia class populations. These findings support the superiority of ATAD in selectively reducing potential human and animal pathogens compared to ALF, which is a typical manure stabilization method used in livestock farms. PMID:20922382

  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).

  17. Morphological and phylogenetic diversity of thermophilic cyanobacteria in Algerian hot springs.

    PubMed

    Amarouche-Yala, Samia; Benouadah, Ali; El Ouahab Bentabet, Abd; López-García, Purificación

    2014-11-01

    Geothermal springs in Algeria have been known since the Roman Empire. They mainly locate in Eastern Algeria and are inhabited by thermophilic organisms, which include cyanobacteria forming mats and concretions. In this work, we have investigated the cyanobacterial diversity of these springs. Cyanobacteria were collected from water, concretions and mats in nine hot springs with water temperatures ranging from 39 to 93 °C. Samples were collected for isolation in culture, microscopic morphological examination, and molecular diversity analysis based on 16S rRNA gene sequences. Nineteen different cyanobacterial morphotypes were identified, the most abundant of which were three species of Leptolyngbya, accompanied by members of the genera Gloeocapsa, Gloeocapsopsis, Stigonema, Fischerella, Synechocystis, Microcoleus, Cyanobacterium, Chroococcus and Geitlerinema. Molecular diversity analyses were in good general agreement with classical identification and allowed the detection of additional species in three springs with temperatures higher than 50 °C. They corresponded to a Synechococcus clade and to relatives of the intracellularly calcifying Candidatus Gloeomargarita lithophora. The hottest springs were dominated by members of Leptolyngbya, Synechococcus-like cyanobacteria and Gloeomargarita, whereas Oscillatoriales other than Leptolyngbya, Chroococcales and Stigonematales dominated lower temperature springs. The isolation of some of these strains sets the ground for future studies on the biology of thermophilic cyanobacteria.

  18. Thermophilic, anaerobic bacteria isolated from a deep borehole in granite in Sweden.

    PubMed

    Szewzyk, U; Szewzyk, R; Stenström, T A

    1994-03-01

    A borehole drilled to a total depth of 6779 m in granitic rock in Gravberg, Sweden, was sampled and examined for the presence of anaerobic, thermophilic, fermenting bacteria and sulfate-reducing bacteria. Growth in enrichment cultures was obtained only from water samples collected from a specific sampling depth in the borehole (3500 m). The hole was cased down to a depth of 5278 m and open to the formation below that level. All the water below 2000 m in depth standing in the borehole at the time of sampling must have entered at the 5278-m level or below, during a prior pumping operation. A strong salinity stratification certifies that no major amount of vertical mixing had taken place. The depth from which bacteria could be enriched was that of a pronounced local minimum of salinity. Pure cultures of thermophilic, anaerobic, fermenting bacteria were obtained with the following substrates: glucose, starch, xylan, ethanol, and lactate. The morphology and physiology of the glucose- and starch-degrading strains indicate a relationship to Thermoanaerobacter and Thermoanaerobium species. All but one of the newly isolated strains differ however from those by lacking acetate as a fermentation product. The glucose-degrading strain Gluc1 is phylogenetically related to Clostridium thermohydrosulfuricum, with an evolutionary distance based upon rRNA sequence comparisons of 3%. No sulfate-reducing or methanogenic bacteria were found.

  19. 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

  20. 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

  1. 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

  2. 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

  3. Quantitative Influences of Butyrate or Propionate on Thermophilic Production of Methane from Biomass †

    PubMed Central

    Henson, J. Michael; Bordeaux, F. M.; Rivard, Christopher J.; Smith, P. H.

    1986-01-01

    Sodium butyrate and sodium propionate were continuously infused into separate 4-liter thermophilic digesters. These digesters were operated at 55°C, had a retention time of 20 days, and had a pH of 7.8. Infusion rates were started at 10 mM day−1 and were increased incrementally when new stable external organic acid pool sizes and new stable gas production rates were observed. Stable conditions were obtained in both digesters at an infusion rate of 15 mM day−1, with methanogenesis elevated over that of control digesters. Calculations based on expected CH4 at this infusion rate and measured CH4 production in the treated and control digesters, however, showed an approximately 25% inhibition of methanogenesis in both digesters. A digester infused with sodium chloride showed little or no inhibition at this infusion rate, but was totally inhibited when its infusion rate was increased to 20 mM day−1, and cumulative added NaCl reached 0.38 M. The butyrate and propionate-amended digesters tolerated addition rates of 20 mM day−1, but both failed when they were increased to 25 mM day−1. These results indicate that the thermophilic digesters could function stably at higher external pool sizes of butyrate or propionate than routinely observed. PMID:16346985

  4. [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.

  5. 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.

  6. [Gradient elevation of temperature startup experiment of thermophilic ASBR treating thermal-hydrolyzed sewage sludge].

    PubMed

    Ouyang, Er-Ming; Wang, Wei; Long, Neng; Li, Huai

    2009-04-15

    Startup experiment was conducted for thermophilic anaerobic sequencing batch reactor (ASBR) treating thermal-hydrolyzed sewage sludge using the strategy of the step-wise temperature increment: 35 degrees C-->40 degrees C-->47 degrees C-->53 degrees C. The results showed that the first step-increase (from 35 degrees C to 40 degrees C) and final step-increase (from 47 degrees C to 53 degrees C) had only a slight effect on the digestion process. The second step-increase (from 40 degrees C to 47 degrees C) resulted in a severe disturbance: the biogas production, methane content, CODeffluent and microorganism all have strong disturbance. At the steady stage of thermophilic ASBR treating thermal-hydrolyzed sewage sludge, the average daily gas production, methane content, specific methane production (CH4/CODinfluent), TCOD removal rate and SCOD removal rate were 2.038 L/d, 72.0%, 188.8 mL/g, 63.8%, 83.3% respectively. The results of SEM and DGGE indicated that the dominant species are obviously different at early stage and steady stage.

  7. 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).

  8. 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

  9. 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

  10. Role of Mn2+ and Compatible Solutes in the Radiation Resistance of Thermophilic Bacteria and Archaea

    PubMed Central

    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 Mn2+-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

  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.

  12. 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.

  13. [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

  14. 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

  15. Bioleaching of a low-grade nickel-copper sulfide by mixture of four thermophiles.

    PubMed

    Li, Shuzhen; Zhong, Hui; Hu, Yuehua; Zhao, Jiancun; He, Zhiguo; Gu, Guohua

    2014-02-01

    This study investigated thermophilic bioleaching of a low grade nickel-copper sulfide using mixture of four acidophilic thermophiles. Effects of 0.2g/L l-cysteine on the bioleaching process were further evaluated. It aimed at offering new alternatives for enhancing metal recoveries from nickel-copper sulfide. Results showed a recovery of 80.4% nickel and 68.2% copper in 16-day bioleaching without l-cysteine; while 83.7% nickel and 81.4% copper were recovered in the presence of l-cysteine. Moreover, nickel recovery was always higher than copper recovery. l-Cysteine was found contributing to lower pH value, faster microbial growth, higher Oxidation-Reduction Potential (ORP), higher zeta potential and absorbing on the sulfide surfaces through amino, carboxyl and sulfhydryl groups. X-ray Diffraction (XRD) patterns of leached residues showed generation of S, jarosite and ammoniojarosite. Denaturing Gradient Gel Electrophoresis (DGGE) results revealed that l-cysteine could have variant impacts on different microorganisms and changed the microbial community composition dramatically during nickel-copper sulfide bioleaching.

  16. Biotransformation of eugenol via protocatechuic acid by thermophilic Geobacillus sp. AY 946034 strain.

    PubMed

    Giedraityte, Gražina; Kalėdienė, Lilija

    2014-04-01

    The metabolic pathway of eugenol degradation by thermophilic Geobacillus sp. AY 946034 strain was analyzed based on the lack of data about eugenol degradation by thermophiles. TLC, GC-MS, and biotransformation with resting cells showed that eugenol was oxidized through coniferyl alcohol, and ferulic and vanillic acids to protocatechuic acid before the aromatic ring was cleaved. The cell-free extract of Geobacillus sp. AY 946034 strain grown on eugenol showed a high activity of eugenol hydroxylase, feruloyl-CoA synthetase, vanillate-O-demethylase, and protocatechuate 3,4-dioxygenase. The key enzyme, protocatechuate 3,4- dioxygenase, which plays a crucial role in the degradation of various aromatic compounds, was purified 135-fold to homogeneity with a 34% overall recovery from Geobacillus sp. AY 946034. The relative molecular mass of the native enzyme was about 450 ± 10 kDa and was composed of the non-identical subunits. The pH and temperature optima for enzyme activity were 8 and 60°C, respectively. The half-life of protocatechuate 3,4-dioxygenase at the optimum temperature was 50 min.

  17. 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.

  18. 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%).

  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.

  20. Detection of anaerobic carbon monoxide-oxidizing thermophiles in hydrothermal environments.

    PubMed

    Yoneda, Yasuko; Kano, Sanae I; Yoshida, Takashi; Ikeda, Eitaro; Fukuyama, Yuto; Omae, Kimiho; Kimura-Sakai, Shigeko; Daifuku, Takashi; Watanabe, Tetsuhiro; Sako, Yoshihiko

    2015-09-01

    Carboxydotrophic anaerobic thermophiles have been isolated from various hydrothermal environments and are considered to be important carbon monoxide (CO) scavengers or primary producers. However, the ecological factors that influence the distribution, abundance and CO-oxidizing activities of these bacteria are poorly understood. A previous study detected the carboxydotrophic bacteria Carboxydothermus spp. in a hot spring sample and found that they constituted up to 10% of the total bacterial cells. In this study, we investigated environmental features, potential microbial CO-oxidation activities and the abundance of Carboxydothermus spp. in various hot springs to determine environmental factors that affect CO oxidizers and to see whether Carboxydothermus spp. are common in these environments. We detected potential microbial CO-oxidation activities in samples that showed relatively high values of total organic carbon, total nitrogen, oxidation-reduction potential and soil-water content. The abundance of Carboxydothermus spp. did not correlate with the presence of potential microbial CO-oxidation activities; however, Carboxydothermus spp. were detected in a wide range of environments, suggesting that these bacteria are widely distributed in spite of the relatively low population size. This study implies that thermophilic CO oxidizers occur in a wide range of environments and oxidize CO in somewhat oxidative environments rich in organic matter.

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

    PubMed

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

    2012-08-01

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

  2. 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

  3. 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.

  4. Production and properties of two novel exopolysaccharides synthesized by a thermophilic bacterium Aeribacillus pallidus 418.

    PubMed

    Radchenkova, Nadja; Vassilev, Spasen; Panchev, Ivan; Anzelmo, Gianluca; Tomova, Iva; Nicolaus, Barbara; Kuncheva, Margarita; Petrov, Kaloyan; Kambourova, Margarita

    2013-09-01

    Synthesis of innovative exocellular polysaccharides (EPSs) was reported for few thermophilic microorganisms as one of the mechanisms for surviving at high temperature. Thermophilic aerobic spore-forming bacteria able to produce exopolysaccharides were isolated from hydrothermal springs in Bulgaria. They were referred to four species, such as Aeribacillus pallidus, Geobacillus toebii, Brevibacillus thermoruber, and Anoxybacillus kestanbolensis. The highest production was established for the strain 418, whose phylogenetic and phenotypic properties referred it to the species A. pallidus. Maltose and NH4Cl were observed to be correspondingly the best carbon and nitrogen sources and production yield was increased more than twofold in the process of culture condition optimization. After purification of the polymer fraction, a presence of two different EPSs, electroneutral EPS 1 and negatively charged EPS 2, in a relative weight ratio 3:2.2 was established. They were heteropolysaccharides consisting of unusual high variety of sugars (six for EPS 1 and seven for EPS 2). Six of the sugars were common for both EPSs. The main sugar in EPS 1 was mannose (69.3 %); smaller quantities of glucose (11.2 %), galactosamine (6.3 %), glucosamine (5.4 %), galactose (4.7 %), and ribose (2.9 %) were also identified. The main sugar in EPS 2 was also mannose (33.9 %), followed by galactose (17.9 %), glucose (15.5 %), galactosamine (11.7 %), glucosamine (8.1 %), ribose (5.3 %), and arabinose (4.9 %). Both polymers showed high molecular weight and high thermostability.

  5. Enrichment of anaerobic syngas-converting bacteria from thermophilic bioreactor sludge.

    PubMed

    Alves, Joana I; Stams, Alfons J M; Plugge, Caroline M; Alves, M Madalena; Sousa, Diana Z

    2013-12-01

    Thermophilic (55 °C) anaerobic microbial communities were enriched with a synthetic syngas mixture (composed of CO, H2 , and CO2 ) or with CO alone. Cultures T-Syn and T-CO were incubated and successively transferred with syngas (16 transfers) or CO (9 transfers), respectively, with increasing CO partial pressures from 0.09 to 0.88 bar. Culture T-Syn, after 4 successive transfers with syngas, was also incubated with CO and subsequently transferred (9 transfers) with solely this substrate - cultures T-Syn-CO. Incubation with syngas and CO caused a rapid decrease in the microbial diversity of the anaerobic consortium. T-Syn and T-Syn-CO showed identical microbial composition and were dominated by Desulfotomaculum and Caloribacterium species. Incubation initiated with CO resulted in the enrichment of bacteria from the genera Thermincola and Thermoanaerobacter. Methane was detected in the first two to three transfers of T-Syn, but production ceased afterward. Acetate was the main product formed by T-Syn and T-Syn-CO. Enriched T-CO cultures showed a two-phase conversion, in which H2 was formed first and then converted to acetate. This research provides insight into how thermophilic anaerobic communities develop using syngas/CO as sole energy and carbon source can be steered for specific end products and subsequent microbial synthesis of chemicals.

  6. 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.

  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.

  8. 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

  9. 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

  10. Characterization of microbial compositions in a thermophilic chemostat of mixed culture fermentation.

    PubMed

    Zhang, Fang; Yang, Jing-Hua; Dai, Kun; Chen, Yun; Li, Qiu-Rong; Gao, Fa-Ming; Zeng, Raymond J

    2016-02-01

    The microbial community compositions of a chemostat enriched in a thermophilic (55 °C) mixed culture fermentation (MCF) for hydrogen production under different operational conditions were revealed in this work by integrating denaturing gradient gel electrophoresis (DGGE), Illumina Miseq high-throughput sequencing, and 16S rRNA clone library sequencing. The results showed that the community structure of the enriched cultures was relatively simple. Clones close to the genera of Thermoanaerobacter and/or Bacillus mainly dominated the bacteria. And homoacetogens and archaea were washed out and not detected even by Illumina Miseq high-throughput sequencing which supported the benefit for hydrogen production. On the other hand, the results revealed that the metabolic shift was clearly associated with the change of dominated bacterial groups. The effects of hydrogen partial pressure (PH2) and pH from 4.0 to 5.5 on the microbial compositions were not notable and Thermoanaerobacter was dominant, thus, the metabolites were also not changed. While Bacillus, Thermoanaerobacter and Propionispora hippei dominated the bacteria communities at neutral pH, or Bacillus and Thermoanaerobacter dominated at high influent glucose concentrations, consequently the main metabolites shifted to acetate, ethanol, propionate, or lactate. Thereby, the effect of microbial composition on the metabolite distribution and shift shall be considered when modeling thermophilic MCF in the future. PMID:26563549

  11. Bioleaching of a low-grade nickel-copper sulfide by mixture of four thermophiles.

    PubMed

    Li, Shuzhen; Zhong, Hui; Hu, Yuehua; Zhao, Jiancun; He, Zhiguo; Gu, Guohua

    2014-02-01

    This study investigated thermophilic bioleaching of a low grade nickel-copper sulfide using mixture of four acidophilic thermophiles. Effects of 0.2g/L l-cysteine on the bioleaching process were further evaluated. It aimed at offering new alternatives for enhancing metal recoveries from nickel-copper sulfide. Results showed a recovery of 80.4% nickel and 68.2% copper in 16-day bioleaching without l-cysteine; while 83.7% nickel and 81.4% copper were recovered in the presence of l-cysteine. Moreover, nickel recovery was always higher than copper recovery. l-Cysteine was found contributing to lower pH value, faster microbial growth, higher Oxidation-Reduction Potential (ORP), higher zeta potential and absorbing on the sulfide surfaces through amino, carboxyl and sulfhydryl groups. X-ray Diffraction (XRD) patterns of leached residues showed generation of S, jarosite and ammoniojarosite. Denaturing Gradient Gel Electrophoresis (DGGE) results revealed that l-cysteine could have variant impacts on different microorganisms and changed the microbial community composition dramatically during nickel-copper sulfide bioleaching. PMID:24374030

  12. An analysis of temperature adaptation in cold active, mesophilic and thermophilic Bacillus α-amylases.

    PubMed

    Mahdavi, Atiyeh; Sajedi, Reza H; Asghari, S Mohsen; Taghdir, Majid; Rassa, Mehdi

    2011-12-01

    A comparative biochemical and structural study was performed on a cold active α-amylase from Bacillus cereus (BCA) and two well-known homologous mesophilic and thermophilic α-amylases from Bacillus amyloliquefaciens (BAA) and Bacillus licheniformis (BLA). In spite of a high degree of sequence and structural similarity, drastic variations were found for T(opt) as 50, 70 and 90°C for BCA, BAA and BLA, respectively. The half-lives of thermoinactivation were 1 and 9 min for BCA and BAA at 80°C respectively, whilst there was no inactivation for BLA at this temperature. Thermodynamic studies on inactivation process suggested that lower thermostability of BCA is due to lower inactivation slope of the Arrhenius plots and subsequently, lower E(a) and ΔH(#). Increased K(m) and accessible surface area for catalytic residues along with a decreased number of internal interactions in this region in BCA compared to BLA suggest that BCA substrate-binding site might be temperature sensitive and is probably more flexible. On the other hand, fewer ion pairs, destructive substitutions and disruption of aromatic interaction networks in structurally critical regions of Bacillus α-amylases result in a severe decrease in BCA thermostability compared to its mesophilic and thermophilic homologues.

  13. [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

  14. Role of loops connecting secondary structure elements in the stabilization of proteins isolated from thermophilic organisms

    PubMed Central

    Balasco, Nicole; Esposito, Luciana; Simone, Alfonso De; Vitagliano, Luigi

    2013-01-01

    It has been recently discovered that the connection of secondary structure elements (ββ-unit, βα- and αβ-units) in proteins follows quite stringent principles regarding the chirality and the orientation of the structural units (Koga et al., Nature 2012;491:222–227). By exploiting these rules, a number of protein scaffolds endowed with a remarkable thermal stability have been designed (Koga et al., Nature 2012;491:222–227). By using structural databases of proteins isolated from either mesophilic or thermophilic organisms, we here investigate the influence of supersecondary associations on the thermal stability of natural proteins. Our results suggest that β-hairpins of proteins from thermophilic organisms are very frequently characterized by shortenings of the loops. Interestingly, this shortening leads to states that display a very strong preference for the most common connectivity of the strands observed in native protein hairpins. The abundance of selective states in these proteins suggests that they may achieve a high stability by adopting a strategy aimed to reduce the possible conformations of the unfolded ensemble. In this scenario, our data indicate that the shortening is effective if it increases the adherence to these rules. We also show that this mechanism may operate in the stabilization of well-known protein folds (thioredoxin and RNase A). These findings suggest that future investigations aimed at defining mechanism of protein stabilization should also consider these effects. PMID:23661276

  15. The crystal structure of a hyper-thermophilic carboxylesterase from the archaeon Archaeoglobus fulgidus.

    PubMed

    De Simone, G; Menchise, V; Manco, G; Mandrich, L; Sorrentino, N; Lang, D; Rossi, M; Pedone, C

    2001-11-30

    The crystal structure of AFEST, a novel hyper-thermophilic carboxylesterase from the archaeon Archaeoglobus fulgidus, complexed with a sulphonyl derivative, has been determined and refined to 2.2 A resolution. This enzyme, which has recently been classified as a member of the hormone- sensitive-lipase (H) group of the esterase/lipase superfamily, presents a canonical alpha/beta hydrolase core, shielded on the C-terminal side by a cap region composed of five alpha-helices. It contains the catalytic triad Ser160, His285 and Asp255, whereby the nucleophile is covalently modified and the oxyanion hole formed by Gly88, Gly89 and Ala161. A structural comparison of AFEST with its mesophilic and thermophilic homologues, Brefeldin A esterase from Bacillus subtilis (BFAE) and EST2 from Alicyclobacillus acidocaldarius, reveals an increase in the number of intramolecular ion pairs and secondary structure content, as well as a significant reduction in loop extensions and ratio of hydrophobic to charged surface area. The variety of structural differences suggests possible strategies for thermostabilization of lipases and esterases with potential industrial applications.

  16. Modification of progesterone and testosterone by a food-borne thermophile Geobacillus kaustophilus.

    PubMed

    Al-Tamimi, Safa; Al-Awadi, Sameera; Oommen, Sosamma; Afzal, Mohammad

    2010-02-01

    The present work was carried out to study structural modification of steroids by Geobacillus kaustophilus, a bacterial thermophile present in milk and the environment. Incubation of progesterone and testosterone with G. kaustophilus at 65 degrees C resulted in oxygenated steroid nuclei. The oxygenation of the steroid molecule was stereo specific. Seven metabolites of progesterone horizontal line 6beta/6alpha-hydroxytestosterone, 20-hydroxyprogesterone, 6beta-/6alpha-20-dihydroxyprogesterone, 5alpha-pregnane-3,6,20-trione, and 3beta-hydroxy-5alpha-pregnane-6,20-dione horizontal line were identified. Four compounds horizontal line namely, 66-/6--hydroxytestosterone and 6beta/6alpha-hydroxyandrostenedione horizontal line and androst-4-en-3,17-dione were identified as testosterone metabolites. This shows that G. kaustophilus is capable of modifying steroid nuclei at elevated temperatures. G. kaustophilus is a stable thermophile first isolated from milk. Our results show that endogenous steroids present in milk can be modified by G. kaustophilus, causing detrimental effect on human health.

  17. 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.

  18. 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.

  19. 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)

  20. Moderate-resolution holographic spectrograph

    NASA Astrophysics Data System (ADS)

    Muslimov, E. R.; Pavlycheva, N. K.; Valyavin, G. G.; Fabrika, S. N.

    2016-07-01

    We present a new scheme of a moderate-resolution spectrograph based on a cascade of serial holographic gratings each of which produces an individual spectrum with a resolution of about 6000 and a bandwidth of 80 nm. The gratings ensure centering of each part of the spectrum they produce so as to provide uniform coverage of the broadest possible wavelength interval. In this study we manage to simultaneously cover the 430-680 nm interval without gaps using three gratings. Efficiency of the spectrograph optical system itself from the entrance slit to the CCD detector is typically of about 60% with a maximum of 75%. We discuss the advantages and drawbacks of the new spectrograph scheme as well as the astrophysical tasks for which the instrument can be used.

  1. Digestion of cattle manure under mesophilic and thermophilic conditions: characterization of organic matter applying thermal analysis and 1H NMR.

    PubMed

    Gómez, X; Blanco, D; Lobato, A; Calleja, A; Martínez-Núñez, F; Martin-Villacorta, J

    2011-06-01

    Digestion of cattle manure collected from a livestock farm together with bedding material (straw) has been studied under mesophilic and thermophilic conditions in batch reactors. The digestion was carried out for a prolonged period with the aim of evaluating the changes undergone by the organic matter. The mesophilic digestion carried out revealed a greater capacity to produce gas and transform organic matter, while a higher conversion rate, but a lower gas yield, was obtained under thermophilic conditions. Degradation of the organic matter was evaluated by means of thermal analysis and (1)H NMR. Stabilisation through anaerobic digestion (either mesophilic or thermophilic) resulted in an increase in the quality of the organic matter, as characterised by an enrichment in thermostable compounds, and an accumulation of long chain aliphatic materials. The experiments performed demonstrated the transformation of organic matter into complex materials under anaerobic conditions with an accumulation of aliphatic components under both types of conditions tested. Degradation through mesophilic digestion, in comparison to the thermophilic process, resulted in a greater destruction of straw particles.

  2. Isolation and polyphasic characterization of a novel hyper catalase producing thermophilic bacterium for the degradation of hydrogen peroxide.

    PubMed

    Sooch, Balwinder Singh; Kauldhar, Baljinder Singh; Puri, Munish

    2016-11-01

    A newly isolated microbial strain of thermophilic genus Geobacillus has been described with emphasis on polyphasic characterization and its application for degradation of hydrogen peroxide. The validation of this thermophilic strain of genus Geobacillus designated as BSS-7 has been demonstrated by polyphasic taxonomy approaches through its morphological, biochemical, fatty acid methyl ester profile and 16S rDNA sequencing. This thermophilic species of Geobacillus exhibited growth at broad pH and temperature ranges coupled with production of extraordinarily high quantities of intracellular catalase, the latter of which as yet not been reported in any member of this genus. The isolated thermophilic bacterial culture BSS-7 exhibited resistance against a variety of organic solvents. The immobilized whole cells of the bacterium successfully demonstrated the degradation of hydrogen peroxide (H2O2) in a packed bed reactor. This strain has potential application in various analytical and diagnostic methods in the form of biosensors and biomarkers in addition to applications in the textile, paper, food and pharmaceutical industries. PMID:27450069

  3. Community analysis of hydrogen-producing extreme thermophilic anaerobic microflora enriched from cow manure with five substrates.

    PubMed

    Yokoyama, Hiroshi; Moriya, Naoko; Ohmori, Hideyuki; Waki, Miyoko; Ogino, Akifumi; Tanaka, Yasuo

    2007-11-01

    The present study analyzed the community structures of anaerobic microflora producing hydrogen under extreme thermophilic conditions by two culture-independent methods: denaturing gradient gel electrophoresis (DGGE) and clone library analyses. Extreme thermophilic microflora (ETM) was enriched from cow manure by repeated batch cultures at 75 degrees C, using a substrate of xylose, glucose, lactose, cellobiose, or soluble starch, and produced hydrogen at yields of 0.56, 2.65, 2.17, 2.68, and 1.73 mol/mol-monosaccharide degraded, respectively. The results from the DGGE and clone library analyses were consistent and demonstrated that the community structures of ETM enriched with the four hexose-based substrates (glucose, lactose, cellobiose, and soluble starch) consisted of a single species, closely related to a hydrogen-producing extreme thermophile, Caldoanaerobacter subterraneus, with diversity at subspecies levels. The ETM enriched with xylose was more diverse than those enriched with the other substrates, and contained the bacterium related to C. subterraneus and an unclassified bacterium, distantly related to a xylan-degrading and hydrogen-producing extreme thermophile, Caloramator fervidus.

  4. Isolation and characterization of a thermophilic Bacillus sp. with protease activity isolated from hot spring of Tarabalo, Odisha, India

    PubMed Central

    Panda, Mrunmaya Kumar; Sahu, Mahesh Kumar; Tayung, Kumananda

    2013-01-01

    Background and Objectives Thermophilic bacteria are less studied but important group of microorganisms due to their ability to produce industrial enzymes. Materials and Methods In this study, thermophilic bacteria were isolated from hot spring of Tarabalo, India. A bacterium that could tolerate high temperatures was characterized by morphology, biochemistry and sequencing of its 16S rRNA gene. The isolate was screened for protease and amylase activity. Phylogenetic affiliations and G+C content of the isolate was studied. Results The bacterium with the ability to tolerate high temperatures was identified as Bacillus sp. both by morphology, biochemistry and sequencing of its 16S rRNA gene. BLAST search analysis of the sequence showed maximum identity with Bacillus amyloliquefaciens (99% similarity). Strain exhibited considerable protease activity. Phylogenetic analysis of the isolate revealed close affiliation with thermophilic Bacillus species. The G+C content was found to be 54.7%. Conclusion The study confirmed that the isolated Bacillus sp. to be a true thermophile and could be a source of thermostable protease which can be exploited for pharmaceutical and industrials applications. PMID:23825735

  5. Solubilization of Waste Activated Sludge and Nitrogenous Compounds Transformation During Solubilization by Thermophilic Enzyme (S-TE) Process.

    PubMed

    Yang, Qi; Luo, Kun; Li, Xiao-ming; Zhong, Yu; Chen, Hong-bo; Yang, Guo-jing; Shi, Yan-wei; Zeng, Guang-ming

    2015-06-01

    A representative thermophilic bacterial strain (AT06-1) capable of secreting protease was isolated from thermophilic aerobic digestion reactor, and 16S rRNA gene analysis indicated that it was Bacillus sp. The isolated strain was inoculated in waste activated sludge (WAS) to evaluate the performance of solubilization by thermophilic enzyme (S-TE) process under aerobic or microaerobic conditions at different temperatures (55-70 °C). Results showed that the inoculation of specific thermophilic strain significantly affected the volatile suspended solids (VSS) removal. At the optimal temperature of 65 °C, the maximum VSS removal of 43.6 % and highest SCOD of 4475 mg/L was achieved during microaerobic S-TE process. Compared to the noninoculation, more soluble protein was released during S-TE process due to the higher protease activity associated with the protein hydrolysis originated from cell lysis. The protease activity at aerobic and microaerobic S-TE process was respectively 1.73 and 1.88 times that of the noninoculation. Ammonia was the end nitrogenous compound of protein hydrolysis during S-TE process, which was stripped from the digestion system through continuous aeration.

  6. 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.

  7. Digestion of cattle manure under mesophilic and thermophilic conditions: characterization of organic matter applying thermal analysis and 1H NMR.

    PubMed

    Gómez, X; Blanco, D; Lobato, A; Calleja, A; Martínez-Núñez, F; Martin-Villacorta, J

    2011-06-01

    Digestion of cattle manure collected from a livestock farm together with bedding material (straw) has been studied under mesophilic and thermophilic conditions in batch reactors. The digestion was carried out for a prolonged period with the aim of evaluating the changes undergone by the organic matter. The mesophilic digestion carried out revealed a greater capacity to produce gas and transform organic matter, while a higher conversion rate, but a lower gas yield, was obtained under thermophilic conditions. Degradation of the organic matter was evaluated by means of thermal analysis and (1)H NMR. Stabilisation through anaerobic digestion (either mesophilic or thermophilic) resulted in an increase in the quality of the organic matter, as characterised by an enrichment in thermostable compounds, and an accumulation of long chain aliphatic materials. The experiments performed demonstrated the transformation of organic matter into complex materials under anaerobic conditions with an accumulation of aliphatic components under both types of conditions tested. Degradation through mesophilic digestion, in comparison to the thermophilic process, resulted in a greater destruction of straw particles. PMID:21082330

  8. Studies on the composition and properties of thermophilic Bacillus populations in subterranean thermal waters of Budapest (Hungary).

    PubMed

    Martin, A M

    1997-01-01

    A collection of 148 thermophilic Bacillus isolates were obtained from two thermal wells (water temperature at about 70 degrees C) in the area of Budapest (Hungary). These isolates, furthermore seven authentic thermophilic reference strains of Bacillus spp. were submitted to detailed comparative biochemical-physiological studies. Overall similarities of these strains for 134 unit characters were determined by the SSM coefficient and clustering achieved using the UPGMA algorithm. According to the results of our computer aided numerical taxonomic analyses, numerous metabolic types, varieties and mutants of a single Bacillus sp. not identical with the known thermophilic Bacillus species constitute the major fraction of the indigenous thermophilic microbial communities in the thermal waters of the Pascal and Széchenyi wells. The data also revealed, (1) that the most adapted varieties of this Bacillus sp. can form the most dense local populations in these subterranean ecological systems. Furthermore, it seems to be probable (2) that the cells of the most commonly occurring varieties of this Bacillus sp. may be able to migrate in the underground thermal water systems between diverse subterranean regions located more or less far from one another.

  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. 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.

  12. 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.

  13. 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.

  14. Moderators of youth exercise intention and behavior.

    PubMed

    Ellis, Rebecca; Kosma, Maria; Symons Downs, Danielle

    2013-06-01

    This study tested moderators of the theory of planned behavior (TPB) based on geographical region, gender, race, and income among adolescents in an exercise context using multigroup path analyses. Participants were eighth- and ninth-grade students from Louisiana (LA; N = 448, M age = 14.37 years) and Pennsylvania (PA; N = 681, M age = 14.28 years). They completed measures of intention, attitude, subjective norm, perceived behavioral control, and exercise behavior. Based on two path analyses, geographical region was a significant moderator (p < .001); therefore, the moderating effects of gender, race, and income were analyzed separately for each state. Gender was a significant moderator for LA (p < .001) but not for PA (p = .90). Race and income did not moderate the TPB relationships within each state. Findings support the moderating effect of geographical region on the TPB construct relationships and indicate that gender moderates the TPB construct relationships in LA youth.

  15. 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

  16. 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

  17. Methods for integrating moderation and mediation: a general analytical framework using moderated path analysis.

    PubMed

    Edwards, Jeffrey R; Lambert, Lisa Schurer

    2007-03-01

    Studies that combine moderation and mediation are prevalent in basic and applied psychology research. Typically, these studies are framed in terms of moderated mediation or mediated moderation, both of which involve similar analytical approaches. Unfortunately, these approaches have important shortcomings that conceal the nature of the moderated and the mediated effects under investigation. This article presents a general analytical framework for combining moderation and mediation that integrates moderated regression analysis and path analysis. This framework clarifies how moderator variables influence the paths that constitute the direct, indirect, and total effects of mediated models. The authors empirically illustrate this framework and give step-by-step instructions for estimation and interpretation. They summarize the advantages of their framework over current approaches, explain how it subsumes moderated mediation and mediated moderation, and describe how it can accommodate additional moderator and mediator variables, curvilinear relationships, and structural equation models with latent variables.

  18. 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.

  19. 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

  20. Distribution of methanogenic potential in fractions of turf grass used as inoculum for the start-up of thermophilic anaerobic digestion.

    PubMed

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

    2012-08-01

    This study aims to investigate thermophilic methanogens in turf used as an inoculum. Results showed that Methanoculleus sp. regarded as hydrogenotrophic and Methanosarcina sp. regarded as acetoclastic methanogens were present in turf tested. However, active acetoclastic methanogens were present in turf soil only. The current study showed that thermophilic methanogens were present in various turf grass species: Stenotaphrum secundatum, Cynodon dactylon, and Zoysia japonica. Severe treatments of grass leaves under oxic conditions, including blending, drying and pulverizing did not affect the thermophilic hydrogenotrophic methanogenic activity of the grass. A dried and pulverized grass extract could be generated that can serve as a readily storable methanogenic inoculum for thermophilic anaerobic digestion. The methanogens could also be physically extracted into an aqueous suspension, suitable as an inoculum. The possible contribution of the presence of methanogens on grass plants to global greenhouse emissions is briefly discussed.

  1. 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.

  2. 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

  3. Survival of thermophilic campylobacters on fingertips and their elimination by washing and disinfection.

    PubMed Central

    Coates, D.; Hutchinson, D. N.; Bolton, F. J.

    1987-01-01

    A simple impression-plate technique has been used to investigate the survival of four thermophilic campylobacter strains applied to fingertips. Campylobacters suspended in 0.1% peptone water and dried on the fingertips survived for different periods of time ranging from less than 1 to greater than or equal to 4 min. However, campylobacters suspended in chicken liquor or blood survived for much longer periods. The most resilient organism was Campylobacter jejuni NCTC 11392 which, when suspended in 50% horse blood, survived for an hour. Suspensions containing 10(6)-10(7) organisms prepared in 50% blood and dried on to fingertips were removed by thorough hand washing with either soap and water or water alone followed by drying on paper towels, but persisted on wet hands. The organisms were also eliminated by wiping the hands with a tissue saturated with 70% isopropyl alcohol for 15 sec. PMID:3678391

  4. The effects of a thermophile metabolite, tryptophol, upon protecting shrimp against white spot syndrome virus.

    PubMed

    Zhu, Fei; Jin, Min

    2015-12-01

    White spot syndrome virus (WSSV) is a shrimp pathogen responsible for significant economic loss in commercial shrimp farms and until now, there has been no effective approach to control this disease. In this study, tryptophol (indole-3-ethanol) was identified as a metabolite involved in bacteriophage-thermophile interactions. The dietary addition of tryptophol reduced the mortality in shrimp Marsupenaeus japonicus when orally challenged with WSSV. Our results revealed that 50 mg/kg tryptophol has a better protective effect in shrimp than 10 or 100 mg/kg tryptophol. WSSV copies in shrimp were reduced significantly (P < 0.01) when supplemented with 50 mg/kg tryptophol, indicating that virus replication was inhibited by tryptophol. Consequently, tryptophol represents an effective antiviral dietary supplement for shrimp, and thus holds significant promise as a novel and efficient therapeutic approach to control WSSV in shrimp aquaculture.

  5. High thermal stability and unique trimer formation of cytochrome c' from thermophilic Hydrogenophilus thermoluteolus.

    PubMed

    Fujii, Sotaro; Masanari, Misa; Inoue, Hiroki; Yamanaka, Masaru; Wakai, Satoshi; Nishihara, Hirofumi; Sambongi, Yoshihiro

    2013-01-01

    Sequence analysis indicated that thermophilic Hydrogenophilus thermoluteolus cytochrome c' (PHCP) and its mesophilic homolog, Allochromatium vinosum cytochrome c' (AVCP), closely resemble each other in a phylogenetic tree of the cytochrome c' family, with 55% sequence identity. The denaturation temperature of PHCP was 87 °C, 35 °C higher than that of AVCP. Furthermore, PHCP exhibited a larger enthalpy change value during its thermal denaturation than AVCP. While AVCP was dimeric, as observed previously, PHCP was trimeric, and this was the first observation as a cytochrome c'. Dissociation of trimeric PHCP and its protein denaturation reversibly occurred at the same time in a two-state transition manner. Therefore, PHCP is enthalpically more stable than AVCP, perhaps due to its unique trimeric form, in addition to the lower number of Gly residues in its putative α-helical regions.

  6. Temperature-Driven Changeover in the Electron-Transfer Mechanism of a Thermophilic Plastocyanin.

    PubMed

    Olloqui-Sariego, José Luis; Moreno-Beltrán, Blas; Díaz-Quintana, Antonio; De la Rosa, Miguel A; Calvente, Juan José; Andreu, Rafael

    2014-03-01

    Electron-transfer kinetics of the thermophilic protein Plastocyanin from Phormidium laminosum adsorbed on 1,ω-alkanedithiol self-assembled monolayers (SAMs) deposited on gold have been investigated. The standard electron-transfer rate constant has been determined as a function of electrode-protein distance and solution viscosity over a broad temperature range (0-90 °C). For either thin or thick SAMs, the electron-transfer regime remains invariant with temperature, whereas for the 1,11-undecanethiol SAM of intermediate chain length, a kinetic regime changeover from a gated or friction-controlled mechanism at low temperature (0-30 °C) to a nonadiabatic mechanism above 40 °C is observed. To the best of our knowledge, this is the first time a thermal-induced transition between these two kinetic regimes is reported for a metalloprotein.

  7. Thermophilic biohydrogen production by an anaerobic heat treated-hot spring culture.

    PubMed

    Karadag, Dogan; Mäkinen, Annukka E; Efimova, Elena; Puhakka, Jaakko A

    2009-12-01

    Batch experiments were conducted to investigate the thermophilic biohydrogen production using an enrichment culture from a Turkish hot spring. Following the enrichment, the culture was heat treated at 100 degrees C for 10 min to select for spore-forming bacteria. H(2) production was accompanied by production of acetate, butyrate, lactate and ethanol. H(2) production was associated by acetate-butyrate type fermentation while accumulation of lactate and ethanol negatively affected the H(2) yield. H(2) production was highest in the temperature range from 49.6 to 54.8 degrees C and optimum values for initial pH and concentrations of iron, yeast extract and glucose were 6.5, 40 mg/l, 4-13.5 g/l, respectively. PCR-DGGE profiling showed that the heat treated culture consisted of species closely affiliated to genus Thermoanaerobacterium.

  8. 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

  9. Crystal structure of TTC0263, a thermophilic TPR protein from Thermus thermophilus HB27.

    PubMed

    Lim, Hyosun; Kim, Kyunggon; Han, Dohyun; Oh, Jongkil; Kim, Youngsoo

    2007-08-31

    The hypothetical protein TTC0263 of Thermus thermophilus HB27 is a thermophilic tetratricopeptide repeat (TPR)-containing protein. In the present study, the TPR region (residues 26-230) was resolved at 2.5 A with R-factors of R/Rfree = 23.6%/28.6%. TTC0263 consists of 11 helices that form five TPR units. Uniquely, it contains one atypical "extended" TPR (eTPR) unit. This comprises extended helical residues near the loop region of TTC0263, such that the helical length of eTPR is longer than that of the canonical TPR sequence. In addition, the hybrid TPR domain of TTC0263 possesses oligomer-forming characteristics. TPR domains are generally involved in forming multi-subunit complexes by interacting with each other or with other subunit proteins. The dynamic structure of TTC0263 described here goes some way to explaining how TPR domains mediate the formation of multi-subunit complexes.

  10. 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.

  11. The Geobacillus paradox: why is a thermophilic bacterial genus so prevalent on a mesophilic planet?

    PubMed

    Zeigler, Daniel R

    2014-01-01

    The genus Geobacillus comprises endospore-forming obligate thermophiles. These bacteria have been isolated from cool soils and even cold ocean sediments in anomalously high numbers, given that the ambient temperatures are significantly below their minimum requirement for growth. Geobacilli are active in environments such as hot plant composts, however, and examination of their genome sequences reveals that they are endowed with a battery of sensors, transporters and enzymes dedicated to hydrolysing plant polysaccharides. Although they appear to be relatively minor members of the plant biomass-degrading microbial community, Geobacillus bacteria have achieved a significant population with a worldwide distribution, probably in large part due to adaptive features of their spores. First, their morphology and resistance properties enable them to be mobilized in the atmosphere and transported long distances. Second, their longevity, which in theory may be extreme, enables them to lie quiescent but viable for long periods of time, accumulating gradually over time to achieve surprisingly high population densities.

  12. 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.

  13. 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.

  14. 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

  15. 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.

  16. Pyrophosphate as a central energy carrier in the hydrogen-producing extremely thermophilic Caldicellulosiruptor saccharolyticus.

    PubMed

    Bielen, Abraham A M; Willquist, Karin; Engman, Jakob; van der Oost, John; van Niel, Ed W J; Kengen, Servé W M

    2010-06-01

    The role of inorganic pyrophosphate (PPi) as an energy carrier in the central metabolism of the extremely thermophilic bacterium Caldicellulosiruptor saccharolyticus was investigated. In agreement with its annotated genome sequence, cell extracts were shown to exhibit PPi-dependent phosphofructokinase and pyruvate phosphate dikinase activity. In addition, membrane-bound pyrophosphatase activity was demonstrated, while no significant cytosolic pyrophosphatase activity was detected. During the exponential growth phase, high PPi levels (approximately 4 +/- 2 mM) and relatively low ATP levels (0.43 +/- 0.07 mM) were found, and the PPi/ATP ratio decreased 13-fold when the cells entered the stationary phase. Pyruvate kinase activity appeared to be allosterically affected by PPi. Altogether, these findings suggest an important role for PPi in the central energy metabolism of C. saccharolyticus.

  17. 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.

  18. Anaerobic thermophilic digestion of sewage sludge with a thickened sludge recycle.

    PubMed

    Vanyushina, A Ya; Nikolaev, Yu A; Agarev, A M; Kevbrina, M V; Kozlov, M N

    2012-01-01

    The process of anaerobic thermophilic digestion of municipal wastewater sludge with a recycled part of thickened digested sludge, was studied in semi-continuous laboratory digesters. This modified recycling process resulted in increased solids retention time (SRT) with the same hydraulic retention time (HRT) as compared with traditional digestion without recycling. Increased SRT without increasing of HRT resulted in the enhancement of volatile substance reduction by up to 68% in the reactor with the recycling process compared with 34% in a control conventional reactor. Biogas production was intensified from 0.3 L/g of influent volatile solids (VS) in the control reactor up to 0.35 L/g VS. In addition, the recycling process improved the dewatering properties of digested sludge. PMID:22258668

  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. 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

  1. Isolation, purification and characterization of the ATPase complex from the thermophilic cyanobacterium Synechococcus 6716.

    PubMed

    Lubberding, H J; Zimmer, G; van Walraven, H S; Schrickx, J; Kraayenhof, R

    1983-12-01

    The ATPase complex is isolated and purified from membrane vesicles of the thermophilic cyanobacterium Synechococcus 6716 by octyl glucoside and cholic acid by a modification of the procedure for its extraction from spinach chloroplasts. The complex is purified by differential centrifugation and ammonium sulfate precipitation and by gel filtration on Sepharose 6B. The purified fraction, without any phycocyanin contamination, shows ATP hydrolysis activity and Pi/ATP exchange activity of 1564 and 350 nmol X min-1 X mg protein-1, respectively. N,N'-Dicyclohexylcarbodiimide inhibits the ATP hydrolysis activity of this purified fraction. On polyacrylamide gels most typical F1 ATPase polypeptides are identified, but the low-molecular weight polypeptides visible cannot be ascribed to the F0 part of the complex with certainty; non-identified bands around 30 kDa are also present.

  2. 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

  3. 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

  4. 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

  5. Production and properties of an alkaline, thermophilic lipase from Pseudomonas fluorescens NS2W.

    PubMed

    Kulkarni, N; Gadre, R V

    2002-06-01

    Eighteen bacterial strains were isolated from soil samples and screened for alkaline, thermophilic lipase production. Pseudomonas fluorescens NS2W was selected and its production of lipase was optimized in shake flasks using a statistical experimental design. Cell growth and lipase production were studied in shake flasks and in a 1-l fermenter in the optimized medium. Maximum lipase yields were 69.7 and 68.7 U ml(-1), respectively. The optimized medium resulted in about a five-fold increase in the enzyme production, compared to that obtained in the basal medium. The lipase had an optimal activity at pH 9.0 and was stable over a wide pH range of 3-11 with more than 70% activity retention. The lipase had an optimal activity at 55 degrees C and was stable up to 60 degrees C with more than 70% activity retention for at least 2 h. PMID:12032808

  6. Temperature adaptations in psychrophilic, mesophilic and thermophilic chloride-dependent alpha-amylases.

    PubMed

    Cipolla, Alexandre; Delbrassine, François; Da Lage, Jean-Luc; Feller, Georges

    2012-09-01

    The functional and structural adaptations to temperature have been addressed in homologous chloride-dependent α-amylases from a psychrophilic Antarctic bacterium, the ectothermic fruit fly, the homeothermic pig and from a thermophilic actinomycete. This series covers nearly all temperatures encountered by living organisms. We report a striking continuum in the functional properties of these enzymes coupled to their structural stability and related to the thermal regime of the source organism. In particular, thermal stability recorded by intrinsic fluorescence, circular dichroism and differential scanning calorimetry appears to be a compromise between the requirement for a stable native state and the proper structural dynamics to sustain the function at the environmental/physiological temperatures. The thermodependence of activity, the kinetic parameters, the activations parameters and fluorescence quenching support these activity-stability relationships in the investigated α-amylases.

  7. Thermophilic Fe(III)-reducing bacteria from the deep subsurface: The evolutionary implications

    SciTech Connect

    Liu, Shi V.; Zhou, Jizhong; Phelps, T.J.

    1997-08-22

    Thermophilic (45{degrees} to 75{degrees}C) bacteria that reduce amorphous Fe(III)-oxyhydroxide magnetic iron oxides have been discovered in two geologically and hydrologically isolated Cretaceous- and Triassic-age sedimentary basins in the deep (>860 meters below land surface) terrestrial subsurface. Molecular analyses based on 16S ribosomal RNA (rRNA) gene sequences revealed that some of these bacteria represent unrecognized phylogenetic group of dissimilatory Fe(III)-reducing bacteria. This covery adds another dimension to the study of microbial Fe(III)-reduction and biogenic magnetism. It also provides examples for understanding the history of Fe(III)-reducing microorganisms and for assessing possible roles of such microorganisms on hot primitive planets.

  8. Proteolysin, a Novel Highly Thermostable and Cosolvent-Compatible Protease from the Thermophilic Bacterium Coprothermobacter proteolyticus

    PubMed Central

    Toplak, Ana; Wu, Bian; Fusetti, Fabrizia; Quaedflieg, Peter J. L. M.

    2013-01-01

    Through genome mining, we identified a gene encoding a putative serine protease of the thermitase subgroup of subtilases (EC 3.4.21.66) in the thermophilic bacterium Coprothermobacter proteolyticus. The gene was functionally expressed in Escherichia coli, and the enzyme, which we called proteolysin, was purified to near homogeneity from crude cell lysate by a single heat treatment step. Proteolysin has a broad pH tolerance and is active at temperatures of up to 80°C. In addition, the enzyme shows good activity and stability in the presence of organic solvents, detergents, and dithiothreitol, and it remains active in 6 M guanidinium hydrochloride. Based on its stability and activity profile, proteolysin can be an excellent candidate for applications where resistance to harsh process conditions is required. PMID:23851086

  9. 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.

  10. 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

  11. Fecal shedding of thermophilic Campylobacter in a dairy herd producing raw milk for direct human consumption.

    PubMed

    Merialdi, Giuseppe; Giacometti, Federica; Bardasi, Lia; Stancampiano, Laura; Taddei, Roberta; Serratore, Patrizia; Serraino, Andrea

    2015-03-01

    Factors affecting the fecal shedding of thermophilic Campylobacter in Italian dairy farms were investigated in a 12-month longitudinal study performed on a dairy farm authorized to sell raw milk in Italy. Fifty animals were randomly selected from 140 adult and young animals, and fecal samples were collected six times at 2-month intervals. At each sampling time, three trough water samples and two trough feed samples also were collected for both adult and young animals. Samples were analyzed with real-time PCR assay and culture examination. Overall, 33 samples (9.7%) were positive for thermophilic Campylobacter by real-time PCR: 26 (9.2%) of 280 fecal samples, 6 (16.6%) of 36 water samples, and 1 (4.2%) of 24 feed samples. Campylobacter jejuni was isolated from 6 of 280 samples; no other Campylobacter species was isolated. A higher (but not significantly) number of positive fecal samples were found in younger animals (11.33 versus 6.92% of adult animals), and a significantly higher number of positive water samples were collected from the water troughs of young animals. A distinct temporal trend was observed during the study period for both cows and calves, with two prevalence peaks between November and December and between May and July. Several factors such as calving, housing practices, herd size, management practices forcing together a higher number of animals, and variations in feed or water sources (previously reported as a cause of temporal variation in different farming conditions) were excluded as the cause of the two seasonal peaks in this study. The factors affecting the seasonality of Campylobacter shedding in the dairy herds remain unclear and warrant further investigation. The results of the present study indicate that special attention should be paid to farm hygiene management on farms authorized to produce and sell raw milk, with increased surveillance by the authorities at certain times of the year.

  12. 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

  13. Structure and Stability of the Dimeric Triosephosphate Isomerase from the Thermophilic Archaeon Thermoplasma acidophilum.

    PubMed

    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.

  14. Screening of thermophilic anaerobic bacteria for solid substrate cultivation on lignocellulosic substrates.

    PubMed

    Chinn, Mari S; Nokes, Sue E; Strobel, Herbert J

    2006-01-01

    Interest in solid substrate cultivation (SSC) techniques is gaining for biochemical production from renewable resources; however, heat and mass transfer problems may limit application of this technique. The use of anaerobic thermophiles in SSC offers a unique solution to overcoming these challenges. The production potential of nine thermophilic anaerobic bacteria was examined on corn stover, sugar cane bagasse, paper pulp sludge, and wheat bran in submerged liquid cultivation (SmC) and SSC. Production of acetate, ethanol, and lactate was measured over a 10 day period, and total product concentrations were used to compare the performance of different organism-substrate combinations using the two cultivation methods. Overall microbial activity in SmC and SSC was dependent on the organism and growth substrate. Clostridium thermocellum strains JW20, LQRI, and 27405 performed significantly better in SSC when grown on sugar cane bagasse and paper pulp sludge, producing at least 70 and 170 mM of total products, respectively. Growth of C. thermocellum strains in SSC on paper pulp sludge proved to be most favorable, generating at least twice the concentration of total products produced in SmC (p-value < 0.05). Clostridium thermolacticum TC21 demonstrated growth on all substrates producing 30-80 and 60-116 mM of total product in SmC and SSC, respectively. Bacterial species with optimal growth temperatures of 70 degrees C grew best on wheat bran in SmC, producing total product concentrations of 45-75 mM. For some of the organism-substrate combinations total end product concentrations in SSC exceeded those in SmC, indicating that SSC may be a promising alternative for microbial activity and value-added biochemical production.

  15. Purification and properties of thermostable tryptophanase from an obligately symbiotic thermophile, Symbiobacterium thermophilum.

    PubMed

    Suzuki, S; Hirahara, T; Horinouchi, S; Beppu, T

    1991-12-01

    A thermostable tryptophanase was extracted from a thermophilic bacterium, Symbiobacterium thermophilum strain T, which is obligately symbiotic with the thermophilic Bacillus strain S. The enzyme was purified 21-fold to homogeneity with 19% recovery by a series of chromatographies using anion-exchange, hydroxylapatite, hydrophobic interaction, and MonoQ anion-exchange columns. The molecular weight of the purified enzyme was estimated to be approximately 210,000 by gel filtration, while the molecular weight of its subunit was 46,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, which indicates that the native enzyme is composed of four homologous subunits. The isoelectric point of the enzyme was 4.9. The tryptophanase was stable to heating at 65 degrees C for 20 min and the optimum temperature for the enzyme activity for 20 min reaction was 70 degrees C. The optimum pH was 7.0. The NH2-terminal amino acid sequence of this tryptophanase shows similarity to that of Escherichia coli K-12, despite a great difference in the thermostability of these two enzymes. The purified enzyme catalyzed the degradation (alpha, beta-elimination) of L-tryptophan into indole, pyruvate, and ammonia in the presence of pyridoxal-5'-phosphate. The Km value for L-tryptophan was 1.47 mM. 5-Hydroxy-L-tryptophan, 5-methyl-DL-tryptophan, L-cysteine, S-methyl-L-cysteine, and L-serine were also used as substrates and converted to pyruvate. The reverse reaction of alpha, beta-elimination of this tryptophanase produced L-tryptophan from indole and pyruvate in the presence of a high concentration of ammonium acetate.

  16. 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.

  17. Silver Sulfidation in Thermophilic Anaerobic Digesters and Effects on Antibiotic Resistance Genes

    SciTech Connect

    Kim, Bojeong; Miller, Jennifer H.; Monsegue, Niven; Levard, Clément; Hong, Yanjuan; Hull, Matthew S.; Murayama, Mitsuhiro; Brown, Gordon E.; Vikesland, Peter J.; Knocke, William R.; Pruden, Amy; Hochella, Michael F.

    2015-12-15

    Physical and chemical transformations and biological responses of silver nanoparticles (AgNPs) in wastewater treatment systems are of particular interest because of the extensive existing and continually growing uses of AgNPs in consumer products. In this study, we investigated the transformation of AgNPs and AgNO3 during thermophilic anaerobic digestion and effects on selection or transfer of antibiotic resistance genes (ARGs). Ag2S-NPs, sulfidation products of both AgNPs and AgNO3, were recovered from raw and digested sludges and were analyzed by analytical transmission electron microscopy (TEM) and X-ray absorption spectroscopy (XAS). TEM and XAS revealed rapid (≤20 min) Ag sulfidation for both Ag treatments. Once transformed, Ag2S-NPs (as individual NPs or an NP aggregate) persisted for the duration of the batch digestion. The digestion process produced Ag2S-NPs that were strongly associated with sludge organics and/or other inorganic precipitates. Ag treatments (up to 1,000 mg Ag/kg) did not have an impact on the performance of thermophilic anaerobic digesters or ARG response, as indicated by quantitative polymerase chain reaction measurements of sul1, tet(W), and tet(O) and also intI1, an indicator of horizontal gene transfer of ARGs. Thus, rapid Ag sulfidation and stabilization with organics effectively sequester Ag and prevent biological interactions with the digester microbial community that could induce horizontal gene transfer or adversely impact digester performance through antimicrobial activity. This finding suggests that sulfide-rich anaerobic environments, such as digesters, likely have a high buffer capacity to mitigate the biological effects of AgNPs.

  18. 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.

  19. 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

  20. A novel ecological role of the Firmicutes identified in thermophilic microbial fuel cells.

    PubMed

    Wrighton, Kelly C; Agbo, Peter; Warnecke, Falk; Weber, Karrie A; Brodie, Eoin L; DeSantis, Todd Z; Hugenholtz, Philip; Andersen, Gary L; Coates, John D

    2008-11-01

    Significant effort is currently focused on microbial fuel cells (MFCs) as a source of renewable energy. Most studies concentrate on operation at mesophilic temperatures. However, anaerobic digestion studies have reported on the superiority of thermophilic operation and demonstrated a net energy gain in terms of methane yield. As such, our studies focused on MFC operation and microbiology at 55 degrees C. Over a 100-day operation, these MFCs were stable and achieved a power density of 37 mW m(-2) with a coulombic efficiency of 89%. To infer activity and taxonomic identity of dominant members of the electricity-producing community, we performed phylogenetic microarray and clone library analysis with small subunit ribosomal RNA (16S rRNA) and ribosomal RNA gene (16S rDNA). The results illustrated the dominance (80% of clone library sequences) of the Firmicutes in electricity production. Similarly, rRNA sequences from Firmicutes accounted for 50% of those taxa that increased in relative abundance from current-producing MFCs, implying their functional role in current production. We complemented these analyses by isolating the first organisms from a thermophilic MFC. One of the isolates, a Firmicutes Thermincola sp. strain JR, not only produced more current than known organisms (0.42 mA) in an H-cell system but also represented the first demonstration of direct anode reduction by a member of this phylum. Our research illustrates the importance of using a variety of molecular and culture-based methods to reliably characterize bacterial communities. Consequently, we revealed a previously unidentified functional role for Gram-positive bacteria in MFC current generation. PMID:18769460