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Sample records for anaerobic protein-degrading hyperthermophilic

  1. Complete genome sequence of the anaerobic, protein-degrading hyperthermophilic crenarchaeon Desulfurococcus kamchatkensis.

    Science.gov (United States)

    Ravin, Nikolai V; Mardanov, Andrey V; Beletsky, Alexey V; Kublanov, Ilya V; Kolganova, Tatiana V; Lebedinsky, Alexander V; Chernyh, Nikolai A; Bonch-Osmolovskaya, Elizaveta A; Skryabin, Konstantin G

    2009-04-01

    Desulfurococcus kamchatkensis is an anaerobic organotrophic hyperthermophilic crenarchaeon isolated from a terrestrial hot spring. Its genome consists of a single circular chromosome of 1,365,223 bp with no extrachromosomal elements. A total of 1,474 protein-encoding genes were annotated, among which 205 are exclusive for D. kamchatkensis. The search for a replication origin site revealed a single region coinciding with a global extreme of the nucleotide composition disparity curve and containing a set of crenarchaeon-type origin recognition boxes. Unlike in most archaea, two genes encoding homologs of the eukaryotic initiator proteins Orc1 and Cdc6 are located distantly from this site. A number of mobile elements are present in the genome, including seven transposons representing IS607 and IS200/IS605 families and multiple copies of miniature inverted repeat transposable elements. Two large clusters of regularly interspaced repeats are present; none of the spacer sequences matches known archaeal extrachromosomal elements, except one spacer matches the sequence of a resident gene of D. kamchatkensis. Many of the predicted metabolic enzymes are associated with the fermentation of peptides and sugars, including more than 30 peptidases with diverse specificities, a number of polysaccharide degradation enzymes, and many transporters. Consistently, the genome encodes both enzymes of the modified Embden-Meyerhof pathway of glucose oxidation and a set of enzymes needed for gluconeogenesis. The genome structure and content reflect the organism's nutritionally diverse, competitive natural environment, which is periodically invaded by viruses and other mobile elements.

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

    DEFF Research Database (Denmark)

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

    2008-01-01

    The present study focuses on a two-step process for treatment and stabilisation of primary sludge. The process consists of a hyperthermophilic hydrolysis step operated at 70 degrees C and a hydraulic retention time (HRT) of 2 clays followed by a thermophilic (55 degrees C) anaerobic digestion step...... at a HRT of 13 days. A one-step anaerobic digester operated at 55 degrees C and 15 days HRT Was used as a reference process. The two-step process was characterized by a 12% higher organic suspended solids removal efficiency and better pathogen reduction effect than the conventional one-step digestion....... The microbial community of the digester fed with pre-treated sludge was characterised by it higher activity compared to that of the digester treating raw sludge. Moreover, the pre-treatment of the primary sludge resulted up to 48% increase of the methane potential (20.09 and 13.56 mmol CH4 g(-)VS(-1...

  3. Thermogladius calderae gen. nov., sp. nov., an anaerobic, hyperthermophilic crenarchaeote from a Kamchatka hot spring.

    Science.gov (United States)

    Kochetkova, Tatiana V; Kublanov, Ilya V; Toshchakov, Stepan V; Osburn, Magdalena R; Novikov, Andrei A; Bonch-Osmolovskaya, Elizaveta A; Perevalova, Anna A

    2016-01-21

    An obligately anaerobic, hyperthermophilic, organoheterotrophic archaeon, strain 1633T, was isolated from a terrestrial hot spring of the Uzon Caldera (Kamchatka Peninsula, Russia). Cells were regular cocci, 0.5-0.9 μm in diameter, with one flagellum. The temperature range for growth was 80-95°C, with an optimum at 84°C. Strain 1633T grew on yeast extract, beef extract, peptone, cellulose and cellobiose. No growth was detected on other sugars or carbohydrates, organic acids, or under autotrophic conditions. The only detected growth products were CO2, acetate, and H2. Growth rate was stimulated by elemental sulfur, which was reduced to hydrogen sulfide. In silico calculated G+C content of strain 1633T genomic DNA was 55.64 mol%. 16S rRNA gene sequence analysis placed the strain 1633T together with the non-validly published "Thermogladius shockii" strain WB1 in a separate genus-level cluster within the Desulfurococcaceae family. ANI results revealed 75.72% identity between 1633T and WB1. Based on these results we propose a novel genus and species, for which the name Thermogladius calderae gen. nov., sp. nov. (type strain 1633T=DSM 22663T=VKM B-2946T) is proposed.

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

    Science.gov (United States)

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

    2016-09-01

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

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

    DEFF Research Database (Denmark)

    Hartmann, H.; Ahring, Birgitte Kiær

    2005-01-01

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

  6. Metabolism of hyperthermophiles.

    Science.gov (United States)

    Schönheit, P; Schäfer, T

    1995-01-01

    Hyperthermophiles are characterized by a temperature optimum for growth between 80 and 110°C. They are considered to represent the most ancient phenotype of living organisms and thus their metabolic design might reflect the situation at an early stage of evolution. Their modes of metabolism are diverse and include chemolithoautotrophic and chemoorganoheterotrophic. No extant phototrophic hyperthermophiles are known. Lithotrophic energy metabolism is mostly anaerobic or microaerophilic and based on the oxidation of H2 or S coupled to the reduction of S, SO inf4 (sup2-) , CO2 and NO inf3 (sup-) but rarely to O2. the substrates are derived from volcanic activities in hyperthermophilic habitats. The lithotrophic energy metabolism of hyperthermophiles appears to be similar to that of mesophiles. Autotrophic CO2 fixation proceeds via the reductive citric acid cycle, considered to be one of the first metabolic cycles, and via the reductive acetyl-CoA/carbon monoxide dehydrogenase pathway. The Calvin cycle has not been found in hyperthermophiles (or any Archaea). Organotrophic metabolism mainly involves peptides and sugars as substrates, which are either oxidized to CO2 by external electron acceptors or fermented to acetate and other products. Sugar catabolism in hyperthermophiles involves non-phosphorylated versions of the Entner-Doudoroff pathway and modified versions of the Embden-Meyerhof pathway. The 'classical' Embden-Meyerhof pathway is present in hyperthermophilic Bacteria (Thermotoga) but not in Archaea. All hyperthermophiles (and Archaea) tested so far utilize pyruvate:ferredoxin oxidoreductase for acetyl-CoA formation from pyruvate. Acetyl-CoA oxidation in anaerobic sulphur-reducing and aerobic hyperthermophiles proceeds via the citric acid cycle; in the hyperthermophilic sulphate-reducer Archaeoglobus an oxidative acetyl-CoA/carbon monoxide dehydrogenase pathway is operative. Acetate formation from acetyl-CoA in Archaea, including hyperthermophiles, is

  7. Rat myocardial protein degradation.

    Science.gov (United States)

    Steer, J H; Hopkins, B E

    1981-07-01

    1. Myocardial protein degradation rates were determined by following tyrosine release from rat isolated left hemi-atria in vitro. 2. After two 20 min preincubations the rate of tyrosine release from hemi-atria was constant for 4 h. 3. Skeletal muscle protein degradation was determined by following tyrosine release from rat isolated hemi-diaphragm (Fulks, Li & Goldberg, 1975). 4. Insulin (10(-7) M) inhibited tyrosine release from hemi-atria and hemi-diaphragm to a similar extent. A 48 h fast increased tyrosine release rate from hemi-diaphragm and decreased tyrosine release rate from hemi-atria. Hemi-diaphragm tyrosine release was inhibited by 15 mmol/l D-glucose but a variety of concentrations of D-glucose (0, 5, 15 mmol/l) had no effect on tyrosine release from hemi-atria. Five times the normal plasma levels of the branched-chain amino acids leucine, isoleucine and valine had no effect on tyrosine release from either hemi-atria or hemi-diaphragm.

  8. POLYPEPTIDE AND POLYSACCHARIDE PROCESSING IN HYPERTHERMOPHILIC MICROORGANISMS

    Energy Technology Data Exchange (ETDEWEB)

    KELLY, ROBERT M.

    2008-12-22

    This project focused on the microbial physiology and biochemistry of heterotrophic hyperthermophiles with respect to mechanisms by which these organisms process polypeptides and polysaccharides under normal and stressed conditions. Emphasis is on two model organisms, for which completed genome sequences are available: Pyrococcus furiosus (growth Topt of 98°C), an archaeon, and Thermotoga maritima (growth Topt of 80°C), a bacterium. Both organisms are obligately anaerobic heterotrophs that reduce sulfur facultatively. Whole genome cDNA spotted microarrays were used to follow transcriptional response to a variety of environmental conditions in order to identify genes encoding proteins involved in the acquisition, synthesis, processing and utilization of polypeptides and polysaccharides. This project provided new insights into the physiological aspects of hyperthermophiles as these relate to microbial biochemistry and biological function in high temperature habitats. The capacity of these microorganisms to produce biohydrogen from renewable feedstocks makes them important for future efforts to develop biofuels.

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

    Science.gov (United States)

    Cha, Minseok; Chung, Daehwan; Westpheling, Janet

    2016-02-01

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

  10. A First Analysis of Metallome Biosignatures of Hyperthermophilic Archaea

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    Vyllinniskii Cameron

    2012-01-01

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

  11. Stability of Hyperthermophilic Proteins

    DEFF Research Database (Denmark)

    Stiefler-Jensen, Daniel

    cheaper products. One aspect that can have a large impact on the efficiency of an enzyme is its stability. By increasing the enzyme stability production cost and time can be reduced, and consumers will have a better product with longer activity. In the past it was only possible to increasing enzymes...... stability by randomly generate mutants and lengthy screening processes to identify the best new mutants. However, with the increase in available genomic sequences of thermophilic or hyperthermophilic organisms a world of enzymes with intrinsic high stability are now available. As these organisms are adapted...... to life at high temperatures so are their enzymes, as a result the high stability is accompanied by low activity at moderate temperatures. Thus, much effort had been put into decoding the mechanisms behind the high stability of the thermophilic enzymes. The hope is to enable scientist to design enzymes...

  12. Habitability of Mars: hyperthermophiles in permafrost

    Science.gov (United States)

    Gilichinsky, David; Rivkina, Elizaveta; Vishnivetskaya, Tatiana; Felipe, Gomez; Mironov, Vasilii; Blamey, Jenny; Ramos, Miguel; Ángel de Pablo, Miguel; Castro, Miguel; Boehmwald, Freddy

    This is a first microbiological study of volcanic permafrost carried out on Kluchevskaya volcano group (Kamchatka Peninsula) and Deception Island (Antarctica). By culture-and culture-independent methods we showed the presence of viable hyper(thermophilic) microorganisms and their genes within volcanic permafrost. The optimal temperature for sulfide producing bacteria was 65, whereas acetogens and methanogens were able to produce acetate and methane at temperatures up to 75o C, while sulphur-reducers showed optimal growth at 85-92o C. Hy-per(thermophiles) were never found in permafrost outside the volcanic areas before. The only way they are to appear within a frozen material is a concurrent deposition during the eruption, together with products associated with volcano heated subsurface geothermal oases. The elo-quent evidence to the hypothesis is the presence among clones of the sequences affiliated with (hyper)thermophilic bacteria, both, aerobic and anaerobic, in the environmental DNA derived from ashes freshly deposited on snow in close proximity to volcano Shiveluch (Kamchatka) and aerobic bacteria incubated at 80o C from ashes freshly deposited on the top of Llaima Vol-cano glacier (Andes). Thus, in the areas of active volcanism the catastrophic geological events transports the life from the depths to the surface and this life from high-temperature ecological niches might survive in permafrost over a long period of time. The results obtained give insights for habitability of Mars. Terrestrial permafrost represents a possible ecosystem for Mars as an Earth-like cryogenic planet. But permafrost on Earth and Mars vary in age, from a few million years on Earth to a few billion years on Mars. Because such difference in age, the longevity of life forms preserved within terrestrial permafrost may only serve as an approximate model for Mars. On the other hand, numerous ancient extinct volcanoes are known on Mars. Their past eruptions periodically burn-through the

  13. Inhibitors and pathways of hepatocytic protein degradation.

    Science.gov (United States)

    Seglen, P O; Gordon, P B; Grinde, B; Solheim, A; Kovács, A L; Poli, A

    1981-01-01

    On the basis of experiments using amino acids and various inhibitors (lysosomotropic amines, leupeptin, chymostatin, vanadate, vinblastine, anoxia, methylaminopurines), five different modes of endogenous protein degradation in isolated rat hepatocytes can be distinguished. The two non-lysosomal (amine-resistant) mechanisms preferentially degrade relatively labile (short-lived) proteins: one of these mechanisms is energy-dependent and chymostatin-sensitive, the other is not. Of the three lysosomal (amine-sensitive) mechanisms, one--quantitatively minor--is amino acid-resistant and preferentially degrades labile proteins. The two amino acid-sensitive mechanisms each seen account for about one-half of the degradation of relatively stable (long-lived) proteins; one of them is suppressed by leucine and apparently corresponds to the formation of electron microscopically visible autophagosomes; the other may represent a different type of autophagy, inhibited by asparagine and glutamine. A new class of inhibitors, the purine derivatives (methylated 6-aminopurines, and 6-mercaptopurines) appear to specifically suppress autophagic/lysosomal protein degradation, and may help to further elucidate the mechanisms of autophagy.

  14. Complete genome sequence of Desulfurococcus fermentans, a hyperthermophilic cellulolytic crenarchaeon isolated from a freshwater hot spring in Kamchatka, Russia.

    Science.gov (United States)

    Susanti, Dwi; Johnson, Eric F; Rodriguez, Jason R; Anderson, Iain; Perevalova, Anna A; Kyrpides, Nikos; Lucas, Susan; Han, James; Lapidus, Alla; Cheng, Jan-Fang; Goodwin, Lynne; Pitluck, Sam; Mavrommatis, Konstantinos; Peters, Lin; Land, Miriam L; Hauser, Loren; Gopalan, Venkat; Chan, Patricia P; Lowe, Todd M; Atomi, Haruyuki; Bonch-Osmolovskaya, Elizaveta A; Woyke, Tanja; Mukhopadhyay, Biswarup

    2012-10-01

    Desulfurococcus fermentans is the first known cellulolytic archaeon. This hyperthermophilic and strictly anaerobic crenarchaeon produces hydrogen from fermentation of various carbohydrates and peptides without inhibition by accumulating hydrogen. The complete genome sequence reported here suggested that D. fermentans employs membrane-bound hydrogenases and novel glycohydrolases for hydrogen production from cellulose.

  15. Complete Genome Sequence of Desulfurococcus fermentans, a Hyperthermophilic Cellulolytic Crenarchaeon Isolated from a Freshwater Hot Spring in Kamchatka, Russia

    Energy Technology Data Exchange (ETDEWEB)

    Susanti, Dwi [Virginia Polytechnic Institute and State University (Virginia Tech); Johnson, Eric F [Virginia Polytechnic Institute and State University (Virginia Tech); Rodriquez, Jason [Virginia Polytechnic Institute and State University (Virginia Tech); Anderson, Iain [U.S. Department of Energy, Joint Genome Institute; Perevalova, Anna [Virginia Polytechnic Institute and State University (Virginia Tech); Kyrpides, Nikos C [U.S. Department of Energy, Joint Genome Institute; Lucas, Susan [U.S. Department of Energy, Joint Genome Institute; Han, James [U.S. Department of Energy, Joint Genome Institute; Lapidus, Alla L. [U.S. Department of Energy, Joint Genome Institute; Cheng, Jan-Fang [U.S. Department of Energy, Joint Genome Institute; Goodwin, Lynne A. [Los Alamos National Laboratory (LANL); Pitluck, Sam [U.S. Department of Energy, Joint Genome Institute; Mavromatis, K [U.S. Department of Energy, Joint Genome Institute; Peters, Lin [U.S. Department of Energy, Joint Genome Institute; Land, Miriam L [ORNL; Hauser, Loren John [ORNL; Gopapan, Venkay [Ohio State University; Chan, Patricia [University of California, Santa Cruz; Atomi, Haruyuki [Kyoto University, Japan; Bonch-Osmolovskaya, Elizaveta [Russian Academy of Sciences, Moscow; Woyke, Tanja [U.S. Department of Energy, Joint Genome Institute; Mukhopadhyay, Biswarup [Virginia Polytechnic Institute and State University (Virginia Tech)

    2012-01-01

    Desulfurococcus fermentans is the first known cellulolytic archaeon. This hyperthermophilic and strictly anaerobic crenarchaeon produces hydrogen from fermentation of various carbohydrates and peptides without inhibition by accumulating hydrogen. The complete genome sequence reported here suggested that D. fermentans employs membrane-bound hydrogenases and novel glycohydrolases for hydrogen production from cellulose.

  16. A dicarboxylate/4-hydroxybutyrate autotrophic carbon assimilation cycle in the hyperthermophilic Archaeum Ignicoccus hospitalis

    OpenAIRE

    Huber, Harald; Gallenberger, Martin; Jahn, Ulrike; Eylert, Eva; Berg, Ivan A.; Kockelkorn, Daniel; Eisenreich, Wolfgang; Fuchs, Georg

    2008-01-01

    Ignicoccus hospitalis is an anaerobic, autotrophic, hyperthermophilic Archaeum that serves as a host for the symbiotic/parasitic Archaeum Nanoarchaeum equitans. It uses a yet unsolved autotrophic CO2 fixation pathway that starts from acetyl-CoA (CoA), which is reductively carboxylated to pyruvate. Pyruvate is converted to phosphoenol-pyruvate (PEP), from which glucogenesis as well as oxaloacetate formation branch off. Here, we present the complete metabolic cycle by which the primary CO2 acce...

  17. Anaerobic Thermophiles

    Directory of Open Access Journals (Sweden)

    Francesco Canganella

    2014-02-01

    Full Text Available 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

  18. Enhancing Heat Tolerance of the Little Dogwood Cornus canadensis L. f. with Introduction of a Superoxide Reductase Gene from the Hyperthermophilic Archaeon Pyrococcus furiosus.

    Science.gov (United States)

    Geng, Xing-Min; Liu, Xiang; Ji, Mikyoung; Hoffmann, William A; Grunden, Amy; Xiang, Qiu-Yun J

    2016-01-01

    Production of reactive oxygen species (ROS) can be accelerated under various biotic and abiotic stresses causing lipid peroxidation, protein degradation, enzyme inactivation, and DNA damage. Superoxide reductase (SOR) is a novel antioxidant enzyme from Pyrococcus furiosus and is employed by this anaerobic hyperthermophilic archaeon for efficient detoxification of ROS. In this study, SOR was introduced into a flowering plant Cornus canadensis to enhance its heat tolerance and reduce heat induced damage. A fusion construct of the SOR gene and Green Fluorescent Protein gene (GFP) was introduced into C. canadensis using Agrobacterium-mediated transformation. Heat tolerance of the GFP-SOR expressing transgenic plants was investigated by observing morphological symptoms of heat injury and by examining changes in photosynthesis, malondialdehyde (MDA), and proline levels in the plants. Our results indicate that the expression of the P. furiosus SOR gene in the transgenic plants alleviated lipid peroxidation of cell membranes and photoinhibition of PS II, and decreased the accumulation of proline at 40°C. After a series of exposures to increasing temperatures, the SOR transgenic plants remained healthy and green whereas most of the non-transgenic plants dried up and were unable to recover. While it had previously been reported that expression of SOR in Arabidopsis enhanced heat tolerance, this is the first report of the successful demonstration of improved heat tolerance in a non-model plant resulting from the introduction of P. furiosus SOR. The study demonstrates the potential of SOR for crop improvement and that inherent limitations of plant heat tolerance can be ameliorated with P. furiosus SOR.

  19. Enhancing heat tolerance of the little dogwood Cornus canadensis L. f. with introduction of a superoxide reductase gene from the hyperthermophilic archaeon Pyrococcus furiosus

    Directory of Open Access Journals (Sweden)

    Xinmin eGeng

    2016-01-01

    Full Text Available Production of reactive oxygen species (ROS can be accelerated under various biotic and abiotic stresses causing lipid peroxidation, protein degradation, enzyme inactivation, and DNA damage. Superoxide reductase (SOR is a novel antioxidant enzyme from Pyrococcus furiosus and is employed by this anaerobic hyperthermophilic archaeon for efficient detoxification of ROS. In this study, SOR was introduced into a flowering plant Cornus canadensis to enhance its heat tolerance and reduce heat induced damage. A fusion construct of the SOR gene and Green Fluorescent Protein gene (GFP was introduced into C. canadensis using Agrobacterium-mediated transformation. Heat tolerance of the GFP-SOR expressing transgenic plants was investigated by observing morphological symptoms of heat injury and by examining changes in photosynthesis, malondialdehyde (MDA, and proline levels in the plants. Our results indicate that the expression of the P. furiosus SOR gene in the transgenic plants alleviated lipid peroxidation of cell membranes and photoinhibition of PS II, and decreased the accumulation of proline at 40°C. After a series of exposures to increasing temperatures, the SOR transgenic plants remained healthy and green whereas most of the non-transgenic plants dried up and were unable to recover. While it had previously been reported that expression of SOR in Arabidopsis enhanced heat tolerance, this is the first report of the successful demonstration of improved heat tolerance in a non-model plant resulting from the introduction of P. furiosus SOR. The study demonstrates the potential of SOR for crop improvement and that inherent limitations of plant heat tolerance can be ameliorated with P. furiosus SOR.

  20. Molecular characterization of hydrolytic enzymes from hyperthermophilic archaea.

    OpenAIRE

    Voorhorst, W.G.B.

    1998-01-01

    Hyperthermophiles are recently discovered microorganisms which are able to grow optimally above 85 °C. Most hyperthermophiles belong to the Archaea, the third domain of life. One of the main interests in hyperthermophiles to deepen the insight in the way their proteins are stabilized and how to apply this knowledge to improve the stability of biotechnologically relevant enzymes. In this thesis attention has been focused on hydrolytic enzymes from hyperthermophilic archaea to provide insight i...

  1. Molecular characterization of hydrolytic enzymes from hyperthermophilic archaea.

    NARCIS (Netherlands)

    Voorhorst, W.G.B.

    1998-01-01

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

  2. Molecular characterization of hydrolytic enzymes from hyperthermophilic archaea

    NARCIS (Netherlands)

    Voorhorst, W.G.B.

    1998-01-01

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

  3. Protein degradation during reconsolidation as a mechanism for memory reorganization

    Directory of Open Access Journals (Sweden)

    Bong-Kiun Kaang

    2011-02-01

    Full Text Available Memory is a reference formed from a past experience that is used to respond to present situations. However, the world is dynamic and situations change, so it is important to update the memory with new information each time it is reactivated in order to adjust the response in the future. Recent researches indicate that memory may undergo a dynamic process that could work as an updating mechanism. This process which is called reconsolidation involves destabilization of the memory after it is reactivated, followed by restabilization. Recently, it has been demonstrated that the initial destabilization process of reconsolidation requires protein degradation. Using protein degradation inhibition as a method to block reconsolidation, recent researches suggest that reconsolidation, especially the protein degradation-dependent destabilization process is necessary for memory reorganization.

  4. Slow Unfolding of Monomeric Proteins from Hyperthermophiles with Reversible Unfolding

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    Atsushi Mukaiyama

    2009-03-01

    Full Text Available Based on the differences in their optimal growth temperatures microorganisms can be classified into psychrophiles, mesophiles, thermophiles, and hyperthermophiles. Proteins from hyperthermophiles generally exhibit greater stability than those from other organisms. In this review, we collect data about the stability and folding of monomeric proteins from hyperthermophilies with reversible unfolding, from the equilibrium and kinetic aspects. The results indicate that slow unfolding is a general strategy by which proteins from hyperthermophiles adapt to higher temperatures. Hydrophobic interaction is one of the factors in the molecular mechanism of the slow unfolding of proteins from hyperthermophiles.

  5. Bioenergetic and physiological studies of hyperthermophilic archaea. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Kelly, R.M.

    1999-03-01

    This project focuses on physiological and bioenergetic characteristics of two representative hyperthermophilic archaea: Thermococcus litoralis (T{sub opt} 88 C) and Pyrococcus furiosus (T{sub opt} 98 C). Both are obligately anaerobic heterotrophs which grow in the presence or absence of reducible sulfur compounds. T. litoralis was studied in relation to information previously developed for P. furiosus: effect of sulfur reduction on bioenergetics, preferred fermentation patterns, tungsten requirement, etc. A defined medium was developed for T. litoralis consisting of amino acids, vitamins and nucleotides. This serves as the basis for continuous culture studies probing metabolic response to media changes. P. furiosus and T. litoralis have also been found to produce a polysaccharide in the presence of maltose and yeast extract. The composition and chemical structure of this polysaccharide was investigated as well as the metabolic motivation for its production. A novel and, perhaps, primitive intracellular proteolytic complex (previously designated as protease S66) in P. furiosus was isolated and the gene encoding the subunit of the complex was cloned, sequenced and the protease expressed in active form in Eschericia coli. Among other issues, the role of this complex in protein turnover and stress response was examined in the context of this organism in addition to comparing it to other complexes in eubacterial and eukaryotic cells. Biochemical characteristics of the protease have been measured in addition to examining other proteolytic species in P. furiosus.

  6. Light-induced protein degradation in human-derived cells.

    Science.gov (United States)

    Sun, Wansheng; Zhang, Wenyao; Zhang, Chao; Mao, Miaowei; Zhao, Yuzheng; Chen, Xianjun; Yang, Yi

    2017-05-27

    Controlling protein degradation can be a valuable tool for posttranslational regulation of protein abundance to study complex biological systems. In the present study, we designed a light-switchable degron consisting of a light oxygen voltage (LOV) domain of Avena sativa phototropin 1 (AsLOV2) and a C-terminal degron. Our results showed that the light-switchable degron could be used for rapid and specific induction of protein degradation in HEK293 cells by light in a proteasome-dependent manner. Further studies showed that the light-switchable degron could also be utilized to mediate the degradation of secreted Gaussia princeps luciferase (GLuc), demonstrating the adaptability of the light-switchable degron in different types of protein. We suggest that the light-switchable degron offers a robust tool to control protein levels and may serves as a new and significant method for gene- and cell-based therapies. Copyright © 2017 Elsevier Inc. All rights reserved.

  7. Gene Transfer & Hybridization Studies in Hyperthermophilic Species

    Energy Technology Data Exchange (ETDEWEB)

    Nelson, Karen E.

    2005-10-14

    A. ABSTRACT The importance of lateral gene transfer (LGT) in the evolution of microbial species has become increasingly evident with each completed microbial genome sequence. Most significantly, the genome of Thermotoga maritima MSB8, a hyperthermophilic bacterium isolated by Karl Stetter and workers from Vulcano Italy in 1986, and sequenced at The Institute for Genomic Research (TIGR) in Rockville Maryland in 1999, revealed extensive LGT between % . this bacterium and members of the archaeal domain (in particular Archaeoglobus fulgidus, and Pyracoccus frcriosus species). Based on whole genome comparisons, it was estimated that 24% of the genetic information in this organism was acquired by genetic exchange with archaeal species, Independent analyses including periodicity analysis of the T. maritimu genomic DNA sequence, phylogenetic reconstruction based on genes that appear archaeal-like, and codon and amino acid usage, have provided additional evidence for LGT between T. maritima and the archaea. More recently, DiRuggiero and workers have identified a very recent LGT event between two genera of hyperthermophilic archaea, where a nearly identical DNA fragment of 16 kb in length flanked by insertion sequence (IS) elements, exists. Undoubtedly, additional examples of LGT will be identified as more microbial genomes are completed. For the present moment however, the genome sequence of T. maritima and other hyperthermophiles including P. furiosus, Pyrococcus horikoshii, Pyrococcus abyssi, A. fulgidus, and Aquifex aeolicus, have significantly increased out awareness of evolution being a web of life rather than a tree of life, as suggested by single gene phylogenies. In this proposal, we will aim to determine the extent of LGT across the hyperthemophiles, employing iY maritima as the model organism. A variety of biochemical techniques and phylogenetic reconstructions will allow for a detailed and thorough characterization of the extent of LGT in this species. The

  8. Decarboxylation of Pyruvate to Acetaldehyde for Ethanol Production by Hyperthermophiles

    Directory of Open Access Journals (Sweden)

    Mohammad S. Eram

    2013-08-01

    Full Text Available Pyruvate decarboxylase (PDC encoded by pdc is a thiamine pyrophosphate (TPP-containing enzyme responsible for the conversion of pyruvate to acetaldehyde in many mesophilic organisms. However, no pdc/PDC homolog has yet been found in fully sequenced genomes and proteomes of hyper/thermophiles. The only PDC activity reported in hyperthermophiles was a bifunctional, TPP- and CoA-dependent pyruvate ferredoxin oxidoreductase (POR/PDC enzyme from the hyperthermophilic archaeon Pyrococcus furiosus. Another enzyme known to be involved in catalysis of acetaldehyde production from pyruvate is CoA-acetylating acetaldehyde dehydrogenase (AcDH encoded by mhpF and adhE. Pyruvate is oxidized into acetyl-CoA by either POR or pyruvate formate lyase (PFL, and AcDH catalyzes the reduction of acetyl-CoA to acetaldehyde in mesophilic organisms. AcDH is present in some mesophilic (such as clostridia and thermophilic bacteria (e.g., Geobacillus and Thermoanaerobacter. However, no AcDH gene or protein homologs could be found in the released genomes and proteomes of hyperthermophiles. Moreover, no such activity was detectable from the cell-free extracts of different hyperthermophiles under different assay conditions. In conclusion, no commonly-known PDCs was found in hyperthermophiles. Instead of the commonly-known PDC, it appears that at least one multifunctional enzyme is responsible for catalyzing the non-oxidative decarboxylation of pyruvate to acetaldehyde in hyperthermophiles.

  9. Hyperthermophilic endoglucanase for in planta lignocellulose conversion

    Directory of Open Access Journals (Sweden)

    Klose Holger

    2012-08-01

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

  10. Hyperthermophilic endoglucanase for in planta lignocellulose conversion

    Science.gov (United States)

    2012-01-01

    Background The enzymatic conversion of lignocellulosic plant biomass into fermentable sugars is a crucial step in the sustainable and environmentally friendly production of biofuels. However, a major drawback of enzymes from mesophilic sources is their suboptimal activity under established pretreatment conditions, e.g. high temperatures, extreme pH values and high salt concentrations. Enzymes from extremophiles are better adapted to these conditions and could be produced by heterologous expression in microbes, or even directly in the plant biomass. Results Here we show that a cellulase gene (sso1354) isolated from the hyperthermophilic archaeon Sulfolobus solfataricus can be expressed in plants, and that the recombinant enzyme is biologically active and exhibits the same properties as the wild type form. Since the enzyme is inactive under normal plant growth conditions, this potentially allows its expression in plants without negative effects on growth and development, and subsequent heat-inducible activation. Furthermore we demonstrate that the recombinant enzyme acts in high concentrations of ionic liquids and can therefore degrade α-cellulose or even complex cell wall preparations under those pretreatment conditions. Conclusion The hyperthermophilic endoglucanase SSO1354 with its unique features is an excellent tool for advanced biomass conversion. Here we demonstrate its expression in planta and the possibility for post harvest activation. Moreover the enzyme is suitable for combined pretreatment and hydrolysis applications. PMID:22928996

  11. Formate hydrogenlyase in the hyperthermophilic archaeon, Thermococcus litoralis

    Directory of Open Access Journals (Sweden)

    Rákhely Gábor

    2008-06-01

    Full Text Available Abstract Background Thermococcus litoralis is a heterotrophic facultative sulfur dependent hyperthermophilic Archaeon, which was isolated from a shallow submarine thermal spring. It has been successfully used in a two-stage fermentation system, where various keratinaceous wastes of animal origin were converted to biohydrogen. In this system T. litoralis performed better than its close relative, P. furiosus. Therefore, new alternative enzymes involved in peptide and hydrogen metabolism were assumed in T. litoralis. Results An about 10.5 kb long genomic region was isolated and sequenced from Thermococcus litoralis. In silico analysis revealed that the region contained a putative operon consisting of eight genes: the fdhAB genes coding for a formate dehydrogenase and the mhyCDEFGH genes encoding a [NiFe] hydrogenase belonging to the group of the H2-evolving, energy-conserving, membrane-bound hydrogenases. Reverse transcription linked quantitative Real-Time PCR and Western blotting experiments showed that the expression of the fdh-mhy operon was up-regulated during fermentative growth on peptides and down-regulated in cells cultivated in the presence of sulfur. Immunoblotting and protein separation experiments performed on cell fractions indicated that the formate dehydrogenase part of the complex is associated to the membrane-bound [NiFe] hydrogenase. Conclusion The formate dehydrogenase together with the membrane-bound [NiFe] hydrogenase formed a formate hydrogenlyase (formate dehydrogenase coupled hydrogenase, FDH-MHY complex. The expression data suggested that its physiological role is linked to the removal of formate likely generated during anaerobic peptide fermentation.

  12. Effect of protein degradability on milk production of dairy ewes.

    Science.gov (United States)

    Mikolayunas-Sandrock, C; Armentano, L E; Thomas, D L; Berger, Y M

    2009-09-01

    The objective of this experiment was to determine the effect of protein degradability of dairy sheep diets on milk yield and protein utilization across 2 levels of milk production. Three diets were formulated to provide similar energy concentrations and varying concentrations of rumen-degradable protein (RDP) and rumen-undegradable protein (RUP): 12% RDP and 4% RUP (12-4) included basal levels of RDP and RUP, 12% RDP and 6% RUP (12-6) included additional RUP, and 14% RDP and 4% RUP (14-4) included additional RDP. Diets were composed of alfalfa-timothy cubes, whole and ground corn, whole oats, dehulled soybean meal, and expeller soybean meal (SoyPlus, West Central, Ralston, IA). Estimates of RDP and RUP were based on the Small Ruminant Nutrition System model (2008) and feed and orts were analyzed for Cornell N fractions. Eighteen multiparous dairy ewes in midlactation were divided by milk yield (low and high) into 2 blocks of 9 ewes each and were randomly assigned within block (low and high) to 3 pens of 3 ewes each. Dietary treatments were arranged in a 3 x 3 Latin square within each block and applied to pens for 14-d periods. We hypothesized that pens consuming high-RUP diets (12-6) would produce more milk and milk protein than the basal diet (12-4) and pens consuming high-RDP diets (14-4) would not produce more milk than the basal diet (12-4). Ewes in the high-milk-yield square consumed more dry matter and produced more milk, milk fat, and milk protein than ewes in the low-milk-yield square. There was no effect of dietary treatment on dry matter intake. Across both levels of milk production, the 12-6 diet increased milk yield by 14%, increased milk fat yield by 14%, and increased milk protein yield by 13% compared with the 14-4 and 12-4 diets. Gross N efficiency (milk protein N/intake protein N) was 11 and 15% greater in the 12-6 and 12-4 diets, respectively, compared with the 14-4 diet. Milk urea N concentration was greater in the 12-6 diet and tended to be

  13. Hyperthermophilic Archaeal Viruses as Novel Nanoplatforms

    DEFF Research Database (Denmark)

    Uldahl, Kristine Buch

    of a broad range of genetic and chemical engineering methods, viral research has expanded. Viruses are now emerging as nanoplatforms with applications in materials science and medicine. A great challenge in biomedicine is the targeting of therapeutics to specific locations in the body in order to increase...... nanoplatforms than mammalian viruses because they cannot proliferate in humans and hence are less likely to trigger adverse effects. Another group of viruses that fits this criterion is archaeal viruses yet their potential remains untapped. As a group, archaeal viruses offer distinct advantages such as unique...... hyperthermophilic archaeal viruses, SMV1 and SSV2 and cells of human origin. This chapter provides the first results demonstrating that archaeal viruses can be taken up and internalized by human cells, thus indicating a potential as intracellular delivery agents. Chapter III investigates SMV1 particles as potential...

  14. Protein Degradation Rate in Arabidopsis thaliana Leaf Growth and Development.

    Science.gov (United States)

    Li, Lei; Nelson, Clark J; Trösch, Josua; Castleden, Ian; Huang, Shaobai; Millar, A Harvey

    2017-02-01

    We applied (15)N labeling approaches to leaves of the Arabidopsis thaliana rosette to characterize their protein degradation rate and understand its determinants. The progressive labeling of new peptides with (15)N and measuring the decrease in the abundance of >60,000 existing peptides over time allowed us to define the degradation rate of 1228 proteins in vivo. We show that Arabidopsis protein half-lives vary from several hours to several months based on the exponential constant of the decay rate for each protein. This rate was calculated from the relative isotope abundance of each peptide and the fold change in protein abundance during growth. Protein complex membership and specific protein domains were found to be strong predictors of degradation rate, while N-end amino acid, hydrophobicity, or aggregation propensity of proteins were not. We discovered rapidly degrading subunits in a variety of protein complexes in plastids and identified the set of plant proteins whose degradation rate changed in different leaves of the rosette and correlated with leaf growth rate. From this information, we have calculated the protein turnover energy costs in different leaves and their key determinants within the proteome.

  15. Muscle protein degradation and amino acid metabolism during prolonged knee-extensor exercise in humans

    DEFF Research Database (Denmark)

    Van Hall, Gerrit; Saltin, B; Wagenmakers, A J

    1999-01-01

    to a substantial increase in net muscle protein degradation, and that a lowering of the starting muscle glycogen content leads to a further increase. The carbon atoms of the branched-chain amino acids (BCAA), glutamate, aspartate and asparagine, liberated by protein degradation, and the BCAA and glutamate...

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

    NARCIS (Netherlands)

    Lebbink, J.H.G.

    1999-01-01

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

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

    NARCIS (Netherlands)

    Lebbink, J.H.G.

    1999-01-01

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

  18. Histones and chromatin structure in hyperthermophilic Archaea.

    Science.gov (United States)

    Grayling, R A; Sandman, K; Reeve, J N

    1996-05-01

    HMf is a histone from the hyperthermophile Methanothermus fervidus. It is the archetype and most studied member of a family of archaeal histones that have primary sequences and three-dimensional structures in common with the eukaryal nucleosome core histones and that bind and compact DNA molecules into nucleosome-like structures (NLS). HMf preparations are mixtures of two similar, small (approximately 7.5 kDa) polypeptides designated HMfA and HMfB that in vivo form both homodimers and heterodimers. HMfA synthesis predominates during exponential growth but the relative amount of HMfB increases as M. fervidus cells enter the stationary growth phase. Analyses of homogeneous preparations of recombinant (r) (HMfA)2 and (rHMfB)2 have demonstrated that these proteins have different DNA-binding and compaction properties in vitro, consistent with different roles in vivo for the (HMfA)2, (HMfB)2 and HMfA. HmfB dimers, and for the NLS that they form, in regulating gene expression and in genome compaction and stability.

  19. Hydrogenases from the Hyperthermophilic Archaeon Pyrococcus furiosus

    NARCIS (Netherlands)

    van Haaster, D.J.

    2007-01-01

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

  20. Hydrogenases from the Hyperthermophilic Archaeon Pyrococcus furiosus

    NARCIS (Netherlands)

    van Haaster, D.J.

    2007-01-01

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

  1. Complete Genome Sequence of the Hyperthermophilic Archaeon Pyrococcus sp. Strain ST04, Isolated from a Deep-Sea Hydrothermal Sulfide Chimney on the Juan de Fuca Ridge

    Science.gov (United States)

    Jung, Jong-Hyun; Lee, Ju-Hoon; Holden, James F.; Seo, Dong-Ho; Shin, Hakdong; Kim, Hae-Yeong; Kim, Wooki; Ryu, Sangryeol

    2012-01-01

    Pyrococcus sp. strain ST04 is a hyperthermophilic, anaerobic, and heterotrophic archaeon isolated from a deep-sea hydrothermal sulfide chimney on the Endeavour Segment of the Juan de Fuca Ridge in the northeastern Pacific Ocean. To further understand the distinct characteristics of this archaeon at the genome level (polysaccharide utilization at high temperature and ATP generation by a Na+ gradient), the genome of strain ST04 was completely sequenced and analyzed. Here, we present the complete genome sequence analysis results of Pyrococcus sp. ST04 and report the major findings from the genome annotation, with a focus on its saccharolytic and metabolite production potential. PMID:22843576

  2. Pac-Man for biotechnology: co-opting degrons for targeted protein degradation to control and alter cell function.

    Science.gov (United States)

    Yu, Geng; Rosenberg, Julian N; Betenbaugh, Michael J; Oyler, George A

    2015-12-01

    Protein degradation in normal living cells is precisely regulated to match the cells' physiological requirements. The selectivity of protein degradation is determined by an elaborate degron-tagging system. Degron refers to an amino acid sequence that encodes a protein degradation signal, which is oftentimes a poly-ubiquitin chain that can be transferred to other proteins. Current understanding of ubiquitination dependent and independent protein degradation processes has expanded the application of degrons for targeted protein degradation and novel cell engineering strategies. Recent findings suggest that small molecules inducing protein association can be exploited to create degrons that target proteins for degradation. Here, recent applications of degron-based targeted protein degradation in eukaryotic organisms are reviewed. The degron mediated protein degradation represents a rapidly tunable methodology to control protein abundance, which has broad application in therapeutics and cellular function control and monitoring.

  3. Hyperthermophilic archaea produce membrane vesicles that can transfer DNA

    NARCIS (Netherlands)

    Gaudin, M.; Gauliard, E.; Schouten, S.; Houel-Renault, L.; Lenormand, P.; Marguet, E.; Forterre, P.

    2013-01-01

    Thermococcales are hyperthermophilic archaea found in deep-sea hydrothermal vents. They have been recently reported to produce membrane vesicles (MVs) into their culture medium. Here, we have characterized the mode of production and determined the biochemical composition of MVs from two species of

  4. Similarity and divergence between the RNA polymerase alpha subunits from hyperthermophilic Thermotoga maritima and mesophilic Escherichia coli bacteria.

    Science.gov (United States)

    Braun, Frederique; Marhuenda, Fanny B; Morin, Amelie; Guevel, Laetitia; Fleury, Fabrice; Takahashi, Masayuki; Sakanyan, Vehary

    2006-10-01

    The alpha subunit (alphaTm) of Thermotoga maritima RNA polymerase has been characterized to investigate its role in transcriptional regulation in one of the few known anaerobic hyperthermophilic bacteria. The highly thermostable alphaTm shares 54% similarity with its Escherichia coli analogue (alphaEc). The T. maritima rpoA gene coding the alpha subunit does not complement the thermosensitive rpoA112 mutation of E. coli. However, alphaTm and alphaEc show similar folding patterns as determined by circular dichroism. Purified alphaTm binds to the T. maritima PargGo promoter region (probably to a UP-element) and Arg282 appears to be crucial for DNA binding. The thermostable protein is also able to interact with transcription regulatory proteins, like ArgR from T. neapolitana or CRP from E. coli. These data indicate that the RNA polymerase alpha subunit might play a crucial role in the modulation of gene expression in hyperthermophiles.

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

    Directory of Open Access Journals (Sweden)

    Juan Pablo Cardenas

    2016-11-01

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

  6. Anaerobic lipid degradation through acidification and methanization.

    Science.gov (United States)

    Kim, Ijung; Kim, Sang-Hyoun; Shin, Hang-Sik; Jung, Jin-Young

    2010-01-01

    In biological wastewater treatment high lipid concentration is known to inhibit microorganisms and cause active biomass flotation. To reduce lipid inhibition, a two-phase anaerobic system, consisting of an anaerobic sequencing batch reactor (ASBR) and an upflow anaerobic sludge blanket (UASB) reactor, was applied to synthetic dairy wastewater. During 153 days of operation, the two-phase system showed stable performance in lipid degradation. In the ASBR, a 13% lipid removal efficiency and 10% double bond removal efficiency were maintained. In the UASB, the chemical oxygen demand (COD), lipid and volatile fatty acid (VFA) removal efficiencies were more than 80%, 70% and 95%, respectively, up to organic loading rate 6.5 g COD/L/day. There were no operational problems such as serious scum formation or sludge washout. Protein degradation occurred prior to degradation during acidogenesis.

  7. Protease Activities of Semi-purified Pseudomonas fluorescensin Protein Degradation of Pasteurized Milk at Storage

    Directory of Open Access Journals (Sweden)

    Tatik Khusniati

    2012-10-01

    Full Text Available Protein on stored milks spoiled due to protease activities of Pseudomonas fluorescens. To know protease effect on milks, protease activities of semi-purified P. fluorescens on protein degradation in stored pasteurized milks were detected. Protease was semi-purified by ethanol 70%. Protease activities were detected by modified Lowry method, protein degradation by formol titration, and protein content by Kjeldahl method. Milk storage' times were conducted on 0 day (4 days before expired date up to 12 days (8 days after expired date. The results show that the longer the storage times the higher protease activities and protein degradation of milks. At storage 12 days, protease activities on control were 0.2556 Unit/mL (skim and 0.2116 Unit/mL (whole, and on inoculated milk (crude was 2.2044 Unit/mL (skim and 1.5378 Unit/mL (whole; while on inoculated milk (semi-purified was 3.5355 Unit/mL (skim and 1.6778 Unit/mL (whole, respectively. The decrease of inoculated milk' homogeneous were faster than that of control. Protein degradation on control, inoculated skim milks (crude and semi-purified on 12 days were 4.48%, 7.28% and 7.62%, while that on whole milks were 3.81%, 7.28% and 6.04%, respectively. Based on protease, protein degradation and homogeneous, it can be concluded that skim milk spoiled faster than whole milk.

  8. Protein degradation and protein synthesis in long-term memory formation

    Directory of Open Access Journals (Sweden)

    Timothy J Jarome

    2014-06-01

    Full Text Available Long-term memory (LTM formation requires transient changes in the activity of intracellular signaling cascades that are thought to regulate new gene transcription and de novo protein synthesis in the brain. Consistent with this, protein synthesis inhibitors impair LTM for a variety of behavioral tasks when infused into the brain around the time of training or following memory retrieval, suggesting that protein synthesis is a critical step in LTM storage in the brain. However, evidence suggests that protein degradation mediated by the ubiquitin-proteasome system may also be a critical regulator of LTM formation and stability following retrieval. This requirement for increased protein degradation has been shown in the same brain regions in which protein synthesis is required for LTM storage. Additionally, increases in the phosphorylation of proteins involved in translational control parallel increases in protein polyubiquitination and the increased demand for protein degradation is regulated by intracellular signaling molecules thought to regulate protein synthesis during LTM formation. In some cases inhibiting proteasome activity can rescue memory impairments that result from pharmacological blockade of protein synthesis, suggesting that protein degradation may control the requirement for protein synthesis during the memory storage process. Results such as these suggest that protein degradation and synthesis are both critical for LTM formation and may interact to properly consolidate and store memories in the brain. Here, we review the evidence implicating protein synthesis and degradation in LTM storage and highlight the areas of overlap between these two opposing processes. We also discuss evidence suggesting these two processes may interact to properly form and store memories. LTM storage likely requires a coordinated regulation between protein degradation and synthesis at multiple sites in the mammalian brain.

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

    Directory of Open Access Journals (Sweden)

    Taku Amo

    2002-01-01

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

  10. Minimal sulfur requirement for growth and sulfur-dependent metabolism of the hyperthermophilic archaeon Staphylothermus marinus

    Directory of Open Access Journals (Sweden)

    Xiaolei Hao

    2003-01-01

    Full Text Available Staphylothermus marinus is an anaerobic hyperthermophilic archaeon that uses peptides as carbon and energy sources. Elemental sulfur (S° is obligately required for its growth and is reduced to H2S. The metabolic functions and mechanisms of S° reduction were explored by examining S°-dependent growth and activities of key enzymes present in this organism. All three forms of S° tested—sublimed S°, colloidal S° and polysulfide—were used by S. marinus, and no other sulfur-containing compounds could replace S°. Elemental sulfur did not serve as physical support but appeared to function as an electron acceptor. The minimal S° concentration required for optimal growth was 0.05% (w/v. At this concentration, there appeared to be a metabolic transition from H2 production to S° reduction. Some enzymatic activities related to S°-dependent metabolism, including sulfur reductase, hydrogenase, glutamate dehydrogenase and electron transfer activities, were detected in cell-free extracts of S. marinus. These results indicate that S° plays an essential role in the heterotrophic metabolism of S. marinus. Reducing equivalents generated by the oxidation of amino acids from peptidolysis may be transferred to sulfur reductase and hydrogenase, which then catalyze the production of H2S and H2, respectively.

  11. Alpha-amylase from the Hyperthermophilic Archaeon Thermococcus thioreducens

    Science.gov (United States)

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

    2003-01-01

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

  12. Alpha-amylase from the Hyperthermophilic Archaeon Thermococcus thioreducens

    Science.gov (United States)

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

    2003-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Gerrit Jan Schut

    2016-01-01

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

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

    Science.gov (United States)

    Schut, Gerrit J.; Lipscomb, Gina L.; Nguyen, Diep M. N.; Kelly, Robert M.; Adams, Michael W. W.

    2016-01-01

    Carbon monoxide (CO) is an important intermediate in anaerobic carbon fixation pathways in acetogenesis and methanogenesis. In addition, some anaerobes can utilize CO as an energy source. In the hyperthermophilic archaeon Thermococcus onnurineus, which grows optimally at 80°C, CO oxidation and energy conservation is accomplished by a respiratory complex encoded by a 16-gene cluster containing a CO dehydrogenase, a membrane-bound [NiFe]-hydrogenase and a Na+/H+ antiporter module. This complex oxidizes CO, evolves CO2 and H2, and generates a Na+ motive force that is used to conserve energy by a Na+-dependent ATP synthase. Herein we used a bacterial artificial chromosome to insert the 13.2 kb gene cluster encoding the CO-oxidizing respiratory complex of T. onnurineus into the genome of the heterotrophic archaeon, Pyrococcus furiosus, which grows optimally at 100°C. P. furiosus is normally unable to utilize CO, however, the recombinant strain readily oxidized CO and generated H2 at 80°C. Moreover, CO also served as an energy source and allowed the P. furiosus strain to grow with a limiting concentration of sugar or with peptides as the carbon source. Moreover, CO oxidation by P. furiosus was also coupled to the re-utilization, presumably for biosynthesis, of acetate generated by fermentation. The functional transfer of CO utilization between Thermococcus and Pyrococcus species demonstrated herein is representative of the horizontal gene transfer of an environmentally relevant metabolic capability. The transfer of CO utilizing, hydrogen-producing genetic modules also has applications for biohydrogen production and a CO-based industrial platform for various thermophilic organisms. PMID:26858706

  15. A dicarboxylate/4-hydroxybutyrate autotrophic carbon assimilation cycle in the hyperthermophilic Archaeum Ignicoccus hospitalis.

    Science.gov (United States)

    Huber, Harald; Gallenberger, Martin; Jahn, Ulrike; Eylert, Eva; Berg, Ivan A; Kockelkorn, Daniel; Eisenreich, Wolfgang; Fuchs, Georg

    2008-06-03

    Ignicoccus hospitalis is an anaerobic, autotrophic, hyperthermophilic Archaeum that serves as a host for the symbiotic/parasitic Archaeum Nanoarchaeum equitans. It uses a yet unsolved autotrophic CO(2) fixation pathway that starts from acetyl-CoA (CoA), which is reductively carboxylated to pyruvate. Pyruvate is converted to phosphoenol-pyruvate (PEP), from which glucogenesis as well as oxaloacetate formation branch off. Here, we present the complete metabolic cycle by which the primary CO(2) acceptor molecule acetyl-CoA is regenerated. Oxaloacetate is reduced to succinyl-CoA by an incomplete reductive citric acid cycle lacking 2-oxoglutarate dehydrogenase or synthase. Succinyl-CoA is reduced to 4-hydroxybutyrate, which is then activated to the CoA thioester. By using the radical enzyme 4-hydroxybutyryl-CoA dehydratase, 4-hydroxybutyryl-CoA is dehydrated to crotonyl-CoA. Finally, beta-oxidation of crotonyl-CoA leads to two molecules of acetyl-CoA. Thus, the cyclic pathway forms an extra molecule of acetyl-CoA, with pyruvate synthase and PEP carboxylase as the carboxylating enzymes. The proposal is based on in vitro transformation of 4-hydroxybutyrate, detection of all enzyme activities, and in vivo-labeling experiments using [1-(14)C]4-hydroxybutyrate, [1,4-(13)C(2)], [U-(13)C(4)]succinate, or [1-(13)C]pyruvate as tracers. The pathway is termed the dicarboxylate/4-hydroxybutyrate cycle. It combines anaerobic metabolic modules to a straightforward and efficient CO(2) fixation mechanism.

  16. Involvement of protein degradation by the ubiquitin proteasome system in opiate addictive behaviors.

    Science.gov (United States)

    Massaly, Nicolas; Dahan, Lionel; Baudonnat, Mathieu; Hovnanian, Caroline; Rekik, Khaoula; Solinas, Marcello; David, Vincent; Pech, Stéphane; Zajac, Jean-Marie; Roullet, Pascal; Mouledous, Lionel; Frances, Bernard

    2013-03-01

    Plastic changes in the nucleus accumbens (NAcc), a structure occupying a key position in the neural circuitry related to motivation, are among the critical cellular processes responsible for drug addiction. During the last decade, it has been shown that memory formation and related neuronal plasticity may rely not only on protein synthesis but also on protein degradation by the ubiquitin proteasome system (UPS). In this study, we assess the role of protein degradation in the NAcc in opiate-related behaviors. For this purpose, we coupled behavioral experiments to intra-accumbens injections of lactacystin, an inhibitor of the UPS. We show that protein degradation in the NAcc is mandatory for a full range of animal models of opiate addiction including morphine locomotor sensitization, morphine conditioned place preference, intra-ventral tegmental area morphine self-administration and intra-venous heroin self-administration but not for discrimination learning rewarded by highly palatable food. This study provides the first evidence of a specific role of protein degradation by the UPS in addiction.

  17. APC/C activity during the cell cycle. Shifting gears in protein degradation

    NARCIS (Netherlands)

    Boekhout, M.

    2015-01-01

    For correct cell division to take place, many different mechanisms ensure genomic integrity and formation healthy daughter cells. One mechanism that has evolved to provide a safe passage from one cell cycle phase into the next, is protein degradation. With our work we provide new insights into activ

  18. Tannin content and rate of ruminal protein degradation of legume hays

    Science.gov (United States)

    This work evaluated ruminal protein degradation rates of legume hays that varied in tannin content. Two cuttings of 5 varieties of birdsfoot trefoil, (Lotus corniculatus), selected for different tannin contents but similar NDF and CP contents, and Spredor 4 alfalfa (control) were conserved as hay. S...

  19. Role of Proteasome-Dependent Protein Degradation in Long-Term Operant Memory in "Aplysia"

    Science.gov (United States)

    Lyons, Lisa C.; Gardner, Jacob S.; Gandour, Catherine E.; Krishnan, Harini C.

    2017-01-01

    We investigated the in vivo role of protein degradation during intermediate (ITM) and long-term memory (LTM) in "Aplysia" using an operant learning paradigm. The proteasome inhibitor MG-132 inhibited the induction and molecular consolidation of LTM with no effect on ITM. Remarkably, maintenance of steady-state protein levels through…

  20. Role of Proteasome-Dependent Protein Degradation in Long-Term Operant Memory in "Aplysia"

    Science.gov (United States)

    Lyons, Lisa C.; Gardner, Jacob S.; Gandour, Catherine E.; Krishnan, Harini C.

    2017-01-01

    We investigated the in vivo role of protein degradation during intermediate (ITM) and long-term memory (LTM) in "Aplysia" using an operant learning paradigm. The proteasome inhibitor MG-132 inhibited the induction and molecular consolidation of LTM with no effect on ITM. Remarkably, maintenance of steady-state protein levels through…

  1. Leucine and isoleucine reduce protein degradation in rainbow trout (Oncorhynchus mykiss) primary myoblast cultures

    Science.gov (United States)

    Myogenic precursor cells were isolated from rainbow trout skeletal muscle and incubated in media containing 10% fetal bovine serum for 7 days, thereby differentiating into myoblasts. Rates of protein degradation were determined in response to minimal essential media (MEM) of various amino acid (AA)...

  2. Anaerobic bacteria

    Science.gov (United States)

    Brook I, Goldstein EJ. Diseases caused by non-spore forming anaerobic bacteria. In: Goldman L, Schafer AI, eds. Goldman's Cecil Medicine . 25th ed. Philadelphia, PA: Elsevier Saunders; 2015:chap 297. Stedman's Online ...

  3. A Role of Protein Degradation in Memory Consolidation after Initial Learning and Extinction Learning in the Honeybee ("Apis mellifera")

    Science.gov (United States)

    Felsenberg, Johannes; Dombrowski, Vincent; Eisenhardt, Dorothea

    2012-01-01

    Protein degradation is known to affect memory formation after extinction learning. We demonstrate here that an inhibitor of protein degradation, MG132, interferes with memory formation after extinction learning in a classical appetitive conditioning paradigm. In addition, we find an enhancement of memory formation when the same inhibitor is…

  4. A Role of Protein Degradation in Memory Consolidation after Initial Learning and Extinction Learning in the Honeybee ("Apis mellifera")

    Science.gov (United States)

    Felsenberg, Johannes; Dombrowski, Vincent; Eisenhardt, Dorothea

    2012-01-01

    Protein degradation is known to affect memory formation after extinction learning. We demonstrate here that an inhibitor of protein degradation, MG132, interferes with memory formation after extinction learning in a classical appetitive conditioning paradigm. In addition, we find an enhancement of memory formation when the same inhibitor is…

  5. Energetic and hydrogen limitations of thermophilic and hyperthermophilic methanogens

    Science.gov (United States)

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

    2013-12-01

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

  6. Lysine suppresses protein degradation through autophagic-lysosomal system in C2C12 myotubes.

    Science.gov (United States)

    Sato, Tomonori; Ito, Yoshiaki; Nedachi, Taku; Nagasawa, Takashi

    2014-06-01

    Muscle mass is determined between protein synthesis and protein degradation. Reduction of muscle mass leads to bedridden condition and attenuation of resistance to diseases. Moreover, bedridden condition leads to additional muscle loss due to disuse muscle atrophy. In our previous study (Sato et al. 2013), we showed that administered lysine (Lys), one of essential amino acid, suppressed protein degradation in skeletal muscle. In this study, we investigated that the mechanism of the suppressive effects of Lys on skeletal muscle proteolysis in C2C12 cell line. C2C12 myotubes were incubated in the serum-free medium containing 10 mM Lys or 20 mM Lys, and myofibrillar protein degradation was determined by the rates of 3-methylhistidine (MeHis) release from the cells. The mammalian target of rapamycin (mTOR) activity from the phosphorylation levels of p70-ribosormal protein S6 kinase 1 and eIF4E-binding protein 1 and the autophagic-lysosomal system activity from the ratio of LC3-II/I in C2C12 myotubes stimulated by 10 mM Lys for 0-3 h were measured. The rates of MeHis release were markedly reduced by addition of Lys. The autophagic-lysosomal system activity was inhibited upon 30 min of Lys supplementation. The activity of mTOR was significantly increased upon 30 min of Lys supplementation. The suppressive effect of Lys on the proteolysis by the autophagic-lysosomal system was maintained partially when mTOR activity was inhibited by 100 nM rapamycin, suggesting that some regulator other than mTOR signaling, for example, Akt, might also suppress the autophagic-lysosomal system. From these results, we suggested that Lys suppressed the activity of the autophagic-lysosomal system in part through activation of mTOR and reduced myofibrillar protein degradation in C2C12 myotubes.

  7. The relationship between protein synthesis and protein degradation in object recognition memory.

    Science.gov (United States)

    Furini, Cristiane R G; Myskiw, Jociane de C; Schmidt, Bianca E; Zinn, Carolina G; Peixoto, Patricia B; Pereira, Luiza D; Izquierdo, Ivan

    2015-11-01

    For decades there has been a consensus that de novo protein synthesis is necessary for long-term memory. A second round of protein synthesis has been described for both extinction and reconsolidation following an unreinforced test session. Recently, it was shown that consolidation and reconsolidation depend not only on protein synthesis but also on protein degradation by the ubiquitin-proteasome system (UPS), a major mechanism responsible for protein turnover. However, the involvement of UPS on consolidation and reconsolidation of object recognition memory remains unknown. Here we investigate in the CA1 region of the dorsal hippocampus the involvement of UPS-mediated protein degradation in consolidation and reconsolidation of object recognition memory. Animals with infusion cannulae stereotaxically implanted in the CA1 region of the dorsal hippocampus, were exposed to an object recognition task. The UPS inhibitor β-Lactacystin did not affect the consolidation and the reconsolidation of object recognition memory at doses known to affect other forms of memory (inhibitory avoidance, spatial learning in a water maze) while the protein synthesis inhibitor anisomycin impaired the consolidation and the reconsolidation of the object recognition memory. However, β-Lactacystin was able to reverse the impairment caused by anisomycin on the reconsolidation process in the CA1 region of the hippocampus. Therefore, it is possible to postulate a direct link between protein degradation and protein synthesis during the reconsolidation of the object recognition memory.

  8. Draft Genome Sequence of an Anaerobic and Extremophilic Bacterium, Caldanaerobacter yonseiensis, Isolated from a Geothermal Hot Stream

    OpenAIRE

    2013-01-01

    Caldanaerobacter yonseiensis is a strictly anaerobic, thermophilic, spore-forming bacterium, which was isolated from a geothermal hot stream in Indonesia. This bacterium utilizes xylose and produces a variety of proteases. Here, we report the draft genome sequence of C. yonseiensis, which reveals insights into the pentose phosphate pathway and protein degradation metabolism in thermophilic microorganisms.

  9. Genomic analysis of hyperthermophilic archaea; Chokonetsusei kosaikin no genomu kaiseki

    Energy Technology Data Exchange (ETDEWEB)

    Kato, C. [Japan Marine Science and Technology Center, Kanagawa (Japan)

    1997-05-20

    Whole genome sequences of five strains of microorganisms have been reported up to the present and many genome analysis projects are in progress in the world. Among archaea (archaebacteria), the genome analysis of Methanococcus jannaschii have been completed and the sequencing data are opened to public. While 134 regulatory genes were identified in Synechocystis sp. PCC 6803 (eubacteria, 3.6 genome size), only 7 regulatory genes were identified in M. jannaschii (1.7Mb). Difference of the genome size is believed to correspond to the quantity of the environmental stresses. In Japan, the genome analysis project on a new hyperthermophilic archaeon, Pyrococcus horikoshii is in progress. P. horikoshii was isolated in a deep sea hydrothermal vent. It shows barophilic growth at maximum high temperature of 103degC under pressure of 30MPa. Thus, the genome analysis of barophilic hyperthermophilic archaea is expected to contribute to the understanding of the origin of life and evolution. 19 refs., 4 figs., 1 tab.

  10. Thermostable enzymes of hyperthermophilic archaea; Chokonetsukin no tainetsusei koso

    Energy Technology Data Exchange (ETDEWEB)

    Imanaka, T. [Kyoto Univ., Kyoto (Japan). Graduate School; Fujiwara, S. [Osaka Univ., Osaka (Japan). Graduate School of Engineering

    1997-05-20

    All known organisms are divided into three groups, eukaryote, eubacteria and archaea (archaebacteria) according to the phylogenic tree based on 16SrRNA or protein sequences. Archaea is considered to be closer to eukaryote than eubacteria, and particularly, hyperthermophilic archaea evolved most slowly in archaea domain remaining ancestral features of eukaryote. Pyrococcus sp. KOD 1, a hyperthermophilic archaeon were isolated from a solfatara at a wharf of Kodakara island, Kagoshima, Japan with 95degC of the optimum temperature for cell growth. Unusual characteristics of enzymes produced by KOD 1 are discussed. The structural features of protease, glutamate dehydrogenase, glutamine synthase and amylase of KOD 1 are close to corresponding eubacterial enzymes. DNA polymerase from KOD 1 shows a high rate of DNA synthesis and is used in PCR practically. Molecular chaperonin of KOD 1 has high similarity to TCP-1 of eukaryote and is expected for application to in vitro enzyme stabilization and in vivo solubilization of foreign proteins. 26 refs., 8 figs., 2 tabs.

  11. Aerobic Lineage of the Oxidative Stress Response Protein Rubrerythrin Emerged in an Ancient Microaerobic, (Hyper)Thermophilic Environment

    Science.gov (United States)

    Cardenas, Juan P.; Quatrini, Raquel; Holmes, David S.

    2016-01-01

    Rubrerythrins (RBRs) are non-heme di-iron proteins belonging to the ferritin-like superfamily. They are involved in oxidative stress defense as peroxide scavengers in a wide range of organisms. The vast majority of RBRs, including classical forms of this protein, contain a C-terminal rubredoxin-like domain involved in electron transport that is used during catalysis in anaerobic conditions. Rubredoxin is an ancient and large protein family of short length (Fe-S center involved in electron transfer. However, functional forms of the enzyme lacking the rubredoxin-like domain have been reported (e.g., sulerythrin and ferriperoxin). In this study, phylogenomic evidence is presented that suggests that a complete lineage of rubrerythrins, lacking the rubredoxin-like domain, arose in an ancient microaerobic and (hyper)thermophilic environments in the ancestors of the Archaea Thermoproteales and Sulfolobales. This lineage (termed the “aerobic-type” lineage) subsequently evolved to become adapted to environments with progressively lower temperatures and higher oxygen concentrations via the acquisition of two co-localized genes, termed DUF3501 and RFO, encoding a conserved protein of unknown function and a predicted Fe-S oxidoreductase, respectively. Proposed Horizontal Gene Transfer events from these archaeal ancestors to Bacteria expanded the opportunities for further evolution of this RBR including adaption to lower temperatures. The second lineage (termed the cyanobacterial lineage) is proposed to have evolved in cyanobacterial ancestors, maybe in direct response to the production of oxygen via oxygenic photosynthesis during the Great Oxygen Event (GOE). It is hypothesized that both lineages of RBR emerged in a largely anaerobic world with “whiffs” of oxygen and that their subsequent independent evolutionary trajectories allowed microorganisms to transition from this anaerobic world to an aerobic one. PMID:27917155

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

    Science.gov (United States)

    Ramos, Lucas Rodrigues; Silva, Edson Luiz

    2016-12-16

    The objective of this study was to investigate the effects of hydraulic retention time (HRT) (8 to 0.5 h) and temperature (55 to 75 °C) in two anaerobic fluidized bed reactors (AFBR) using cheese whey (AFBR-CW = 10,000 mg sugars L(-1)) and vinasse (AFBR-V = 10,000 mg COD L(-1)) as substrates. Decreasing the HRT to 0.5 h increased the hydrogen production rates in both reactors, with maximum values of 5.36 ± 0.81 L H2 h(-1) L(-1) in AFBR-CW and 0.71 ± 0.16 L H2 h(-1) L(-1) in AFBR-V. The optimal conditions for hydrogen production were the HRT of 4 h and temperature of 65 °C in AFBR-CW, observing maximum hydrogen yield (HY) of 5.51 ± 0.37 mmol H2 g COD(-1). Still, the maximum HY in AFBR-V was 1.64 ± 0.22 mmol H2 g COD(-1) at 4 h and 55 °C. However, increasing the temperature to 75 °C reduced the hydrogen production in both reactors. Methanol and butyric, acetic, and lactic acids were the main metabolites at temperatures of 55 and 65 °C, favoring the butyric and acetic metabolic pathways of hydrogen production. The increased productions of lactate, propionate, and methanol at 75 °C indicate that the hydrogen-producing bacteria in the thermophilic inoculum were inhibited under hyperthermophilic conditions.

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

    NARCIS (Netherlands)

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

    2010-01-01

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

  14. Exceptionally diverse morphotypes and genomes of crenarchaeal hyperthermophilic viruses

    DEFF Research Database (Denmark)

    Prangishvili, D; Garrett, R A

    2004-01-01

    The remarkable diversity of the morphologies of viruses found in terrestrial hydrothermal environments with temperatures >80 degrees C is unprecedented for aquatic ecosystems. The best-studied viruses from these habitats have been assigned to novel viral families: Fuselloviridae, Lipothrixviridae...... no significant matches to sequences in public databases. This suggests that these hyperthermophilic viruses have exceptional biochemical solutions for biological functions. Specific features of genome organization, as well as strategies for DNA replication, suggest that phylogenetic relationships exist between...... crenarchaeal rudiviruses and the large eukaryal DNA viruses: poxviruses, the African swine fever virus and Chlorella viruses. Sequence patterns at the ends of the linear genome of the lipothrixvirus AFV1 are reminiscent of the telomeric ends of linear eukaryal chromosomes and suggest that a primitive telomeric...

  15. Anaerobic biodegradability of macropollutants

    DEFF Research Database (Denmark)

    Angelidaki, Irini

    2002-01-01

    A variety of test procedures for determination of anaerobic biodegradability has been reported. This paper reviews the methods developed for determination of anaerobic biodegradability of macro-pollutants. Anaerobic biodegradability of micro-pollutants is not included. Furthermore, factors...

  16. LOV Domain-Containing F-Box Proteins:Light-Dependent Protein Degradation Modules in Arabidopsis

    Institute of Scientific and Technical Information of China (English)

    Shogo Ito; Young Hun Song; Takato Imaizumi

    2012-01-01

    Plants constantly survey the surrounding environment using several sets of photoreceptors.They can sense changes in the quantity (=intensity) and quality (=wavelength) of light and use this information to adjust their physiological responses,growth,and developmental patterns.In addition to the classical photoreceptors,such as phytochromes,cryptochromes,and phototropins,ZEITLUPE (ZTL),FLAVIN-BINDING,KELCH REPEAT,F-BOX 1 (FKF1),and LOV KELCH PROTEIN 2 (LKP2) proteins have been recently identified as blue-light photoreceptors that are important for regulation of the circadian clock and photoperiodic flowering.The ZTL/FKF1/LKP2 protein family possesses a unique combination of domains:a blue-light-absorbing LOV (Light,Oxygen,or Voltage) domain along with domains involved in protein degradation.Here,we summarize recent advances in our understanding of the function of the Arabidopsis ZTL/FKF1/LKP2 proteins.We summarize the distinct photochemical properties of their LOV domains and discuss the molecular mechanisms by which the ZTL/FKF1/LKP2 proteins regulate the circadian clock and photoperiodic flowering by controlling blue-light-dependent protein degradation.

  17. Benefits of heat treatment to the protease packed neutrophil for proteome analysis: halting protein degradation.

    Science.gov (United States)

    Kennedy, Susan A; Scaife, Caitríona; Dunn, Michael J; Wood, Alfred E; Watson, R William G

    2011-06-01

    Neutrophils, cells of the innate immune system, contain an array of proteases and reactive oxygen species-generating enzymes that assist in controlling the invasion of bacteria and pathogens. The high content of intracellular proteolytic enzymes makes them difficult cells to work with as they can degrade proteins of potential interest. Here, we describe the benefits of heat treatment of neutrophils in reducing protein degradation for subsequent proteome analysis. Neutrophils isolated from four healthy volunteers were each divided into three aliquots and subjected to different preparation methods for 2-DE: (i) Heat treatment, (ii) resuspension in NP40 lysis buffer and (iii) resuspension in standard 2-DE lysis buffer. Representative spots found to be statistically significant between groups (pHeat-treated samples contained proteins in the high-molecular-weight range that were absent from NP40-treated samples. Moreover, NP40-treated samples showed an increase in spot number and volume at lower molecular weights suggestive of protein degradation. Incorporating heat treatment into sample preparation resulted in the identification of proteins that may not have previously been detected due to sample degradation, thus leading to a more comprehensive 2-DE map of the human neutrophil proteome.

  18. Anaerobic Digestion.

    Science.gov (United States)

    Liebetrau, Jan; Sträuber, Heike; Kretzschmar, Jörg; Denysenko, Velina; Nelles, Michael

    2017-04-09

    The term anaerobic digestion usually refers to the microbial conversion of organic material to biogas, which mainly consists of methane and carbon dioxide. The technical application of the naturally-occurring process is used to provide a renewable energy carrier and - as the substrate is often waste material - to reduce the organic matter content of the substrate prior to disposal.Applications can be found in sewage sludge treatment, the treatment of industrial and municipal solid wastes and wastewaters (including landfill gas utilization), and the conversion of agricultural residues and energy crops.For biorefinery concepts, the anaerobic digestion (AD) process is, on the one hand, an option to treat organic residues from other production processes. Concomitant effects are the reduction of organic carbon within the treated substance, the conversion of nitrogen and sulfur components, and the production of an energy-rich gas - the biogas. On the other hand, the multistep conversion of complex organic material offers the possibility of interrupting the conversion chain and locking out intermediates for utilization as basic material within the chemical industry.

  19. Multisite phosphorylation provides an effective and flexible mechanism for switch-like protein degradation.

    Science.gov (United States)

    Varedi K, S Marjan; Ventura, Alejandra C; Merajver, Sofia D; Lin, Xiaoxia Nina

    2010-12-13

    Phosphorylation-triggered degradation is a common strategy for elimination of regulatory proteins in many important cell signaling processes. Interesting examples include cyclin-dependent kinase inhibitors such as p27 in human and Sic1 in yeast, which play crucial roles during the G1/S transition in the cell cycle. In this work, we have modeled and analyzed the dynamics of multisite-phosphorylation-triggered protein degradation systematically. Inspired by experimental observations on the Sic1 protein and a previous intriguing theoretical conjecture, we develop a model to examine in detail the degradation dynamics of a protein featuring multiple phosphorylation sites and a threshold site number for elimination in response to a kinase signal. Our model explains the role of multiple phosphorylation sites, compared to a single site, in the regulation of protein degradation. A single-site protein cannot convert a graded input of kinase increase to much sharper output, whereas multisite phosphorylation is capable of generating a highly switch-like temporal profile of the substrate protein with two characteristics: a temporal threshold and rapid decrease beyond the threshold. We introduce a measure termed temporal response coefficient to quantify the extent to which a response in the time domain is switch-like and further investigate how this property is determined by various factors including the kinase input, the total number of sites, the threshold site number for elimination, the order of phosphorylation, the kinetic parameters, and site preference. Some interesting and experimentally verifiable predictions include that the non-degradable fraction of the substrate protein exhibits a more switch-like temporal profile; a sequential system is more switch-like, while a random system has the advantage of increased robustness; all the parameters, including the total number of sites, the threshold site number for elimination and the kinetic parameters synergistically

  20. Effects of Dietary Supplementation with Hainanmycin on Protein Degradation and Populations of Ammonia-producing Bacteria

    Directory of Open Access Journals (Sweden)

    Z. B. Wang

    2013-05-01

    Full Text Available An in vitro fermentation was conducted to determine the effects of hainanmycin on protein degradation and populations of ammonia-producing bacteria. The substrates (DM basis for in vitro fermentation consisted of alfalfa hay (31.7%, Chinese wild rye grass hay (28.3%, ground corn grain (24.5%, soybean meal (15.5% with a forage: concentrate of 60:40. Treatments were the control (no additive and hainanmycin supplemented at 0.1 (H0.1, 1 (H1, 10 (H10, and 100 mg/kg (H100 of the substrates. After 24 h of fermentation, the highest addition level of hainanmycin decreased total VFA concentration and increased the final pH. The high addition level of hainanmycin (H1, H10, and H100 reduced (p0.05. After 24 h of fermentation, H10 and H100 increased (p<0.05 concentrations of peptide nitrogen and AA nitrogen and proteinase activity, and decreased (p<0.05 NH3-N concentration and deaminase activity compared with control. Peptidase activitives were not affected by hainanmycin. Hainanmycin supplementation only inhibited the growth of Butyrivibrio fibrisolvens, which is one of the species of low deaminative activity. Hainanmycin supplementation also decreased (p<0.05 relative population sizes of hyper-ammonia-producing species, except for H0.1 on Clostridium aminophilum. It was concluded that dietary supplementation with hainanmycin could improve ruminal fermentation and modify protein degradation by changing population size of ammonia-producing bacteria in vitro; and the addition level of 10 mg/kg appeared to achieve the best results.

  1. Protein Degradation in a TX-TL Cell-free Expression System Using ClpXP Protease

    Science.gov (United States)

    2014-07-14

    1! Protein degradation in a TX-TL cell-free expression system using ClpXP protease AUTHORS: Zachary Z. Sun1, Jongmin Kim1, Vipul Singhal2...COVERED 00-00-2014 to 00-00-2014 4. TITLE AND SUBTITLE Protein Degradation in a TX-TL Cell-free Expression System Using ClpXP Protease 5a...equipment and expertise or demand lower reaction throughput. We explored the possibility of supplementing TX-TL with ClpXP, an AAA+ protease

  2. Identifiability study of the proteins degradation model, based on ADM1, using simultaneous batch experiments

    DEFF Research Database (Denmark)

    Flotats, X.; Palatsi, J.; Ahring, Birgitte Kiær

    2006-01-01

    The objective of the present study is to analyse kinetic and stoichiometric parameter values of gelatine anaerobic degradation at thermophilic range, based on an experiment designed to elucidate if volatile fatty acids (VFA) are inhibitors of the hydrolysis process. Results showed that VFA are no...

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

    Directory of Open Access Journals (Sweden)

    Geraldine V. Dennett

    2016-02-01

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

  4. Characterization of malate dehydrogenase from the hyperthermophilic archaeon Pyrobaculum islandicum.

    Science.gov (United States)

    Yennaco, Lynda J; Hu, Yajing; Holden, James F

    2007-09-01

    Native and recombinant malate dehydrogenase (MDH) was characterized from the hyperthermophilic, facultatively autotrophic archaeon Pyrobaculum islandicum. The enzyme is a homotetramer with a subunit mass of 33 kDa. The activity kinetics of the native and recombinant proteins are the same. The apparent K ( m ) values of the recombinant protein for oxaloacetate (OAA) and NADH (at 80 degrees C and pH 8.0) were 15 and 86 microM, respectively, with specific activity as high as 470 U mg(-1). Activity decreased more than 90% when NADPH was used. The catalytic efficiency of OAA reduction by P. islandicum MDH using NADH was significantly higher than that reported for any other archaeal MDH. Unlike other archaeal MDHs, specific activity of the P. islandicum MDH back-reaction also decreased more than 90% when malate and NAD(+) were used as substrates and was not detected with NADP(+). A phylogenetic tree of 31 archaeal MDHs shows that they fall into 5 distinct groups separated largely along taxonomic lines suggesting minimal lateral mdh transfer between Archaea.

  5. Statistical Optimization of Medium Components for Improved Product Ion of Recombinant Hyperthermophilic Esterase

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    The optimization of nutrient levels for the production of recombinant hyperthermophilic esterase by E. coli was carried out with response surface methodology(RSM) based on the central composite rotatable design(CCRD). A 24central composite rotatable design was used to study the combined effect of the nutritional constituents like yeast extract, peptone, mineral salt and trace metals. The P-value of the coefficient for the linear effect of peptone concentration was 0. 0081 and trace metals solution was less than 0. 0001, suggesting that these were the principal variables with significant effect on the hyperthermophilic esterase production. The predicted optimal hyperthermophilic esterase yield was 269. 17 U/mL, whereas an actual experimental value of 284. 58 U/mL was obtained.

  6. Crystallization and preliminary X-ray diffraction analysis of the hyperthermophilic Sulfolobus solfataricus phosphotriesterase

    Energy Technology Data Exchange (ETDEWEB)

    Elias, Mikael [Laboratoire de Cristallographie et Modélisation des Matériaux Minéraux et Biologiques, CNRS-Université Henri Poincaré, 54506 Vandoeuvre-lès-Nancy (France); Dupuy, Jérôme [Laboratoire de Cristallogenèse et Cristallographie des Protéines, Institut de Biologie Structurale J.-P. Ebel, 38027 Grenoble (France); Merone, Luigia [Istituto di Biochimica delle Proteine, Consiglio Nazionale delle Ricerche, Via P. Castellino 111, 80131 Napoli (Italy); Lecomte, Claude [Laboratoire de Cristallographie et Modélisation des Matériaux Minéraux et Biologiques, CNRS-Université Henri Poincaré, 54506 Vandoeuvre-lès-Nancy (France); Rossi, Mosè [Istituto di Biochimica delle Proteine, Consiglio Nazionale delle Ricerche, Via P. Castellino 111, 80131 Napoli (Italy); Masson, Patrick [Unité d’Enzymologie, Département de Toxicologie, Centre de Recherches du Service de Santé des Armées, 38702 La Tronche (France); Manco, Giuseppe [Istituto di Biochimica delle Proteine, Consiglio Nazionale delle Ricerche, Via P. Castellino 111, 80131 Napoli (Italy); Chabriere, Eric, E-mail: eric.chabriere@lcm3b.uhp-nancy.fr [Laboratoire de Cristallographie et Modélisation des Matériaux Minéraux et Biologiques, CNRS-Université Henri Poincaré, 54506 Vandoeuvre-lès-Nancy (France); Unité d’Enzymologie, Département de Toxicologie, Centre de Recherches du Service de Santé des Armées, 38702 La Tronche (France)

    2007-07-01

    A phosphotriesterase (PTE) from the hyperthermophilic archaeon S. solfataricus has been crystallized. Combined with biochemical and bioengineering studies, it is expected that the structure of this protein will provide insight into the natural function of the PTE family and provide important data for achieving an efficient organophosphate biodecontaminant. Organophosphates constitute the largest class of insecticides used worldwide and some of them are potent nerve agents. Consequently, organophosphate-degrading enzymes are of paramount interest as they could be used as bioscavengers and biodecontaminants. Phosphotriesterases (PTEs) are capable of hydrolyzing these toxic compounds with high efficiency. A distant and hyperthermophilic representative of the PTE family was cloned from the archeon Sulfolobus solfataricus MT4, overexpressed in Escherichia coli and crystallized; the crystals diffracted to 2.54 Å resolution. Owing to its exceptional thermostability, this PTE may be an excellent candidate for obtaining an efficient organophosphate biodecontaminant. Here, the crystallization conditions and data collection for the hyperthermophilic S. solfataricus PTE are reported.

  7. Protein degradation and post-deboning tenderization in broiler breast meat with different degrees of muscle shortening

    Science.gov (United States)

    Deboning broiler breast fillets prior to rigor mortis negatively influences tenderness due to sarcomere shortening. The effects of sarcomere shortening on muscle protein degradation and breast meat tenderization during post-deboning aging are not well understood. The objective of this study was to m...

  8. Effect of feed deprivation and insulin-like growth hormone on indices of protein degradation in rainbow trout (Oncorhynchus mykiss)

    Science.gov (United States)

    Insulin-like growth factor-I (IGF-I) is a hormone that promotes growth by both increasing protein synthesis and decreasing protein degradation. This study utilizes a comparative slaughter approach to determine the effect of feed deprivation and IGF-I treatment on weight loss and indices of protein ...

  9. Anaerobic Digestion: Process

    DEFF Research Database (Denmark)

    Angelidaki, Irini; Batstone, Damien J.

    2011-01-01

    Organic waste may degrade anaerobically in nature as well as in engineered systems. The latter is called anaerobic digestion or biogasification. Anaerobic digestion produces two main outputs: An energy-rich gas called biogas and an effluent. The effluent, which may be a solid as well as liquid...... with very little dry matter may also be called a digest. The digest should not be termed compost unless it specifically has been composted in an aerated step. This chapter describes the basic processes of anaerobic digestion. Chapter 9.5 describes the anaerobic treatment technologies, and Chapter 9.......6 addresses the mass balances and environmental aspects of anaerobic digestion....

  10. Targeted Protein Degradation by Salmonella under Phagosome-Mimicking Culture Conditions Investigated Using Comparative Peptidomics

    Energy Technology Data Exchange (ETDEWEB)

    Manes, Nathan P.; Gustin, Jean K.; Rue, Joanne; Mottaz, Heather M.; Purvine, Samuel O.; Norbeck, Angela D.; Monroe, Matthew E.; Zimmer, Jennifer S.; Metz, Thomas O.; Adkins, Joshua N.; Smith, Richard D.; Heffron, Fred

    2007-04-01

    The pathogen Salmonella enterica is known to cause both food poisoning and typhoid fever. Due to the emergence of antibiotic-resistant isolates and the threat of bioterrorism (e.g., contamination of the food supply), there is a growing need to study this bacterium. In this investigation, comparative peptidomics was used to study Salmonella enterica serovar Typhimurium cultured in either a rich medium or in an acidic, low magnesium, and minimal nutrient medium designed to roughly mimic the macrophage phagosomal compartment (within which Salmonella are known to survive). Native peptides from cleared cell lysates were enriched by using isopropanol extraction and analyzed by using both LC-MS/MS and LC-FTICR-MS. We identified 5,163 distinct peptides originating from 682 proteins and the data clearly indicated that compared to cells cultured in the rich medium, Salmonella cultured in the phagosome-mimicking medium had dramatically higher abundances of a wide variety of protein degradation products, especially from ribosomal proteins. Salmonella from the same cultures were also analyzed by using bottom-up proteomics, and when the peptidomic and proteomic data were analyzed together, two clusters of proteins targeted for proteolysis were tentatively identified. Possible roles of targeted proteolysis by phagocytosed Salmonella are discussed.

  11. Stromal protein degradation is incomplete in Arabidopsis thaliana autophagy mutants undergoing natural senescence

    Directory of Open Access Journals (Sweden)

    Lee Travis A

    2013-01-01

    Full Text Available Abstract Background Degradation of highly abundant stromal proteins plays an important role in the nitrogen economy of the plant during senescence. Lines of evidence supporting proteolysis within the chloroplast and outside the chloroplast have been reported. Two extra-plastidic degradation pathways, chlorophagy and Rubisco Containing Bodies, rely on cytoplasmic autophagy. Results In this work, levels of three stromal proteins (Rubisco large subunit, chloroplast glutamine synthetase and Rubisco activase and one thylakoid protein (the major light harvesting complex protein of photosystem II were measured during natural senescence in WT and in two autophagy T-DNA insertion mutants (atg5 and atg7. Thylakoid-localized protein decreased similarly in all genotypes, but stromal protein degradation was incomplete in the two atg mutants. In addition, degradation of two stromal proteins was observed in chloroplasts isolated from mid-senescence leaves. Conclusions These data suggest that autophagy does contribute to the complete proteolysis of stromal proteins, but does not play a major degenerative role. In addition, support for in organello degradation is provided.

  12. Postmortem muscle protein degradation in humans as a tool for PMI delimitation.

    Science.gov (United States)

    Pittner, Stefan; Ehrenfellner, Bianca; Monticelli, Fabio C; Zissler, Angela; Sänger, Alexandra M; Stoiber, Walter; Steinbacher, Peter

    2016-11-01

    Forensic estimation of time since death relies on diverse approaches, including measurement and comparison of environmental and body core temperature and analysis of insect colonization on a dead body. However, most of the applied methods have practical limitations or provide insufficient results under certain circumstances. Thus, new methods that can easily be implemented into forensic routine work are required to deliver more and discrete information about the postmortem interval (PMI). Following a previous work on skeletal muscle degradation in the porcine model, we analyzed human postmortem skeletal muscle samples of 40 forensic cases by Western blotting and casein zymography. Our results demonstrate predictable protein degradation processes in human muscle that are distinctly associated with temperature and the PMI. We provide information on promising degradation markers for certain periods of time postmortem, which can be useful tools for time since death delimitation. In addition, we discuss external influencing factors such as age, body mass index, sex, and cause of death that need to be considered in future routine application of the method in humans.

  13. The Role of Lectin-Carbohydrate Interactions in the Regulation of ER-Associated Protein Degradation

    Directory of Open Access Journals (Sweden)

    Monika Słomińska-Wojewódzka

    2015-05-01

    Full Text Available Proteins entering the secretory pathway are translocated across the endoplasmic reticulum (ER membrane in an unfolded form. In the ER they are restricted to a quality control system that ensures correct folding or eventual degradation of improperly folded polypeptides. Mannose trimming of N-glycans on newly synthesized proteins plays an important role in the recognition and sorting of terminally misfolded glycoproteins for ER-associated protein degradation (ERAD. In this process misfolded proteins are retrotranslocated into the cytosol, polyubiquitinated, and eventually degraded by the proteasome. The mechanism by which misfolded glycoproteins are recognized and recruited to the degradation machinery has been extensively studied during last decade. In this review, we focus on ER degradation-enhancing α-mannosidase-like protein (EDEM family proteins that seem to play a key role in the discrimination between proteins undergoing a folding process and terminally misfolded proteins directed for degradation. We describe interactions of EDEM proteins with other components of the ERAD machinery, as well as with various protein substrates. Carbohydrate-dependent interactions together with N-glycan-independent interactions seem to regulate the complex process of protein recognition and direction for proteosomal degradation.

  14. DBC2 resistance is achieved by enhancing 26S proteasome-mediated protein degradation.

    Science.gov (United States)

    Collado, Denise; Yoshihara, Takashi; Hamaguchi, Masaaki

    2007-08-31

    Tumor suppressor gene DBC2 stops growth of tumor cells through regulation of CCND1. Interference of CCND1 down-regulation prevented growth arrest caused by DBC2 [T. Yoshihara, D. Collado, M. Hamaguchi, Cyclin D1 down-regulation is essential for DBC2's tumor suppressor function, Biochemical and biophysical research communications 358 (2007) 1076-1079]. It was also noted that DBC2 resistant cells eventually arose after repeated induction of DBC2 with muristerone A treatment [M. Hamaguchi, J.L. Meth, C. Von Klitzing, W. Wei, D. Esposito, L. Rodgers, T. Walsh, P. Welcsh, M.C. King, M.H. Wigler, DBC2, a candidate for a tumor suppressor gene involved in breast cancer, Proc. Natl. Acad. Sci. USA 99 (2002) 13647-13652]. In order to elucidate the mechanism of resistance acquisition, we analyzed DBC2 sensitive and resistant cells derived from the same progenitor cells (T-47D). We discovered that DBC2 protein was abundantly expressed in the sensitive cells when DBC2 was induced. In contrast, it was undetectable by western blot analysis in the resistant cells. We confirmed that the inducible gene expression system was responsive in both cells by detecting induced GFP. Additionally, inhibition of 26S proteasome by MG132 revealed production of DBC2 protein in the resistant cells. These findings indicate that the resistant T-47D cells survive DBC2 induction by rapid destruction of DBC2 through 26S proteasome-mediated protein degradation.

  15. Metabolic evolution of a deep-branching hyperthermophilic chemoautotrophic bacterium.

    Directory of Open Access Journals (Sweden)

    Rogier Braakman

    Full Text Available Aquifex aeolicus is a deep-branching hyperthermophilic chemoautotrophic bacterium restricted to hydrothermal vents and hot springs. These characteristics make it an excellent model system for studying the early evolution of metabolism. Here we present the whole-genome metabolic network of this organism and examine in detail the driving forces that have shaped it. We make extensive use of phylometabolic analysis, a method we recently introduced that generates trees of metabolic phenotypes by integrating phylogenetic and metabolic constraints. We reconstruct the evolution of a range of metabolic sub-systems, including the reductive citric acid (rTCA cycle, as well as the biosynthesis and functional roles of several amino acids and cofactors. We show that A. aeolicus uses the reconstructed ancestral pathways within many of these sub-systems, and highlight how the evolutionary interconnections between sub-systems facilitated several key innovations. Our analyses further highlight three general classes of driving forces in metabolic evolution. One is the duplication and divergence of genes for enzymes as these progress from lower to higher substrate specificity, improving the kinetics of certain sub-systems. A second is the kinetic optimization of established pathways through fusion of enzymes, or their organization into larger complexes. The third is the minimization of the ATP unit cost to synthesize biomass, improving thermodynamic efficiency. Quantifying the distribution of these classes of innovations across metabolic sub-systems and across the tree of life will allow us to assess how a tradeoff between maximizing growth rate and growth efficiency has shaped the long-term metabolic evolution of the biosphere.

  16. Anaerobes in pleuropulmonary infections

    Directory of Open Access Journals (Sweden)

    De A

    2002-01-01

    Full Text Available A total of 76 anaerobes and 122 aerobes were isolated from 100 patients with pleuropulmonary infections, e.g. empyema (64, pleural effusion (19 and lung abscess (13. In 14% of the patients, only anaerobes were recovered, while a mixture of aerobes and anaerobes was encountered in 58%. From all cases of lung abscess, anaerobic bacteria were isolated, alone (04 or along with aerobic bacteria (13. From empyema and pleural effusion cases, 65.6% and 68.4% anaerobes were recovered respectively. Amongst anaerobes, gram negative anaerobic bacilli predominated (Prevotella melaninogenicus 16, Fusobacterium spp. 10, Bacteroides spp. 9, followed by gram positive anaerobic cocci (Peptostreptococcus spp. 31. Coliform bacteria (45 and Pseudomonas aeruginosa (42 were the predominant aerobic isolates.

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

    DEFF Research Database (Denmark)

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

    2005-01-01

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

  18. Model for a novel membrane envelope in a filamentous hyperthermophilic virus

    NARCIS (Netherlands)

    Kasson, P.; DiMaio, F.; Yu, X.; Lucas-Staat, S.; Krupovic, M.; Schouten, S.; Prangishvili, D.; Egelman, E.H.

    2017-01-01

    Biological membranes create compartments, and are usually formed by lipid bilayers.However, in hyperthermophilic archaea that live optimally at temperatures above 80°C themembranes are monolayers which resemble fused bilayers. Many double-stranded DNA viruseswhich parasitize such hosts, including

  19. Development of a Simvastatin Selection Marker for a Hyperthermophilic Acidophile, Sulfolobus islandicus

    DEFF Research Database (Denmark)

    Zheng, Tao; Huang, Qihong; Zhang, Changyi;

    2012-01-01

    We report here a novel selectable marker for the hyperthermophilic crenarchaeon Sulfolobus islandicus. The marker cassette is composed of the sac7d promoter and the hmg gene coding for the 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase (P(sac7d)-hmg), which confers simvastatin resistance...

  20. Production and characterization of a thermostable L-threonine dehydrogenase from the hyperthermophilic archaeon Pyrococcus furiosus

    NARCIS (Netherlands)

    Machielsen, M.P.; Oost, van der J.

    2006-01-01

    The gene encoding a threonine dehydrogenase (TDH) has been identified in the hyperthermophilic archaeon Pyrococcus furiosus. The Pf-TDH protein has been functionally produced in Escherichia coli and purified to homogeneity. The enzyme has a tetrameric conformation with a molecular mass of ¿ 155 kDa.

  1. UV-inducible cellular aggregation of the hyperthermophilic archaeon Sulfolobus solfataricus is mediated by pili formation

    NARCIS (Netherlands)

    Froels, Sabrina; Ajon, Malgorzata; Wagner, Michaela; Teichmann, Daniela; Zolghadr, Behnam; Folea, Mihaela; Boekema, Egbert J.; Driessen, Arnold J. M.; Schleper, Christa; Albers, Sonja-Verena

    2008-01-01

    The hyperthermophilic archaeon Sulfolobus solfataricus has been shown to exhibit a complex transcriptional response to UV irradiation involving 55 genes. Among the strongest UV-induced genes was a putative pili biogenesis operon encoding a potential secretion ATPase, two pre-pilins, a putative trans

  2. Functional and structural characterization of the minimal Sec translocase of the hyperthermophile Thermotoga maritima

    NARCIS (Netherlands)

    Pretz, MG; Remigy, H; Swaving, J; Albers, SV; Garrido, VG; Chami, M; Engel, A; Driessen, AJM; Pretz, Monika G.; Garrido, Victoria G.

    2005-01-01

    The genome of the hyperthermophilic bacterium Thermotoga maritima contains the genes that encode core subunits of the protein translocase, a complex consisting of the molecular motor SecA and the protein conducting pore SecYE. In addition, we identified an erroneous sequence in the genome encoding f

  3. UV-inducible DNA exchange in hyperthermophilic archaea mediated by type IV pili

    NARCIS (Netherlands)

    Ajon, Malgorzata; Froels, Sabrina; van Wolferen, Marleen; Stoecker, Kilian; Teichmann, Daniela; Driessen, Arnold J. M.; Grogan, Dennis W.; Albers, Sonja-Verena; Schleper, Christa; Ajon, Małgorzata

    2011-01-01

    Archaea, like bacteria and eukaryotes, contain proteins involved in various mechanisms of DNA repair, highlighting the importance of these processes for all forms of life. Species of the order Sulfolobales of hyperthermophilic crenarchaeota are equipped with a strongly UV-inducible type IV pilus sys

  4. The Geoglobus acetivorans genome: Fe(III) reduction, acetate utilization, autotrophic growth, and degradation of aromatic compounds in a hyperthermophilic archaeon.

    Science.gov (United States)

    Mardanov, Andrey V; Slododkina, Galina B; Slobodkin, Alexander I; Beletsky, Alexey V; Gavrilov, Sergey N; Kublanov, Ilya V; Bonch-Osmolovskaya, Elizaveta A; Skryabin, Konstantin G; Ravin, Nikolai V

    2015-02-01

    Geoglobus acetivorans is a hyperthermophilic anaerobic euryarchaeon of the order Archaeoglobales isolated from deep-sea hydrothermal vents. A unique physiological feature of the members of the genus Geoglobus is their obligate dependence on Fe(III) reduction, which plays an important role in the geochemistry of hydrothermal systems. The features of this organism and its complete 1,860,815-bp genome sequence are described in this report. Genome analysis revealed pathways enabling oxidation of molecular hydrogen, proteinaceous substrates, fatty acids, aromatic compounds, n-alkanes, and organic acids, including acetate, through anaerobic respiration linked to Fe(III) reduction. Consistent with the inability of G. acetivorans to grow on carbohydrates, the modified Embden-Meyerhof pathway encoded by the genome is incomplete. Autotrophic CO2 fixation is enabled by the Wood-Ljungdahl pathway. Reduction of insoluble poorly crystalline Fe(III) oxide depends on the transfer of electrons from the quinone pool to multiheme c-type cytochromes exposed on the cell surface. Direct contact of the cells and Fe(III) oxide particles could be facilitated by pilus-like appendages. Genome analysis indicated the presence of metabolic pathways for anaerobic degradation of aromatic compounds and n-alkanes, although an ability of G. acetivorans to grow on these substrates was not observed in laboratory experiments. Overall, our results suggest that Geoglobus species could play an important role in microbial communities of deep-sea hydrothermal vents as lithoautotrophic producers. An additional role as decomposers would close the biogeochemical cycle of carbon through complete mineralization of various organic compounds via Fe(III) respiration.

  5. Previously unknown role for the ubiquitin ligase Ubr1 in endoplasmic reticulum-associated protein degradation.

    Science.gov (United States)

    Stolz, Alexandra; Besser, Stefanie; Hottmann, Heike; Wolf, Dieter H

    2013-09-17

    Quality control and degradation of misfolded proteins are essential processes of all cells. The endoplasmic reticulum (ER) is the entry site of proteins into the secretory pathway in which protein folding occurs and terminally misfolded proteins are recognized and retrotranslocated across the ER membrane into the cytosol. Here, proteins undergo polyubiquitination by one of the membrane-embedded ubiquitin ligases, in yeast Hrd1/Der3 (HMG-CoA reductase degradation/degradation of the ER) and Doa10 (degradation of alpha), and are degraded by the proteasome. In this study, we identify cytosolic Ubr1 (E3 ubiquitin ligase, N-recognin) as an additional ubiquitin ligase that can participate in ER-associated protein degradation (ERAD) in yeast. We show that two polytopic ERAD substrates, mutated transporter of the mating type a pheromone, Ste6* (sterile), and cystic fibrosis transmembrane conductance regulator, undergo Ubr1-dependent degradation in the presence and absence of the canonical ER ubiquitin ligases. Whereas in the case of Ste6* Ubr1 is specifically required under stress conditions such as heat or ethanol or in the absence of the canonical ER ligases, efficient degradation of human cystic fibrosis transmembrane conductance regulator requires function of Ubr1 already in wild-type cells under standard growth conditions. Together with the Hsp70 (heat shock protein) chaperone Ssa1 (stress-seventy subfamily A) and the AAA-type ATPase Cdc48 (cell division cycle), Ubr1 directs the substrate to proteasomal degradation. These data unravel another layer of complexity in ERAD.

  6. Protein degradation by ubiquitin–proteasome system in formation and labilization of contextual conditioning memory

    Science.gov (United States)

    Sol Fustiñana, María; de la Fuente, Verónica; Federman, Noel; Freudenthal, Ramiro

    2014-01-01

    The ubiquitin–proteasome system (UPS) of protein degradation has been evaluated in different forms of neural plasticity and memory. The role of UPS in such processes is controversial. Several results support the idea that the activation of this system in memory consolidation is necessary to overcome negative constrains for plasticity. In this case, the inhibition of the UPS during consolidation impairs memory. Similar results were reported for memory reconsolidation. However, in other cases, the inhibition of UPS had no effect on memory consolidation and reconsolidation but impedes the amnesic action of protein synthesis inhibition after retrieval. The last finding suggests a specific action of the UPS inhibitor on memory labilization. However, another interpretation is possible in terms of the synthesis/degradation balance of positive and negative elements in neural plasticity, as was found in the case of long-term potentiation. To evaluate these alternative interpretations, other reconsolidation-interfering drugs than translation inhibitors should be tested. Here we analyzed initially the UPS inhibitor effect in contextual conditioning in crabs. We found that UPS inhibition during consolidation impaired long-term memory. In contrast, UPS inhibition did not affect memory reconsolidation after contextual retrieval but, in fact, impeded memory labilization, blocking the action of drugs that does not affect directly the protein synthesis. To extend these finding to vertebrates, we performed similar experiments in contextual fear memory in mice. We found that the UPS inhibitor in hippocampus affected memory consolidation and blocked memory labilization after retrieval. These findings exclude alternative interpretations to the requirement of UPS in memory labilization and give evidence of this mechanism in both vertebrates and invertebrates. PMID:25135196

  7. Anaerobic Digestion: Process

    DEFF Research Database (Denmark)

    Angelidaki, Irini; Batstone, Damien J.

    2011-01-01

    Organic waste may degrade anaerobically in nature as well as in engineered systems. The latter is called anaerobic digestion or biogasification. Anaerobic digestion produces two main outputs: An energy-rich gas called biogas and an effluent. The effluent, which may be a solid as well as liquid...... with very little dry matter may also be called a digest. The digest should not be termed compost unless it specifically has been composted in an aerated step. This chapter describes the basic processes of anaerobic digestion. Chapter 9.5 describes the anaerobic treatment technologies, and Chapter 9...

  8. Two waves of proteasome-dependent protein degradation in the hippocampus are required for recognition memory consolidation.

    Science.gov (United States)

    Figueiredo, Luciana S; Dornelles, Arethuza S; Petry, Fernanda S; Falavigna, Lucio; Dargél, Vinicius A; Köbe, Luiza M; Aguzzoli, Cristiano; Roesler, Rafael; Schröder, Nadja

    2015-04-01

    Healthy neuronal function and synaptic modification require a concert of synthesis and degradation of proteins. Increasing evidence indicates that protein turnover mediated by proteasome activity is involved in long-term synaptic plasticity and memory. However, its role in different phases of memory remains debated, and previous studies have not examined the possible requirement of protein degradation in recognition memory. Here, we show that the proteasome inhibitor, lactacystin (LAC), infused into the CA1 area of the hippocampus at two specific time points during consolidation, impairs 24-retention of memory for object recognition in rats. Administration of LAC after retrieval did not affect retention. These findings provide the first evidence for a requirement of proteasome activity in recognition memory, indicate that protein degradation in the hippocampus is necessary during selective time windows of memory consolidation, and further our understanding of the role of protein turnover in memory formation. Copyright © 2015 Elsevier Inc. All rights reserved.

  9. Inhibition of Hsp70 by Methylene Blue Affects Signaling Protein Function and Ubiquitination and Modulates Polyglutamine Protein Degradation*

    OpenAIRE

    Wang, Adrienne M; Morishima, Yoshihiro; Clapp, Kelly M.; Peng, Hwei-Ming; Pratt, William B.; Gestwicki, Jason E.; Osawa, Yoichi; Lieberman, Andrew P.

    2010-01-01

    The Hsp90/Hsp70-based chaperone machinery regulates the activity and degradation of many signaling proteins. Cycling with Hsp90 stabilizes client proteins, whereas Hsp70 interacts with chaperone-dependent E3 ubiquitin ligases to promote protein degradation. To probe these actions, small molecule inhibitors of Hsp70 would be extremely useful; however, few have been identified. Here we test the effects of methylene blue, a recently described inhibitor of Hsp70 ATPase activity, in three well est...

  10. Adsorption of an endoglucanase from the hyperthermophilic Pyrococcus furiosus of hydrophobic (polysterene) and hydrophilic (silica) surfaces increases protein heat stability

    NARCIS (Netherlands)

    Koutsopoulos, S.; Oost, van der J.; Norde, W.

    2004-01-01

    The interaction of an endoglucanase from the hyperthermophilic microorganism Pyrococcus furiosus with two types of surfaces, that is, hydrophobic polystyrene and hydrophilic silica, was investigated, and the adsorption isotherms were determined. The adsorbed hyperthermostable enzyme did not undergo

  11. Adsorption of an endoglucanase from the hyperthermophilic Pyrococcus furiosus on hydrophobic (polystyrene) and hydrophilic (silica) surfaces increases protein heat stability

    NARCIS (Netherlands)

    Koutsopoulos, S.; van der Oost, J.; Norde, Willem

    2004-01-01

    The interaction of an endoglucanase from the hyperthermophilic microorganism Pyrococcus furiosus with two types of surfaces, that is, hydrophobic polystyrene and hydrophilic silica, was investigated, and the adsorption isotherms were determined. The adsorbed hyperthermostable enzyme did not undergo

  12. Activity dependent protein degradation is critical for the formation and stability of fear memory in the amygdala.

    Directory of Open Access Journals (Sweden)

    Timothy J Jarome

    Full Text Available Protein degradation through the ubiquitin-proteasome system [UPS] plays a critical role in some forms of synaptic plasticity. However, its role in memory formation in the amygdala, a site critical for the formation of fear memories, currently remains unknown. Here we provide the first evidence that protein degradation through the UPS is critically engaged at amygdala synapses during memory formation and retrieval. Fear conditioning results in NMDA-dependent increases in degradation-specific polyubiquitination in the amygdala, targeting proteins involved in translational control and synaptic structure and blocking the degradation of these proteins significantly impairs long-term memory. Furthermore, retrieval of fear memory results in a second wave of NMDA-dependent polyubiquitination that targets proteins involved in translational silencing and synaptic structure and is critical for memory updating following recall. These results indicate that UPS-mediated protein degradation is a major regulator of synaptic plasticity necessary for the formation and stability of long-term memories at amygdala synapses.

  13. Regulation of protein degradation pathways by amino acids and insulin in skeletal muscle of neonatal pigs

    Institute of Scientific and Technical Information of China (English)

    Agus Suryawan; Teresa ADavis

    2014-01-01

    Background:The rapid gain in lean mass in neonates requires greater rates of protein synthesis than degradation. We previously delineated the molecular mechanisms by which insulin and amino acids, especially leucine, modulate skeletal muscle protein synthesis and how this changes with development. In the current study, we identified mechanisms involved in protein degradation regulation. In experiment 1, 6-and 26-d-old pigs were studied during 1) euinsulinemic-euglycemic-euaminoacidemic, 2) euinsulinemic-euglycemic-hyperaminoacidemic, and 3) hyperinsulinemic-euglycemic-euaminoacidemic clamps for 2 h. In experiment 2, 5-d-old pigs were studied during 1) euinsulinemic-euglycemic-euaminoacidemic-euleucinemic, 2) euinsulinemic-euglycemic-hypoaminoacidemic-hyperleucinemic, and 3) euinsulinemic-euglycemic-euaminoacidemic-hyperleucinemic clamps for 24 h. We determined in muscle indices of ubiquitin-proteasome, i.e., atrogin-1 (MAFbx) and muscle RING-finger protein-1 (MuRF1) and autophagy-lysosome systems, i.e., unc51-like kinase 1 (UKL1), microtubule-associated protein light chain 3 (LC3), and lysosomal-associated membrane protein 2 (Lamp-2). For comparison, we measured ribosomal protein S6 (rpS6) and eukaryotic initiation factor 4E (eIF4E) activation, components of translation initiation. Results:Abundance of atrogin-1, but not MuRF1, was greater in 26-than 6-d-old pigs and was not affected by insulin, amino acids, or leucine. Abundance of ULK1 and LC3 was higher in younger pigs and not affected by treatment. The LC3-II/LC3-I ratio was reduced and ULK1 phosphorylation increased by insulin, amino acids, and leucine. These responses were more profound in younger pigs. Abundance of Lamp-2 was not affected by treatment or development. Abundance of eIF4E, but not rpS6, was higher in 6-than 26-d-old-pigs but unaffected by treatment. Phosphorylation of eIF4E was not affected by treatment, however, insulin, amino acids, and leucine stimulated rpS6 phosphorylation, and the

  14. Central Metabolic Pathways of Hyperthermophiles: Important Clues on how Metabolism Gives Rise to Life

    Science.gov (United States)

    Ronimus, R. S.; Morgan, H. W.

    2004-06-01

    Vital clues on life's origins within the galaxy exist here on present day Earth. Life is currently divided into the three domains Bacteria, Archaea and Eukarya based on the phylogeny of small ribosomal subunit RNA (16S/18S) gene sequences. The domains are presumed to share a ``last universal common ancestor'' (LUCA). Hyperthermophilic bacteria and archaea, which are able to thrive at 80^{circ}C or higher, dominate the bottom of the tree of life and are thus suggested to be the least evolved, or most ``ancient''. Geochemical data indicates that life first appeared on Earth approximately 3.8 billion years ago in a hot environment. Due to these considerations, hyperthermophiles represent the most appropriate microorganisms to investigate the origins of metabolism. The central biochemical pathway of gluconeogenesis/glycolysis (the Embden-Meyerhof pathway) which produces six carbon sugars from three carbon compounds is present in all organisms and can provide important hints concerning the early development of metabolism. Significantly, there are a number of striking deviations from the textbook canonical reaction sequence that are found, particularly in hyperthermophilic archaea. In this paper the phylogenetic istribution of enzymes of the pathway is detailed; overall, the distribution pattern provides strong evidence for the pathway to have developed from the bottom-up.

  15. Characterization of protein degradation in serum-based lubricants during simulation wear testing of metal-on-metal hip prostheses.

    Science.gov (United States)

    Maskiewicz, Victoria K; Williams, Paul A; Prates, Sarah J; Bowsher, John G; Clarke, Ian C

    2010-08-01

    A size exclusion high performance liquid chromatography (SEC-HPLC) method has been developed which is capable of separation and quantitation of bovine serum albumin (BSA) and bovine serum globulin (BSG) components of serum-based lubricant (SBL) solutions. This allowed characterization of the stability profiles of these proteins when acting as lubricants during hip wear simulation, and identification of wear-specific mechanisms of degradation. Using cobalt-chromium metal-on-metal (MOM) hip joints, it was observed that BSA remained stable for up to 3 days (215K cycles) of wear testing after which the protein degraded in a fairly linear fashion. BSG on the other hand, began to degrade immediately and in a linear fashion with a rate constant of 5% per day. Loss of both proteins occurred via the formation of high molecular weight aggregates which precipitated out of solution. No fragmentation of the polypeptide backbone of either protein was observed. Data obtained suggest that protein degradation was not due to microbial contamination, denaturation at the air-water interface, or frictional heating of articulating joint surfaces in these studies. We conclude that the primary source of protein degradation during MOM simulation testing occurs via high shear rates experienced by SBL solutions at articulating surfaces, possibly coupled with metal-protein interactions occurring as new and reactive metal surfaces are generated during wear testing. The development of this analytical methodology will allow new studies to clarify the role of SBL solutions in wear simulation studies and the interactions and lubricating properties of serum proteins with prosthetic surfaces other than MOM.

  16. Perspectives for anaerobic digestion

    DEFF Research Database (Denmark)

    Ahring, Birgitte Kiær

    2003-01-01

    to the soil. Anaerobic digestion (AD) is one way of achieving this goal and it will furthermore, reduce energy consumption or may even be net energy producing. This chapter aims at provide a basic understanding of the world in which anaerobic digestion is operating today. The newest process developments...

  17. Dry anaerobic digestion of lignocellulosic and protein residues

    Directory of Open Access Journals (Sweden)

    Maryam M Kabir

    2015-12-01

    Full Text Available Utilisation of wheat straw and wool textile waste in dry anaerobic digestion (AD process was investigated. Dry-AD of the individual substrates as well as co-digestion of those were evaluated using different total solid (TS contents ranging between 6 to 30%. Additionally, the effects of the addition of nutrients and cellulose- or protein-degrading enzymes on the performance of the AD process were also investigated. Dry-AD of the wheat straw resulted in methane yields of 0.081 – 0.200 Nm3CH4/kgVS with the lowest and highest values obtained at 30 and 21% TS, respectively. The addition of the cellulolytic enzymes could significantly increase the yield in the reactor containing 13% TS (0.231 Nm3CH4/kg VS. Likewise, degradation of wool textile waste was enhanced significantly at TS of 13% with the addition of the protein-degrading enzyme (0.131 Nm3CH4/kg VS. Furthermore, the co-digestion of these two substrates showed higher methane yields compared with the methane potentials calculated for the individual fractions at all the investigated TS contents due to synergetic effects and better nutritional balance.

  18. A novel role for ATM in regulating proteasome-mediated protein degradation through suppression of the ISG15 conjugation pathway.

    Directory of Open Access Journals (Sweden)

    Laurence M Wood

    Full Text Available Ataxia Telangiectasia (A-T is an inherited immunodeficiency disorder wherein mutation of the ATM kinase is responsible for the A-T pathogenesis. Although the precise role of ATM in A-T pathogenesis is still unclear, its function in responding to DNA damage has been well established. Here we demonstrate that in addition to its role in DNA repair, ATM also regulates proteasome-mediated protein turnover through suppression of the ISG15 pathway. This conclusion is based on three major pieces of evidence: First, we demonstrate that proteasome-mediated protein degradation is impaired in A-T cells. Second, we show that the reduced protein turnover is causally linked to the elevated expression of the ubiquitin-like protein ISG15 in A-T cells. Third, we show that expression of the ISG15 is elevated in A-T cells derived from various A-T patients, as well as in brain tissues derived from the ATM knockout mice and A-T patients, suggesting that ATM negatively regulates the ISG15 pathway. Our current findings suggest for the first time that proteasome-mediated protein degradation is impaired in A-T cells due to elevated expression of the ISG15 conjugation pathway, which could contribute to progressive neurodegeneration in A-T patients.

  19. Anaerobic Digestion and its Applications

    Science.gov (United States)

    Anaerobic digestion is a natural biological process. The initials "AD" may refer to the process of anaerobic digestion, or the built systems of anaerobic digesters. While there are many kinds of digesters, the biology is basically the same for all. Anaerobic digesters are built...

  20. Anaerobic Digestion and its Applications

    Science.gov (United States)

    Anaerobic digestion is a natural biological process. The initials "AD" may refer to the process of anaerobic digestion, or the built systems of anaerobic digesters. While there are many kinds of digesters, the biology is basically the same for all. Anaerobic digesters are built...

  1. Anaerobic Digestion Foaming Causes

    OpenAIRE

    Ganidi, Nafsika

    2008-01-01

    Anaerobic digestion foaming has been encountered in several sewage treatment plants in the UK. Foaming has raised major concerns for the water utilities due to significant impacts on process efficiency and operational costs. Several foaming causes have been suggested over the past few years by researchers. However, the supporting experimental information is limited and in some cases site specific. The present report aimed to provide a better understanding of the anaerobic di...

  2. Thermococcus piezophilus sp. nov., a novel hyperthermophilic and piezophilic archaeon with a broad pressure range for growth, isolated from a deepest hydrothermal vent at the Mid-Cayman Rise.

    Science.gov (United States)

    Dalmasso, Cécile; Oger, Philippe; Selva, Gwendoline; Courtine, Damien; L'Haridon, Stéphane; Garlaschelli, Alexandre; Roussel, Erwan; Miyazaki, Junichi; Reveillaud, Julie; Jebbar, Mohamed; Takai, Ken; Maignien, Lois; Alain, Karine

    2016-10-01

    A novel strictly anaerobic, hyperthermophilic archaeon, designated strain CDGS(T), was isolated from a deep-sea hydrothermal vent in the Cayman Trough at 4964m water depth. The novel isolate is obligate anaerobe and grows chemoorganoheterotrophically with stimulation of growth by sulphur containing compounds. Its growth is optimal at 75°C, pH 6.0 and under a pressure of 50MPa. It possesses the broadest hydrostatic pressure range for growth that has ever been described for a microorganism. Its genomic DNA G+C content is 51.11mol%. The novel isolate belongs to the genus Thermococcus. Phylogenetic analyses indicated that it is most closely related to Thermococcus barossii DSM17882(T) based on its 16S rRNA gene sequence, and to 'Thermococcus onnurineus' NA1 based on its whole genome sequence. The average nucleotide identity scores with these strains are 77.66% for T. barossii and 84.84% for 'T. onnurineus', respectively. Based on the draft whole genome sequence and phenotypic characteristics, strain CDGS(T) is suggested to be separated into a novel species within the genus Thermococcus, with proposed name Thermococcus piezophilus (type strain CDGS(T)=ATCC TSD-33(T)=UBOCC 3296(T)). Copyright © 2016 Elsevier GmbH. All rights reserved.

  3. Oxidative modifications, mitochondrial dysfunction, and impaired protein degradation in Parkinson's disease: how neurons are lost in the Bermuda triangle

    Directory of Open Access Journals (Sweden)

    Malkus Kristen A

    2009-06-01

    Full Text Available Abstract While numerous hypotheses have been proposed to explain the molecular mechanisms underlying the pathogenesis of neurodegenerative diseases, the theory of oxidative stress has received considerable support. Although many correlations have been established and encouraging evidence has been obtained, conclusive proof of causation for the oxidative stress hypothesis is lacking and potential cures have not emerged. Therefore it is likely that other factors, possibly in coordination with oxidative stress, contribute to neuron death. Using Parkinson's disease (PD as the paradigm, this review explores the hypothesis that oxidative modifications, mitochondrial functional disruption, and impairment of protein degradation constitute three interrelated molecular pathways that execute neuron death. These intertwined events are the consequence of environmental exposure, genetic factors, and endogenous risks and constitute a "Bermuda triangle" that may be considered the underlying cause of neurodegenerative pathogenesis.

  4. Oxidative modifications, mitochondrial dysfunction, and impaired protein degradation in Parkinson's disease: how neurons are lost in the Bermuda triangle.

    Science.gov (United States)

    Malkus, Kristen A; Tsika, Elpida; Ischiropoulos, Harry

    2009-06-05

    While numerous hypotheses have been proposed to explain the molecular mechanisms underlying the pathogenesis of neurodegenerative diseases, the theory of oxidative stress has received considerable support. Although many correlations have been established and encouraging evidence has been obtained, conclusive proof of causation for the oxidative stress hypothesis is lacking and potential cures have not emerged. Therefore it is likely that other factors, possibly in coordination with oxidative stress, contribute to neuron death. Using Parkinson's disease (PD) as the paradigm, this review explores the hypothesis that oxidative modifications, mitochondrial functional disruption, and impairment of protein degradation constitute three interrelated molecular pathways that execute neuron death. These intertwined events are the consequence of environmental exposure, genetic factors, and endogenous risks and constitute a "Bermuda triangle" that may be considered the underlying cause of neurodegenerative pathogenesis.

  5. Inhibition of Hsp70 by Methylene Blue Affects Signaling Protein Function and Ubiquitination and Modulates Polyglutamine Protein Degradation*

    Science.gov (United States)

    Wang, Adrienne M.; Morishima, Yoshihiro; Clapp, Kelly M.; Peng, Hwei-Ming; Pratt, William B.; Gestwicki, Jason E.; Osawa, Yoichi; Lieberman, Andrew P.

    2010-01-01

    The Hsp90/Hsp70-based chaperone machinery regulates the activity and degradation of many signaling proteins. Cycling with Hsp90 stabilizes client proteins, whereas Hsp70 interacts with chaperone-dependent E3 ubiquitin ligases to promote protein degradation. To probe these actions, small molecule inhibitors of Hsp70 would be extremely useful; however, few have been identified. Here we test the effects of methylene blue, a recently described inhibitor of Hsp70 ATPase activity, in three well established systems of increasing complexity. First, we demonstrate that methylene blue inhibits the ability of the purified Hsp90/Hsp70-based chaperone machinery to enable ligand binding by the glucocorticoid receptor and show that this effect is due to specific inhibition of Hsp70. Next, we establish that ubiquitination of neuronal nitric-oxide synthase by the native ubiquitinating system of reticulocyte lysate is dependent upon both Hsp70 and the E3 ubiquitin ligase CHIP and is blocked by methylene blue. Finally, we demonstrate that methylene blue impairs degradation of the polyglutamine expanded androgen receptor, an Hsp90 client mutated in spinal and bulbar muscular atrophy. In contrast, degradation of an amino-terminal fragment of the receptor, which lacks the ligand binding domain and, therefore, is not a client of the Hsp90/Hsp70-based chaperone machinery, is enhanced through homeostatic induction of autophagy that occurs when Hsp70-dependent proteasomal degradation is inhibited by methylene blue. Our data demonstrate the utility of methylene blue in defining Hsp70-dependent functions and reveal divergent effects on polyglutamine protein degradation depending on whether the substrate is an Hsp90 client. PMID:20348093

  6. Inhibition of hsp70 by methylene blue affects signaling protein function and ubiquitination and modulates polyglutamine protein degradation.

    Science.gov (United States)

    Wang, Adrienne M; Morishima, Yoshihiro; Clapp, Kelly M; Peng, Hwei-Ming; Pratt, William B; Gestwicki, Jason E; Osawa, Yoichi; Lieberman, Andrew P

    2010-05-21

    The Hsp90/Hsp70-based chaperone machinery regulates the activity and degradation of many signaling proteins. Cycling with Hsp90 stabilizes client proteins, whereas Hsp70 interacts with chaperone-dependent E3 ubiquitin ligases to promote protein degradation. To probe these actions, small molecule inhibitors of Hsp70 would be extremely useful; however, few have been identified. Here we test the effects of methylene blue, a recently described inhibitor of Hsp70 ATPase activity, in three well established systems of increasing complexity. First, we demonstrate that methylene blue inhibits the ability of the purified Hsp90/Hsp70-based chaperone machinery to enable ligand binding by the glucocorticoid receptor and show that this effect is due to specific inhibition of Hsp70. Next, we establish that ubiquitination of neuronal nitric-oxide synthase by the native ubiquitinating system of reticulocyte lysate is dependent upon both Hsp70 and the E3 ubiquitin ligase CHIP and is blocked by methylene blue. Finally, we demonstrate that methylene blue impairs degradation of the polyglutamine expanded androgen receptor, an Hsp90 client mutated in spinal and bulbar muscular atrophy. In contrast, degradation of an amino-terminal fragment of the receptor, which lacks the ligand binding domain and, therefore, is not a client of the Hsp90/Hsp70-based chaperone machinery, is enhanced through homeostatic induction of autophagy that occurs when Hsp70-dependent proteasomal degradation is inhibited by methylene blue. Our data demonstrate the utility of methylene blue in defining Hsp70-dependent functions and reveal divergent effects on polyglutamine protein degradation depending on whether the substrate is an Hsp90 client.

  7. Glutamine supplementation stimulates protein-synthetic and inhibits protein-degradative signaling pathways in skeletal muscle of diabetic rats.

    Directory of Open Access Journals (Sweden)

    Adriana C Lambertucci

    Full Text Available In this study, we investigated the effect of glutamine (Gln supplementation on the signaling pathways regulating protein synthesis and protein degradation in the skeletal muscle of rats with streptozotocin (STZ-induced diabetes. The expression levels of key regulatory proteins in the synthetic pathways (Akt, mTOR, GSK3 and 4E-BP1 and the degradation pathways (MuRF-1 and MAFbx were determined using real-time PCR and Western blotting in four groups of male Wistar rats; 1 control, non-supplemented with glutamine; 2 control, supplemented with glutamine; 3 diabetic, non-supplemented with glutamine; and 4 diabetic, supplemented with glutamine. Diabetes was induced by the intravenous injection of 65 mg/kg bw STZ in citrate buffer (pH 4.2; the non-diabetic controls received only citrate buffer. After 48 hours, diabetes was confirmed in the STZ-treated animals by the determination of blood glucose levels above 200 mg/dL. Starting on that day, a solution of 1 g/kg bw Gln in phosphate buffered saline (PBS was administered daily via gavage for 15 days to groups 2 and 4. Groups 1 and 3 received only PBS for the same duration. The rats were euthanized, and the soleus muscles were removed and homogenized in extraction buffer for the subsequent measurement of protein and mRNA levels. The results demonstrated a significant decrease in the muscle Gln content in the diabetic rats, and this level increased toward the control value in the diabetic rats receiving Gln. In addition, the diabetic rats exhibited a reduced mRNA expression of regulatory proteins in the protein synthesis pathway and increased expression of those associated with protein degradation. A reduction in the skeletal muscle mass in the diabetic rats was observed and was alleviated partially with Gln supplementation. The data suggest that glutamine supplementation is potentially useful for slowing the progression of muscle atrophy in patients with diabetes.

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

    Science.gov (United States)

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

    1990-01-01

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

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

    Science.gov (United States)

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

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

  10. Reactor-Scale Cultivation of the Hyperthermophilic Methanarchaeon Methanococcus jannaschii to High Cell Densities

    OpenAIRE

    Mukhopadhyay, Biswarup; Johnson, Eric F.; Wolfe, Ralph S.

    1999-01-01

    For the hyperthermophilic and barophilic methanarchaeon Methanococcus jannaschii, we have developed a medium and protocols for reactor-scale cultivation that improved the final cell yield per liter from ∼0.5 to ∼7.5 g of packed wet cells (∼1.8 g dry cell mass) under autotrophic growth conditions and to ∼8.5 g of packed wet cells (∼2 g dry cell mass) with yeast extract (2 g liter−1) and tryptone (2 g liter−1) as medium supplements. For growth in a sealed bottle it was necessary to add Se to th...

  11. Anaerobic Biodegradation of Detergent Surfactants

    Directory of Open Access Journals (Sweden)

    Erich Jelen

    2009-03-01

    Full Text Available Detergent surfactantscan be found in wastewater in relevant concentrations. Most of them are known as ready degradable under aerobic conditions, as required by European legislation. Far fewer surfactants have been tested so far for biodegradability under anaerobic conditions. The natural environment is predominantly aerobic, but there are some environmental compartments such as river sediments, sub-surface soil layer and anaerobic sludge digesters of wastewater treatment plants which have strictly anaerobic conditions. This review gives an overview on anaerobic biodegradation processes, the methods for testing anaerobic biodegradability, and the anaerobic biodegradability of different detergent surfactant types (anionic, nonionic, cationic, amphoteric surfactants.

  12. Increased susceptibility of ß-glucosidase from the hyperthermophile Pyrococcus furiosus to thermal inactivation at higher pressures

    NARCIS (Netherlands)

    Bruins, M.E.; Meersman, F.; Janssen, A.E.M.; Heremans, K.; Boom, R.M.

    2009-01-01

    The stability of ß-glucosidase from the hyperthermophile Pyrococcus furiosus was studied as a function of pressure, temperature and pH. The conformational stability was monitored using FTIR spectroscopy, and the functional enzyme stability was monitored by inactivation studies. The enzyme proved to

  13. Responses of Wild-Type and Resistant Strains of the Hyperthermophilic Bacterium Thermotoga maritima to Chloramphenicol Challenge▿ †

    Science.gov (United States)

    Montero, Clemente I.; Johnson, Matthew R.; Chou, Chung-Jung; Conners, Shannon B.; Geouge, Sarah G.; Tachdjian, Sabrina; Nichols, Jason D.; Kelly, Robert M.

    2007-01-01

    Transcriptomes and growth physiologies of the hyperthermophile Thermotoga maritima and an antibiotic-resistant spontaneous mutant were compared prior to and following exposure to chloramphenicol. While the wild-type response was similar to that of mesophilic bacteria, reduced susceptibility of the mutant was attributed to five mutations in 23S rRNA and phenotypic preconditioning to chloramphenicol. PMID:17557852

  14. Biochemical evidence for the presence of two α-glucoside ABC-transport systems in the hyperthermophilic archaeon Pyrococcus furiosus

    NARCIS (Netherlands)

    Koning, Sonja M.; Konings, Wil N.; Driessen, Arnold J.M.

    2002-01-01

    The hyperthermophilic archaeon Pyrococcus furiosus can utilize different carbohydrates, such as starch, maltose and trehalose. Uptake of α-glucosides is mediated by two different, binding protein-dependent, ATP-binding cassette (ABC)-type transport systems. The maltose transporter also transports tr

  15. Responses of wild-type and resistant strains of the hyperthermophilic bacterium Thermotoga maritima to chloramphenicol challenge.

    Science.gov (United States)

    Montero, Clemente I; Johnson, Matthew R; Chou, Chung-Jung; Conners, Shannon B; Geouge, Sarah G; Tachdjian, Sabrina; Nichols, Jason D; Kelly, Robert M

    2007-08-01

    Transcriptomes and growth physiologies of the hyperthermophile Thermotoga maritima and an antibiotic-resistant spontaneous mutant were compared prior to and following exposure to chloramphenicol. While the wild-type response was similar to that of mesophilic bacteria, reduced susceptibility of the mutant was attributed to five mutations in 23S rRNA and phenotypic preconditioning to chloramphenicol.

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

    DEFF Research Database (Denmark)

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

    2006-01-01

    A novel virus, ATV, of the hyperthermophilic archaeal genus Acidianus has the unique property of undergoing a major morphological development outside of, and independently of, the host cell. Virions are extruded from host cells as lemon-shaped tail-less particles, after which they develop long...

  17. Bypassing rRNA methylation by RsmA/Dim1during ribosome maturation in the hyperthermophilic archaeon Nanoarchaeum equitans

    DEFF Research Database (Denmark)

    Seistrup, Kenneth H; Rose, Simon; Birkedal, Ulf

    2017-01-01

    In all free-living organisms a late-stage checkpoint in the biogenesis of the small ribosomal subunit involves rRNA modification by an RsmA/Dim1 methyltransferase. The hyperthermophilic archaeon Nanoarchaeum equitans, whose existence is confined to the surface of a second archaeon, Ignicoccus hos...

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

    Directory of Open Access Journals (Sweden)

    Mart Krupovic

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

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

    Energy Technology Data Exchange (ETDEWEB)

    Nakabayashi, Makoto; Kataoka, Misumi; Mishima, Yumiko; Maeno, Yuka; Ishikawa, Kazuhiko, E-mail: kazu-ishikawa@aist.go.jp [National Institute of Advanced Industrial Science, 3-11-32, Kagamiyama, Higashi-Hiroshima, Hiroshima 739-0046 (Japan)

    2014-03-01

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

  20. Variation of the virus-related elements within syntenic genomes of the hyperthermophilic archaeon aeropyrum

    DEFF Research Database (Denmark)

    Daifuku, Takashi; Yoshida, Takashi; Kitamura, Takayuki

    2013-01-01

    The increasing number of genome sequences of archaea and bacteria show their adaptation to different environmental conditions at the genomic level. Aeropyrum spp. are aerobic and hyperthermophilic archaea. Aeropyrum camini was isolated from a deep-sea hydrothermal vent, and Aeropyrum pernix was i...... the genomes of A. camini and A. pernix were conserved, we observed nonsynteny that was attributed primarily to virus-related elements. Our findings indicated that the genomic diversification of Aeropyrum spp. is substantially caused by viruses.......The increasing number of genome sequences of archaea and bacteria show their adaptation to different environmental conditions at the genomic level. Aeropyrum spp. are aerobic and hyperthermophilic archaea. Aeropyrum camini was isolated from a deep-sea hydrothermal vent, and Aeropyrum pernix...... having stable genomes, interference of synteny occurred with two proviruses, A. pernix spindle-shaped virus 1 (APSV1) and A. pernix ovoid virus 1 (APOV1), and clustered regularly interspaced short palindromic repeat (CRISPR) elements. Spacer sequences derived from the A. camini CRISPR showed significant...

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

    Science.gov (United States)

    Krupovic, Mart; Gonnet, Mathieu; Hania, Wajdi Ben; Forterre, Patrick; Erauso, Gaël

    2013-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Yoon-Jung Moon

    2015-04-01

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

  3. Preliminary characterization of two different crystal forms of acylphosphatase from the hyperthermophile archaeon Sulfolobus solfataricus

    Energy Technology Data Exchange (ETDEWEB)

    Zuccotti, Simone [Department of Physics-INFM and Center of Excellence for Biomedical Research, University of Genova, Via Dodecaneso 33, 16132 Genova (Italy); Rosano, Camillo [National Institute for Cancer Research (IST), X-ray Structural Biology Unit, Largo R. Benzi 10, 16132 Genova (Italy); Bemporad, Francesco [Department of Biochemical Sciences, University of Firenze, Viale Morgagni 50, 50134 Florence (Italy); Stefani, Massimo [Department of Biochemical Sciences, University of Firenze, Viale Morgagni 50, 50134 Florence (Italy); Centro di Ricerca, Trasferimento e Alta Formazione MCIDNENT, University of Firenze, Viale Morgagni 50, 50134 Florence (Italy); Bolognesi, Martino, E-mail: bolognes@fisica.unige.it [Department of Physics-INFM and Center of Excellence for Biomedical Research, University of Genova, Via Dodecaneso 33, 16132 Genova (Italy)

    2005-01-01

    S. solfataricus acylphosphatase has been expressed, purified and crystallized in two different crystal forms. Preliminary characterization of a triclinic and a monoclinic crystal form is reported and data were collected to 1.27 and 1.90 Å, respectively. Acylphosphatase is a ubiquitous small enzyme that was first characterized in mammals. It is involved in the hydrolysis of carboxyl-phosphate bonds in several acylphosphate substrates, such as carbamoylphosphate and 1,3-biphosphoglycerate; however, a consensus on acylphosphatase action in vivo has not yet been reached. Recent investigations have focused on acylphosphatases from lower phyla, such as Drosophila melanogaster and Escherichia coli, in view of the application of these small proteins as models in the study of folding, misfolding and aggregation processes. An acylphosphatase from the hyperthermophilic archaeon Sulfolobus solfataricus has been cloned, expressed and purified. Here, the growth and characterization of a triclinic and a monoclinic crystal form of the hyperthermophilic enzyme are reported; X-ray diffraction data have been collected to 1.27 and 1.90 Å resolution, respectively.

  4. Postpartum anaerobic wound infection

    Directory of Open Access Journals (Sweden)

    N. A. Korobkov

    2013-01-01

    Full Text Available The objective of the study was to elucidate the role of nonclostridial anaerobic microflora in the pathogenesis of postpartum infectious and inflammatory diseases, and to choose an effective combination of antimicrobial chemotherapy. Inclusion of hyperbaric oxygen and of chemotherapy of the directed action on the obligate anaerobic flora contributed to rapid disappearance of the systemic inflammation signs and to proper involution of the uterus in 56 (86.2 % parturients in the main group. Successful treatment in the control group was observed in 44 (67.7 % patients (P=0.12.

  5. Effect of prolonged intravenous glucose and essential amino acid infusion on nitrogen balance, muscle protein degradation and ubiquitin-conjugating enzyme gene expression in calves

    Directory of Open Access Journals (Sweden)

    Scaife Jes R

    2008-02-01

    Full Text Available Abstract Background Intravenous infusions of glucose and amino acids increase both nitrogen balance and muscle accretion. We hypothesised that co-infusion of glucose (to stimulate insulin and essential amino acids (EAA would act additively to improve nitrogen balance by decreasing muscle protein degradation in association with alterations in muscle expression of components of the ubiquitin-proteasome proteolytic pathway. Methods We examined the effect of a 5 day intravenous infusions of saline, glucose, EAA and glucose + EAA, on urinary nitrogen excretion and muscle protein degradation. We carried out the study in 6 restrained calves since ruminants offer the advantage that muscle protein degradation can be assessed by excretion of 3 methyl-histidine and multiple muscle biopsies can be taken from the same animal. On the final day of infusion blood samples were taken for hormone and metabolite measurement and muscle biopsies for expression of ubiquitin, the 14-kDa E2 ubiquitin conjugating enzyme, and proteasome sub-units C2 and C8. Results On day 5 of glucose infusion, plasma glucose, insulin and IGF-1 concentrations were increased while urea nitrogen excretion and myofibrillar protein degradation was decreased. Co-infusion of glucose + EAA prevented the loss of urinary nitrogen observed with EAA infusions alone and enhanced the increase in plasma IGF-1 concentration but there was no synergistic effect of glucose + EAA on the decrease in myofibrillar protein degradation. Muscle mRNA expression of the ubiquitin conjugating enzyme, 14-kDa E2 and proteasome sub-unit C2 were significantly decreased, after glucose but not amino acid infusions, and there was no further response to the combined infusions of glucose + EAA. Conclusion Prolonged glucose infusion decreases myofibrillar protein degradation, prevents the excretion of infused EAA, and acts additively with EAA to increase plasma IGF-1 and improve net nitrogen balance. There was no evidence of

  6. Heat-induced protein structure and subfractions in relation to protein degradation kinetics and intestinal availability in dairy cattle.

    Science.gov (United States)

    Doiron, K; Yu, P; McKinnon, J J; Christensen, D A

    2009-07-01

    The objectives of this study were to reveal protein structures of feed tissues affected by heat processing at a cellular level, using the synchrotron-based Fourier transform infrared microspectroscopy as a novel approach, and quantify protein structure in relation to protein digestive kinetics and nutritive value in the rumen and intestine in dairy cattle. The parameters assessed included 1) protein structure alpha-helix to beta-sheet ratio; 2) protein subfractions profiles; 3) protein degradation kinetics and effective degradability; 4) predicted nutrient supply using the intestinally absorbed protein supply (DVE)/degraded protein balance (OEB) system for dairy cattle. In this study, Vimy flaxseed protein was used as a model feed protein and was autoclave-heated at 120 degrees C for 20, 40, and 60 min in treatments T1, T2, and T3, respectively. The results showed that using the synchrotron-based Fourier transform infrared microspectroscopy revealed and identified the heat-induced protein structure changes. Heating at 120 degrees C for 40 and 60 min increased the protein structure alpha-helix to beta-sheet ratio. There were linear effects of heating time on the ratio. The heating also changed chemical profiles, which showed soluble CP decreased upon heating with concomitant increases in nonprotein nitrogen, neutral, and acid detergent insoluble nitrogen. The protein subfractions with the greatest changes were PB1, which showed a dramatic reduction, and PB2, which showed a dramatic increase, demonstrating a decrease in overall protein degradability. In situ results showed a reduction in rumen-degradable protein and in rumen-degradable dry matter without differences between the treatments. Intestinal digestibility, determined using a 3-step in vitro procedure, showed no changes to rumen undegradable protein. Modeling results showed that heating increased total intestinally absorbable protein (feed DVE value) and decreased degraded protein balance (feed OEB value

  7. Heat-induced Protein Structure and Subfractions in Relation to Protein Degradation Kinetics and Intestinal Availability in Dairy Cattle

    Energy Technology Data Exchange (ETDEWEB)

    Doiron, K.; Yu, P; McKinnon, J; Christensen, D

    2009-01-01

    The objectives of this study were to reveal protein structures of feed tissues affected by heat processing at a cellular level, using the synchrotron-based Fourier transform infrared microspectroscopy as a novel approach, and quantify protein structure in relation to protein digestive kinetics and nutritive value in the rumen and intestine in dairy cattle. The parameters assessed included (1) protein structure a-helix to e-sheet ratio; (2) protein subfractions profiles; (3) protein degradation kinetics and effective degradability; (4) predicted nutrient supply using the intestinally absorbed protein supply (DVE)/degraded protein balance (OEB) system for dairy cattle. In this study, Vimy flaxseed protein was used as a model feed protein and was autoclave-heated at 120C for 20, 40, and 60 min in treatments T1, T2, and T3, respectively. The results showed that using the synchrotron-based Fourier transform infrared microspectroscopy revealed and identified the heat-induced protein structure changes. Heating at 120C for 40 and 60 min increased the protein structure a-helix to e-sheet ratio. There were linear effects of heating time on the ratio. The heating also changed chemical profiles, which showed soluble CP decreased upon heating with concomitant increases in nonprotein nitrogen, neutral, and acid detergent insoluble nitrogen. The protein subfractions with the greatest changes were PB1, which showed a dramatic reduction, and PB2, which showed a dramatic increase, demonstrating a decrease in overall protein degradability. In situ results showed a reduction in rumen-degradable protein and in rumen-degradable dry matter without differences between the treatments. Intestinal digestibility, determined using a 3-step in vitro procedure, showed no changes to rumen undegradable protein. Modeling results showed that heating increased total intestinally absorbable protein (feed DVE value) and decreased degraded protein balance (feed OEB value), but there were no differences

  8. Draft Genome Sequence of Pyrodictium occultum PL19T, a Marine Hyperthermophilic Species of Archaea That Grows Optimally at 105°C.

    Science.gov (United States)

    Utturkar, Sagar M; Huber, Harald; Leptihn, Sebastian; Loh, Belinda; Brown, Steven D; Stetter, Karl O; Podar, Mircea

    2016-02-25

    We report here the draft genome sequence of Pyrodictium occultum PL19(T), a marine hyperthermophilic archaeon. The genome provides insights into molecular and cellular adaptation mechanisms to life in extreme environments and the evolution of early organisms on Earth.

  9. Anaerobic digestion without biogas?

    NARCIS (Netherlands)

    Kleerebezem, R.; Joosse, B.; Rozendaal, R.; Van Loosdrecht, M.C.M.

    2015-01-01

    Anaerobic digestion for the production of methane containing biogas is the classic example of a resource recovery process that combines stabilization of particulate organic matter or wastewater treatment with the production of a valuable end-product. Attractive features of the process include the pr

  10. The anaerobic digestion process

    Energy Technology Data Exchange (ETDEWEB)

    Rivard, C.J. [National Renewable Energy Lab., Golden, CO (United States); Boone, D.R. [Oregon Graduate Inst., Portland, OR (United States)

    1996-01-01

    The microbial process of converting organic matter into methane and carbon dioxide is so complex that anaerobic digesters have long been treated as {open_quotes}black boxes.{close_quotes} Research into this process during the past few decades has gradually unraveled this complexity, but many questions remain. The major biochemical reactions for forming methane by methanogens are largely understood, and evolutionary studies indicate that these microbes are as different from bacteria as they are from plants and animals. In anaerobic digesters, methanogens are at the terminus of a metabolic web, in which the reactions of myriads of other microbes produce a very limited range of compounds - mainly acetate, hydrogen, and formate - on which the methanogens grow and from which they form methane. {open_quotes}Interspecies hydrogen-transfer{close_quotes} and {open_quotes}interspecies formate-transfer{close_quotes} are major mechanisms by which methanogens obtain their substrates and by which volatile fatty acids are degraded. Present understanding of these reactions and other complex interactions among the bacteria involved in anaerobic digestion is only now to the point where anaerobic digesters need no longer be treated as black boxes.

  11. Anaerobic digestion without biogas?

    NARCIS (Netherlands)

    Kleerebezem, R.; Joosse, B.; Rozendaal, R.; Van Loosdrecht, M.C.M.

    2015-01-01

    Anaerobic digestion for the production of methane containing biogas is the classic example of a resource recovery process that combines stabilization of particulate organic matter or wastewater treatment with the production of a valuable end-product. Attractive features of the process include the

  12. Anaerobic digestion without biogas?

    NARCIS (Netherlands)

    Kleerebezem, R.; Joosse, B.; Rozendaal, R.; Van Loosdrecht, M.C.M.

    2015-01-01

    Anaerobic digestion for the production of methane containing biogas is the classic example of a resource recovery process that combines stabilization of particulate organic matter or wastewater treatment with the production of a valuable end-product. Attractive features of the process include the pr

  13. Hydrogen production using amino acids obtained by protein degradation in waste biomass by combined dark- and photo-fermentation.

    Science.gov (United States)

    Cheng, Jun; Ding, Lingkan; Xia, Ao; Lin, Richen; Li, Yuyou; Zhou, Junhu; Cen, Kefa

    2015-03-01

    The biological hydrogen production from amino acids obtained by protein degradation was comprehensively investigated to increase heating value conversion efficiency. The five amino acids (i.e., alanine, serine, aspartic acid, arginine, and leucine) produced limited hydrogen (0.2-16.2 mL/g) but abundant soluble metabolic products (40.1-84.0 mM) during dark-fermentation. The carbon conversion efficiencies of alanine (85.3%) and serine (94.1%) during dark-fermentation were significantly higher than those of other amino acids. Residual dark-fermentation solutions treated with zeolite for NH4(+) removal were inoculated with photosynthetic bacteria to further produce hydrogen during photo-fermentation. The hydrogen yields of alanine and serine through combined dark- and photo-fermentation were 418.6 and 270.2 mL/g, respectively. The heating value conversion efficiency of alanine to hydrogen was 25.1%, which was higher than that of serine (21.2%).

  14. TAILS N-Terminomics and Proteomics Show Protein Degradation Dominates over Proteolytic Processing by Cathepsins in Pancreatic Tumors

    Directory of Open Access Journals (Sweden)

    Anna Prudova

    2016-08-01

    Full Text Available Deregulated cathepsin proteolysis occurs across numerous cancers, but in vivo substrates mediating tumorigenesis remain ill-defined. Applying 8-plex iTRAQ terminal amine isotopic labeling of substrates (TAILS, a systems-level N-terminome degradomics approach, we identified cathepsin B, H, L, S, and Z in vivo substrates and cleavage sites with the use of six different cathepsin knockout genotypes in the Rip1-Tag2 mouse model of pancreatic neuroendocrine tumorigenesis. Among 1,935 proteins and 1,114 N termini identified by TAILS, stable proteolytic products were identified in wild-type tumors compared with one or more different cathepsin knockouts (17%–44% of 139 cleavages. This suggests a lack of compensation at the substrate level by other cathepsins. The majority of neo-N termini (56%–83% for all cathepsins was consistent with protein degradation. We validated substrates, including the glycolytic enzyme pyruvate kinase M2 associated with the Warburg effect, the ER chaperone GRP78, and the oncoprotein prothymosin-alpha. Thus, the identification of cathepsin substrates in tumorigenesis improves the understanding of cathepsin functions in normal physiology and cancer.

  15. Experimental Priapism is Associated with Increased Oxidative Stress and Activation of Protein Degradation Pathways in Corporal Tissue

    Science.gov (United States)

    Kanika, Nirmala D.; Melman, Arnold; Davies, Kelvin P.

    2010-01-01

    Priapism is a debilitating disease for which there is at present no clinically accepted pharmacologic intervention. It has been estimated that priapism lasting more than 24 hours in patients is associated with a 44–90% rate of erectile dysfunction (ED). In this investigation we determined in two animal models of priapism (opiorpin-induced priapism in the rat and priapism in a mouse model of sickle cell disease) if there is evidence for an increase in markers of oxidative stress in corporal tissue. In both animal models we demonstrate that priapism results in increased levels of lipid peroxidation, glutathione S-transferase activity, and oxidatively damaged proteins in corporal tissue. Using Western blot analysis we demonstrated there is up regulation of the ubiquitination ligase proteins, Nedd-4 and Mdm-2, and the lysososomal autophage protein, LC3. The anti-apoptotic protein, Bcl-2, was also up regulated. Overall, we demonstrate that priapism is associated with increased oxidative stress in corporal tissue and the activation of protein degradation pathways. Since oxidative stress is known to mediate the development of ED resulting from several etiologies (for example ED resulting from diabetes and aging) we suggest that damage to erectile tissue resulting from priapism might be prevented by treatments targeting oxidative stress. PMID:21085184

  16. Effects of rapid or slow body weight reduction on intramuscular protein degradation pathways during equivalent weight loss on rats.

    Science.gov (United States)

    Nonaka, Y; Urashima, S; Inai, M; Nishimura, S; Higashida, K; Terada, S

    2017-07-18

    The purpose of this study was to compare the effects of short-term fasting-induced rapid weight loss with those of slower but equivalent body weight loss induced by daily calorie restriction on muscle protein degradation pathways and muscle protein content. Male Fischer rats were subjected to either 30 % calorie restriction for 2 weeks to slowly decrease body weight (Slow) or 3-day fasting to rapidly decrease body weight by a comparable level of that of the Slow group (Rapid). The final body weights were about 15 % lower in both the Slow and Rapid groups than in the Con group (pweight of fast-twitch plantaris muscle, but not slow-twitch soleus muscle, were significantly lower in the Rapid group compared with the control rats fed ad libitum. Substantial increases in the expression ratio of autophagosomal membrane proteins (LC3-II/-I ratio) and polyubiquitinated protein concentration, used as biomarkers of autophagy-lysosome and ubiquitin-proteasome activities, respectively, were observed in the plantaris muscle of the Rapid group. Moreover, the LC3-II/-I ratio and polyubiquitinated protein concentration were negatively correlated with the total protein content and wet weight of plantaris muscle. These results suggest that short-term fasting-induced rapid body weight loss activates autophagy-lysosome and ubiquitin-proteasome systems more strongly than calorie restriction-induced slower weight reduction, resulting in muscular atrophy in fast-twitch muscle.

  17. Genes Related to Mitochondrial Functions, Protein Degradation, and Chromatin Folding Are Differentially Expressed in Lymphomonocytes of Rett Syndrome Patients

    Science.gov (United States)

    Leoni, Guido; Cervellati, Franco; Canali, Raffaella; Cortelazzo, Alessio; De Felice, Claudio; Ciccoli, Lucia; Hayek, Joussef

    2013-01-01

    Rett syndrome (RTT) is mainly caused by mutations in the X-linked methyl-CpG binding protein (MeCP2) gene. By binding to methylated promoters on CpG islands, MeCP2 protein is able to modulate several genes and important cellular pathways. Therefore, mutations in MeCP2 can seriously affect the cellular phenotype. Today, the pathways that MeCP2 mutations are able to affect in RTT are not clear yet. The aim of our study was to investigate the gene expression profiles in peripheral blood lymphomonocytes (PBMC) isolated from RTT patients to try to evidence new genes and new pathways that are involved in RTT pathophysiology. LIMMA (Linear Models for MicroArray) and SAM (Significance Analysis of Microarrays) analyses on microarray data from 12 RTT patients and 7 control subjects identified 482 genes modulated in RTT, of which 430 were upregulated and 52 were downregulated. Functional clustering of a total of 146 genes in RTT identified key biological pathways related to mitochondrial function and organization, cellular ubiquitination and proteosome degradation, RNA processing, and chromatin folding. Our microarray data reveal an overexpression of genes involved in ATP synthesis suggesting altered energy requirement that parallels with increased activities of protein degradation. In conclusion, these findings suggest that mitochondrial-ATP-proteasome functions are likely to be involved in RTT clinical features. PMID:24453408

  18. Genes Related to Mitochondrial Functions, Protein Degradation, and Chromatin Folding Are Differentially Expressed in Lymphomonocytes of Rett Syndrome Patients

    Directory of Open Access Journals (Sweden)

    Alessandra Pecorelli

    2013-01-01

    Full Text Available Rett syndrome (RTT is mainly caused by mutations in the X-linked methyl-CpG binding protein (MeCP2 gene. By binding to methylated promoters on CpG islands, MeCP2 protein is able to modulate several genes and important cellular pathways. Therefore, mutations in MeCP2 can seriously affect the cellular phenotype. Today, the pathways that MeCP2 mutations are able to affect in RTT are not clear yet. The aim of our study was to investigate the gene expression profiles in peripheral blood lymphomonocytes (PBMC isolated from RTT patients to try to evidence new genes and new pathways that are involved in RTT pathophysiology. LIMMA (Linear Models for MicroArray and SAM (Significance Analysis of Microarrays analyses on microarray data from 12 RTT patients and 7 control subjects identified 482 genes modulated in RTT, of which 430 were upregulated and 52 were downregulated. Functional clustering of a total of 146 genes in RTT identified key biological pathways related to mitochondrial function and organization, cellular ubiquitination and proteosome degradation, RNA processing, and chromatin folding. Our microarray data reveal an overexpression of genes involved in ATP synthesis suggesting altered energy requirement that parallels with increased activities of protein degradation. In conclusion, these findings suggest that mitochondrial-ATP-proteasome functions are likely to be involved in RTT clinical features.

  19. Antioxidant properties of neohesperidin dihydrochalcone: inhibition of hypochlorous acid-induced DNA strand breakage, protein degradation, and cell death.

    Science.gov (United States)

    Choi, Je-Min; Yoon, Byoung-Seok; Lee, Sang-Kyou; Hwang, Jae-Kwan; Ryang, Ryung

    2007-02-01

    Neohesperidin dihydrochalcone (NHDC), a non-nutritive sweetening agent, is simply produced by hydrogenation of neohesperidin. The aim of this study is to evaluate the antioxidant and radical scavenging properties of neohesperidin dihydrochalcone and other structurally related compounds (phloridzin, neohesperidin) toward different reactive radical and oxygen species including .ABTS+, .O2-, .OH, H2O2, and HOCl in vitro. NHDC showed remarkable radical scavenging activity against stable radical and reactive oxygen species (ROS) in concentration dependent manner. Especially, NHDC was the most potent inhibitor of H2O2 and HOCl. NHDC showed HOCl scavenging activity of 93.5% and H2O2 scavenging property of 73.5% which was more than those of all the tested compounds including ascorbic acid and BHT. Moreover, NHDC could inhibit protein degradation, plasmid DNA strand cleavage and HIT-T15, HUVEC cell death from HOCl attack while mannitol, BHT, and ascorbic acid could not protect them effectively. These results suggest that NHDC is a potent antioxidant, especially it is evaluated as a novel HOCl scavenger. This study implies the possibility of therapeutic effect of NHDC on ROS-related inflammatory diseases.

  20. [Microbial diversity of deep-sea extremophiles--Piezophiles, Hyperthermophiles, and subsurface microorganisms].

    Science.gov (United States)

    Kato, C; Takai, K

    2000-12-01

    Knowledge of our Planet's biosphere has increased tremendously during the last 10 to 20 years. In the field of Microbiology in particular, scientists have discovered novel "extremophiles", microorganisms capable of living in extreme environments such as highly acidic or alkaline conditions, at high salt concentration, with no oxygen, extreme temperatures (as low as -20 degrees C and as high as 300 degrees C), at high concentrations of heavy metals and in high pressure environments such as the deep-sea. It is apparent that microorganisms can exist in any extreme environment of the Earth, yet already scientists have started to look for life on other planets; the so-called "Exobiology" project. But as yet we have little knowledge of the deep-sea and subsurface biosphere of our own planet. We believe that we should elucidate the Biodiversity of Earth more thoroughly before exploring life on other planets, and these attempts would provide deeper insight into clarifying the existence of extraterrestrial life. We focused on two deep-sea extremophiles in this article; one is "Piezophiles", and another is "Hyperthermophiles". Piezophiles are typical microorganisms adapted to high-pressure and cold temperature environments, and located in deep-sea bottom. Otherwise, hyperthermophiles are living in high temperature environment, and located at around the hydrothermal vent systems in deep-sea. They are not typical deep-sea microorganisms, but they can grow well at high-pressure condition, just like piezophiles. Deming and Baross mentioned that most of the hyperthermophilic archaea isolated from deep-sea hydrothermal vents are able to grow under conditions of high temperature and pressure, and in most cases their optimal pressure for growth was greater than the environmental pressure they were isolated from. It is possible that originally their native environment may have been deeper than the sea floor and that there had to be a deeper biosphere. This implication suggests that

  1. Effect of DNA binding protein Ssh12 from hyperthermophilic archaeon Sulfolobus shibatae on DNA supercoiling

    Institute of Scientific and Technical Information of China (English)

    楼慧强; 黄力; VietQ.Mai

    1999-01-01

    An 11.5-ku DNA binding protein, designated as Sshl2, was purified from the hyperthermophilic archaeon Sulfolobus shibatae by column chromatography in SP Sepharose, DNA cellulose and phosphocellulose. Sshl2 accounts for about 4 % of the total cellular protein. The protein is capable of binding to both negatively supercoiled and relaxed DNAs. Nick closure analysis revealed that Sshl2 constrains negative supercoils upon binding to DNA. While the ability of the protein to constrain supercoils is weak at 22℃ , it is enhanced substantially at temperatures higher than 37℃ . Both the cellular content and supercoil-constraining ability of Sshl2 suggest that the protein may play an important role in the organization and stabilization of the chromosome of S. shibatae.

  2. Biological effects of DNA damage in the hyperthermophilic archaeon Sulfolobus acidocaldarius.

    Science.gov (United States)

    Reilly, Michelle S; Grogan, Dennis W

    2002-02-19

    To investigate the generality of efficient double-strand break repair and damage-induced mutagenesis in hyperthermophilic archaea, we systematically measured the effects of five DNA-damaging agents on Sulfolobus acidocaldarius and compared the results to those obtained for Escherichia coli under corresponding conditions. The observed lethality of gamma-radiation was very similar for S. acidocaldarius and E. coli, arguing against unusually efficient double-strand break repair in S. acidocaldarius. In addition, DNA-strand-breaking agents (gamma-radiation or bleomycin), as well as DNA-cross-linking agents (mechlorethamine, butadiene diepoxide or cisplatin) stimulated forward mutation, reverse mutation, and formation of recombinants via conjugation in Sulfolobus cells. Although two of the five DNA-damaging agents failed to revert the E. coli auxotrophs under these conditions, all five reverted S. acidocaldarius auxotrophs.

  3. Influence of osmotic stress on desiccation and irradiation tolerance of (hyper)-thermophilic microorganisms.

    Science.gov (United States)

    Beblo-Vranesevic, Kristina; Galinski, Erwin A; Rachel, Reinhard; Huber, Harald; Rettberg, Petra

    2017-01-01

    This study examined the influence of prior salt adaptation on the survival rate of (hyper)-thermophilic bacteria and archaea after desiccation and UV or ionizing irradiation treatment. Survival rates after desiccation of Hydrogenothermus marinus and Archaeoglobus fulgidus increased considerably when the cells were cultivated at higher salt concentrations before drying. By doubling the concentration of NaCl, a 30 times higher survival rate of H. marinus after desiccation was observed. Under salt stress, the compatible solute diglycerol phosphate in A. fulgidus and glucosylglycerate in H. marinus accumulated in the cytoplasm. Several different compatible solutes were added as protectants to A. fulgidus and H. marinus before desiccation treatment. Some of these had similar effects as intracellularly produced compatible solutes. The survival rates of H. marinus and A. fulgidus after exposure to UV-C (254 nm) or ionizing X-ray/gamma radiation were irrespective of the salt-induced synthesis or the addition of compatible solutes.

  4. Identification and characterization of small RNAs in the hyperthermophilic archaeon Sulfolobus solfataricus.

    Directory of Open Access Journals (Sweden)

    Ning Xu

    Full Text Available The term RNA silencing (RNA interference, RNAi describes a set of mechanisms that regulate gene expression in eukaryotes. Small interfering RNAs (siRNA and microRNAs (miRNAs are two major types of RNAi-associated small RNAs (smRNAs found in most eukaryotic organisms. Despite the presence of a plethora of non-coding RNAs longer than 50-nucleotide (nt in length in various species of Archaea, little is known about smRNAs in archaea that resemble the 20-24-nt long smRNAs found in eukaryotes, which have been implicated in the post-transcriptional control of gene expression. Here, we report the finding of a large number of smRNAs approximatelly 20-nt in length, including phased smRNAs and potential miRNAs, from the hyperthermophilic archaeon Sulfolobus solfataricus p2 (Ssp2 based on deep sequencing. The expression of some of the miRNA candidates in Ssp2 was confirmed. Consistent with the Ssp2 hyperthermophilic properties, we found that higher temperatures more efficiently induced the production of the miRNA candidates in an in vitro system using the putative foldback precursor transcripts incubated with Ssp2 extract. Although we initially predicted putative target genes of some miRNA candidates, further analysis mapped the cleavage sites downstream of the miRNA candidate complementary regions, similar to those involved in plant miRNA-mediated TAS transcript cleavage. We also identified smRNAs from clustered, regularly interspaced, short palindromic repeat (CRISPR loci, which play important roles in prokaryotic microbial defense systems. Archaea represent a unique life form next to Bacteria and Eukarya, and our results may provide a useful resource for further in-depth study on the regulation and evolution of smRNAs in this special organism.

  5. Crystal structure of T state aspartate carbamoyltransferase of the hyperthermophilic archaeon Sulfolobus acidocaldarius.

    Science.gov (United States)

    De Vos, Dirk; Van Petegem, Filip; Remaut, Han; Legrain, Christianne; Glansdorff, Nicolas; Van Beeumen, Jozef J

    2004-06-11

    Aspartate carbamoyltransferase (ATCase) is a model enzyme for understanding allosteric effects. The dodecameric complex exists in two main states (T and R) that differ substantially in their quaternary structure and their affinity for various ligands. Many hypotheses have resulted from the structure of the Escherichia coli ATCase, but so far other crystal structures to test these have been lacking. Here, we present the tertiary and quaternary structure of the T state ATCase of the hyperthermophilic archaeon Sulfolobus acidocaldarius (SaATC(T)), determined by X-ray crystallography to 2.6A resolution. The quaternary structure differs from the E.coli ATCase, by having altered interfaces between the catalytic (C) and regulatory (R) subunits, and the presence of a novel C1-R2 type interface. Conformational differences in the 240 s loop region of the C chain and the C-terminal region of the R chain affect intersubunit and interdomain interfaces implicated previously in the allosteric behavior of E.coli ATCase. The allosteric-zinc binding domain interface is strengthened at the expense of a weakened R1-C4 type interface. The increased hydrophobicity of the C1-R1 type interface may stabilize the quaternary structure. Catalytic trimers of the S.acidocaldarius ATCase are unstable due to a drastic weakening of the C1-C2 interface. The hyperthermophilic ATCase presents an interesting example of how an allosteric enzyme can adapt to higher temperatures. The structural rearrangement of this thermophilic ATCase may well promote its thermal stability at the expense of changes in the allosteric behavior.

  6. Functional analysis of hyperthermophilic endocellulase from Pyrococcus horikoshii by crystallographic snapshots.

    Science.gov (United States)

    Kim, Han-Woo; Ishikawa, Kazuhiko

    2011-07-15

    A hyperthermophilic membrane-related β-1,4-endoglucanase (family 5, cellulase) of the archaeon Pyrococcus horikoshii was found to be capable of hydrolysing cellulose at high temperatures. The hyperthermophilic cellulase has promise for applications in biomass utilization. To clarify its detailed function, we determined the crystal structures of mutants of the enzyme in complex with either the substrate or product ligands. We were able to resolve different kinds of complex structures at 1.65-2.01 Å (1 Å=0.1 nm). The structural analysis of various mutant enzymes yielded a sequence of crystallographic snapshots, which could be used to explain the catalytic process of the enzyme. The substrate position is fixed by the alignment of one cellobiose unit between the two aromatic amino acid residues at subsites +1 and +2. During the enzyme reaction, the glucose structure of cellulose substrates is distorted at subsite -1, and the β-1,4-glucoside bond between glucose moieties is twisted between subsites -1 and +1. Subsite -2 specifically recognizes the glucose residue, but recognition by subsites +1 and +2 is loose during the enzyme reaction. This type of recognition is important for creation of the distorted boat form of the substrate at subsite -1. A rare enzyme-substrate complex was observed within the low-activity mutant Y299F, which suggested the existence of a trapped ligand structure before the formation by covalent bonding of the proposed intermediate structure. Analysis of the enzyme-substrate structure suggested that an incoming water molecule, essential for hydrolysis during the retention process, might be introduced to the cleavage position after the cellobiose product at subsites +1 and +2 was released from the active site.

  7. 3-Phosphoglycerate is an allosteric activator of pyruvate kinase from the hyperthermophilic archaeon Pyrobaculum aerophilum.

    Science.gov (United States)

    Solomons, J T Graham; Johnsen, Ulrike; Schönheit, Peter; Davies, Christopher

    2013-08-27

    Pyruvate kinase (PK) is a highly regulated enzyme that catalyzes the final step of glycolysis. PK from the hyperthermophilic archaeon Pyrobaculum aerophilum (PaPK) is distinguished from most PK enzymes of eukarya and bacteria by not responding to any known allosteric effectors and apparently exhibiting only cooperative regulation. We determined the crystal structure of PaPK to 2.2 Å resolution and, in a manner consistent with the lack of a response to conventional effectors, observed that the canonical allosteric site is occluded by a tyrosine. Unexpectedly, though, a bound sulfate was observed at a position equivalent to the 6'-phosphate of sugar effectors, suggesting an allosteric site, but for an unknown effector and sharing only the phosphate position. A search of three-carbon intermediates of glycolysis revealed 3-phosphoglycerate (3PG) as a potent allosteric activator of PaPK. The response was abolished by mutation of residues that contact the sulfate and of an arginine proposed to interact with the 3PG carboxylate group. Regulation of PK by 3PG is consistent with the ancestral glycolysis of hyperthermophilic archaea in which this intermediate is produced by an irreversible enzyme, glyceraldehyde 3-phosphate ferredoxin oxidoreductase. Coordinated regulation within the lower half of glycolysis contrasts sharply with conventional glycolysis in which 3PG is produced reversibly and PK is regulated by fructose 1,6-bisphosphate, the product of phosphofructokinase, an irreversible enzyme in the upper half of the pathway. Regulation of PaPK by a carboxylate molecule rather than a sugar phosphate may reflect a step in the evolution of glycolysis that predates the dominance of sugars in metabolism.

  8. Protein degradation in a LAMP-2-deficient B-lymphoblastoid cell line from a patient with Danon disease.

    Science.gov (United States)

    Sánchez-Lanzas, Raul; Alvarez-Castelao, Beatriz; Bermejo, Teresa; Ayuso, Teresa; Tuñón, Teresa; Castaño, José G

    2016-08-01

    Danon disease, a condition characterized by cardiomyopathy, myopathy, and intellectual disability, is caused by mutations in the LAMP-2 gene. Lamp-2A protein, generated by alternative splicing from the Lamp-2 pre-mRNA, is reported to be the lysosomal membrane receptor essential for the chaperone-mediated autophagic pathway (CMA) aimed to selective protein targeting and translocation into the lysosomal lumen for degradation. To study the relevance of Lamp-2 in protein degradation, a lymphoblastoid cell line was obtained by EBV transformation of B-cells from a Danon patient. The derived cell line showed no significant expression of Lamp-2 protein. The steady-state mRNA and protein levels of alpha-synuclein, IΚBα, Rcan1, and glyceraldehyde-3-phosphate dehydrogenase, four proteins reported to be selective substrates of the CMA pathway, were similar in control and Lamp-2-deficient cells. Inhibition of protein synthesis showed that the half-life of alpha-synuclein, IΚBα, and Rcan1 was similar in control and Lamp-2-deficient cells, and its degradation prevented by proteasome inhibitors. Both in control and Lamp-2-deficient cells, induction of CMA and macroautophagy by serum and aminoacid starvation of cells for 8h produced a similar decrease in IΚBα and Rcan1 protein levels and was prevented by the addition of lysosome and autophagy inhibitors. In conclusion, the results presented here showed that Lamp-2 deficiency in human lymphoblastoid cells did not modify the steady-state levels or the degradation of several protein substrates reported as selective substrates of the CMA pathway.

  9. Dysregulation of protein degradation pathways may mediate the liver injury and phospholipidosis associated with a cationic amphiphilic antibiotic drug

    Energy Technology Data Exchange (ETDEWEB)

    Mosedale, Merrie [Hamner-University of North Carolina Institute for Drug Safety Sciences, The Hamner Institutes for Health Sciences, Research Triangle Park, NC 27709 (United States); Wu, Hong [Drug Safety Research and Development, Pfizer Global Research and Development, Groton, CT06340 (United States); Kurtz, C. Lisa [Hamner-University of North Carolina Institute for Drug Safety Sciences, The Hamner Institutes for Health Sciences, Research Triangle Park, NC 27709 (United States); Schmidt, Stephen P. [Drug Safety Research and Development, Pfizer Global Research and Development, Groton, CT06340 (United States); Adkins, Karissa, E-mail: Karissa.Adkins@pfizer.com [Drug Safety Research and Development, Pfizer Global Research and Development, Groton, CT06340 (United States); Harrill, Alison H. [Hamner-University of North Carolina Institute for Drug Safety Sciences, The Hamner Institutes for Health Sciences, Research Triangle Park, NC 27709 (United States); University of Arkansas for Medical Sciences, Little Rock, AR72205 (United States)

    2014-10-01

    A large number of antibiotics are known to cause drug-induced liver injury in the clinic; however, interpreting clinical risk is not straightforward owing to a lack of predictivity of the toxicity by standard preclinical species and a poor understanding of the mechanisms of toxicity. An example is PF-04287881, a novel ketolide antibiotic that caused elevations in liver function tests in Phase I clinical studies. In this study, a mouse diversity panel (MDP), comprised of 34 genetically diverse, inbred mouse strains, was utilized to model the toxicity observed with PF-04287881 treatment and investigate potential mechanisms that may mediate the liver response. Significant elevations in serum alanine aminotransferase (ALT) levels in PF-04287881-treated animals relative to vehicle-treated controls were observed in the majority (88%) of strains tested following a seven day exposure. The average fold elevation in ALT varied by genetic background and correlated with microscopic findings of hepatocellular hypertrophy, hepatocellular single cell necrosis, and Kupffer cell vacuolation (confirmed as phospholipidosis) in the liver. Global liver mRNA expression was evaluated in a subset of four strains to identify transcript and pathway differences that distinguish susceptible mice from resistant mice in the context of PF-04287881 treatment. The protein ubiquitination pathway was highly enriched among genes associated with PF-04287881-induced hepatocellular necrosis. Expression changes associated with PF-04287881-induced phospholipidosis included genes involved in drug transport, phospholipid metabolism, and lysosomal function. The findings suggest that perturbations in genes involved in protein degradation leading to accumulation of oxidized proteins may mediate the liver injury induced by this drug. - Highlights: • Identified susceptible and resistant mouse strains to liver injury induced by a CAD • Liver injury characterized by single cell necrosis, and phospholipidosis

  10. The ubiquitin+proteasome protein degradation pathway as a therapeutic strategy in the treatment of solid tumor malignancies.

    Science.gov (United States)

    Driscoll, James J; Minter, Alex; Driscoll, Daniel A; Burris, Jason K

    2011-02-01

    A concept that currently steers the development of cancer therapies has been that agents directed against specific proteins that facilitate tumorigenesis or maintain a malignant phenotype will have greater efficacy, less toxicity and a more sustained response relative to traditional cytotoxic chemotherapeutic agents. The clinical success of the targeted agent Imatinib mesylate as an inhibitor of the tyrosine kinase associated with the breakpoint cluster region-Abelson oncogene locus (BCR-ABL) in the treatment of Philadelphia-positive chronic myelogenous leukemia (CML) has served as a paradigm. While intellectually gratifying, the selective targeting of a single driver event by a small molecule, e.g., kinase inhibitor, to dampen a tumor-promoting pathway in the treatment of solid tumors is limited by many factors. Focus can alternatively be placed on targeting fundamental cellular processes that regulate multiple events, e.g., protein degradation, through the Ubiquitin (Ub)+Proteasome System (UPS). The UPS plays a critical role in modulating numerous cellular proteins to regulate cellular processes such as signal transduction, growth, proliferation, differentiation and apoptosis. Clinical success with the proteasome inhibitor bortezomib revolutionized treatment of B-cell lineage malignancies such as Multiple Myeloma (MM). However, many patients harbor primary resistance and do not respond to bortezomib and those that do respond inevitably develop resistance (secondary resistance). The lack of clinical efficacy of proteasome inhibitors in the treatment of solid tumors may be linked mechanistically to the resistance detected during treatment of hematologic malignancies. Potential mechanisms of resistance and means to improve the response to proteasome inhibitors in solid tumors are discussed.

  11. Effects of vanillin, quillaja saponin, and essential oils on in vitro fermentation and protein-degrading microorganisms of the rumen.

    Science.gov (United States)

    Patra, Amlan K; Yu, Zhongtang

    2014-01-01

    This study investigated the effects of vanillin on methanogenesis and rumen fermentation, and the responses of ruminal protein-degrading bacteria to vanillin (at concentrations of 0, 0.76 and 1.52 g/L), essential oils (clove oil, 1 g/L; origanum oil, 0.50 g/L, and peppermint oil, 1 g/L), and quillaja saponin (at concentration of 0 and 6 g/L) in vitro. Methane production, degradabilities of feed substrate, and ammonia concentration decreased linearly with increasing doses of vanillin. Concentration of total volatile fatty acids also decreased, whereas proportion of butyrate tended to increase linearly with increasing doses of vanillin. Protozoa population decreased, but abundances of Ruminococcus flavefaciens, Prevotella bryantii, Butyrivibrio fibrisolvens, Prevotella ruminicola, Clostridium aminophilum, and Ruminobacter amylophilus increased with increasing doses of vanillin. Origanum and clove oils resulted in lower ammonia concentrations compared to control and peppermint oil. All the tested essential oils decreased abundances of protozoa, Selenomonas ruminantium, R. amylophilus, P. ruminicola and P. bryantii, with the largest decrease resulted from origanum oil followed by clove oil and peppermint oil. The abundances of Megasphaera elsdenii, C. aminophilum, and Clostridium sticklandii were deceased by origanum oil while that of B. fibrisolvens was lowered by both origanum and clove oils. Saponin decreased ammonia concentration and protozoal population, but increased the abundances of S. ruminantium, R. amylophilus, P. ruminicola, and P. bryantii, though the magnitude was small (less than one log unit). The results suggest that reduction of ammonia production by vanillin and saponin may not be caused by direct inhibition of major known proteolytic bacteria, and essential oils can have different inhibitory effects on different proteolytic bacteria, resulting in varying reduction in ammonia production.

  12. Cadmium inhibits the protein degradation of Sml1 by inhibiting the phosphorylation of Sml1 in Saccharomyces cerevisiae

    Energy Technology Data Exchange (ETDEWEB)

    Baek, In-Joon; Kang, Hyun-Jun; Chang, Miwha; Choi, Il-Dong; Kang, Chang-Min [School of Life Sciences and Biotechnology, Korea University, Anam-dong, Sungbuk-gu, Seoul (Korea, Republic of); Yun, Cheol-Won, E-mail: cheolwony@korea.ac.kr [School of Life Sciences and Biotechnology, Korea University, Anam-dong, Sungbuk-gu, Seoul (Korea, Republic of)

    2012-08-03

    Highlights: Black-Right-Pointing-Pointer Cd inhibits Sml1-p formation. Black-Right-Pointing-Pointer Cd affects cell cycle. Black-Right-Pointing-Pointer Cd inhibits Sml1 ubiquitination. -- Abstract: Cadmium is a toxic metal, and the mechanism of cadmium toxicity in living organisms has been well studied. Here, we used Saccharomyces cerevisiae as a model system to examine the detailed molecular mechanism of cell growth defects caused by cadmium. Using a plate assay of a yeast deletion mutant collection, we found that deletion of SML1, which encodes an inhibitor of Rnr1, resulted in cadmium resistance. Sml1 protein levels increased when cells were treated with cadmium, even though the mRNA levels of SML1 remained unchanged. Using northern and western blot analyses, we found that cadmium inhibited Sml1 degradation by inhibiting Sml1 phosphorylation. Sml1 protein levels increased when cells were treated with cadmium due to disruption of the dependent protein degradation pathway. Furthermore, cadmium promoted cell cycle progression into the G2 phase. The same result was obtained using cells in which SML1 was overexpressed. Deletion of SML1 delayed cell cycle progression. These results are consistent with Sml1 accumulation and with growth defects caused by cadmium stress. Interestingly, although cadmium treatment led to increase Sml1 levels, intracellular dNTP levels also increased because of Rnr3 upregulation due to cadmium stress. Taken together, these results suggest that cadmium specifically affects the phosphorylation of Sml1 and that Sml1 accumulates in cells.

  13. Using Multiple Phenotype Assays and Epistasis Testing to Enhance the Reliability of RNAi Screening and Identify Regulators of Muscle Protein Degradation

    Directory of Open Access Journals (Sweden)

    Nathaniel J. Szewczyk

    2012-11-01

    Full Text Available RNAi is a convenient, widely used tool for screening for genes of interest. We have recently used this technology to screen roughly 750 candidate genes, in C. elegans, for potential roles in regulating muscle protein degradation in vivo. To maximize confidence and assess reproducibility, we have only used previously validated RNAi constructs and have included time courses and replicates. To maximize mechanistic understanding, we have examined multiple sub-cellular phenotypes in multiple compartments in muscle. We have also tested knockdowns of putative regulators of degradation in the context of mutations or drugs that were previously shown to inhibit protein degradation by diverse mechanisms. Here we discuss how assaying multiple phenotypes, multiplexing RNAi screens with use of mutations and drugs, and use of bioinformatics can provide more data on rates of potential false positives and negatives as well as more mechanistic insight than simple RNAi screening.

  14. Adaptive engineering of a hyperthermophilic archaeon on CO and discovering the underlying mechanism by multi-omics analysis.

    Science.gov (United States)

    Lee, Seong Hyuk; Kim, Min-Sik; Lee, Jae-Hak; Kim, Tae Wan; Bae, Seung Seob; Lee, Sung-Mok; Jung, Hae Chang; Yang, Tae-Jun; Choi, Ae Ran; Cho, Yong-Jun; Lee, Jung-Hyun; Kwon, Kae Kyoung; Lee, Hyun Sook; Kang, Sung Gyun

    2016-03-15

    The hyperthermophilic archaeon Thermococcus onnurineus NA1 can grow and produce H2 on carbon monoxide (CO) and its H2 production rates have been improved through metabolic engineering. In this study, we applied adaptive evolution to enhance H2 productivity. After over 150 serial transfers onto CO medium, cell density, CO consumption rate and H2 production rate increased. The underlying mechanism for those physiological changes could be explained by using multi-omics approaches including genomic, transcriptomic and epigenomic analyses. A putative transcriptional regulator was newly identified to regulate the expression levels of genes related to CO oxidation. Transcriptome analysis revealed significant changes in the transcript levels of genes belonging to the categories of transcription, translation and energy metabolism. Our study presents the first genome-scale methylation pattern of hyperthermophilic archaea. Adaptive evolution led to highly enhanced H2 productivity at high CO flow rates using synthesis gas produced from coal gasification.

  15. Phylogenetic analysis of hyperthermophilic natural populations using ribosomal RNA sequences. Final report, July 15, 1995--July 14, 1996

    Energy Technology Data Exchange (ETDEWEB)

    Pace, N.R. [Univ. of California, Berkeley, CA (United States)

    1997-06-01

    It has become clear over the past few decades that substantial microbial diversity occurs at very high temperatures. Hyperthermophilic organisms (temperature optima > 80{degrees}C) promise a wealth of unknown biochemistry and biotechnological potential, and challenge our comprehension of biomolecular structure. Nonetheless, relatively little is known about the diversity of life at high temperatures because of a traditional problem in microbial ecology; inability to cultivate naturally occurring organisms. Molecular techniques have recently been developed, however, that allow the detection and some characterization of organisms without cultivation. Limited surveys of hyperthermophilic communities using such techniques have revealed the existence of an unexpected plethora of organisms, some profoundly different from known ones. The main objective of the proposed program is to continue to characterize, phylogenetically and quantitatively, without cultivation, the constituents of ecosystems that are known to be associated with particular high-temperature sites. Main focus is on the Yellowstone geothermal system.

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

    Directory of Open Access Journals (Sweden)

    Bossa Francesco

    2008-02-01

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

  17. New perspectives in anaerobic digestion

    DEFF Research Database (Denmark)

    van Lier, J.B.; Tilche, A.; Ahring, Birgitte Kiær

    2001-01-01

    The IWA specialised group on anaerobic digestion (AD) is one of the oldest working groups of the former IAWQ organisation. Despite the fact that anaerobic technology dates back more than 100 years, the technology is still under development, adapting novel treatment systems to the modern...... requirements. In fact, most advances were achieved during the last three decades, when high-rate reactor systems were developed and a profound insight was obtained in the microbiology of the anaerobic communities. This insight led to a better understanding of anaerobic treatment and, subsequently, to a broader...

  18. Anaerobic digestion of solid material

    DEFF Research Database (Denmark)

    Vavilin, V.A.; Lokshina, L.Y.; Flotats, X.

    2007-01-01

    A new multidimensional (3 and 2D) anaerobic digestion model for cylindrical reactor with non-uniform influent concentration distributions was developed to study the way in which mixing intensity affects the efficiency of continuous-flow anaerobic digestion. Batch experiments reported and simulate...

  19. Anaerobic Digestion of Piggery Waste

    NARCIS (Netherlands)

    Velsen, van A.F.M.

    1981-01-01

    Anaerobic digestion is a biological process by which organic matter is converted to methane and carbon dioxide by microbes in the absence of air (oxygen). In nature, anaerobic conversions occur at all places where organic material accumulates and the supply of oxygen is deficient, e.g. in marshes an

  20. Economic viability of anaerobic digestion

    Energy Technology Data Exchange (ETDEWEB)

    Wellinger, A. [INFOENERGIE, Ettenhausen (Switzerland)

    1996-01-01

    The industrial application of anaerobic digestion is a relatively new, yet proven waste treatment technology. Anaerobic digestion reduces and upgrades organic waste, and is a good way to control air pollution as it reduces methane and nitrous gas emissions. For environmental and energy considerations, anaerobic digestion is a nearly perfect waste treatment process. However, its economic viability is still in question. A number of parameters - type of waste (solid or liquid), digester system, facility size, product quality and end use, environmental requirements, cost of alternative treatments (including labor), and interest rates - define the investment and operating costs of an anaerobic digestion facility. Therefore, identical facilities that treat the same amount and type of waste may, depending on location, legislation, and end product characteristics, reveal radically different costs. A good approach for evaluating the economics of anaerobic digestion is to compare it to treatment techniques such as aeration or conventional sewage treatment (for industrial wastewater), or composting and incineration (for solid organic waste). For example, the cost (per ton of waste) of in-vessel composting with biofilters is somewhat higher than that of anaerobic digestion, but the investment costs 1 1/2 to 2 times more than either composting or anaerobic digestion. Two distinct advantages of anaerobic digestion are: (1) it requires less land than either composting or incinerating, which translates into lower costs and milder environmental and community impacts (especially in densely populated areas); and (2) it produces net energy, which can be used to operate the facility or sold to nearby industries.

  1. New perspectives in anaerobic digestion

    DEFF Research Database (Denmark)

    van Lier, J.B.; Tilche, A.; Ahring, Birgitte Kiær

    2001-01-01

    The IWA specialised group on anaerobic digestion (AD) is one of the oldest working groups of the former IAWQ organisation. Despite the fact that anaerobic technology dates back more than 100 years, the technology is still under development, adapting novel treatment systems to the modern requireme...

  2. New perspectives in anaerobic digestion.

    NARCIS (Netherlands)

    Lier, van J.B.; Tilche, A.; Ahring, B.K.; Macarie, H.; Moletta, R.; Dohanyos, M.; Hulshoff Pol, L.W.; Lens, P.N.L.; Verstraete, W.

    2001-01-01

    The IWA specialised group on anaerobic digestion (AD) is one of the oldest working groups of the former IAWQ organisation. Despite the fact that anaerobic technology dates back more than 100 years, the technology is still under development, adapting novel treatment systems to the modern

  3. Differential Effects of Hydrophobic Core Packing Residues for Thermodynamic and Mechanical Stability of a Hyperthermophilic Protein.

    Science.gov (United States)

    Tych, Katarzyna M; Batchelor, Matthew; Hoffmann, Toni; Wilson, Michael C; Hughes, Megan L; Paci, Emanuele; Brockwell, David J; Dougan, Lorna

    2016-07-26

    Proteins from organisms that have adapted to environmental extremes provide attractive systems to explore and determine the origins of protein stability. Improved hydrophobic core packing and decreased loop-length flexibility can increase the thermodynamic stability of proteins from hyperthermophilic organisms. However, their impact on protein mechanical stability is not known. Here, we use protein engineering, biophysical characterization, single-molecule force spectroscopy (SMFS), and molecular dynamics (MD) simulations to measure the effect of altering hydrophobic core packing on the stability of the cold shock protein TmCSP from the hyperthermophilic bacterium Thermotoga maritima. We make two variants of TmCSP in which a mutation is made to reduce the size of aliphatic groups from buried hydrophobic side chains. In the first, a mutation is introduced in a long loop (TmCSP L40A); in the other, the mutation is introduced on the C-terminal β-strand (TmCSP V62A). We use MD simulations to confirm that the mutant TmCSP L40A shows the most significant increase in loop flexibility, and mutant TmCSP V62A shows greater disruption to the core packing. We measure the thermodynamic stability (ΔGD-N) of the mutated proteins and show that there is a more significant reduction for TmCSP L40A (ΔΔG = 63%) than TmCSP V62A (ΔΔG = 47%), as might be expected on the basis of the relative reduction in the size of the side chain. By contrast, SMFS measures the mechanical stability (ΔG*) and shows a greater reduction for TmCSP V62A (ΔΔG* = 8.4%) than TmCSP L40A (ΔΔG* = 2.5%). While the impact on the mechanical stability is subtle, the results demonstrate the power of tuning noncovalent interactions to modulate both the thermodynamic and mechanical stability of a protein. Such understanding and control provide the opportunity to design proteins with optimized thermodynamic and mechanical properties.

  4. Three multihaem cytochromes c from the hyperthermophilic archaeon Ignicoccus hospitalis: purification, properties and localization.

    Science.gov (United States)

    Naß, Bastian; Pöll, Uwe; Langer, Julian David; Kreuter, Lydia; Küper, Ulf; Flechsler, Jennifer; Heimerl, Thomas; Rachel, Reinhard; Huber, Harald; Kletzin, Arnulf

    2014-06-01

    Three different multihaem cytochromes c were purified from cell extracts of the hyperthermophilic archaeon Ignicoccus hospitalis. One tetrahaem cytochrome, locus tag designation Igni_0530, was purified from membrane fractions together with the iron-sulfur protein Igni_0529. Two octahaem cytochromes, Igni_0955 and Igni_1359, were purified from soluble fractions but were also present in the membrane fraction. N-terminal sequencing showed that three of the four proteins had their signal peptides cleaved off, while results were ambiguous for Igni_0955. In contrast, mass spectrometry of Igni_0955 and Igni_1359 resulted in single mass peaks including the signal sequences and eight haems per subunit and so both forms might be present in the cell. Igni_0955 and Igni_1359 belong to the hydroxylamine dehydrogenase (HAO) family (29 % mutual identity). HAO or reductase activities with inorganic sulfur compounds were not detected. Igni_0955 was reduced by enriched I. hospitalis hydrogenase at a specific activity of 243 nmol min(-1) (mg hydrogenase)(-1) while activity was non-existent for Igni_0530 and low for Igni_1359. Immuno-electron microscopy of ultra-thin sections showed that Igni_0955 and Igni_1359 are located in both I. hospitalis membranes and also in the intermembrane compartment. We concluded that these cytochromes might function as electron shuttles between the hydrogenase in the outer cellular membrane and cellular reductases, whereas Igni_0530 might be part of the sulfur-reducing mechanism. © 2014 The Authors.

  5. Ligation reaction specificities of an NAD(+)-dependent DNA ligase from the hyperthermophile Aquifex aeolicus.

    Science.gov (United States)

    Tong, J; Barany, F; Cao, W

    2000-03-15

    An NAD(+)-dependent DNA ligase from the hyperthermophilic bacterium Aquifex aeolicus was cloned, expressed in Escherichia coli and purified to homogeneity. The enzyme is most active in slightly alkaline pH conditions with either Mg(2+)or Mn(2+)as the metal cofactor. Ca(2+)and Ni(2+)mainly support formation of DNA-adenylate intermediates. The catalytic cycle is characterized by a low k (cat)value of 2 min(-1)with concomitant accumulation of the DNA - adenylate intermediate when Mg(2+)is used as the metal cofactor. The ligation rates of matched substrates vary by up to 4-fold, but exhibit a general trend of T/A ligation reaction is reaffirmed by results from 1 nt insertions on either the 3'- or 5'-side of the nick. Furthermore, most of the unligatable 3' mismatched base pairs prohibit formation of the DNA-adenylate intermediate, indicating that the substrate adenylation step is also a control point for ligation fidelity. Unlike previously studied ATP ligases, gapped substrates cannot be ligated and intermediate accumulation is minimal, suggesting that complete elimination of base pair complementarity on one side of the nick affects substrate adenylation on the 5'-side of the nick junction. Relationships among metal cofactors, ligation products and intermediate, and ligation fidelity are discussed.

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

    Science.gov (United States)

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

    2016-03-01

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

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

    Science.gov (United States)

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

    2004-01-01

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

  8. Modeling DNA Repair: Approaching In Vivo Techniques in the Hyperthermophile Sulfolobus Solfataricus

    Energy Technology Data Exchange (ETDEWEB)

    Blanton, J.; Fuss, J.; Yannone, S.M.; Tainer, J.A.; Cooper, P.K.

    2005-01-01

    Archaea are found in some of the most extreme environments on earth and represent a third domain of life distinct from Eukarya and Eubacteria. The hyperthermophilic archaeon Sulfolobus solfataricus, isolated from acidic hot springs (80oC, pH 3) in Yellowstone National Park, has emerged as a potential model system for studying human DNA repair processes. Archaea are more closely related to Eukarya than to Eubacteria, suggesting that archaeal DNA repair machinery may model the complex human system much more closely than that of other prokaryotes. DNA repair requires coordinated protein-protein interactions that are frequently transient. Protein complexes that are transient at extreme temperatures where archaea thrive may be more stable at room temperature, allowing for the characterization of otherwise short-lived complexes. However, characterization of these systems in archaea has been limited by the absence of a stable in vivo transformation and expression system. The work presented here is a pilot study in gene cloning and recombinant protein expression in S. solfataricus. Three genes associated with DNA repair were selected for expression: MRE11, PCNA1, and a putative CSB homologue. Though preparation of these recombinant genes followed standard methods, preparation of a suitable vector proved more challenging. The shuttle vector pSSV64, derived from the SSV1 virus and the E. coli vector pBSSK+, was most successfully isolated from the DH5α E. coli strain. Currently, alternative vectors are being designed for more efficient genetic manipulations in S. solfataricus.

  9. Molecular characteristics of spontaneous deletions in the hyperthermophilic archaeon Sulfolobus acidocaldarius.

    Science.gov (United States)

    Grogan, Dennis W; Hansen, Josh E

    2003-02-01

    Prokaryotic genomes acquire and eliminate blocks of DNA sequence by lateral gene transfer and spontaneous deletion, respectively. The basic parameters of spontaneous deletion, which are expected to influence the course of genome evolution, have not been determined for any hyperthermophilic archaeon. We therefore screened a number of independent pyrimidine auxotrophs of Sulfolobus acidocaldarius for deletions and sequenced those detected. Deletions accounted for only 0.4% of spontaneous pyrE mutations, corresponding to a frequency of about 10(-8) per cell. Nucleotide sequence analysis of five independent deletions showed no significant association of the endpoints with short direct repeats, despite the fact that several such repeats occur within the pyrE gene and that duplication mutations in pyrE reverted at high frequencies. Endpoints of the spontaneous deletions did not coincide with short inverted repeats or potential stem-loop structures. No consensus sequence common to all the deletions could be identified, although two deletions showed the potential of being stabilized by octanucleotide sequences elsewhere in pyrE, and another pair of deletions shared an octanucleotide at their 3' ends. The unusually low frequency and low sequence dependence of spontaneous deletions in the S. acidocaldarius pyrE gene compared to other genetic systems could not be explained in terms of possible constraints imposed by the 5-fluoroorotate selection.

  10. Preparation of linear maltodextrins using a hyperthermophilic amylopullulanase with cyclodextrin- and starch-hydrolysing activities.

    Science.gov (United States)

    Li, Xiaolei; Li, Dan

    2015-03-30

    A novel method for the preparation of linear maltodextrins from cyclodextrins and starch was proposed. To accomplish this process, an amylopullulanase from hyperthermophilic archaeon Caldivirga maquilingensis (CMApu) was characterized and used. CMApu with an estimated molecular mass of 62.7 kDa by SDS-PAGE had a maximal pullulan-hydrolysing activity at 100°C and pH 5.0. It could also hydrolyse amylopectin (AP), starch, β-CD and amylose (AM), in a decreasing order of relative activities from 88.96% to 57.17%. TLC and HPAEC analysis revealed that CMApu catalyzed the debranching and degrading reactions to produce linear malto-oligosaccharides (≤ G8-G1) from G8-β-CD and/or normal CDs, amylodextrins (DP6-96) from AM, and amylodextrins (DP1-76) from AP and potato starch. Our results showed that CMApu had a great potential for the industrial preparation of linear maltodextrins from normal starch instead of waxy starch, malto-oligosaccharides or sucrose. And the high optimal temperature of CMApu facilitated the simultaneous gelatinization and hydrolysis of cereal starch.

  11. Denaturing Effects of Urea and Guanidine Hydrochloride on Hyperthermophilic Esterase from Aeropyrum pernix K1

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    The changes in the activity and the conformation of the hyperthermophilic esterase derived from aerobic thermophilic Aeropyrumpernix K1 (APE1547) were studied during denaturation by guanidine hydrochloride (GdnHCl)and urea. The denaturation course of APE1547 was followed by the steady-state and time resolved fluorescence methods. An increase in the denaturant concentration in the denatured system can significantly enhance the inactivation and unfolding of APE1547. The enzyme can be completely inactivated with a urea concentration of 2. 7 mol/L or a GdnHCl concentration of 7.5 mol/L. The fluorescence emission maximum of the enzyme protein red shifts in magnitude to a maximum value(355 nm) when the concentration of GdnHCl is 5.1 mol/L. The experimental results indicate that APE1547 has a high resistance to urea. Unfolding of APE1547 in GdnHCl(4.2-6.0 mol/L) was shown to be an irreversible process. The present results indicate that the ion pairs in this protein may be a key factor for the stability of this esterase.

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

    Science.gov (United States)

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

    2004-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Anaïs eCario

    2015-10-01

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

  14. Acetaminophen analog N-acetyl-m-aminophenol, but not its reactive metabolite, N-acetyl-p-benzoquinone imine induces CYP3A activity via inhibition of protein degradation.

    Science.gov (United States)

    Santoh, Masataka; Sanoh, Seigo; Ohtsuki, Yuya; Ejiri, Yoko; Kotake, Yaichiro; Ohta, Shigeru

    2017-05-06

    Cytochrome P450 (CYP) 3A subfamily members are known to metabolize various types of drugs, highlighting the importance of understanding drug-drug interactions (DDI) depending on CYP3A induction or inhibition. While transcriptional regulation of CYP3A members is widely understood, post-translational regulation needs to be elucidated. We previously reported that acetaminophen (APAP) induces CYP3A activity via inhibition of protein degradation and proposed a novel DDI concept. N-Acetyl-p-benzoquinone imine (NAPQI), the reactive metabolite of APAP formed by CYP, is known to cause adverse events related to depletion of intracellular reduced glutathione (GSH). We aimed to inspect whether NAPQI rather than APAP itself could cause the inhibitory effects on protein degradation. We found that N-acetyl-l-cysteine, the precursor of GSH, and 1-aminobenzotriazole, a nonselective CYP inhibitor, had no effect on CYP3A1/23 protein levels affected by APAP. Thus, we used APAP analogs to test CYP3A1/23 mRNA levels, protein levels, and CYP3A activity. We found N-acetyl-m-aminophenol (AMAP), a regioisomer of APAP, has the same inhibitory effects of CYP3A1/23 protein degradation, while p-acetamidobenzoic acid (PAcBA), a carboxy-substituted form of APAP, shows no inhibitory effects. AMAP and PAcBA cannot be oxidized to quinone imine forms such as NAPQI, so the inhibitory effects could depend on the specific chemical structure of APAP. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. Anaerobic Origin of Ergothioneine.

    Science.gov (United States)

    Burn, Reto; Misson, Laëtitia; Meury, Marcel; Seebeck, Florian P

    2017-10-02

    Ergothioneine is a sulfur metabolite that occurs in microorganisms, fungi, plants, and animals. The physiological function of ergothioneine is not clear. In recent years broad scientific consensus has formed around the idea that cellular ergothioneine primarily protects against reactive oxygen species. Herein we provide evidence that this focus on oxygen chemistry may be too narrow. We describe two enzymes from the strictly anaerobic green sulfur bacterium Chlorobium limicola that mediate oxygen-independent biosynthesis of ergothioneine. This anoxic origin suggests that ergothioneine is also important for oxygen-independent life. Furthermore, one of the discovered ergothioneine biosynthetic enzymes provides the first example of a rhodanese-like enzyme that transfers sulfur to non-activated carbon. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. The relation of muscle protein degradation and 26S proteasome%肌肉蛋白降解与蛋白酶复合体

    Institute of Scientific and Technical Information of China (English)

    谭银玲

    2004-01-01

    The ubiquitin-dependent 20s/26s proteasome system is the capital pathway of exo-lyso-some proteolysis within eukaryotic cell. Under conditions of denervation, starvation, glucocorticoid, infection,tumor, bum and so on, the proteasome system was stimulated tofast. Glucocorticoid, insulin, thyroid honnone,TNFα and IL-1β degrade protein, which results in muscle lose cle protein degradation and the proteasome system. Inhibition or activation of the proteasome system was approved to be a novel means of treatment with cachexia and negative nitrogen balance.

  17. Isolation and Cultivation of Anaerobes

    DEFF Research Database (Denmark)

    Aragao Börner, Rosa

    2016-01-01

    Anaerobic microorganisms play important roles in different biotechnological processes. Their complex metabolism and special cultivation requirements have led to less isolated representatives in comparison to their aerobic counterparts.In view of that, the isolation and cultivation of anaerobic...... microorganisms is still a promising venture, and conventional methodologies as well as considerations and modifications are presented here. An insight into new methodologies and devices as well as a discussion on future perspectives for the cultivation of anaerobes may open the prospects of the exploitation...

  18. Implementing Livestock Anaerobic Digestion Projects

    Science.gov (United States)

    Page provides information to help make an informed decision about installing an anaerobic digester. Is it a good match for a farm’s organic waste, project financing, development guidelines and permit requirements?

  19. Coupled TLC and MALDI-TOF/MS Analyses of the Lipid Extract of the Hyperthermophilic Archaeon Pyrococcus furiosus

    Directory of Open Access Journals (Sweden)

    Simona Lobasso

    2012-01-01

    Full Text Available The lipidome of the marine hyperthermophilic archaeon Pyrococcus furiosus was studied by means of combined thin-layer chromatography and MALDI-TOF/MS analyses of the total lipid extract. 80–90% of the major polar lipids were represented by archaeol lipids (diethers and the remaining part by caldarchaeol lipids (tetraethers. The direct analysis of lipids on chromatography plate showed the presence of the diphytanylglycerol analogues of phosphatidylinositol and phosphatidylglycerol, the N-acetylglucosamine-diphytanylglycerol phosphate plus some caldarchaeol lipids different from those previously described. In addition, evidence for the presence of the dimeric ether lipid cardiolipin is reported, suggesting that cardiolipins are ubiquitous in archaea.

  20. Anaerobic Biodegradation of Detergent Surfactants

    OpenAIRE

    Erich Jelen; Ute Merrettig-Bruns

    2009-01-01

    Detergent surfactantscan be found in wastewater in relevant concentrations. Most of them are known as ready degradable under aerobic conditions, as required by European legislation. Far fewer surfactants have been tested so far for biodegradability under anaerobic conditions. The natural environment is predominantly aerobic, but there are some environmental compartments such as river sediments, sub-surface soil layer and anaerobic sludge digesters of wastewater treatment plants which have str...

  1. Influence of supplemental vitamin C on postmortem protein degradation and fatty acid profiles of the longissimus thoracis of steers fed varying concentrations of dietary sulfur.

    Science.gov (United States)

    Pogge, Danielle J; Lonergan, Steven M; Hansen, Stephanie L

    2014-02-01

    The objective was to examine the effects of supplemental vitamin C (VC) on postmortem protein degradation and fatty acid profiles of cattle receiving varying concentrations of dietary sulfur (S). A longissimus muscle was collected from 120 Angus-cross steers assigned to a 3 × 2 factorial, evaluating three concentrations of dietary S (0.22, 0.34, and 0.55%) and two concentrations of supplemental VC (0 or 10 g h(-1)d(-1)). Increasing dietary S and VC supplementation (Pacids of steaks. Addition of VC tended to increase (P = 0.09) both Fe and 2-thiobarbituric acid content of longissimus thoracis. Increasing S increased (P = 0.03) the proportion of 80-kDa subunit of μ-calpain. Addition of VC within the high S treatment increased (P = 0.05) the abundance of 76-kDa subunit of μ-calpain. Increasing S decreased troponin T degradation (P = 0.07) and protein carbonylation (P<0.01). Supplemental VC appears to alleviate negative effects of high S on autolysis of μ-calpain and protein degradation. © 2013.

  2. Characterization of a Zinc-Containing Alcohol Dehydrogenase with Stereoselectivity from the Hyperthermophilic Archaeon Thermococcus guaymasensis▿

    Science.gov (United States)

    Ying, Xiangxian; Ma, Kesen

    2011-01-01

    An alcohol dehydrogenase (ADH) from hyperthermophilic archaeon Thermococcus guaymasensis was purified to homogeneity and was found to be a homotetramer with a subunit size of 40 ± 1 kDa. The gene encoding the enzyme was cloned and sequenced; this gene had 1,095 bp, corresponding to 365 amino acids, and showed high sequence homology to zinc-containing ADHs and l-threonine dehydrogenases with binding motifs of catalytic zinc and NADP+. Metal analyses revealed that this NADP+-dependent enzyme contained 0.9 ± 0.03 g-atoms of zinc per subunit. It was a primary-secondary ADH and exhibited a substrate preference for secondary alcohols and corresponding ketones. Particularly, the enzyme with unusual stereoselectivity catalyzed an anti-Prelog reduction of racemic (R/S)-acetoin to (2R,3R)-2,3-butanediol and meso-2,3-butanediol. The optimal pH values for the oxidation and formation of alcohols were 10.5 and 7.5, respectively. Besides being hyperthermostable, the enzyme activity increased as the temperature was elevated up to 95°C. The enzyme was active in the presence of methanol up to 40% (vol/vol) in the assay mixture. The reduction of ketones underwent high efficiency by coupling with excess isopropanol to regenerate NADPH. The kinetic parameters of the enzyme showed that the apparent Km values and catalytic efficiency for NADPH were 40 times lower and 5 times higher than those for NADP+, respectively. The physiological roles of the enzyme were proposed to be in the formation of alcohols such as ethanol or acetoin concomitant to the NADPH oxidation. PMID:21515780

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

    Directory of Open Access Journals (Sweden)

    Liu Liangwei

    2012-06-01

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

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

    Directory of Open Access Journals (Sweden)

    Wirojne Kanoksilapatham

    2004-01-01

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

  5. Metabolism of pentose sugars in the hyperthermophilic archaea Sulfolobus solfataricus and Sulfolobus acidocaldarius.

    Science.gov (United States)

    Nunn, Charlotte E M; Johnsen, Ulrike; Schönheit, Peter; Fuhrer, Tobias; Sauer, Uwe; Hough, David W; Danson, Michael J

    2010-10-29

    We have previously shown that the hyperthermophilic archaeon, Sulfolobus solfataricus, catabolizes d-glucose and d-galactose to pyruvate and glyceraldehyde via a non-phosphorylative version of the Entner-Doudoroff pathway. At each step, one enzyme is active with both C6 epimers, leading to a metabolically promiscuous pathway. On further investigation, the catalytic promiscuity of the first enzyme in this pathway, glucose dehydrogenase, has been shown to extend to the C5 sugars, D-xylose and L-arabinose. In the current paper we establish that this promiscuity for C6 and C5 metabolites is also exhibited by the third enzyme in the pathway, 2-keto-3-deoxygluconate aldolase, but that the second step requires a specific C5-dehydratase, the gluconate dehydratase being active only with C6 metabolites. The products of this pathway for the catabolism of D-xylose and L-arabinose are pyruvate and glycolaldehyde, pyruvate entering the citric acid cycle after oxidative decarboxylation to acetyl-coenzyme A. We have identified and characterized the enzymes, both native and recombinant, that catalyze the conversion of glycolaldehyde to glycolate and then to glyoxylate, which can enter the citric acid cycle via the action of malate synthase. Evidence is also presented that similar enzymes for this pentose sugar pathway are present in Sulfolobus acidocaldarius, and metabolic tracer studies in this archaeon demonstrate its in vivo operation in parallel with a route involving no aldol cleavage of the 2-keto-3-deoxy-pentanoates but direct conversion to the citric acid cycle C5-metabolite, 2-oxoglutarate.

  6. Heteroduplex formation, mismatch resolution, and genetic sectoring during homologous recombination in the hyperthermophilic archaeon sulfolobus acidocaldarius.

    Science.gov (United States)

    Mao, Dominic; Grogan, Dennis W

    2012-01-01

    Hyperthermophilic archaea exhibit certain molecular-genetic features not seen in bacteria or eukaryotes, and their systems of homologous recombination (HR) remain largely unexplored in vivo. We transformed a Sulfolobus acidocaldariuspyrE mutant with short DNAs that contained multiple non-selected genetic markers within the pyrE gene. From 20 to 40% of the resulting colonies were found to contain two Pyr(+) clones with distinct sets of the non-selected markers. The dual-genotype colonies could not be attributed to multiple DNAs entering the cells, or to conjugation between transformed and non-transformed cells. These colonies thus appear to represent genetic sectoring in which regions of heteroduplex DNA formed and then segregated after partial resolution of inter-strand differences. Surprisingly, sectoring was also frequent in cells transformed with single-stranded DNAs. Oligonucleotides produced more sectored transformants when electroporated as single strands than as a duplex, although all forms of donor DNA (positive-strand, negative-strand, and duplex) produced a diversity of genotypes, despite the limited number of markers. The marker patterns in the recombinants indicate that S. acidocaldarius resolves individual mismatches through un-coordinated short-patch excision followed by re-filling of the resulting gap. The conversion events that occur during transformation by single-stranded DNA do not show the strand bias necessary for a system that corrects replication errors effectively; similar events also occur in pre-formed heteroduplex electroporated into the cells. Although numerous mechanistic details remain obscure, the results demonstrate that the HR system of S. acidocaldarius can generate remarkable genetic diversity from short intervals of moderately diverged DNAs.

  7. Heteroduplex formation, mismatch resolution, and genetic sectoring during homologous recombination in the hyperthermophilic archaeon Sulfolobus acidocaldarius

    Directory of Open Access Journals (Sweden)

    Dennis W. Grogan

    2012-06-01

    Full Text Available Hyperthermophilic archaea exhibit certain molecular-genetic features not seen in bacteria or eukaryotes, and their systems of homologous recombination (HR remain largely unexplored in vivo. We transformed a Sulfolobus acidocaldarius pyrE mutant with short DNAs that contained multiple non-selected genetic markers within the pyrE gene. From 20 to 40% of the resulting colonies were found to contain two Pyr+ clones with distinct sets of the non-selected markers. The dual-genotype colonies could not be attributed to multiple DNAs entering the cells or conjugation between transformed and non-transformed cells. These colonies thus appear to represent genetic sectoring in which stretches of heteroduplex DNA formed during HR and segregated without complete resolution of inter-strand differences. Surprisingly, sectoring was also frequent in transformation with single-stranded DNAs. Oligonucleotides, for example, produced somewhat more sectored transformants when electroporated as single strands than as a duplex, although all forms (positive-strand, negative-strand, and duplex produced a diversity of genotypes from the limited number of markers. The marker patterns in the recombinants indicate that S. acidocaldarius resolves individual mismatches through un-coordinated short-patch excision followed by re-filling of the resulting gap. These gene-conversion events exhibit little strand bias, and can occur in pre-formed heteroduplex. These properties suggest that this process does not play a central role in the fidelity of genome replication, but may generate 3’ single-strand tails, and thereby initiate the incorporation of duplex DNA into the recipient chromosome. Regardless of the molecular details of its mechanism, HR between the S. acidocaldarius chromosome and a multiply-marked DNA produces a strikingly high level of genetic diversity in a very short chromosomal interval, and suggests that HR in Sulfolobus has significant mutagenic potential if not

  8. A mutant ('lab strain') of the hyperthermophilic archaeon Pyrococcus furiosus, lacking flagella, has unusual growth physiology.

    Science.gov (United States)

    Lewis, Derrick L; Notey, Jaspreet S; Chandrayan, Sanjeev K; Loder, Andrew J; Lipscomb, Gina L; Adams, Michael W W; Kelly, Robert M

    2015-03-01

    A mutant ('lab strain') of the hyperthermophilic archaeon Pyrococcus furiosus DSM3638 exhibited an extended exponential phase and atypical cell aggregation behavior. Genomic DNA from the mutant culture was sequenced and compared to wild-type (WT) DSM3638, revealing 145 genes with one or more insertions, deletions, or substitutions (12 silent, 33 amino acid substitutions, and 100 frame shifts). Approximately, half of the mutated genes were transposases or hypothetical proteins. The WT transcriptome revealed numerous changes in amino acid and pyrimidine biosynthesis pathways coincidental with growth phase transitions, unlike the mutant whose transcriptome reflected the observed prolonged exponential phase. Targeted gene deletions, based on frame-shifted ORFs in the mutant genome, in a genetically tractable strain of P. furiosus (COM1) could not generate the extended exponential phase behavior observed for the mutant. For example, a putative radical SAM family protein (PF2064) was the most highly up-regulated ORF (>25-fold) in the WT between exponential and stationary phase, although this ORF was unresponsive in the mutant; deletion of this gene in P. furiosus COM1 resulted in no apparent phenotype. On the other hand, frame-shifting mutations in the mutant genome negatively impacted transcription of a flagellar biosynthesis operon (PF0329-PF0338).Consequently, cells in the mutant culture lacked flagella and, unlike the WT, showed minimal evidence of exopolysaccharide-based cell aggregation in post-exponential phase. Electron microscopy of PF0331-PF0337 deletions in P. furiosus COM1 showed that absence of flagella impacted normal cell aggregation behavior and, furthermore, indicated that flagella play a key role, beyond motility, in the growth physiology of P. furiosus.

  9. Reactor-scale cultivation of the hyperthermophilic methanarchaeon Methanococcus jannaschii to high cell densities.

    Science.gov (United States)

    Mukhopadhyay, B; Johnson, E F; Wolfe, R S

    1999-11-01

    For the hyperthermophilic and barophilic methanarchaeon Methanococcus jannaschii, we have developed a medium and protocols for reactor-scale cultivation that improved the final cell yield per liter from approximately 0.5 to approximately 7.5 g of packed wet cells ( approximately 1.8 g dry cell mass) under autotrophic growth conditions and to approximately 8.5 g of packed wet cells ( approximately 2 g dry cell mass) with yeast extract (2 g liter(-1)) and tryptone (2 g liter(-1)) as medium supplements. For growth in a sealed bottle it was necessary to add Se to the medium, and a level of 2 microM for added Se gave the highest final cell yield. In a reactor M. jannaschii grew without added Se in the medium; it is plausible that the cells received Se as a contaminant from the reactor vessel and the H(2)S supply. But, for the optimal performance of a reactor culture, an addition of Se to a final concentration of 50 to 100 microM was needed. Also, cell growth in a reactor culture was inhibited at much higher Se concentrations. These observations and the data from previous work with methanogen cell extracts (B. C. McBride and R. S. Wolfe, Biochemistry 10:4312-4317, 1971) suggested that from a continuously sparged reactor culture Se was lost in the exhaust gas as volatile selenides, and this loss raised the apparent required level of and tolerance for Se. In spite of having a proteinaceous cell wall, M. jannaschii withstood an impeller tip speed of 235.5 cms(-1), which was optimal for achieving high cell density and also was the higher limit for the tolerated shear rate. The organism secreted one or more acidic compounds, which lowered pH in cultures without pH control; this secretion continued even after cessation of growth.

  10. Citric acid cycle in the hyperthermophilic archaeon Pyrobaculum islandicum grown autotrophically, heterotrophically, and mixotrophically with acetate.

    Science.gov (United States)

    Hu, Yajing; Holden, James F

    2006-06-01

    The hyperthermophilic archaeon Pyrobaculum islandicum uses the citric acid cycle in the oxidative and reductive directions for heterotrophic and autotrophic growth, respectively, but the control of carbon flow is poorly understood. P. islandicum was grown at 95 degrees C autotrophically, heterotrophically, and mixotrophically with acetate, H2, and small amounts of yeast extract and with thiosulfate as the terminal electron acceptor. The autotrophic growth rates and maximum concentrations of cells were significantly lower than those in other media. The growth rates on H2 and 0.001% yeast extract with and without 0.05% acetate were the same, but the maximum concentration of cells was fourfold higher with acetate. There was no growth with acetate if 0.001% yeast extract was not present, and addition of H2 to acetate-containing medium greatly increased the growth rates and maximum concentrations of cells. P. islandicum cultures assimilated 14C-labeled acetate in the presence of H2 and yeast extract with an efficiency of 55%. The activities of 11 of 19 enzymes involved in the central metabolism of P. islandicum were regulated under the three different growth conditions. Pyruvate synthase and acetate:coenzyme A (CoA) ligase (ADP-forming) activities were detected only in heterotrophically grown cultures. Citrate synthase activity decreased in autotrophic and acetate-containing cultures compared to the activity in heterotrophic cultures. Acetylated citrate lyase, acetate:CoA ligase (AMP forming), and phosphoenolpyruvate carboxylase activities increased in autotrophic and acetate-containing cultures. Citrate lyase activity was higher than ATP citrate synthase activity in autotrophic cultures. These data suggest that citrate lyase and AMP-forming acetate:CoA ligase, but not ATP citrate synthase, work opposite citrate synthase to control the direction of carbon flow in the citric acid cycle.

  11. Differential Virus Host-Ranges of the Fuselloviridae of Hyperthermophilic Archaea: Implications for Evolution in Extreme Environments

    Directory of Open Access Journals (Sweden)

    Ruben Michael eCeballos

    2012-08-01

    Full Text Available An emerging model for investigating virus-host interactions in hyperthermophilic Archaea is the Fusellovirus-Sulfolobus system. The host, Sulfolobus, is a hyperthermophilic acidophile endemic to sulfuric volcanic-driven hot springs worldwide. The Fuselloviruses, also known as Sulfolobus Spindle-shaped Viruses (SSVs, are lemon or spindle shaped double-stranded DNA viruses that are also found worldwide. Although a few studies have addressed the host-range for the type virus, SSV1, using common Sulfolobus strains, a comprehensive host-range study for SSV-Sulfolobus systems has not been performed. Herein, we examine six bona fide SSV strains (SSV1, SSV2, SSV3, SSVL1, SSVK1, SSVRH and their respective infection characteristics on multiple hosts from the family Sulfolobaceae. A halo assay was used to determine virus infectivity and host susceptibility. Different SSV strains have different host-ranges with SSV1 exhibiting the narrowest host-range and SSVRH exhibiting the broadest host range. There is no correlation between geographic separation of viruses and their hosts and their relative infectivity and susceptibility. In contrast to previous reports, SSVs can infect hosts beyond the genus Sulfolobus. Furthermore, the Fusellovirus-Sulfolobus system appears to exhibit host-advantage. This work provides a foundation for understanding Fusellovirus biology and virus-host co-evolution in extreme ecosystems, a rapidly emerging field of study.

  12. Thermal stability and unfolding pathways of hyperthermophilic and mesophilic periplasmic binding proteins studied by molecular dynamics simulation.

    Science.gov (United States)

    Chen, Lin; Li, Xue; Wang, Ruige; Fang, Fengqin; Yang, Wanli; Kan, Wei

    2016-07-01

    The ribose binding protein (RBP), a sugar-binding periplasmic protein, is involved in the transport and signaling processes in both prokaryotes and eukaryotes. Although several cellular and structural studies have been reported, a description of the thermostability of RBP at the molecular level remains elusive. Focused on the hyperthermophilic Thermoytoga maritima RBP (tmRBP) and mesophilic Escherichia coli homolog (ecRBP), we applied molecular dynamics simulations at four different temperatures (300, 380, 450, and 500 K) to obtain a deeper insight into the structural features responsible for the reduced thermostability of the ecRBP. The simulations results indicate that there are distinct structural differences in the unfolding pathway between the two homologs and the ecRBP unfolds faster than the hyperthermophilic homologs at certain temperatures in accordance with the lower thermal stability found experimentally. Essential dynamics analysis uncovers that the essential subspaces of ecRBP and tmRBP are non-overlapping and these two proteins show different directions of motion within the simulations trajectories. Such an understanding is required for designing efficient proteins with characteristics for a particular application.

  13. X-ray crystalline structures of pyrrolidone carboxyl peptidase from a hyperthermophile, Pyrococcus furiosus, and its cys-free mutant.

    Science.gov (United States)

    Tanaka, H; Chinami, M; Mizushima, T; Ogasahara, K; Ota, M; Tsukihara, T; Yutani, K

    2001-07-01

    In order to elucidate the mechanism of the thermostability of proteins from hyperthermophiles, X-ray crystalline structures of pyrrolidone carboxyl peptidase from a hyperthermophile, Pyrococcus furiosus (PfPCP), and its mutant protein with Ser substituted at Cys142 and Cys188 were determined at 2.2 and 2.7 A resolution, respectively. The obtained structures were compared with those previously reported for pyrrolidone carboxyl peptidases from a hyperthermophilie, Thermococcus litoralis (TlPCP), and from a mesophile, Bacillus amyloliquefaciens (BaPCP). The PfPCP structure is a tetramer of four identical subunits similar to that of the TlPCP and BaPCP. The largest structural changes among the three PCPs were detected in the C-terminal protrusion, which interacts with that of another subunit. A comparison of the three structures indicated that the high stability of PfPCP is caused by increases in hydrophobic interactions and hydrogen bonds, the formation of an intersubunit ion-pair network, and improvement to an ideal conformation. On the basis of the structures of the three proteins, it can be concluded that PfPCP does not have any special factors responsible for its extremely high stability and that the conformational structure of PfPCP is superior in its combination of positive and negative stabilizing factors compared with BaPCP.

  14. Efficient CRISPR-Mediated Post-Transcriptional Gene Silencing in a Hyperthermophilic Archaeon Using Multiplexed crRNA Expression

    Directory of Open Access Journals (Sweden)

    Ziga Zebec

    2016-10-01

    Full Text Available CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats-mediated RNA degradation is catalyzed by a type III system in the hyperthermophilic archaeon Sulfolobus solfataricus. Earlier work demonstrated that the system can be engineered to target specifically mRNA of an endogenous host reporter gene, namely the β-galactosidase in S. solfataricus. Here, we investigated the effect of single and multiple spacers targeting the mRNA of a second reporter gene, α-amylase, at the same, and at different, locations respectively, using a minimal CRISPR (miniCR locus supplied on a viral shuttle vector. The use of increasing numbers of spacers reduced mRNA levels at progressively higher levels, with three crRNAs (CRISPR RNAs leading to ∼ 70–80% reduction, and five spacers resulting in an α-amylase gene knockdown of > 90% measured on both mRNA and protein activity levels. Our results indicate that this technology can be used to increase or modulate gene knockdown for efficient post-transcriptional gene silencing in hyperthermophilic archaea, and potentially also in other organisms.

  15. Coenzyme Engineering of a Hyperthermophilic 6-Phosphogluconate Dehydrogenase from NADP+ to NAD+ with Its Application to Biobatteries

    Science.gov (United States)

    Chen, Hui; Zhu, Zhiguang; Huang, Rui; Zhang, Yi-Heng Percival

    2016-11-01

    Engineering the coenzyme specificity of redox enzymes plays an important role in metabolic engineering, synthetic biology, and biocatalysis, but it has rarely been applied to bioelectrochemistry. Here we develop a rational design strategy to change the coenzyme specificity of 6-phosphogluconate dehydrogenase (6PGDH) from a hyperthermophilic bacterium Thermotoga maritima from its natural coenzyme NADP+ to NAD+. Through amino acid-sequence alignment of NADP+- and NAD+-preferred 6PGDH enzymes and computer-aided substrate-coenzyme docking, the key amino acid residues responsible for binding the phosphate group of NADP+ were identified. Four mutants were obtained via site-directed mutagenesis. The best mutant N32E/R33I/T34I exhibited a ~6.4 × 104-fold reversal of the coenzyme selectivity from NADP+ to NAD+. The maximum power density and current density of the biobattery catalyzed by the mutant were 0.135 mW cm-2 and 0.255 mA cm-2, ~25% higher than those obtained from the wide-type 6PGDH-based biobattery at the room temperature. By using this 6PGDH mutant, the optimal temperature of running the biobattery was as high as 65 °C, leading to a high power density of 1.75 mW cm-2. This study demonstrates coenzyme engineering of a hyperthermophilic 6PGDH and its application to high-temperature biobatteries.

  16. Biology of gut anaerobic fungi.

    Science.gov (United States)

    Bauchop, T

    1989-01-01

    The obligately anaerobic nature of the gut indigenous fungi distinguishes them from other fungi. They are distributed widely in large herbivores, both in the foregut of ruminant-like animals and in the hindgut of hindgut fermenters. Comparative studies indicate that a capacious organ of fermentative digestion is required for their development. These fungi have been assigned to the Neocallimasticaceae, within the chytridiomycete order Spizellomycetales. The anaerobic fungi of domestic ruminants have been studied most extensively. Plant material entering the rumen is rapidly colonized by zoospores that attach and develop into thalli. The anaerobic rumen fungi have been shown to produce active cellulases and xylanases and specifically colonise and grow on plant vascular tissues. Large populations of anaerobic fungi colonise plant fragment in the rumens of cattle and sheep on high-fibre diets. The fungi actively ferment cellulose which results in formation of a mixture of products including acetate, lactate, ethanol, formate, succinate, CO2 and H2. The properties of the anaerobic fungi together with the extent of their populations on plant fragments in animals on high-fibre diets indicates a significant role for the fungi in fibre digestion.

  17. Label-free, real-time detection of the dynamic processes of protein degradation using oblique-incidence reflectivity difference method

    Science.gov (United States)

    Liu, S.; Zhu, J. H.; He, L. P.; Dai, J.; Lu, H. B.; Wu, L.; Jin, K. J.; Yang, G. Z.; Zhu, H.

    2014-04-01

    Based on the requirements for studying the dynamic process of proteinase action substrates in life science, we selected six random proteins including 1L-10, SCGB2A2, CENPQ, GST, HK1, KLHL7, as well as five different concentrations of 1L-10 proteins of 1 mg/ml, 0.5 mg/ml, 0.25 mg/ml, 0.125 mg/ml, and 0.0625 mg/ml, and fabricated two types of substrate protein microarrays, respectively. We detected the dynamic processes of proteins degraded by proteinase K using oblique-incidence reflectivity difference (OIRD) method in a label-free and real-time manner. We obtained the relevant degradation velocities and the degradation time. The experimental results demonstrate that OIRD has the ability to study proteinase action substrates which is out of reach of label methods and is expected to offer opportunities to determine protease-substrate relationships on the systems biology level.

  18. Protein degradation rate is the dominant mechanism accounting for the differences in protein abundance of basal p53 in a human breast and colorectal cancer cell line.

    Science.gov (United States)

    Lakatos, Eszter; Salehi-Reyhani, Ali; Barclay, Michael; Stumpf, Michael P H; Klug, David R

    2017-01-01

    We determine p53 protein abundances and cell to cell variation in two human cancer cell lines with single cell resolution, and show that the fractional width of the distributions is the same in both cases despite a large difference in average protein copy number. We developed a computational framework to identify dominant mechanisms controlling the variation of protein abundance in a simple model of gene expression from the summary statistics of single cell steady state protein expression distributions. Our results, based on single cell data analysed in a Bayesian framework, lends strong support to a model in which variation in the basal p53 protein abundance may be best explained by variations in the rate of p53 protein degradation. This is supported by measurements of the relative average levels of mRNA which are very similar despite large variation in the level of protein.

  19. Mechanical stimulation of C2C12 cells increases m-calpain expression, focal adhesion plaque protein degradation and cell differentiation

    DEFF Research Database (Denmark)

    Grossi, Alberto; Lawson, Moira Ann

    Abstract Mechanical stimulation of C2C12 cells increases m-calpain expression, focal adhesion plaque protein degradation and cell differentiation. A. Grossi, M. A. Lawson; Department of Food Science, Royal Veterinary and Agricultural University, Frederiksberg C, Denmark The process of muscle...... documented and has been shown to affect transcription of specific gene sequences, protein synthesis, the immune system and increase in Ca2+ influx. The past 10 years has seen a dramatic increase in the understanding of how proteolytic enzymes such as calpains can affect the growth of muscle. In vivo studies...... have shown that m-calpain is necessary for myoblast fusion leading to the formation of muscle fibers and that inhibition of this enzyme restricts myotube formation. Whether there is a link between stretchor load induced signaling and m-calpain expression and activation is not known. Using a magnetic...

  20. Mechanical stimulation of C2C12 cells increases m-calpain expression, focal adhesion plaque protein degradation and cell differentiation

    DEFF Research Database (Denmark)

    Grossi, Alberto; Lawson, Moira Ann

    Abstract Mechanical stimulation of C2C12 cells increases m-calpain expression, focal adhesion plaque protein degradation and cell differentiation. A. Grossi, M. A. Lawson; Department of Food Science, Royal Veterinary and Agricultural University, Frederiksberg C, Denmark The process of muscle...... development and growth is a complex sequence of events whereby muscle cells respond to a number of stimuli in order to form organised muscle tissue. Increase in muscle mass is greatly influenced by the rate of skeletal muscle protein synthesis and degradation, processes that can be altered by mechanical...... forces. Stretch- or load-induced signaling is now beginning to be understood as a factor which affects the mass and phenotype of muscles as well as the expression of a number of proteins within muscle cells. Use of magnetic field to produce mechanical forces to stimulate cell populations has been well...

  1. Inhibition of ERK1/2 Pathway Suppresses Adiponectin Secretion via Accelerating Protein Degradation by Ubiquitin-Proteasome System: Relevance to Obesity-related Adiponectin Decline

    Science.gov (United States)

    Gu, Dongfang; Wang, Zhigang; Dou, Xiaobing; Zhang, Ximei; Li, Songtao; Vu, Lyndsey; Yao, Tong; Song, Zhenyuan

    2013-01-01

    Objective Predominantly secreted by adipose tissue, adiponectin possesses insulin-sensitizing, anti-atherogenic, anti-inflammatory, and anti-angiogenic properties. Paradoxically, obesity is associated with declined plasma adiponectin levels; however, the underlying mechanisms remain elusive. In this study, we investigated the mechanistic involvement of MEK/ERK1/2 pathway in obesity-related adiponectin decrease. Materials/Methods C57 BL/6 mice exposed to a high-fat diet (HFD) were employed as animal obesity model. Both fully-differentiated 3T3-L1 and mouse primary adipocytes were used in the in vitro experiments. Results Obesity and plasma adiponectin decline induced by prolonged HFD exposure was associated with suppressed ERK1/2 activation in adipose tissue. In adipocytes, specific inhibition of MEK/ERK1/2 pathway decreased intracellular and secretory adiponectin levels, whereas adiponectin gene expression was increased, suggesting that MEK/ERK1/2 inhibition may promote adiponectin protein degradation. Cycloheximide (CHX)-chase assay revealed that MEK/ERK1/2 inhibition accelerated adiponectin protein degradation, which was prevented by MG132, a potent proteasome inhibitor. Immunoprecipitation assay showed that intracellular MEK/ERK1/2 activity was negatively associated with ubiqutinated adiponectin protein levels. Consistently, long-term HFD feeing in mice increased ubiquitinated adiponectin levels in the epididymal fat pads. Conclusions Adipose tissue MEK/ERK1/2 activity can differentially regulates adiponectin gene expression and protein abundance and its suppression in obesity may play a mechanistic role in obesity-related plasma adiponectin decline. PMID:23490586

  2. Acetylsalicylic acid regulates overexpressed small GTPase RhoA in vascular smooth muscle cells through prevention of new synthesis and enhancement of protein degradation.

    Science.gov (United States)

    Li, Dong-Bo; Fu, Zhi-Xuan; Ruan, Shu-Qin; Hu, Shen-Jiang; Li, Xia

    2012-04-01

    RhoA has been shown to play a major role in vascular processes and acetylsalicylic acid (aspirin) is known to exert a cytoprotective effect via multiple mechanisms. In the present study, we aimed at investigating the effect of aspirin on RhoA expression under a stress state in rat VSMCs (vascular smooth muscle cells) and the underlying mechanisms. The expression of iNOS (inducible nitric oxide synthase) and iNOS activity as well as NO concentration was significantly promoted by LPS (lipopolysaccharide) accompanying the elevation of RhoA expression, which was blocked by the addition of the iNOS inhibitor L-NIL [L-N6-(1-iminoethyl)lysine dihydrochloride]. Aspirin (30 μM) significantly attenuated the elevation of RhoA, while indomethacin and salicylate had no similar effect. The sGC (soluble guanylate cyclase) inhibitor ODQ (1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one) showed the same effect as aspirin in down-regulating RhoA but was reversed by the addition of the cGMP analogue 8-Br-PET-cGMP (β-phenyl-1,N2-ethano-8-bromoguanosine 3',5'-cyclic monophosphorothioate). 8-Br-PET-cGMP solely enhanced the RhoA expression that was abrogated by preincubation with aspirin. Degradation analysis indicated that aspirin enhanced the protein degradation rate of RhoA and GDP-bound RhoA seemed to be more susceptible to aspirin-enhanced degradation compared with the GTP-bound form. Our results indicate that aspirin attenuates the LPS-induced overexpression of RhoA both by inhibiting new synthesis and accelerating protein degradation, which may help elucidate the multiple beneficial effects of aspirin.

  3. RISK FACTORS IN NEONATAL ANAEROBIC INFECTIONS

    Directory of Open Access Journals (Sweden)

    M. S. Tabib

    2008-06-01

    Full Text Available Anaerobic bacteria are well known causes of sepsis in adults but there are few studies regarding their role in neonatal sepsis. In an attempt to define the incidence of neonatal anaerobic infections a prospective study was performed during one year period. A total number of 400 neonates under sepsis study were entered this investigation. Anaerobic as well as aerobic cultures were sent. The patients were subjected to comparison in two groups: anaerobic culture positive and anaerobic culture negative and this comparison were analyzed statistically. There were 7 neonates with positive anaerobic culture and 35 neonates with positive aerobic culture. A significant statistical relationship was found between anaerobic infections and abdominal distention and pneumonia. It is recommended for those neonates with abdominal distention and pneumonia refractory to antibiotic treatment to be started on antibiotics with anaerobic coverage.

  4. Potential for anaerobic conversion of xenobiotics

    DEFF Research Database (Denmark)

    Mogensen, Anders Skibsted; Dolfing, J.; Haagensen, Frank

    2003-01-01

    This review covers the latest research on the anaerobic biodegradation of aromatic xenobiotic compounds, with emphasis on surfactants, polycyclic aromatic hydrocarbons, phthalate esters, polychlorinated biphenyls, halogenated phenols, and pesticides. The versatility of anaerobic reactor systems...

  5. Arsenic, Anaerobes, and Astrobiology

    Science.gov (United States)

    Stolz, J. F.; Oremland, R. S.; Switzer Blum, J.; Hoeft, S. E.; Baesman, S. M.; Bennett, S.; Miller, L. G.; Kulp, T. R.; Saltikov, C.

    2013-12-01

    Arsenic is an element best known for its highly poisonous nature, so it is not something one would associate with being a well-spring for life. Yet discoveries made over the past two decades have delineated that not only are some microbes resistant to arsenic, but that this element's primary redox states can be exploited to conserve energy and support prokaryotic growth ('arsenotrophy') in the absence of oxygen. Hence, arsenite [As(III)] can serve as an electron donor for chemo- or photo-autotrophy while arsenate [As(V)] will serve as an electron acceptor for chemo-heterotrophs and chemo-autotrophs. The phylogenetic diversity of these microbes is broad, encompassing many individual species from diverse taxonomic groups in the Domain Bacteria, with fewer representatives in the Domain Archaea. Speculation with regard to the evolutionary origins of the key functional genes in anaerobic arsenic transformations (arrA and arxA) and aerobic oxidation (aioB) has led to a disputation as to which gene and function is the most ancient and whether arsenic metabolism extended back into the Archaean. Regardless of its origin, robust arsenic metabolism has been documented in extreme environments that are rich in their arsenic content, such as hot springs and especially hypersaline soda lakes associated with volcanic regions. Searles Lake, CA is an extreme, salt-saturated end member where vigorous arsenic metabolism occurs, but there is no detectable sulfate-reduction or methanogenesis. The latter processes are too weak bio-energetically to survive as compared with arsenotrophy, and are also highly sensitive to the abundance of borate ions present in these locales. These observations have implications with respect to the search for microbial life elsewhere in the Solar System where volcanic-like processes have been operative. Hence, because of the likelihood of encountering dense brines in the regolith of Mars (formed by evapo-concentration) or beneath the ice layers of Europa

  6. Accelerated anaerobic hydrolysis rates under a combination of intermittent aeration and anaerobic conditions

    DEFF Research Database (Denmark)

    Jensen, T. R.; Lastra Milone, T.; Petersen, G.

    2017-01-01

    Anaerobic hydrolysis in activated return sludge was investigated in laboratory scale experiments to find if intermittent aeration would accelerate anaerobic hydrolysis rates compared to anaerobic hydrolysis rates under strict anaerobic conditions. The intermittent reactors were set up in a 240 h ...

  7. Design of pH sensitive binding proteins from the hyperthermophilic Sso7d scaffold.

    Directory of Open Access Journals (Sweden)

    Nimish Gera

    Full Text Available We have engineered pH sensitive binding proteins for the Fc portion of human immunoglobulin G (hIgG (hFc using two different strategies - histidine scanning and random mutagenesis. We obtained an hFc-binding protein, Sso7d-hFc, through mutagenesis of the Sso7d protein from the hyperthermophilic archaeon Sulfolobus solfataricus; Sso7d-hFc was isolated from a combinatorial library of Sso7d mutants using yeast surface display. Subsequently, we identified a pH sensitive mutant, Sso7d-his-hFc, through systematic evaluation of Sso7d-hFc mutants containing single histidine substitutions. In parallel, we also developed a yeast display screening strategy to isolate a different pH sensitive hFc binder, Sso7d-ev-hFc, from a library of mutants obtained by random mutagenesis of a pool of hFc binders. In contrast to Sso7d-hFc, both Sso7d-his-hFc and Sso7d-ev-hFc have a higher binding affinity for hFc at pH 7.4 than at pH 4.5. The Sso7d-mutant hFc binders can be recombinantly expressed at high yield in E. coli and are monomeric in solution. They bind an epitope in the CH3 domain of hFc that has high sequence homology in all four hIgG isotypes (hIgG(1-4, and recognize hIgG(1-4 as well as deglycosylated hIgG in western blotting assays. pH sensitive hFc binders are attractive candidates for use in chromatography, to achieve elution of IgG under milder pH conditions. However, the surface density of immobilized hFc binders, as well as the avidity effect arising from the multivalent interaction of dimeric hFc with the capture surface, influences the pH dependence of dissociation from the capture surface. Therefore, further studies are needed to evaluate if the Sso7d mutants identified in this study are indeed useful as affinity ligands in chromatography.

  8. The phenomenon of granulation of anaerobic sludge.

    NARCIS (Netherlands)

    Hulshoff Pol, L.W.

    1989-01-01

    Successful high-rate anaerobic wastewater treatment can only be accomplished when the slowgrowing anaerobic biomass is efficiently held back in the anaerobic treatment system. This biomass retention can be achieved in various ways including immobilization of the organisms on fixed materials and immo

  9. 21 CFR 866.2120 - Anaerobic chamber.

    Science.gov (United States)

    2010-04-01

    ... and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Microbiology Devices § 866.2120 Anaerobic chamber. (a) Identification. An anaerobic chamber is a device intended for medical purposes to maintain an anaerobic...

  10. The phenomenon of granulation of anaerobic sludge

    NARCIS (Netherlands)

    Hulshoff Pol, L.

    1989-01-01

    Successful high-rate anaerobic wastewater treatment can only be accomplished when the slowgrowing anaerobic biomass is efficiently held back in the anaerobic treatment system. This biomass retention can be achieved in various ways including immobilization of the organisms on fixed materials

  11. Kinetics and modeling of anaerobic digestion process

    DEFF Research Database (Denmark)

    2003-01-01

    Anaerobic digestion modeling started in the early 1970s when the need for design and efficient operation of anaerobic systems became evident. At that time not only was the knowledge about the complex process of anaerobic digestion inadequate but also there were computational limitations. Thus...

  12. Perspectives of Anaerobic Soil Disinfestation

    NARCIS (Netherlands)

    Lamers, J.G.; Runia, W.T.; Molendijk, L.P.G.; Bleeker, P.O.

    2010-01-01

    Biological soil disinfestation is an environmentally friendly method to disinfest soil. From now on we refer to it as anaerobic soil disinfestation (ASD). With ASD a green manure crop (40 t/ha) is homogeneously incorporated into the topsoil (0-30 cm) after which the field is lightly compacted and ir

  13. Perspectives of Anaerobic Soil Disinfestation

    NARCIS (Netherlands)

    Lamers, J.G.; Runia, W.T.; Molendijk, L.P.G.; Bleeker, P.O.

    2010-01-01

    Biological soil disinfestation is an environmentally friendly method to disinfest soil. From now on we refer to it as anaerobic soil disinfestation (ASD). With ASD a green manure crop (40 t/ha) is homogeneously incorporated into the topsoil (0-30 cm) after which the field is lightly compacted and ir

  14. Perspectives of Anaerobic Soil Disinfestation

    NARCIS (Netherlands)

    Lamers, J.G.; Runia, W.T.; Molendijk, L.P.G.; Bleeker, P.O.

    2010-01-01

    Biological soil disinfestation is an environmentally friendly method to disinfest soil. From now on we refer to it as anaerobic soil disinfestation (ASD). With ASD a green manure crop (40 t/ha) is homogeneously incorporated into the topsoil (0-30 cm) after which the field is lightly compacted and

  15. Anaerobic granular sludge and biofilm reactors

    DEFF Research Database (Denmark)

    Skiadas, Ioannis V.; Gavala, Hariklia N.; Schmidt, Jens Ejbye

    2003-01-01

    by the immobilization of the biomass, which forms static biofilms, particle-supported biofilms, or granules depending on the reactor's operational conditions. The advantages of the high-rate anaerobic digestion over the conventional aerobic wastewater treatment methods has created a clear trend for the change......-rate anaerobic treatment systems based on anaerobic granular sludge and biofilm are described in this chapter. Emphasis is given to a) the Up-flow Anaerobic Sludge Blanket (UASB) systems, b) the main characteristics of the anaerobic granular sludge, and c) the factors that control the granulation process...

  16. Complete genome sequence of the hyperthermophilic methanogen Methanocaldococcus bathoardescens JH146(T) isolated from the basalt subseafloor.

    Science.gov (United States)

    Kim, You-Tae; Jung, Jong-Hyun; Stewart, Lucy C; Kwon, Soon-Wo; Holden, James F; Park, Cheon-Seok

    2015-12-01

    Methanocaldococcus bathoardescens JH146(T) is a hyperthermophilic and obligate hydrogenotrophic methanogen isolated from low-temperature (26 °C) hydrothermal vent fluid at Axial Seamount in the northeastern Pacific Ocean. It is most closely related to the N2-fixing methanogen Methanocaldococcus sp. FS406-22; however, they differ in that JH146 cannot fix N2 or reductively assimilate nitrate. In this study, we present the complete genome sequence of strain JH146(T) (1,607,556 bp) with its 1635 protein coding genes, and 41 RNA genes. Our analysis focuses on its methane production via the acetyl-CoA pathway and its deleted gene clusters related to nitrogen assimilation. This study extends our understanding of methanogenesis at high temperatures and the impact of these organisms on the biogeochemistry of subseafloor hydrothermal environments and the deep sea.

  17. Crystallization and preliminary X-ray diffraction analysis of L-threonine dehydrogenase (TDH) from the hyperthermophilic archaeon Thermococcus kodakaraensis.

    Science.gov (United States)

    Bowyer, A; Mikolajek, H; Wright, J N; Coker, A; Erskine, P T; Cooper, J B; Bashir, Q; Rashid, N; Jamil, F; Akhtar, M

    2008-09-01

    The enzyme L-threonine dehydrogenase catalyses the NAD(+)-dependent conversion of L-threonine to 2-amino-3-ketobutyrate, which is the first reaction of a two-step biochemical pathway involved in the metabolism of threonine to glycine. Here, the crystallization and preliminary crystallographic analysis of L-threonine dehydrogenase (Tk-TDH) from the hyperthermophilic organism Thermococcus kodakaraensis KOD1 is reported. This threonine dehydrogenase consists of 350 amino acids, with a molecular weight of 38 kDa, and was prepared using an Escherichia coli expression system. The purified native protein was crystallized using the hanging-drop vapour-diffusion method and crystals grew in the tetragonal space group P4(3)2(1)2, with unit-cell parameters a = b = 124.5, c = 271.1 A. Diffraction data were collected to 2.6 A resolution and preliminary analysis indicates that there are four molecules in the asymmetric unit of the crystal.

  18. A SEL1L mutation links a canine progressive early-onset cerebellar ataxia to the endoplasmic reticulum-associated protein degradation (ERAD machinery.

    Directory of Open Access Journals (Sweden)

    Kaisa Kyöstilä

    Full Text Available Inherited ataxias are characterized by degeneration of the cerebellar structures, which results in progressive motor incoordination. Hereditary ataxias occur in many species, including humans and dogs. Several mutations have been found in humans, but the genetic background has remained elusive in dogs. The Finnish Hound suffers from an early-onset progressive cerebellar ataxia. We have performed clinical, pathological, and genetic studies to describe the disease phenotype and to identify its genetic cause. Neurological examinations on ten affected dogs revealed rapidly progressing generalized cerebellar ataxia, tremors, and failure to thrive. Clinical signs were present by the age of 3 months, and cerebellar shrinkage was detectable through MRI. Pathological and histological examinations indicated cerebellum-restricted neurodegeneration. Marked loss of Purkinje cells was detected in the cerebellar cortex with secondary changes in other cortical layers. A genome-wide association study in a cohort of 31 dogs mapped the ataxia gene to a 1.5 Mb locus on canine chromosome 8 (p(raw = 1.1x10(-7, p(genome = 7.5x10(-4. Sequencing of a functional candidate gene, sel-1 suppressor of lin-12-like (SEL1L, revealed a homozygous missense mutation, c.1972T>C; p.Ser658Pro, in a highly conserved protein domain. The mutation segregated fully in the recessive pedigree, and a 10% carrier frequency was indicated in a population cohort. SEL1L is a component of the endoplasmic reticulum (ER-associated protein degradation (ERAD machinery and has not been previously associated to inherited ataxias. Dysfunctional protein degradation is known to cause ER stress, and we found a significant increase in expression of nine ER stress responsive genes in the cerebellar cortex of affected dogs, supporting the pathogenicity of the mutation. Our study describes the first early-onset neurodegenerative ataxia mutation in dogs, establishes an ERAD-mediated neurodegenerative

  19. Purification and biochemical properties of a cytochrome bc complex from the aerobic hyperthermophilic archaeon Aeropyrum pernix

    OpenAIRE

    Kabashima Yoshiki; Sakamoto Junshi

    2011-01-01

    Abstract Background The bioenergetics of Archaea with respect to the evolution of electron transfer systems is very interesting. In contrast to terminal oxidases, a canonical bc1 complex has not yet been isolated from Archaea. In particular, c-type cytochromes have been reported only for a limited number of species. Results Here, we isolated a c-type cytochrome-containing enzyme complex from the membranes of the hyperthermophilic archaeon, Aeropyrum pernix, grown aerobically. The redox spectr...

  20. E2/ER β Enhances Calcineurin Protein Degradation and PI3K/Akt/MDM2 Signal Transduction to Inhibit ISO-Induced Myocardial Cell Apoptosis

    Directory of Open Access Journals (Sweden)

    Kuan-Ho Lin

    2017-04-01

    Full Text Available Secretion of multifunctional estrogen and its receptor has been widely considered as the reason for markedly higher frequency of heart disease in men than in women. 17β-Estradiol (E2, for instance, has been reported to prevent development of cardiac apoptosis via activation of estrogen receptors (ERs. In addition, protein phosphatase such as protein phosphatase 1 (PP1 and calcineurin (PP2B are also involved in cardiac hypertrophy and cell apoptosis signaling. However, the mechanism by which E2/ERβ suppresses apoptosis is not fully understood, and the role of protein phosphatase in E2/ERβ action also needs further investigation. In this study, we observed that E2/ERβ inhibited isoproterenol (ISO-induced myocardial cell apoptosis, cytochrome c release and downstream apoptotic markers. Moreover, we found that E2/ERβ blocks ISO-induced apoptosis in H9c2 cells through the enhancement of calcineurin protein degradation through PI3K/Akt/MDM2 signaling pathway. Our results suggest that supplementation with estrogen and/or overexpression of estrogen receptor β gene may prove to be effective means to treat stress-induced myocardial damage.

  1. Epidermal Growth Factor Cytoplasmic Domain Affects ErbB Protein Degradation by the Lysosomal and Ubiquitin-Proteasome Pathway in Human Cancer Cells

    Directory of Open Access Journals (Sweden)

    Aleksandra Glogowska

    2012-05-01

    Full Text Available The cytoplasmic domains of EGF-like ligands, including EGF cytoplasmic domain (EGFcyt, have important biological functions. Using specific constructs and peptides of human EGF cytoplasmic domain, we demonstrate that EGFcyt facilitates lysosomal and proteasomal protein degradation, and this coincided with growth inhibition of human thyroid and glioma carcinoma cells. EGFcyt and exon 22–23-encoded peptide (EGF22.23 enhanced procathepsin B (procathB expression and procathB-mediated lysosomal degradation of EGFR/ErbB1 as determined by inhibitors for procathB and the lysosomal ATPase inhibitor BafA1. Presence of mbEGFctF, EGFcyt, EGF22.23, and exon 23-encoded peptides suppressed the expression of the deubiqitinating enzyme ubiquitin C-terminal hydrolase-L1 (UCH-L1. This coincided with hyperubiquitination of total cellular proteins and ErbB1/2 and reduced proteasome activity. Upon small interfering RNA-mediated silencing of endogenously expressed UCH-L1, a similar hyperubiquitinylation phenotype, reduced ErbB1/2 content, and attenuated growth was observed. The exon 23-encoded peptide region of EGFcyt was important for these biologic actions. Structural homology modeling of human EGFcyt showed that this molecular region formed an exposed surface loop. Peptides derived from this EGFcyt loop structure may aid in the design of novel peptide therapeutics aimed at inhibiting growth of cancer cells.

  2. Proteome changes in tomato fruits prior to visible symptoms of chilling injury are linked to defensive mechanisms, uncoupling of photosynthetic processes and protein degradation machinery.

    Science.gov (United States)

    Sanchez-Bel, Paloma; Egea, Isabel; Sanchez-Ballesta, María Teresa; Sevillano, Laura; Del Carmen Bolarin, Maria; Flores, Francisco B

    2012-02-01

    A comparative proteomic analysis between tomato fruits stored at chilling and non-chilling temperatures was carried out just before the appearance of visible symptoms of chilling injury. At this stage of the stress period it was possible to discriminate between proteins involved in symptoms and proteins implicated in response. To investigate the changes in the tomato fruit proteome under this specific stressful condition, two-dimensional differential in-gel electrophoresis coupled with spot identification by mass spectrometry was applied. This proteomic approach allowed the identification of differentially expressed proteins which are involved in two main biological functions: (i) defensive mechanisms represented by small heat shock and late embryogenesis proteins; and (ii) reaction to the uncoupling of photosynthetic processes and the protein degradation machinery. One of the first changes observed in chilled fruits is the down-regulation of ATP synthase, 26S proteasome subunit RPN11 and aspartic proteinase, whereas the first responses in order to deal with the stress are mainly multifunctional proteins involved not only in metabolism but also in stress regulation such as glyceraldehyde phosphate dehydrogenase, 2-oxoglutarate dehydrogenase and invertase. In addition, our data seem to indicate a possible candidate to be used as a protein marker for further studies on cold stress: aldose-1-epimerase, which seems to have an important role in low temperature tolerance.

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

    DEFF Research Database (Denmark)

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

    2010-01-01

    Two novel viral genomes and four plasmids were assembled from an environmental sample collected from a hot spring at Yellowstone National Park, USA, and maintained anaerobically in a bioreactor at 85°C and pH 6. The double-stranded DNA viral genomes are linear (22.7 kb) and circular (17.7 kb...... respectively. Strategies are considered for assembling genomes of smaller genetic elements from complex environmental samples, and for establishing possible host identities on the basis of sequence similarity to host CRISPR immune systems.......), and derive apparently from archaeal viruses HAV1 and HAV2. Genomic DNA was obtained from samples enriched in filamentous and tadpole-shaped virus-like particles respectively. They yielded few significant matches in public sequence databases reinforcing, further, the wide diversity of archaeal viruses...

  4. Speculations on the origin of life and thermophily: Review of available information on reverse gyrase suggests that hyperthermophilic procaryotes are not so primitive

    Science.gov (United States)

    Forterre, Patrick; Confalonier, Fabrice; Charbonnier, Franck; Duguet, Michel

    1995-06-01

    All present-day hyperthermophiles studied so far (eitherBacteria orArchaea) contain a unique DNA topoisomerase, reverse gyrase, which probably helps to stabilize genomic DNA at high temperature. Herein the data relating this enzyme is reviewed and discussed from the perspective of the nature of the last detectable common ancestor and the origin of life. The sequence of the gene encoding reverse gyrase from an archaeon,Sulfolobus acidocaldarius, suggests that this enzyme contains both a helicase and a topoisomerase domains (Confalonieriet al.,Proc. Natl. Acad. Sci., 1993, 90, 4735). Accordingly, it has been proposed that reverse gyrase originated by the fusion of DNA helicase and DNA topoisomerase genes. If reverse gyrase is essential for life at high temperature, its composite structure suggests that DNA helicases and topoisomerases appeared independently and first evolved in a mesophilic world. Such scenario contradicts the hypothesis that a direct link connects present day hyperthermophiles to a hot origin of life. We discuss different patterns for the early cellular evolution in which reverse gyrase appeared either before the emergence of the last common ancestor ofArchaea, Bacteria andEucarya, or in a lineage common to the two procaryotic domains. The latter scenario could explain why all today hyperthermophiles are procaryotes.

  5. Simple and convenient method for culturing anaerobic bacteria.

    OpenAIRE

    Behbehani, M J; Jordan, H. V.; Santoro, D L

    1982-01-01

    A simple and convenient method for culturing anaerobic bacteria is described. Cultures can be grown in commercially available flasks normally used for preparation of sterile external solutions. A special disposable rubber flask closure maintains anaerobic conditions in the flask after autoclaving. Growth of a variety of anaerobic oral bacteria was comparable to that obtained after anaerobic incubation of broth cultures in Brewer Anaerobic Jars.

  6. Molecular ecology of anaerobic reactor systems

    DEFF Research Database (Denmark)

    Hofman-Bang, H. Jacob Peider; Zheng, D.; Westermann, Peter

    2003-01-01

    Anaerobic reactor systems are essential for the treatment of solid and liquid wastes and constitute a core facility in many waste treatment plants. Although much is known about the basic metabolism in different types of anaerobic reactors, little is known about the microbes responsible for these ...... specific nucleic acid probes are discussed and exemplified by studies of anaerobic granular sludge, biofilm and digester systems......Anaerobic reactor systems are essential for the treatment of solid and liquid wastes and constitute a core facility in many waste treatment plants. Although much is known about the basic metabolism in different types of anaerobic reactors, little is known about the microbes responsible...... and malfunctions of anaerobic digesters occasionally experienced, leading to sub-optimal methane production and wastewater treatment. Using a variety of molecular techniques, we are able to determine which microorganisms are active, where they are active, and when they are active, but we still need to determine...

  7. Application of dynamic membranes in anaerobic membranes in anaerobic membrane bioreactor systems

    NARCIS (Netherlands)

    Erşahin, M.E.

    2015-01-01

    Anaerobic membrane bioreactors (AnMBRs) physically ensure biomass retention by the application of a membrane filtration process. With growing application experiences from aerobic membrane bioreactors (MBRs), the combination of membrane and anaerobic processes has received much attention and become

  8. ASSAYING OF AUTOPHAGIC PROTEIN DEGRADATION

    NARCIS (Netherlands)

    C. Bauvy; A.J. Meijer; P. Codogno

    2009-01-01

    Macroautophagy is a three-step process: (1) autophagosomes form and mature, (2) the autophagosomes fuse with lysosomes, and (3) the autophagic cargo is degraded in the lysosomes. It is this lysosomal degradation of the autophagic cargo that constitutes the autophagic flux. As in the case of metaboli

  9. Evaluation of a prereduced anaerobically sterilized medium (PRAS II) system for identification anaerobic microorganisms.

    Science.gov (United States)

    Beaucage, C M; Onderdonk, A B

    1982-09-01

    A prereduced, anaerobically sterilized system of tubed media (PRAS II; Scott Laboratories, Fiskeville, R.I.) was evaluated for accuracy in the identification of anerobic microorganisms. PRAS II was found to be a rapid and accurate identification system for obligate anaerobes which does not require the use of gas cannula inoculation or incubation in a special anaerobic environment.

  10. Evaluation of a prereduced anaerobically sterilized medium (PRAS II) system for identification anaerobic microorganisms.

    OpenAIRE

    Beaucage, C M; Onderdonk, A B

    1982-01-01

    A prereduced, anaerobically sterilized system of tubed media (PRAS II; Scott Laboratories, Fiskeville, R.I.) was evaluated for accuracy in the identification of anerobic microorganisms. PRAS II was found to be a rapid and accurate identification system for obligate anaerobes which does not require the use of gas cannula inoculation or incubation in a special anaerobic environment.

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

    Directory of Open Access Journals (Sweden)

    Sergey N Gavrilov

    2016-05-01

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

  12. Disulfiram/copper-disulfiram Damages Multiple Protein Degradation and Turnover Pathways and Cytotoxicity is Enhanced by Metformin in Oesophageal Squamous Cell Carcinoma Cell Lines.

    Science.gov (United States)

    Jivan, Rupal; Damelin, Leonard Howard; Birkhead, Monica; Rousseau, Amanda Louise; Veale, Robin Bruce; Mavri-Damelin, Demetra

    2015-10-01

    Disulfiram (DSF), used since the 1950s in the treatment of alcoholism, is reductively activated to diethyldithiocarbamate and both compounds are thiol-reactive and readily complex copper. More recently DSF and copper-DSF (Cu-DSF) have been found to exhibit potent anticancer activity. We have previously shown that the anti-diabetic drug metformin is anti-proliferative and induces an intracellular reducing environment in oesophageal squamous cell carcinoma (OSCC) cell lines. Based on these observations, we investigated the effects of Cu-DSF and DSF, with and without metformin, in this present study. We found that Cu-DSF and DSF caused considerable cytotoxicity across a panel of OSCC cells, and metformin significantly enhanced the effects of DSF. Elevated copper transport contributes to DSF and metformin-DSF-induced cytotoxicity since the cell-impermeable copper chelator, bathocuproinedisulfonic acid, partially reversed the cytotoxic effects of these drugs, and interestingly, metformin-treated OSCC cells contained higher intracellular copper levels. Furthermore, DSF may target cancer cells preferentially due to their high dependence on protein degradation/turnover pathways, and we found that metformin further enhances the role of DSF as a proteasome inhibitor. We hypothesized that the lysosome could be an additional, novel, target of DSF. Indeed, this acid-labile compound decreased lysosomal acidification, and DSF-metformin co-treatment interfered with the progression of autophagy in these cells. In summary, this is the first such report identifying the lysosome as a target of DSF and based on the considerable cytotoxic effects of DSF either alone or in the presence of metformin, in vitro, and we propose these as novel potential chemotherapeutic approaches for OSCC.

  13. Insect peptide CopA3-induced protein degradation of p27Kip1 stimulates proliferation and protects neuronal cells from apoptosis

    Energy Technology Data Exchange (ETDEWEB)

    Nam, Seung Taek; Kim, Dae Hong; Lee, Min Bum; Nam, Hyo Jung; Kang, Jin Ku; Park, Mi Jung; Lee, Ik Hwan [Department of Life Science, College of Natural Science, Daejin University, Pocheon, Gyeonggido 487-711 (Korea, Republic of); Seok, Heon [Department of Biomedical Science, Jungwon University, Goesan, Chungcheongbukdo 367-700 (Korea, Republic of); Lee, Dong Gun [School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University, Daegu 702-701 (Korea, Republic of); Hwang, Jae Sam [Department of Agricultural Biology, National Academy of Agricultural Science, RDA, Suwon 441-707 (Korea, Republic of); Kim, Ho, E-mail: hokim@daejin.ac.kr [Department of Life Science, College of Natural Science, Daejin University, Pocheon, Gyeonggido 487-711 (Korea, Republic of)

    2013-07-19

    Highlights: •CopA3 peptide isolated from the Korean dung beetle has antimicrobial activity. •Our study reported that CopA3 has anticancer and immunosuppressive effects. •We here demonstrated that CopA3 has neurotropic and neuroprotective effects. •CopA3 degrades p27Kip1 protein and this mediates effects of CopA3 on neuronal cells. -- Abstract: We recently demonstrated that the antibacterial peptide, CopA3 (a D-type disulfide dimer peptide, LLCIALRKK), inhibits LPS-induced macrophage activation and also has anticancer activity in leukemia cells. Here, we examined whether CopA3 could affect neuronal cell proliferation. We found that CopA3 time-dependently increased cell proliferation by up to 31 ± 2% in human neuroblastoma SH-SY5Y cells, and up to 29 ± 2% in neural stem cells isolated from neonatal mouse brains. In both cell types, CopA3 also significantly inhibited the apoptosis and viability losses caused by 6-hydroxy dopamine (a Parkinson disease-mimicking agent) and okadaic acid (an Alzheimer’s disease-mimicking agent). Immunoblotting revealed that the p27Kip1 protein (a negative regulator of cell cycle progression) was markedly degraded in CopA3-treated SH-SY5Y cells. Conversely, an adenovirus expressing p27Kip1 significantly inhibited the antiapoptotic effects of CopA3 against 6-hydroxy dopamine- and okadaic acid-induced apoptosis, and decreased the neurotropic effects of CopA3. These results collectively suggest that CopA3-mediated protein degradation of p27Kip1 may be the main mechanism through which CopA3 exerts neuroprotective and neurotropic effects.

  14. In vitro determination of ruminal protein degradability of alfalfa and prairie hay via a commercial protease in the presence or absence of cellulase or driselase.

    Science.gov (United States)

    Abdelgadir, I E; Cochran, R C; Titgemeyer, E C; Vanzant, E S

    1997-08-01

    Ruminal protein degradation of alfalfa (2.62% N, 49.6% NDF, and in vivo undegradable intake protein [UIP] = 16.4% of CP) and prairie hay (.88% N, 69.4% NDF, and in vivo UIP = 44.5% of CP) was estimated using the Streptomyces griseus protease (SGP) in vitro method with or without pretreatment with two carbohydrases: cellulase from Penicillium funiculosum or driselase from Basidiomycetes. Driselase is a broad-spectrum carbohydrase. Incubating forage samples for 48 h with cellulase or driselase at a concentration of 800 mg/g per g of hay nearly maximized ADF and NDF disappearances. This concentration and incubation time then were used to pretreat hay samples. A 2-h pretreatment was included to evaluate the potential for reducing the analysis time. Other sets of samples were or were not pretreated with acetate buffer alone. Following pretreatment, samples were subjected to SGP for .25, .5, 1, 2, 4, 8, 12, 24, and 48 h. Pretreatment altered the sizes of protein pools and their degradation rates. When the UIP contents of the forages were estimated using SGP and a single-pool, first-order, kinetic model, cellulase (48 h) or driselase pretreatments yielded UIP predictions that were more similar to in vivo values. Some carbohydrase and protease combinations also yielded single time-point estimates of UIP that were similar to in vivo values. Similarly, when sufficient time was permitted for protease incubation, single time-point estimates derived from protease alone were similar to in vivo values.

  15. Overexpression of alpha-synuclein at non-toxic levels increases dopaminergic cell death induced by copper exposure via modulation of protein degradation pathways.

    Science.gov (United States)

    Anandhan, Annadurai; Rodriguez-Rocha, Humberto; Bohovych, Iryna; Griggs, Amy M; Zavala-Flores, Laura; Reyes-Reyes, Elsa M; Seravalli, Javier; Stanciu, Lia A; Lee, Jaekwon; Rochet, Jean-Christophe; Khalimonchuk, Oleh; Franco, Rodrigo

    2015-09-01

    Gene multiplications or point mutations in alpha (α)-synuclein are associated with familial and sporadic Parkinson's disease (PD). An increase in copper (Cu) levels has been reported in the cerebrospinal fluid and blood of PD patients, while occupational exposure to Cu has been suggested to augment the risk to develop PD. We aimed to elucidate the mechanisms by which α-synuclein and Cu regulate dopaminergic cell death. Short-term overexpression of wild type (WT) or mutant A53T α-synuclein had no toxic effect in human dopaminergic cells and primary midbrain cultures, but it exerted a synergistic effect on Cu-induced cell death. Cell death induced by Cu was potentiated by overexpression of the Cu transporter protein 1 (Ctr1) and depletion of intracellular glutathione (GSH) indicating that the toxic effects of Cu are linked to alterations in its intracellular homeostasis. Using the redox sensor roGFP, we demonstrated that Cu-induced oxidative stress was primarily localized in the cytosol and not in the mitochondria. However, α-synuclein overexpression had no effect on Cu-induced oxidative stress. WT or A53T α-synuclein overexpression exacerbated Cu toxicity in dopaminergic and yeast cells in the absence of α-synuclein aggregation. Cu increased autophagic flux and protein ubiquitination. Impairment of autophagy by overexpression of a dominant negative Atg5 form or inhibition of the ubiquitin/proteasome system (UPS) with MG132 enhanced Cu-induced cell death. However, only inhibition of the UPS stimulated the synergistic toxic effects of Cu and α-synuclein overexpression. Our results demonstrate that α-synuclein stimulates Cu toxicity in dopaminergic cells independent from its aggregation via modulation of protein degradation pathways.

  16. ANAEROBIC BIODEGRADATION OF A BIODEGRADABLE MATERIAL UNDER ANAEROBIC - THERMOPHILIC DIGESTION

    Directory of Open Access Journals (Sweden)

    RICARDO CAMACHO-MUÑOZ

    2014-12-01

    Full Text Available This paper dertermined the anaerobic biodegradation of a polymer obtained by extrusion process of native cassava starch, polylactic acid and polycaprolactone. Initially a thermophilic - methanogenic inoculum was prepared from urban solid waste. The gas final methane concentration and medium’s pH reached values of 59,6% and 7,89 respectively. The assay assembly was carried out according ASTM D5511 standard. The biodegradation percent of used materials after 15 day of digestion were: 77,49%, 61,27%, 0,31% for cellulose, sample and polyethylene respectively. Due cellulose showed biodegradation levels higher than 70% it’s deduced that the inoculum conditions were appropriate. A biodegradation level of 61,27%, 59,35% of methane concentration in sample’s evolved gas and a medium’s finale pH of 7,71 in sample’s vessels, reveal the extruded polymer´s capacity to be anaerobically degraded under thermophilic- high solid concentration conditions.

  17. Integrated anaerobic and aerobic treatment of sewage

    NARCIS (Netherlands)

    Wang, K.

    1994-01-01

    This thesis describes results of investigations dealing with sequential concept of anaerobic-aerobic treatment of municipal wastewater. The main purposes of the study were 1) to develop a proper anaerobic hydrolytic pretreatment unit, consisting of a Hydrolysis Upflow Sludge Bed (HUSB-) rea

  18. Integrated anaerobic and aerobic treatment of sewage.

    NARCIS (Netherlands)

    Kaijun Wang,

    1994-01-01

    This thesis describes results of investigations dealing with sequential concept of anaerobic-aerobic treatment of municipal wastewater. The main purposes of the study were 1) to develop a proper anaerobic hydrolytic pretreatment unit, consisting of a Hydrolysis Upflow Sludge Bed (HUSB-) reactor and

  19. Anaerobic critical velocity in four swimming techniques.

    Science.gov (United States)

    Neiva, H P; Fernandes, R J; Vilas-Boas, J P

    2011-03-01

    The aim of this study was to assess critical velocity in order to control and evaluate anaerobic swimming training. 51 highly trained male swimmers performed maximal 15, 25, 37.5 and 50 m in the 4 swimming techniques to determine critical velocity from the distance-time relationship. Anaerobic critical velocity was compared with 100 m swimming performance and corresponding partials. Complementarily, 9 swimmers performed a 6×50 m (4 min interval) training series at front crawl individual anaerobic critical velocity, capillary blood lactate concentrations being assessed after each repetition. The mean±SD values of anaerobic critical velocity and its relationship with the 100 m event were: 1.61±0.07 (r=0.60, p=0.037), 1.53±0.05 (r=0.81, p=0.015), 1.33±0.05 (r=0.83, p=0.002), and 1.75±0.05 (r=0.74, p=0.001), for butterfly, backstroke, breaststroke and front crawl, respectively. However, differences between anaerobic critical velocity and performance were observed (with exception of the second half of the 100 m swimming events in breaststroke and butterfly). Lactate concentration values at the end of the series were 14.52±1.06 mmol.l (-1), which suggests that it was indeed an anaerobic training set. In this sense, anaerobic critical velocity can be used to prescribe anaerobic training intensities.

  20. Method for anaerobic fermentation and biogas production

    DEFF Research Database (Denmark)

    2013-01-01

    The present invention relates to a method for biomass processing, anaerobic fermentation of the processed biomass, and the production biogas. In particular, the invention relates to a system and method for generating biogas from anaerobic fermentation of processed organic material that comprises...

  1. Carbon monoxide conversion by anaerobic bioreactor sludges

    NARCIS (Netherlands)

    Sipma, J.; Stams, A.J.M.; Lens, P.N.L.; Lettinga, G.

    2003-01-01

    Seven different anaerobic sludges from wastewater treatment reactors were screened for their ability to convert carbon monoxide (CO) at 30 and 55degreesC
    Seven different anaerobic sludges from wastewater treatment reactors were screened for their ability to convert carbon monoxide (CO) at 30 and

  2. Integrated anaerobic and aerobic treatment of sewage

    NARCIS (Netherlands)

    Wang, K.

    1994-01-01

    This thesis describes results of investigations dealing with sequential concept of anaerobic-aerobic treatment of municipal wastewater. The main purposes of the study were 1) to develop a proper anaerobic hydrolytic pretreatment unit, consisting of a Hydrolysis Upflow Sludge Bed (HUSB-)

  3. Anaerobic degradation of linear alkylbenzene sulfonate

    DEFF Research Database (Denmark)

    Mogensen, Anders Skibsted; Haagensen, Frank; Ahring, Birgitte Kiær

    2003-01-01

    increases during anaerobic stabilization due to transformation of easily degradable organic matter. Hence, LAS is regarded as resistant to biodegradation under anaerobic conditions. We present data from a lab-scale semi-continuously stirred tank reactor (CSTR) spiked with linear dodecylbenzene sulfonate (C...

  4. Prospects of Anaerobic Digestion Technology in China

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    As the world's largest developing country, China must face the problem of managing municipal solid waste, and the challenge of organic waste disposal is even more serious. Considering the characteristics of traditional waste disposal technologies and the subsequent secondary pollution, anaerobic digestion has various advantages such as reduction in the land needed for disposal and preservation of environmental quality. In light of the energy crisis, this paper focuses on the potential production of biogas from biowaste through anaerobic digestion processes, the problems incurred by the waste collection system, and the efficiency of the anaerobic digestion process. Use of biogas in a combined heat and power cogeneration system is also discussed. Finally, the advantages of anaerobic digestion technology for the Chinese market are summarized. The anaerobic digestion is suggested to be a promising treating technology for the organic wastes in China.

  5. Anaerobic biorefinery: Current status, challenges, and opportunities.

    Science.gov (United States)

    Sawatdeenarunat, Chayanon; Nguyen, Duc; Surendra, K C; Shrestha, Shilva; Rajendran, Karthik; Oechsner, Hans; Xie, Li; Khanal, Samir Kumar

    2016-09-01

    Anaerobic digestion (AD) has been in use for many decades. To date, it has been primarily aimed at treating organic wastes, mainly manures and wastewater sludge, and industrial wastewaters. However, with the current advancements, a more open mind is required to look beyond these somewhat restricted original applications of AD. Biorefineries are such concepts, where multiple products including chemicals, fuels, polymers etc. are produced from organic feedstocks. The anaerobic biorefinery concept is now gaining increased attention, utilizing AD as the final disposal step. This review aims at evaluating the potential significance of anaerobic biorefineries, including types of feedstocks, uses for the produced energy, as well as sustainable applications of the generated residual digestate. A comprehensive analysis of various types of anaerobic biorefineries has been developed, including both large-scale and household level applications. Finally, future directives are highlighted showing how anaerobic biorefinery concept could impact the bioeconomy in the near future. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Toxicants inhibiting anaerobic digestion: a review.

    Science.gov (United States)

    Chen, Jian Lin; Ortiz, Raphael; Steele, Terry W J; Stuckey, David C

    2014-12-01

    Anaerobic digestion is increasingly being used to treat wastes from many sources because of its manifold advantages over aerobic treatment, e.g. low sludge production and low energy requirements. However, anaerobic digestion is sensitive to toxicants, and a wide range of compounds can inhibit the process and cause upset or failure. Substantial research has been carried out over the years to identify specific inhibitors/toxicants, and their mechanism of toxicity in anaerobic digestion. In this review we present a detailed and critical summary of research on the inhibition of anaerobic processes by specific organic toxicants (e.g., chlorophenols, halogenated aliphatics and long chain fatty acids), inorganic toxicants (e.g., ammonia, sulfide and heavy metals) and in particular, nanomaterials, focusing on the mechanism of their inhibition/toxicity. A better understanding of the fundamental mechanisms behind inhibition/toxicity will enhance the wider application of anaerobic digestion.

  7. Homology modelling of two subtilisin-like proteases from the hyperthermophilic archaea Pyrococcus furiosus and Thermococcus stetteri.

    Science.gov (United States)

    Voorhorst, W G; Warner, A; de Vos, W M; Siezen, R J

    1997-08-01

    The hyperthermophilic archaeon Pyrococcus furiosus produces an extracellular, glycosylated hyperthermostable subtilisin-like serine protease, termed pyrolysin (Voorhorst,W.G.B., Eggen,R.I.L., Geerling,A.C.M., Platteeuw,C., Siezen,R.J. and de Vos,W.M. (1996) J. Biol. Chem., 271, 20426-20431). Based on the pyrolysin coding sequence, a pyrolysin-like gene fragment was cloned and characterized from the extreme thermophilic archaeon Thermococcus stetteri. Like pyrolysin, the deduced sequence of this serine protease, designated stetterlysin, contains a catalytic domain with high homology with other subtilases, allowing homology modelling starting from known crystal structures. Comparison of the predicted three-dimensional models of the catalytic domain of stetterlysin and pyrolysin with the crystal structure of subtilases from mesophilic and thermophilic origin, i.e. subtilisin BPN' and thermitase, and the homology model of subtilisin S41 from psychrophilic origin, led to the identification of features that could be related to protein stabilization. Higher thermostability was found to be correlated with an increased number of residues involved in pairs and networks of charge-charge and aromatic-aromatic interactions. These highly thermostable proteases have several extra surface loops and inserts with a relatively high frequency of aromatic residues and Asn residues. The latter are often present in putative N-glycosylation sites. Results from modelling of known substrates in the substrate-binding region support the broad substrate range and the autocatalytic activation previously suggested for pyrolysin.

  8. Surface-exposed glycoproteins of hyperthermophilic Sulfolobus solfataricus P2 show a common N-glycosylation profile.

    Science.gov (United States)

    Palmieri, Gianna; Balestrieri, Marco; Peter-Katalinić, Jasna; Pohlentz, Gottfried; Rossi, Mosè; Fiume, Immacolata; Pocsfalvi, Gabriella

    2013-06-07

    Cell surface proteins of hyperthermophilic Archaea actively participate in intercellular communication, cellular uptake, and energy conversion to sustain survival strategies in extreme habitats. Surface (S)-layer glycoproteins, the major component of the S-layers in many archaeal species and the best-characterized prokaryotic glycoproteins, were shown to have a large structural diversity in their glycan compositions. In spite of this, knowledge on glycosylation of proteins other than S-layer proteins in Archaea is quite limited. Here, the N-glycosylation pattern of cell-surface-exposed proteins of Sulfolobus solfataricus P2 were analyzed by lectin affinity purification, HPAEC-PAD, and multiple mass spectrometry-based techniques. Detailed analysis of SSO1273, one of the most abundant ABC transporters present in the cell surface fraction of S. solfataricus, revealed a novel glycan structure composed of a branched sulfated heptasaccharide, Hex4(GlcNAc)2 plus sulfoquinovose where Hex is d-mannose and d-glucose. Having one monosaccharide unit more than the glycan of the S-layer glycoprotein of S. acidocaldarius, this is the most complex archaeal glycan structure known today. SSO1273 protein is heavily glycosylated and all 20 theoretical N-X-S/T (where X is any amino acid except proline) consensus sequence sites were confirmed. Remarkably, we show that several other proteins in the surface fraction of S. solfataricus are N-glycosylated by the same sulfated oligosaccharide and we identified 56 N-glycosylation sites in this subproteome.

  9. N-Linked Glycans Are Assembled on Highly Reduced Dolichol Phosphate Carriers in the Hyperthermophilic Archaea Pyrococcus furiosus.

    Science.gov (United States)

    Chang, Michelle M; Imperiali, Barbara; Eichler, Jerry; Guan, Ziqiang

    2015-01-01

    In all three domains of life, N-glycosylation begins with the assembly of glycans on phosphorylated polyisoprenoid carriers. Like eukaryotes, archaea also utilize phosphorylated dolichol for this role, yet whereas the assembled oligosaccharide is transferred to target proteins from dolichol pyrophosphate in eukaryotes, archaeal N-linked glycans characterized to date are derived from a dolichol monophosphate carrier, apart from a single example. In this study, glycan-charged dolichol phosphate from the hyperthermophile Pyrococcus furiosus was identified and structurally characterized. Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry revealed the existence of dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species. As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. Interestingly, an additional 1-4 of the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate are saturated, making this lipid not only the longest archaeal dolichol phosphate described to date but also the most highly saturated.

  10. N-Linked Glycans Are Assembled on Highly Reduced Dolichol Phosphate Carriers in the Hyperthermophilic Archaea Pyrococcus furiosus.

    Directory of Open Access Journals (Sweden)

    Michelle M Chang

    Full Text Available In all three domains of life, N-glycosylation begins with the assembly of glycans on phosphorylated polyisoprenoid carriers. Like eukaryotes, archaea also utilize phosphorylated dolichol for this role, yet whereas the assembled oligosaccharide is transferred to target proteins from dolichol pyrophosphate in eukaryotes, archaeal N-linked glycans characterized to date are derived from a dolichol monophosphate carrier, apart from a single example. In this study, glycan-charged dolichol phosphate from the hyperthermophile Pyrococcus furiosus was identified and structurally characterized. Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry revealed the existence of dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species. As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. Interestingly, an additional 1-4 of the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate are saturated, making this lipid not only the longest archaeal dolichol phosphate described to date but also the most highly saturated.

  11. Characterization of Di-myo-inositol-1,1{prime}-phosphate in the hyperthermophilic bacterium Thermotoga maritima

    Energy Technology Data Exchange (ETDEWEB)

    Ramakrishnan, V.; Verhagen, M.F.J.M.; Adams, M.W.W. [Univ. of Georgia, Athens, GA (United States)

    1997-01-01

    Di-myo-inositol-1,1{prime}-phosphate (DIP) is present at a significant concentration ({approximately}160 nmol/mg of protein) in the cytoplasm of the hyperthermophilic bacterium Thermotoga maritima. The concentrations of DIP was independent of the pH of the growth medium or the cell growth phase but increased with increasing concentrations of NaCl in the growth medium, reaching a maximum ({approximately}450 nmol/mg of protein) at 0.4 to 0.6 M NaCl. A large-scale purification procedure for DIP which yields approximately 18 g of DIP per kg of cells (wet weight) is described. Purified DIP was stable at 90{degrees}C for at least 5 h. The presence of DIP(50 mM) did not increase the stability at 90{degrees}C of pure forms of the hydrogenase or pyruvate ferredoxin oxidoreductase of T. maritima, suggesting that DIP is not a general thermoprotectant. 25 refs., 4 figs.

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

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    Alexia Comte

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

  13. Broad nucleotide cofactor specificity of DNA ligase from the hyperthermophilic crenarchaeon Hyperthermus butylicus and its evolutionary significance.

    Science.gov (United States)

    Kim, Jun-Hwan; Lee, Kang-Keun; Sun, Younguk; Seo, Gang-Jin; Cho, Sung Suk; Kwon, Suk Hyung; Kwon, Suk-Tae

    2013-05-01

    The nucleotide cofactor specificity of the DNA ligase from the hyperthermophilic crenarchaeon Hyperthermus butylicus (Hbu) was studied to investigate the evolutionary relationship of DNA ligases. The Hbu DNA ligase gene was expressed under control of the T7lac promoter of pTARG in Escherichia coli BL21-CodonPlus(DE3)-RIL. The expressed enzyme was purified using the IMPACT™-CN system (intein-mediated purification with an affinity chitin-binding tag) and cation-ion (Arg-tag) chromatography. The optimal temperature for Hbu DNA ligase activity was 75 °C, and the optimal pH was 8.0 in Tris-HCl. The activity was highly dependent on MgCl2 or MnCl2 with maximal activity above 5 mM MgCl2 and 2 mM MnCl2. Notably, Hbu DNA ligase can use ADP and GTP in addition to ATP. The broad nucleotide cofactor specificity of Hbu DNA ligase might exemplify an undifferentiated ancestral stage in the evolution of DNA ligases. This study provides new evidence for possible evolutionary relationships among DNA ligases.

  14. Unusual starch degradation pathway via cyclodextrins in the hyperthermophilic sulfate-reducing archaeon Archaeoglobus fulgidus strain 7324.

    Science.gov (United States)

    Labes, Antje; Schönheit, Peter

    2007-12-01

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

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

    Directory of Open Access Journals (Sweden)

    Roberta V. Branco

    2015-01-01

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

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

    Science.gov (United States)

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

    2013-06-01

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

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

    Science.gov (United States)

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

    2013-03-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2013-01-24

    A novel sulfate-reducing bacterium designated OPF15T was isolated from Obsidian Pool, Yellowstone National Park, Wyoming. The phylogeny of 16S rRNA and functional genes (dsrAB) placed the organism within the family Thermodesulfobacteriaceae. The organism displayed hyperthermophilic temperature requirements for growth with a range of 70 90 C and an optimum of 83 C. Optimal pH was around 6.5 7.0 and the organism required the presence of H2 or formate as an electron donor and CO2 as a carbon source. Electron acceptors supporting growth included sulfate, thiosulfate, and elemental sulfur. Lactate, acetate, pyruvate, benzoate, oleic acid, and ethanol did not serve as electron donors. Membrane lipid analysis revealed diacyl glycerols and acyl/ether glycerols which ranged from C14:0 to C20:0. Alkyl chains present in acyl/ether and diether glycerol lipids ranged from C16:0 to C18:0. Straight, iso- and anteiso-configurations were found for all lipid types. The presence of OPF15T was also shown to increase cellulose consumption during co-cultivation with Caldicellulosiruptor obsidiansis, a fermentative, cellulolytic extreme thermophile isolated from the same environment. On the basis of phylogenetic, phenotypic, and structural analyses, Thermodesulfobacterium geofontis sp. nov. is proposed as a new species with OPF15T representing the type strain.

  19. Effects of silage protein degradability and fermentation acids on metabolizable protein concentration: a meta-analysis of dairy cow production experiments.

    Science.gov (United States)

    Rinne, M; Nousiainen, J; Huhtanen, P

    2009-04-01

    A meta-analysis was conducted using data from dairy cow production studies to evaluate silage metabolizable protein (MP) concentrations. The data consisted of 397 treatment means in 130 comparisons, in which the effects of silage factors (e.g., date of harvest, wilting, silage additives) were investigated. Within a comparison, a fixed amount of the same concentrate was fed. A prerequisite of data to be included in the analysis was that silage dry matter (DM), crude protein (CP), ammonia N, lactic acid (LA), and total acid (TA) concentrations and digestibility were determined. A smaller data set (n = 248) comprised studies in which silage water-soluble N concentration was also analyzed. The supply of MP was estimated as amino acids absorbed from the small intestine using a model with constant values for ruminal effective protein degradability (EPD) and intestinal digestibility of rumen undegraded protein. Microbial protein was calculated on the basis of digestible carbohydrates and rumen degradable protein (RDP). Alternative models were used to estimate microbial protein formation, assuming the energy values of RDP and TA to be equivalent to 1.00, 0.75, 0.50, 0.25, and 0 times that of digestible carbohydrates. Because EPD values are seldom determined in production trials, they were derived using empirical models that estimate them from other feed components. The goodness of fit of models was compared on the basis of root mean squared error (RMSE) of milk protein yield (MPY) predicted from MP supply (adjusted for random study effect) and Akaike's information criterion. Metabolizable protein supply calculated from basal assumptions predicted MPY precisely within a study (RMSE = 16.2 g/d). Variable contribution of RDP to the energy supply for microbial synthesis influenced the precision of MPY prediction very little, but RMSE for MPY increased markedly when the energy supply of rumen microbes was corrected for TA concentration. Using predicted rather than constant EPD

  20. GPG-NH2 acts via the metabolite αHGA to target HIV-1 Env to the ER-associated protein degradation pathway

    Directory of Open Access Journals (Sweden)

    Vahlne Anders

    2010-03-01

    Full Text Available Abstract Background The synthetic peptide glycyl-prolyl-glycine amide (GPG-NH2 was previously shown to abolish the ability of HIV-1 particles to fuse with the target cells, by reducing the content of the viral envelope glycoprotein (Env in progeny HIV-1 particles. The loss of Env was found to result from GPG-NH2 targeting the Env precursor protein gp160 to the ER-associated protein degradation (ERAD pathway during its maturation. However, the anti-viral effect of GPG-NH2 has been shown to be mediated by its metabolite α-hydroxy-glycineamide (αHGA, which is produced in the presence of fetal bovine serum, but not human serum. In accordance, we wanted to investigate whether the targeting of gp160 to the ERAD pathway by GPG-NH2 was attributed to its metabolite αHGA. Results In the presence of fetal bovine serum, GPG-NH2, its intermediary metabolite glycine amide (G-NH2, and final metabolite αHGA all induced the degradation of gp160 through the ERAD pathway. However, when fetal bovine serum was replaced with human serum only αHGA showed an effect on gp160, and this activity was further shown to be completely independent of serum. This indicated that GPG-NH2 acts as a pro-drug, which was supported by the observation that it had to be added earlier to the cell cultures than αHGA to induce the degradation of gp160. Furthermore, the substantial reduction of Env incorporation into HIV-1 particles that occurs during GPG-NH2 treatment was also achieved by treating HIV-1 infected cells with αHGA. Conclusions The previously observed specificity of GPG-NH2 towards gp160 in HIV-1 infected cells, resulting in the production of Env (gp120/gp41 deficient fusion incompetent HIV-1 particles, was most probably due to the action of the GPG-NH2 metabolite αHGA.

  1. EFFECT OF MUSIC ON ANAEROBIC EXERCISE PERFORMANCE

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    Tülin Atan

    2013-01-01

    Full Text Available For years, mostly the effects of music on cardiorespiratory exercise performance have been studied, but a few studies have examined the effect of music on anaerobic exercise. The purpose of this study was to assess the effect of listening to music and its rhythm on anaerobic exercise: on power output, heart rate and the concentration of blood lactate. 28 male subjects were required to visit the laboratory on 6 occasions, each separated by 48 hours. Firstly, each subject performed the Running-based Anaerobic Sprint Test (RAST under 3 conditions on separate days: while listening to “slow rhythm music”, “fast rhythm music” or “no music”. 48 hours after the subjects completed RAST under 3 conditions, Wingate Anaerobic Power (WAN tests were performed under 3 music conditions. The order of the 3 conditions (slow music, fast music and no music was selected randomly to prevent an order effect. Results showed no significant differences between 3 conditions in anaerobic power assessments, heart rate or blood lactate (p>0.05. On the basis of these results it can be said that music cannot improve anaerobic performance. The type of music had no impact on power outputs during RAST and WAN exercise. As a conclusion, listening to music and its rhythm cannot enhance anaerobic performance and cannot change the physiological response to supramaximal exercise.

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

    Science.gov (United States)

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

    2015-08-01

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

  3. Intraspecific variation in aerobic and anaerobic locomotion

    DEFF Research Database (Denmark)

    Svendsen, Jon Christian; Tirsgård, Bjørn; Cordero, Gerardo A.

    2015-01-01

    to unsteady (i.e., burst-assisted) swimming is associated with anaerobic metabolism evidenced as excess post exercise oxygen consumption (EPOC); (2) variation in swimming performance (critical swimming speed; U crit) correlates with metabolic scope (MS) or anaerobic capacity (i.e., maximum EPOC); (3...... respirometry and video analysis. Results showed that anaerobic swimming costs (i.e., EPOC) increase linearly with the number of bursts in S. aurata, with each burst corresponding to 0.53 mg O2 kg(-1). Data are consistent with a previous study on striped surfperch (Embiotoca lateralis), a labriform swimmer...

  4. Fate of Trace Metals in Anaerobic Digestion.

    Science.gov (United States)

    Fermoso, F G; van Hullebusch, E D; Guibaud, G; Collins, G; Svensson, B H; Carliell-Marquet, C; Vink, J P M; Esposito, G; Frunzo, L

    2015-01-01

    A challenging, and largely uncharted, area of research in the field of anaerobic digestion science and technology is in understanding the roles of trace metals in enabling biogas production. This is a major knowledge gap and a multifaceted problem involving metal chemistry; physical interactions of metal and solids; microbiology; and technology optimization. Moreover, the fate of trace metals, and the chemical speciation and transport of trace metals in environments--often agricultural lands receiving discharge waters from anaerobic digestion processes--simultaneously represents challenges for environmental protection and opportunities to close process loops in anaerobic digestion.

  5. Energy from anaerobic methane production. [Sweden

    Energy Technology Data Exchange (ETDEWEB)

    1982-02-01

    Since 1970 Swedish researchers have been testing the ANAMET (anaerobic-aerobic-methane) process, which involves converting industrial wastewaters via an initial anaerobic microbiological step followed by an aerobic one. Recycling the biomass material in each step allows shorter hydraulic retention times without decreasing stability or solids reduction. Since the first ANAMET plants began operating at a Swedish sugar factory in 1972, 17 more plants have started up or are under construction. Moreover, the ANAMET process has engendered to offshoot BIOMET (biomass-methane) process, a thermophilic anaerobic scheme that can handle sugar-beet pulp as well as grass and other soft, fast-growing biomasses.

  6. Anaerobic degradation of linear alkylbenzene sulfonate

    DEFF Research Database (Denmark)

    Mogensen, Anders Skibsted; Haagensen, Frank; Ahring, Birgitte Kiær

    2003-01-01

    Linear alkylbenzene sulfonate (LAS) found in wastewater is removed in the wastewater treatment facilities by sorption and aerobic biodegradation. The anaerobic digestion of sewage sludge has not been shown to contribute to the removal. The concentration of LAS based on dry matter typically...... increases during anaerobic stabilization due to transformation of easily degradable organic matter. Hence, LAS is regarded as resistant to biodegradation under anaerobic conditions. We present data from a lab-scale semi-continuously stirred tank reactor (CSTR) spiked with linear dodecylbenzene sulfonate (C...

  7. Anaerobic degradation and toxicity of commercial cationic surfactants in anaerobic screening tests.

    Science.gov (United States)

    García, M T; Campos, E; Sánchez-Leal, J; Ribosa, I

    2000-09-01

    Anaerobic biodegradability and toxicity on anaerobic bacteria of di(hydrogenated tallow) dimethyl ammonium chloride (DHTDMAC) and two esterquats have been investigated. A batch test system containing municipal digester solids as a source of anaerobic bacteria, based on the method proposed by the ECETOC, has been applied. To evaluate the potential toxicity of such surfactants on anaerobic sludge, a co-substrate, an easily biodegradable compound in anaerobic conditions, has been added to the samples to test and the effects on biogas production have been determined. For the esterquats studied high biodegradation levels were obtained and no toxic effects on anaerobic bacteria were observed even at the highest concentrations tested, 100 and 200 mg C/l, respectively. On the contrary, DHTDMAC was not degradated at the same test conditions. However, no inhibitory effects on the biogas production were detected for this surfactant at concentrations <100 mg C/l.

  8. Antimicrobial resistance and susceptibility testing of anaerobic bacteria.

    Science.gov (United States)

    Schuetz, Audrey N

    2014-09-01

    Infections due to anaerobic bacteria can be severe and life-threatening. Susceptibility testing of anaerobes is not frequently performed in laboratories, but such testing is important to direct appropriate therapy. Anaerobic resistance is increasing globally, and resistance trends vary by geographic region. An overview of a variety of susceptibility testing methods for anaerobes is provided, and the advantages and disadvantages of each method are reviewed. Specific clinical situations warranting anaerobic susceptibility testing are discussed.

  9. Anaerobe Tolerance to Oxygen and the Potentials of Anaerobic and Aerobic Cocultures for Wastewater Treatment

    Directory of Open Access Journals (Sweden)

    M.T. Kato

    1997-12-01

    Full Text Available The anaerobic treatment processes are considered to be well-established methods for the elimination of easily biodegradable organic matter from wastewaters. Some difficulties concerning certain wastewaters are related to the possible presence of dissolved oxygen. The common belief is that anaerobes are oxygen intolerant. Therefore, the common practice is to use sequencing anaerobic and aerobic steps in separate tanks. Enhanced treatment by polishing off the residual biodegradable oxygen demand from effluents of anaerobic reactors, or the biodegradation of recalcitrant wastewater pollutants, usually requires sequenced anaerobic and aerobic bacteria activities. However, the combined activity of both bacteria can also be obtained in a single reactor. Previous experiments with either pure or mixed cultures showed that anaerobes can tolerate oxygen to a certain extent. The oxygen toxicity to methanogens in anaerobic sludges was quantified in batch experiments, as well as in anaerobic reactors. The results showed that methanogens have a high tolerance to oxygen. In practice, it was confirmed that dissolved oxygen does not constitute any detrimental effect on reactor treatment performance. This means that the coexistence of anaerobic and aerobic bacteria in one single reactor is feasible and increases the potentials of new applications in wastewater treatment

  10. EXPRESSION, PURIFICATION, AND SMALL ANGLE X-RAY SCATTERING OF DNA REPLICATION AND REPAIR PROTEINS FROM THE HYPERTHERMOPHILE SULFOLOBUS SOLFATARICUS

    Energy Technology Data Exchange (ETDEWEB)

    Patterson, S.M.; Hatherill, J.R.; Hammel, M.; Hura, G.L.; Tainer, J.A.; Yannone, S.M.

    2008-01-01

    Vital molecular processes such as DNA replication, transcription, translation, and maintenance occur through transient protein interactions. Elucidating the mechanisms by which these protein complexes and interactions function could lead to treatments for diseases related to DNA damage and cell division control. In the recent decades since its introduction as a third domain, Archaea have shown to be simpler models for complicated eukaryotic processes such as DNA replication, repair, transcription, and translation. Sulfolobus solfataricus is one such model organism. A hyperthermophile with an optimal growth temperature of 80°C, Sulfolobus protein-protein complexes and transient protein interactions should be more stable at moderate temperatures, providing a means to isolate and study their structure and function. Here we provide the initial steps towards characterizing three DNA-related Sulfolobus proteins with small angle X-ray scattering (SAXS): Sso0257, a cell division control and origin recognition complex homolog, Sso0768, the small subunit of the replication factor C, and Sso3167, a Mut-T like protein. SAXS analysis was performed at multiple concentrations for both short and long exposure times. The Sso0257 sample was determined to be either a mixture of monomeric and dimeric states or a population of dynamic monomers in various conformational states in solution, consistent with a fl exible winged helix domain. Sso0768 was found to be a complex mixture of multimeric states in solution. Finally, molecular envelope reconstruction from SAXS data for Sso3167 revealed a novel structural component which may function as a disordered to ordered region in the presence of its substrates and/or protein partners.

  11. New functional sulfide oxidase-oxygen reductase supercomplex in the membrane of the hyperthermophilic bacterium Aquifex aeolicus.

    Science.gov (United States)

    Prunetti, Laurence; Infossi, Pascale; Brugna, Myriam; Ebel, Christine; Giudici-Orticoni, Marie-Thérèse; Guiral, Marianne

    2010-12-31

    Aquifex aeolicus, a hyperthermophilic and microaerophilic bacterium, obtains energy for growth from inorganic compounds alone. It was previously proposed that one of the respiratory pathways in this organism consists of the electron transfer from hydrogen sulfide (H(2)S) to molecular oxygen. H(2)S is oxidized by the sulfide quinone reductase, a membrane-bound flavoenzyme, which reduces the quinone pool. We have purified and characterized a novel membrane-bound multienzyme supercomplex that brings together all the molecular components involved in this bioenergetic chain. Our results indicate that this purified structure consists of one dimeric bc(1) complex (complex III), one cytochrome c oxidase (complex IV), and one or two sulfide quinone reductases as well as traces of the monoheme cytochrome c(555) and quinone molecules. In addition, this work strongly suggests that the cytochrome c oxidase in the supercomplex is a ba(3)-type enzyme. The supercomplex has a molecular mass of about 350 kDa and is enzymatically functional, reducing O(2) in the presence of the electron donor, H(2)S. This is the first demonstration of the existence of such a respirasome carrying a sulfide oxidase-oxygen reductase activity. Moreover, the kinetic properties of the sulfide quinone reductase change slightly when integrated in the supercomplex, compared with the free enzyme. We previously purified a complete respirasome involved in hydrogen oxidation and sulfur reduction from Aquifex aeolicus. Thus, two different bioenergetic pathways (sulfur reduction and sulfur oxidation) are organized in this bacterium as supramolecular structures in the membrane. A model for the energetic sulfur metabolism of Aquifex aeolicus is proposed.

  12. The apt/6-Methylpurine Counterselection System and Its Applications in Genetic Studies of the Hyperthermophilic Archaeon Sulfolobus islandicus

    Science.gov (United States)

    Bi, Hongkai; Whitaker, Rachel J.

    2016-01-01

    ABSTRACT Sulfolobus islandicus serves as a model for studying archaeal biology as well as linking novel biology to evolutionary ecology using functional population genomics. In the present study, we developed a new counterselectable genetic marker in S. islandicus to expand the genetic toolbox for this species. We show that resistance to the purine analog 6-methylpurine (6-MP) in S. islandicus M.16.4 is due to the inactivation of a putative adenine phosphoribosyltransferase encoded by M164_0158 (apt). The application of the apt gene as a novel counterselectable marker was first illustrated by constructing an unmarked α-amylase deletion mutant. Furthermore, the 6-MP counterselection feature was employed in a forward (loss-of-function) mutation assay to reveal the profile of spontaneous mutations in S. islandicus M.16.4 at the apt locus. Moreover, the general conservation of apt genes in the crenarchaea suggests that the same strategy can be broadly applied to other crenarchaeal model organisms. These results demonstrate that the apt locus represents a new tool for genetic manipulation and sequence analysis of the hyperthermophilic crenarchaeon S. islandicus. IMPORTANCE Currently, the pyrEF/5-fluoroorotic acid (5-FOA) counterselection system remains the sole counterselection marker in crenarchaeal genetics. Since most Sulfolobus mutants constructed by the research community were derived from genetic hosts lacking the pyrEF genes, the pyrEF/5-FOA system is no longer available for use in forward mutation assays. Demonstration of the apt/6-MP counterselection system for the Sulfolobus model renders it possible to again study the mutation profiles in mutants that have already been constructed by the use of strains with a pyrEF-deficient background. Furthermore, additional counterselectable markers will allow us to conduct more sophisticated genetic studies, i.e., investigate mechanisms of chromosomal DNA transfer and quantify recombination frequencies among S

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

    Directory of Open Access Journals (Sweden)

    Wittinghofer Alfred

    2005-03-01

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

  14. Characteristics of a Novel Highly Thermostable and Extremely Thermophilic Alkalitolerant Amylase from Hyperthermophilic Bacillus Strain HUTBS71

    Directory of Open Access Journals (Sweden)

    Hazem Akel

    2009-01-01

    Full Text Available Problem statement: This study reported the purification and characterization of a novel highly thermostable alkaline amylase from a newly isolated Bacillus strain HUTBS71. Approach: The enzyme was purified using ammonium sulfate precipitation, ion exchange and gel filtration chromatography. Results: Maximum amylase activity (72 U mL-1 was obtained at 100°C after 10 min of incubation. The enzyme was purified 24 fold with 12.5% yield and showed a monomer band with a molecular weight of 58.8 kDa by SDS-PAGE. This enzyme exhibited maximum activity at pH and temperature, 7.8 and 100°C, respectively. It performed stability over a broad range of pH and temperature, 5.2-10.0 and 80-115°C, respectively. The half-life of the enzyme at 90 and 100°C was estimated to be 3 h. The activation energy of denaturation of purified enzyme was 2.53 kJ moL-1. The enzyme was activated by 5 mM of CoCl2, MgSO4, MnCl2, ZnSO4 and MnSO4 (relative activity was 133, 126, 133, 106.6 and 103%, respectively. It was strongly inhibited by CuSO4 and CdCl2 but less affected by NaCl, CaCl2, FeCl3, ZnCl2 and EDTA. Conclusion: The present purified amylase therefore could be defined as a highly thermostable, extremely hyperthermophilic and alkalitolerant with new properties make the present enzyme applicable for many starch processing and food industries.

  15. The elusive third subunit IIa of the bacterial B-type oxidases: the enzyme from the hyperthermophile Aquifex aeolicus.

    Directory of Open Access Journals (Sweden)

    Laurence Prunetti

    Full Text Available The reduction of molecular oxygen to water is catalyzed by complicated membrane-bound metallo-enzymes containing variable numbers of subunits, called cytochrome c oxidases or quinol oxidases. We previously described the cytochrome c oxidase II from the hyperthermophilic bacterium Aquifex aeolicus as a ba(3-type two-subunit (subunits I and II enzyme and showed that it is included in a supercomplex involved in the sulfide-oxygen respiration pathway. It belongs to the B-family of the heme-copper oxidases, enzymes that are far less studied than the ones from family A. Here, we describe the presence in this enzyme of an additional transmembrane helix "subunit IIa", which is composed of 41 amino acid residues with a measured molecular mass of 5105 Da. Moreover, we show that subunit II, as expected, is in fact longer than the originally annotated protein (from the genome and contains a transmembrane domain. Using Aquifex aeolicus genomic sequence analyses, N-terminal sequencing, peptide mass fingerprinting and mass spectrometry analysis on entire subunits, we conclude that the B-type enzyme from this bacterium is a three-subunit complex. It is composed of subunit I (encoded by coxA(2 of 59000 Da, subunit II (encoded by coxB(2 of 16700 Da and subunit IIa which contain 12, 1 and 1 transmembrane helices respectively. A structural model indicates that the structural organization of the complex strongly resembles that of the ba(3 cytochrome c oxidase from the bacterium Thermus thermophilus, the IIa helical subunit being structurally the lacking N-terminal transmembrane helix of subunit II present in the A-type oxidases. Analysis of the genomic context of genes encoding oxidases indicates that this third subunit is present in many of the bacterial oxidases from B-family, enzymes that have been described as two-subunit complexes.

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

    Directory of Open Access Journals (Sweden)

    Koh Eunhee

    2007-07-01

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

  17. Anaerobes in Industrial- and Environmental Biotechnology.

    Science.gov (United States)

    Hatti-Kaul, Rajni; Mattiasson, Bo

    Anaerobic microorganisms present in diverse ecological niches employ alternative strategies for energy conservation in the absence of oxygen which enables them to play a key role in maintaining the global cycles of carbon, nitrogen, and sulfur, and the breakdown of persistent compounds. Thereby they become useful tools in industrial and environmental biotechnology. Although anaerobes have been relatively neglected in comparison to their aerobic counterparts, with increasing knowledge about their diversity and metabolic potential and the development of genetic tools and process technologies to utilize them, we now see a rapid expansion of their applications in the society. This chapter summarizes some of the developments in the use of anaerobes as tools for biomass valorization, in production of energy carriers and chemicals, wastewater treatment, and the strong potential in soil remediation. The ability of several autotrophic anaerobes to reduce carbon dioxide is attracting growing attention as a means for developing a platform for conversion of waste gases to chemicals, materials, and biofuels.

  18. An anaerobic mitochondrion that produces hydrogen

    NARCIS (Netherlands)

    Boxma, Brigitte; Graaf, Rob M. de; Staay, Georg W.M. van der; Alen, Theo A. van; Ricard, Guenola; Gabaldón, Toni; Hoek, Angela H.A.M. van; Moon-van der Staay, Seung Yeo; Koopman, Werner J.H.; Hellemond, Jaap J. van; Tielens, Aloysius G.M.; Friedrich, Thorsten; Veenhuis, Marten; Huynen, Martijn A.; Hackstein, Johannes H.P.

    2005-01-01

    Hydrogenosomes are organelles that produce ATP and hydrogen, and are found in various unrelated eukaryotes, such as anaerobic flagellates, chytridiomycete fungi and ciliates. Although all of these organelles generate hydrogen, the hydrogenosomes from these organisms are structurally and metabolicall

  19. Anaerobic Toxicity of Cationic Silver Nanoparticles

    Data.gov (United States)

    U.S. Environmental Protection Agency — Toxicity data for the impact of nano-silver on anaerobic degradation. This dataset is associated with the following publication: Gitipour, A., S. Thiel, K. Scheckel,...

  20. Exocellular electron transfer in anaerobic microbial communities

    NARCIS (Netherlands)

    Stams, A.J.M.; Bok, de F.A.M.; Plugge, C.M.; Eekert, van M.H.A.; Dolfing, J.; Schraa, G.

    2006-01-01

    Exocellular electron transfer plays an important role in anaerobic microbial communities that degrade organic matter. Interspecies hydrogen transfer between microorganisms is the driving force for complete biodegradation in methanogenic environments. Many organic compounds are degraded by obligatory

  1. Anaerobic membrane bioreactor under extreme conditions (poster)

    NARCIS (Netherlands)

    Munoz Sierra, J.D.; De Kreuk, M.K.; Spanjers, H.; Van Lier, J.B.

    2013-01-01

    Membrane bioreactors ensure biomass retention by the application of micro or ultrafiltration processes. This allows operation at high sludge concentrations. Previous studies have shown that anaerobic membrane bioreactors is an efficient way to retain specialist microorganisms for treating wastewater

  2. Exocellular electron transfer in anaerobic microbial communities

    NARCIS (Netherlands)

    Stams, A.J.M.; Bok, de F.A.M.; Plugge, C.M.; Eekert, van M.H.A.; Dolfing, J.; Schraa, G.

    2006-01-01

    Exocellular electron transfer plays an important role in anaerobic microbial communities that degrade organic matter. Interspecies hydrogen transfer between microorganisms is the driving force for complete biodegradation in methanogenic environments. Many organic compounds are degraded by obligatory

  3. An anaerobic mitochondrion that produces hydrogen

    NARCIS (Netherlands)

    Boxma, Brigitte; Graaf, Rob M. de; Staay, Georg W.M. van der; Alen, Theo A. van; Ricard, Guenola; Gabaldón, Toni; Hoek, Angela H.A.M. van; Moon-van der Staay, Seung Yeo; Koopman, Werner J.H.; Hellemond, Jaap J. van; Tielens, Aloysius G.M.; Friedrich, Thorsten; Veenhuis, Marten; Huynen, Martijn A.; Hackstein, Johannes H.P.

    2005-01-01

    Hydrogenosomes are organelles that produce ATP and hydrogen, and are found in various unrelated eukaryotes, such as anaerobic flagellates, chytridiomycete fungi and ciliates. Although all of these organelles generate hydrogen, the hydrogenosomes from these organisms are structurally and metabolicall

  4. EFFECT OF MUSIC ON ANAEROBIC EXERCISE PERFORMANCE

    OpenAIRE

    Tülin Atan

    2013-01-01

    For years, mostly the effects of music on cardiorespiratory exercise performance have been studied, but a few studies have examined the effect of music on anaerobic exercise. The purpose of this study was to assess the effect of listening to music and its rhythm on anaerobic exercise: on power output, heart rate and the concentration of blood lactate. 28 male subjects were required to visit the laboratory on 6 occasions, each separated by 48 hours. Firstly, each subject performed the Running-...

  5. Anaerobic digestion foaming causes – A review

    OpenAIRE

    Ganidi, Nafsika; Tyrrel, Sean F.; Cartmell, Elise

    2009-01-01

    Anaerobic digestion foaming has been encountered in several sewage treatment plants in the UK. Foaming has raised major concerns for the water companies due to significant impacts on process efficiency and operational costs. Several foaming causes have been identified over the past few years by researchers. However, the supporting experimental information is limited and in some cases absent. The present report aims to provide a detailed review of the current anaerobic digestion foaming proble...

  6. EFFECT OF MUSIC ON ANAEROBIC EXERCISE PERFORMANCE

    OpenAIRE

    Atan, T.

    2013-01-01

    For years, mostly the effects of music on cardiorespiratory exercise performance have been studied, but a few studies have examined the effect of music on anaerobic exercise. The purpose of this study was to assess the effect of listening to music and its rhythm on anaerobic exercise: on power output, heart rate and the concentration of blood lactate. 28 male subjects were required to visit the laboratory on 6 occasions, each separated by 48 hours. Firstly, each subject performed the Running-...

  7. Anaerobic digester for treatment of organic waste

    Energy Technology Data Exchange (ETDEWEB)

    Sharma, V. K. [Indian Insitute of Technology, Delhi (India)]|[ENEA, Centro Ricerche Trisaia, Matera (Italy); Fortuna, F.; Canditelli, M.; Cornacchia, G. [ENEA, Centro Ricerche Trisaia, Matera (Italy). Dipt. Ambiente; Farina, R. [ENEA, centro Ricerche ``Ezio Clementel``, Bologna (Italy). Dipt. Ambiente

    1997-09-01

    The essential features of both new and more efficient reactor systems and their appropriate applications for various organic waste management situations, description of several working plants are discussed in the present communication. It is hoped that significant development reported here would be useful in opening a new vista to the application of anaerobic biotechnology for the waste treatment of both low/high organic strength and specialized treatment for toxic substances, using appropriate anaerobic methods.

  8. Biological phosphorus removal during high-rate, low-temperature, anaerobic digestion of wastewater

    Directory of Open Access Journals (Sweden)

    Ciara eKeating

    2016-03-01

    Full Text Available We report, for the first time, extensive biologically-mediated phosphate removal from wastewater during high-rate anaerobic digestion (AD. A hybrid sludge bed/fixed-film (packed pumice stone reactor was employed for low-temperature (12°C anaerobic treatment of synthetic sewage wastewater. Successful phosphate removal from the wastewater (up to 78% of influent phosphate was observed, mediated by biofilms in the reactor. Scanning electron microscopy and energy dispersive X-ray analysis revealed the accumulation of elemental phosphorus (~2% within the sludge bed and fixed-film biofilms. 4’, 6-diamidino-2-phenylindole (DAPI staining indicated phosphorus accumulation was biological in nature and mediated through the formation of intracellular inorganic polyphosphate (polyP granules within these biofilms. DAPI staining further indicated that polyP accumulation was rarely associated with free cells. Efficient and consistent chemical oxygen demand (COD removal was recorded, throughout the 732-day trial, at applied organic loading rates between 0.4-1.5 kg COD m-3 d-1 and hydraulic retention times of 8-24 hours, while phosphate removal efficiency ranged from 28-78% on average per phase. Analysis of protein hydrolysis kinetics and the methanogenic activity profiles of the biomass revealed the development, at 12˚C, of active hydrolytic and methanogenic populations. Temporal microbial changes were monitored using Illumina Miseq analysis of bacterial and archaeal 16S rRNA gene sequences. The dominant bacterial phyla present in the biomass at the conclusion of the trial were the Proteobacteria and Firmicutes and the dominant archaeal genus was Methanosaeta. Trichococcus and Flavobacterium populations, previously associated with low temperature protein degradation, developed in the reactor biomass. The presence of previously characterised polyphosphate accumulating organisms (PAOs such as Rhodocyclus, Chromatiales, Actinobacter and Acinetobacter was

  9. Biological Phosphorus Removal During High-Rate, Low-Temperature, Anaerobic Digestion of Wastewater.

    Science.gov (United States)

    Keating, Ciara; Chin, Jason P; Hughes, Dermot; Manesiotis, Panagiotis; Cysneiros, Denise; Mahony, Therese; Smith, Cindy J; McGrath, John W; O'Flaherty, Vincent

    2016-01-01

    We report, for the first time, extensive biologically mediated phosphate removal from wastewater during high-rate anaerobic digestion (AD). A hybrid sludge bed/fixed-film (packed pumice stone) reactor was employed for low-temperature (12°C) anaerobic treatment of synthetic sewage wastewater. Successful phosphate removal from the wastewater (up to 78% of influent phosphate) was observed, mediated by biofilms in the reactor. Scanning electron microscopy and energy dispersive X-ray analysis revealed the accumulation of elemental phosphorus (∼2%) within the sludge bed and fixed-film biofilms. 4', 6-diamidino-2-phenylindole (DAPI) staining indicated phosphorus accumulation was biological in nature and mediated through the formation of intracellular inorganic polyphosphate (polyP) granules within these biofilms. DAPI staining further indicated that polyP accumulation was rarely associated with free cells. Efficient and consistent chemical oxygen demand (COD) removal was recorded, throughout the 732-day trial, at applied organic loading rates between 0.4 and 1.5 kg COD m(-3) d(-1) and hydraulic retention times of 8-24 h, while phosphate removal efficiency ranged from 28 to 78% on average per phase. Analysis of protein hydrolysis kinetics and the methanogenic activity profiles of the biomass revealed the development, at 12°C, of active hydrolytic and methanogenic populations. Temporal microbial changes were monitored using Illumina MiSeq analysis of bacterial and archaeal 16S rRNA gene sequences. The dominant bacterial phyla present in the biomass at the conclusion of the trial were the Proteobacteria and Firmicutes and the dominant archaeal genus was Methanosaeta. Trichococcus and Flavobacterium populations, previously associated with low temperature protein degradation, developed in the reactor biomass. The presence of previously characterized polyphosphate accumulating organisms (PAOs) such as Rhodocyclus, Chromatiales, Actinobacter, and Acinetobacter was recorded

  10. Anaerobic electron acceptor chemotaxis in Shewanella putrefaciens

    Science.gov (United States)

    Nealson, K. H.; Moser, D. P.; Saffarini, D. A.

    1995-01-01

    Shewanella putrefaciens MR-1 can grow either aerobically or anaerobically at the expense of many different electron acceptors and is often found in abundance at redox interfaces in nature. Such redox interfaces are often characterized by very strong gradients of electron acceptors resulting from rapid microbial metabolism. The coincidence of S. putrefaciens abundance with environmental gradients prompted an examination of the ability of MR-1 to sense and respond to electron acceptor gradients in the laboratory. In these experiments, taxis to the majority of the electron acceptors that S. putrefaciens utilizes for anaerobic growth was seen. All anaerobic electron acceptor taxis was eliminated by the presence of oxygen, nitrate, nitrite, elemental sulfur, or dimethyl sulfoxide, even though taxis to the latter was very weak and nitrate and nitrite respiration was normal in the presence of dimethyl sulfoxide. Studies with respiratory mutants of MR-1 revealed that several electron acceptors that could not be used for anaerobic growth nevertheless elicited normal anaerobic taxis. Mutant M56, which was unable to respire nitrite, showed normal taxis to nitrite, as well as the inhibition of taxis to other electron acceptors by nitrite. These results indicate that electron acceptor taxis in S. putrefaciens does not conform to the paradigm established for Escherichia coli and several other bacteria. Carbon chemo-taxis was also unusual in this organism: of all carbon compounds tested, the only positive response observed was to formate under anaerobic conditions.

  11. Infected neonatal cephalohematomas caused by anaerobic bacteria.

    Science.gov (United States)

    Brook, Itzhak

    2005-01-01

    To present the microbiological and clinical features of six children with infected cephalohematomas (IC) caused by anaerobic bacteria. Presentation of a case series. Polymicrobial infection was present in all instances, where the number of isolates varied from two to four. Two patients had anaerobes only and the other four had mixed flora of strict anaerobes and facultatives. There were 16 bacterial isolates (12 anaerobic, 4 aerobic). The anaerobic isolates were Peptostreptococcus spp. (5 isolates), Prevotella spp. (4), Bacteroides fragilis group (2), and Propionibacterium acnes (1). The aerobic isolates were E. coli (2), Staphylococcus aureus (1) and group B streptococci (1). Blood cultures were positive for three patients. The most common predisposing conditions were vacuum extraction and amnionitis (4 instances of each), instrumental delivery (3), electronic fetal monitoring (2), prolonged delivery (1), and premature rupture of membranes (1). All patients underwent drainage, and four also had surgical incision and drainage of the IC. Osteomyelitis developed in one instance and scalp abscess developed in two patients, both of whom had electronic fetal monitoring. All patients eventually recovered from infection after receiving parenteral and subsequent oral antibiotic therapy for a total of 14-38 days. This study highlights the polymicrobial nature and potential importance of anaerobic bacteria in IC in newborns.

  12. SLEEP DEPRIVATION INDUCED ANXIETY AND ANAEROBIC PERFORMANCE

    Directory of Open Access Journals (Sweden)

    Selma Arzu Vardar

    2007-12-01

    Full Text Available The aim of this study was to investigate the effects of sleep deprivation induced anxiety on anaerobic performance. Thirteen volunteer male physical education students completed the Turkish version of State Anxiety Inventory and performed Wingate anaerobic test for three times: (1 following a full-night of habitual sleep (baseline measurements, (2 following 30 hours of sleep deprivation, and (3 following partial-night sleep deprivation. Baseline measurements were performed the day before total sleep deprivation. Measurements following partial sleep deprivation were made 2 weeks later than total sleep deprivation measurements. State anxiety was measured prior to each Wingate test. The mean state anxiety following total sleep deprivation was higher than the baseline measurement (44.9 ± 12.9 vs. 27.6 ± 4.2, respectively, p = 0.02 whereas anaerobic performance parameters remained unchanged. Neither anaerobic parameters nor state anxiety levels were affected by one night partial sleep deprivation. Our results suggest that 30 hours continuous wakefulness may increase anxiety level without impairing anaerobic performance, whereas one night of partial sleep deprivation was ineffective on both state anxiety and anaerobic performance

  13. The thermophilic biomass-degrading fungus Thielavia terrestris Co3Bag1 produces a hyperthermophilic and thermostable β-1,4-xylanase with exo- and endo-activity.

    Science.gov (United States)

    García-Huante, Yolanda; Cayetano-Cruz, Maribel; Santiago-Hernández, Alejandro; Cano-Ramírez, Claudia; Marsch-Moreno, Rodolfo; Campos, Jorge E; Aguilar-Osorio, Guillermo; Benitez-Cardoza, Claudia G; Trejo-Estrada, Sergio; Hidalgo-Lara, María Eugenia

    2017-01-01

    A hyperthermophilic and thermostable xylanase of 82 kDa (TtXynA) was purified from the culture supernatant of T. terrestris Co3Bag1, grown on carboxymethyl cellulose (CMC), and characterized biochemically. TtXynA showed optimal xylanolytic activity at pH 5.5 and at 85 °C, and retained more than 90% of its activity at a broad pH range (4.5-10). The enzyme is highly thermostable with a half-life of 23.1 days at 65 °C, and active in the presence of several metal ions. Circular dichroism spectra strongly suggest the enzyme gains secondary structures when temperature increases. TtXynA displayed higher substrate affinity and higher catalytic efficiency towards beechwood xylan than towards birchwood xylan, oat-spelt xylan, and CMC. According to its final hydrolysis products, TtXynA displays endo-/exo-activity, yielded xylobiose, an unknown oligosaccharide containing about five residues of xylose and a small amount of xylose on beechwood xylan. Finally, this report represents the description of the first fungal hyperthermophilic xylanase which is produced by T. terrestris Co3Bag1. Since TtXynA displays relevant biochemical properties, it may be a suitable candidate for biotechnological applications carried out at high temperatures, like the enzymatic pretreatment of plant biomass for the production of bioethanol.

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

    Science.gov (United States)

    Ronimus, Ron S; Morgan, Hugh W

    2003-10-01

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

  15. Anaerobic Nitrogen Fixers on Mars

    Science.gov (United States)

    Lewis, B. G.

    2000-07-01

    The conversion of atmospheric nitrogen gas to the protein of living systems is an amazing process of nature. The first step in the process is biological nitrogen fixation, the transformation of N2 to NH3. The phenomenon is crucial for feeding the billions of our species on Earth. On Mars, the same process may allow us to discover how life can adapt to a hostile environment, and render it habitable. Hostile environments also exist on Earth. For example, nothing grows in coal refuse piles due to the oxidation of pyrite and marcasite to sulfuric acid. Yet, when the acidity is neutralized, alfalfa and soybean plants develop root nodules typical of symbiotic nitrogen fixation with Rhizobium species possibly living in the pyritic material. When split open, these nodules exhibited the pinkish color of leghemoglobin, a protein in the nodule protecting the active nitrogen-fixing enzyme nitrogenase against the toxic effects of oxygen. Although we have not yet obtained direct evidence of nitrogenase activity in these nodules (reduction of acetylene to ethylene, for example), these findings suggested the possibility that nitrogen fixation was taking place in this hostile, non-soil material. This immediately raises the possibility that freeliving anaerobic bacteria which fix atmospheric nitrogen on Earth, could do the same on Mars.

  16. Application of dynamic membranes in anaerobic membranes in anaerobic membrane bioreactor systems

    NARCIS (Netherlands)

    Erşahin, M.E.

    2015-01-01

    Anaerobic membrane bioreactors (AnMBRs) physically ensure biomass retention by the application of a membrane filtration process. With growing application experiences from aerobic membrane bioreactors (MBRs), the combination of membrane and anaerobic processes has received much attention and become m

  17. The effect of outside conditions on anaerobic ammonia oxidation reaction

    Institute of Scientific and Technical Information of China (English)

    YANG Min; WANG Shu-bo

    2016-01-01

    Organic carbon, inorganic carbon, temperature, pH and ORP are all to have a certain influence on the anaerobic ammonia oxidation reaction. We can draw some conclusions on the optimum conditions of anaerobic ammonia oxidation reaction. The optimum temperature of the anaerobic ammonia oxidation reaction is 30-35℃. And the optimum pH of the anaerobic ammonia reaction is 7.5-8.3. The presence of organic matters can affect the anaerobic ammonia reaction, and different organic matters have different influence on it. The concentration of the inorganic carbon also exist great influence on the reaction. High inorganic carbon concentration also can inhibit anaerobic ammonia oxidation reaction.

  18. ANAEROBIC BIOLOGICAL TREATMENT OF PRODUCED WATER

    Energy Technology Data Exchange (ETDEWEB)

    John R. Gallagher

    2001-07-31

    During the production of oil and gas, large amounts of water are brought to the surface and must be disposed of in an environmentally sensitive manner. This is an especially difficult problem in offshore production facilities where space is a major constraint. The chief regulatory criterion for produced water is oil and grease. Most facilities have little trouble meeting this criterion using conventional oil-water separation technologies. However, some operations have significant amounts of naphthenic acids in the water that behave as oil and grease but are not well removed by conventional technologies. Aerobic biological treatment of naphthenic acids in simulated-produced water has been demonstrated by others; however, the system was easily overloaded by the large amounts of low-molecular-weight organic acids often found in produced waters. The objective of this research was to determine the ability of an anaerobic biological system to treat these organic acids in a simulated produced water and to examine the potential for biodegradation of the naphthenic acids in the anaerobic environment. A small fixed-film anaerobic biological reactor was constructed and adapted to treat a simulated produced water. The bioreactor was tubular, with a low-density porous glass packing material. The inocula to the reactor was sediment from a produced-water holding pond from a municipal anaerobic digester and two salt-loving methanogenic bacteria. During start-up, the feed to the reactor contained glucose as well as typical produced-water components. When glucose was used, rapid gas production was observed. However, when glucose was eliminated and the major organic component was acetate, little gas was generated. Methane production from acetate may have been inhibited by the high salt concentrations, by sulfide, or because of the lack, despite seeding, of microbes capable of converting acetate to methane. Toluene, a minor component of the produced water (0.1 g/L) was removed in the

  19. Signature lipids and stable carbon isotope analyses of Octopus Spring hyperthermophilic communities compared with those of Aquificales representatives

    Science.gov (United States)

    Jahnke, L. L.; Eder, W.; Huber, R.; Hope, J. M.; Hinrichs, K. U.; Hayes, J. M.; Des Marais, D. J.; Cady, S. L.; Summons, R. E.

    2001-01-01

    The molecular and isotopic compositions of lipid biomarkers of cultured Aquificales genera have been used to study the community and trophic structure of the hyperthermophilic pink streamers and vent biofilm from Octopus Spring. Thermocrinis ruber, Thermocrinis sp. strain HI 11/12, Hydrogenobacter thermophilus TK-6, Aquifex pyrophilus, and Aquifex aeolicus all contained glycerol-ether phospholipids as well as acyl glycerides. The n-C(20:1) and cy-C(21) fatty acids dominated all of the Aquificales, while the alkyl glycerol ethers were mainly C(18:0). These Aquificales biomarkers were major constituents of the lipid extracts of two Octopus Spring samples, a biofilm associated with the siliceous vent walls, and the well-known pink streamer community (PSC). Both the biofilm and the PSC contained mono- and dialkyl glycerol ethers in which C(18) and C(20) alkyl groups were prevalent. Phospholipid fatty acids included both the Aquificales n-C(20:1) and cy-C(21), plus a series of iso-branched fatty acids (i-C(15:0) to i-C(21:0)), indicating an additional bacterial component. Biomass and lipids from the PSC were depleted in (13)C relative to source water CO(2) by 10.9 and 17.2 per thousand, respectively. The C(20-21) fatty acids of the PSC were less depleted than the iso-branched fatty acids, 18.4 and 22.6 per thousand, respectively. The biomass of T. ruber grown on CO(2) was depleted in (13)C by only 3.3 per thousand relative to C source. In contrast, biomass was depleted by 19.7 per thousand when formate was the C source. Independent of carbon source, T. ruber lipids were heavier than biomass (+1.3 per thousand). The depletion in the C(20-21) fatty acids from the PSC indicates that Thermocrinis biomass must be similarly depleted and too light to be explained by growth on CO(2). Accordingly, Thermocrinis in the PSC is likely to have utilized formate, presumably generated in the spring source region.

  20. Oxygen Effects in Anaerobic Digestion - II

    Directory of Open Access Journals (Sweden)

    Deshai Botheju

    2010-04-01

    Full Text Available Standard models describing bio-gasification using anaerobic digestion do not include necessary processes to describe digester dynamics under the conditions of oxygen presence. Limited oxygenation in anaerobic digestion can sometimes be beneficial. The oxygen effects included anaerobic digestion model, ADM 1-Ox, was simulated against experimental data obtained from laboratory scale anaerobic digesters operated under different oxygenation conditions. ADM 1-Ox predictions are generally in good agreement with the trends of the experimental data. ADM 1-Ox simulations suggest the existence of an optimum oxygenation level corresponding to a peak methane yield. The positive impact of oxygenation on methane yield is more pronounced at conditions characterized by low hydrolysis rate coefficients (slowly degradable feed and low biomass concentrations. The optimum oxygenation point moves towards zero when the hydrolysis rate coefficient and the biomass concentration increase. Accordingly, the impact of oxygenation on methane yield can either be positive or negative depending on the digestion system characteristics. The developed ADM 1-Ox model can therefore be a valuable tool for recognizing suitable operating conditions for achieving the maximum benefits from partial aeration in anaerobic digestion.

  1. Biochemistry and physiology of anaerobic bacteria

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2000-05-18

    We welcome you to The Power of Anaerobes. This conference serves two purposes. One is to celebrate the life of Harry D. Peck, Jr.,who was born May 18, 1927 and would have celebrated his 73rd birthday at this conference. He died November 20, 1998. The second is to gather investigators to exchange views within the realm of anaerobic microbiology, an area in which tremendous progress has been seen during recent years. It is sufficient to mention discoveries of a new form of life (the archaea), hyper or extreme thermophiles, thermophilic alkaliphiles and anaerobic fungi. With these discoveries has come a new realization about physiological and metabolic properties of microorganisms, and this in turn has demonstrated their importance for the development, maintenance and sustenance of life on Earth.

  2. Anaerobic Digestion: Mass Balances and Products

    DEFF Research Database (Denmark)

    Møller, Jacob; Christensen, Thomas Højlund; Jansen, Jes la Cour

    2011-01-01

    While the basic processes involved in anaerobic digestion of waste are described in Chapter 9.4 and the main digestion technologies are presented in Chapter 9.5, this chapter focuses on mass balances, gas production and energy aspects, environmental emissions and unit process inventories. Underst......While the basic processes involved in anaerobic digestion of waste are described in Chapter 9.4 and the main digestion technologies are presented in Chapter 9.5, this chapter focuses on mass balances, gas production and energy aspects, environmental emissions and unit process inventories....... Understanding these issues and being able to account for them is a prerequisite in digestion engineering and for establishing and running a successful anaerobic digestion facility. Of specific importance is the final use of the digestate. Use in agriculture as a fertilizer is described in Chapter 9.10 and use...

  3. Multivariate monitoring of anaerobic co-digestion

    DEFF Research Database (Denmark)

    Madsen, Michael; Holm-Nielsen, Jens Bo

    fertiliser and renewable energy. Meanwhile, in order for the biogas sector to become a significant player in the energy supply chain, the anaerobic digestion process has to be controlled to a greater extent than what is implemented as state-of-the-art today. Through application of the philosophy behind......Anaerobic digestion processes for production of renewable energy in the form of biogas, and in the future hydrogen, are becoming increasingly important worldwide. Sustainable solutions for renewable energy production systems are given high political priority, amongst other things due to global...... warming and environmental concerns. Anaerobic digestion applied in agriculture can simultaneously convert heterogeneous biomasses and wastes from the primary agricultural sector and from the bio processing industries, for instance food processing, pharma, and biofuel production, into valuable organic...

  4. Succession of lignocellulolytic bacterial consortia bred anaerobically from lake sediment

    NARCIS (Netherlands)

    Korenblum, Elisa; Jiménez Avella, Diego; van Elsas, Jan

    2016-01-01

    Anaerobic bacteria degrade lignocellulose in various anoxic and organically rich environments, often in a syntrophic process. Anaerobic enrichments of bacterial communities on a recalcitrant lignocellulose source were studied combining polymerase chain reaction–denaturing gradient gel electrophoresi

  5. The effect of tannic compounds on anaerobic wastewater treatment

    NARCIS (Netherlands)

    Field, J.A.

    1989-01-01

    Anaerobic wastewater treatment is an alternative to the conventional aerobic treatment processes for the removal of easily biodegradable organic matter in medium to high strength industrial wastestreams. Anaerobic treatment has several advantages, however one important disadvantage is the

  6. The effect of tannic compounds on anaerobic wastewater treatment.

    NARCIS (Netherlands)

    Field, J.A.

    1989-01-01

    Anaerobic wastewater treatment is an alternative to the conventional aerobic treatment processes for the removal of easily biodegradable organic matter in medium to high strength industrial wastestreams. Anaerobic treatment has several advantages, however one important disadvantage is the high sensi

  7. Optimization of up-flow anaerobic sludge blanket reactor for ...

    African Journals Online (AJOL)

    Optimization of up-flow anaerobic sludge blanket reactor for treatment of composite fermentation and distillation wastewater. ... treatment, biogas, granulated anaerobic sludge, industrial wastewater. African Journal of Biotechnology, Vol.

  8. Metabolic interactions between methanogenic consortia and anaerobic respiring bacteria

    DEFF Research Database (Denmark)

    Stams, A.J.; Oude Elferink, S.J.; Westermann, Peter

    2003-01-01

    Most types of anaerobic respiration are able to outcompete methanogenic consortia for common substrates if the respective electron acceptors are present in sufficient amounts. Furthermore, several products or intermediate compounds formed by anaerobic respiring bacteria are toxic to methanogenic...

  9. Improving Project Outcomes and Growing the Anaerobic Digestion Industy Report

    Science.gov (United States)

    Anaerobic digestion ombudsmen assist with project development, ensure the long-term sustainability of projects, and help advance the industry. This report explores the benefits of anaerobic digestion ombudsmen and provides guidance for implementing them.

  10. Robust regulation of anaerobic digestion processes.

    Science.gov (United States)

    Mailleret, L; Bernard, O; Steyer, J P

    2003-01-01

    This paper deals with the problem of controlling anaerobic digestion processes. A two-step (i.e. acidogenesis-methanization) mass balance model is considered for a 1 m3 fixed bed digester treating industrial wine distillery wastewater. The control law aims at regulating the organic pollution level while avoiding washout of biomass. To this end, a simple output feedback controller is considered which regulates a variable strongly related to the Chemical Oxygen Demand (COD). Numerical simulations assuming noisy measurements first illustrate the robustness of this control procedure. Then, the regulating procedure is implemented on the considered anaerobic digestion process in order to validate and demonstrate its efficiency in real life experiments.

  11. Anaerobic bacteria, the colon and colitis.

    Science.gov (United States)

    Roediger, W E

    1980-02-01

    Anaerobic bacteria constitute more than 90% of the bacteria in the colon. An anaerobic environment is needed to maintain their growth and the production of short-chain fatty acids by these bacteria from carbohydrates. Short-chain fatty acids are rapidly absorbed and essential for metabolic as well as functional welfare of the colonic mucosa. The importance of these acids in water absorption and in the patogenesis of colitis is discussed in relation to the concept of "energy deficiency diseases" of the colonic mucosa.

  12. The Pasteur effect in facultative anaerobic metazoa.

    Science.gov (United States)

    Schmidt, H; Kamp, G

    1996-05-15

    The existence and the regulatory mechanisms of the Pasteur effect in facultative anaerobic metazoa are discussed. There are three reasons for the controversy surrounding this phenomenon. 1) The different definitions of the Pasteur effect, 2) the antagonistic effect of metabolic depression and its species specific response to hypoxia, as well as 3) the laboratory-specific differences in the experimental procedures for analyzing the Pasteur effect and its regulation. This review aims to clarify the confusion about the existence of the Pasteur effect in facultative anaerobic metazoa and to offer possible molecular mechanisms.

  13. The Influence of Hydration on Anaerobic Performance: A Review

    Science.gov (United States)

    Kraft, Justin A.; Green, James M.; Bishop, Phillip A.; Richardson, Mark T.; Neggers, Yasmin H.; Leeper, James D.

    2012-01-01

    This review examines the influence of dehydration on muscular strength and endurance and on single and repeated anaerobic sprint bouts. Describing hydration effects on anaerobic performance is difficult because various exercise modes are dominated by anaerobic energy pathways, but still contain inherent physiological differences. The critical…

  14. Stability of anaerobic reactors under micro-aeration conditions

    Energy Technology Data Exchange (ETDEWEB)

    Fernandez-Polanco, M.; Perez, S.; Diaz, I.; Fernandez-Polanco, F.

    2009-07-01

    Oxidation of sulphide in anaerobic bioreactors by introducing limited amounts of oxygen provides a relatively simple strategy for reducing the levels of sulphite in anaerobic digesters (biogas and effluent). The introduction of limited amounts of air is a general practice in agricultural anaerobic digesters, it is estimated that worldwide over 3.000 units are operated under such conditions. (Author)

  15. Anaerobic Digestion. Student Manual. Biological Treatment Process Control.

    Science.gov (United States)

    Carnegie, John W., Ed.

    This student manual contains the textual material for a four-lesson unit on anaerobic digestion control. Areas addressed include: (1) anaerobic sludge digestion (considering the nature of raw sludge, purposes of anaerobic digestion, the results of digestion, types of equipment, and other topics); (2) digester process control (considering feeding…

  16. The IWA Anaerobic Digestion Model No 1 (ADM1)

    NARCIS (Netherlands)

    Batstone, D.J.; Keller, J.; Angelidaki, I.; Kalyuzhnyi, S.V.; Pavalostathis, S.G.; Rozzi, A.; Sanders, W.T.M.; Siegrist, H.; Vavilin, V.A.

    2002-01-01

    The IWA Anaerobic Digestion Modelling Task Group was established in 1997 at the 8th World Congress on Anaerobic Digestion (Sendai, Japan) with the goal of developing a generalised anaerobic digestion model. The structured model includes multiple steps describing biochemical as well as

  17. The IWA Anaerobic digestion model no 1. (ADM1)

    DEFF Research Database (Denmark)

    Batstone, Damien J.; Keller, J.; Angelidaki, Irini

    2002-01-01

    The IWA Anaerobic Digestion Modelling Task Group was established in 1997 at the 8th World Congress on Anaerobic Digestion (Sendai, Japan) with the goal of developing a generalised anaerobic digestion model. The structured model includes multiple steps describing biochemical as well...

  18. High-throughput Screening of the Enantioselectivity of Hyperthermophilic Mutant Esterases from Archaeon Aeropyrum pernix K1 for Resolution of (R,S)-2-Octanol Acetate

    Institute of Scientific and Technical Information of China (English)

    ZHANG Gui-rong; GAO Ren-jun; ZHANG Ai-jun; RAO Lang; CAO Shu-gui

    2007-01-01

    To identify the desired hyperthermophilic variants within a mutant esterase library for the resolution of (R,S)-2-octanol acetate, a simple, reliable, and versatile method was developed in this study. We built a screening strategy including two steps, first we selected agar plate with substrate to screen the enzymatic activity; secondly we used a pH indicator to screen the enantioselectivity. This method could rapidly detect favorable mutants with high activity and enantioselectivity. A total of 96.2% of tedious screening work can be precluded using this screening strategy. It is an effective screening for alkyl ester and can be applied to relative screening researches. The four improved mutants were screened from the mutant esterase library. Their enantioselectivities, activities, and structures were investigated at different temperatures.

  19. Dynamics in Thermotoga neapolitana adenylate kinase: 15N relaxation and hydrogen-deuterium exchange studies of a hyperthermophilic enzyme highly active at 30 degrees C.

    Science.gov (United States)

    Krishnamurthy, Harini; Munro, Kim; Yan, Honggao; Vieille, Claire

    2009-03-31

    Backbone conformational dynamics of Thermotoga neapolitana adenylate kinase in the free form (TNAK) and inhibitor-bound form (TNAK*Ap5A) were investigated at 30 degrees C using (15)N NMR relaxation measurements and NMR monitored hydrogen-deuterium exchange. With kinetic parameters identical to those of Escherichia coli AK (ECAK) at 30 degrees C, TNAK is a unique hyperthermophilic enzyme. These catalytic properties make TNAK an interesting and novel model to study the interplay between protein rigidity, stability, and activity. Comparison of fast time scale dynamics (picosecond to nanosecond) in the open and closed states of TNAK and ECAK at 30 degrees C reveals a uniformly higher rigidity across all domains of TNAK. Within this framework of a rigid TNAK structure, several residues located in the AMP-binding domain and in the core-lid hinge regions display high picosecond to nanosecond time scale flexibility. Together with the recent comparison of ECAK dynamics with those of hyperthermophilic Aquifex aeolicus AK (AAAK), our results provide strong evidence for the role of picosecond to nanosecond time scale fluctuations in both stability and activity. In the slow time scales, TNAK's increased rigidity is not uniform but localized in the AMP-binding and lid domains. The core domain amides of ECAK and TNAK in the open and closed states show comparable protection against exchange. Significantly, the hinges framing the lid domain show similar exchange data in ECAK and TNAK open and closed forms. Our NMR relaxation and hydrogen-deuterium exchange studies therefore suggest that TNAK maintains high activity at 30 degrees C by localizing flexibility to the hinge regions that are key to facilitating conformational changes.

  20. Environmental impacts of anaerobic digestion and the use of anaerobic residues as soil amendment

    Energy Technology Data Exchange (ETDEWEB)

    Mosey, F.E. [VFA Services Ltd., Herts (United Kingdom)

    1996-01-01

    This paper defines the environmental role of anaerobic digestion within the overall objective of recovering energy from renewable biomass resources. Examples and opportunities for incorporating anaerobic digestion into biomass-to-energy schemes are discussed, together with environmental aspects of anaerobic digestion plants. These include visual, public amenity, pathogens and public health, odor control, and gaseous emissions. Digestate disposal and the benefits of restrictions on recycling organic wastes and biomass residues back to the land are discussed, particularly as they relate to American and European codes of practice and environmental legislation. The paper concludes that anaerobic digestion, if performed in purpose-designed reactors that efficiently recover and use biogas, is an environmentally benign process that can enhance energy recovery and aid the beneficial land use of plant residues in many biomass-to-energy schemes.

  1. Studies on upflow anaerobic filter

    Science.gov (United States)

    Varandani, Nanik Sobhraj

    The thesis presents a critical review of the available literature on the various studies carried out on various aspects of Upflow Anaerobic Filter (UAF) throughout the world. Young and McCarty (1969) did the pioneering work in developing UAF in 1969, since then several studies have been carried out by different researchers using different substrates under different operating conditions and variety of supporting media. However, the most significant modification of the original reactor developed by Young and McCarty (1968), has been the development and use of high porosity media. The use of high porosity media, in fact, has changed the character of the reactor, from basically a fixed film reactor to a fixed film reactor in which the contribution by the suspended bio-solids, entrapped in the numerous media pores, in the substrate removal is quite significant that is to say that the reactor no longer remains a biological reactor which can be modeled and designed on the basis of biofilm kinetics only. The thesis presents an attempt to validate the developed mathematical model(s) by using the laboratory scale reactor performance data and the calculated values of reaction kinetic and bio-kinetic constants. To simplify the verification process, computer programmes have been prepared using the "EXCELL" software and C language. The results of the "EXCELL" computer program runs are tabulated at table no. 7.1 to 7.5. The verification of various mathematical models indicate that the model III B, i.e. Non ideal plug flow model assumed to consist of Complete Mix Reactors in series based on reaction kinetics, gives results with least deviation from the real situation. An interesting observation being that the model offers least deviation or nearly satisfies the real situation for a particular COD removal efficiency, for a particular OLR, eg. the least deviations are obtained at COD removal efficiency of 89% for OLR 2, 81.5% for OLR 4, 78.5% for OLR 6 . However, the use of the

  2. A simple and sensitive quality control method of the anaerobic atmosphere for identification and antimicrobial susceptibility testing of anaerobic bacteria

    DEFF Research Database (Denmark)

    Justesen, Tage; Justesen, Ulrik Stenz

    2013-01-01

    The maintenance of a strict anaerobic atmosphere is essential for the culture of strict anaerobic bacteria. We describe a simple and sensitive quality control method of the anaerobic atmosphere, based on the measurement of the zone diameter around a 5-μg metronidazole disk when testing an aerotol...

  3. Sulfate-reducing bacteria in anaerobic bioreactors

    NARCIS (Netherlands)

    Oude Elferink, S.J.W.H.

    1998-01-01

    The treatment of industrial wastewaters containing high amounts of easily degradable organic compounds in anaerobic bioreactors is a well-established process. Similarly, wastewaters which in addition to organic compounds also contain sulfate can be treated in this way. For a long time, the

  4. Anaerobic work capacity in elite wheelchair athletes

    NARCIS (Netherlands)

    van der Woude, L H; Bakker, W H; Elkhuizen, J W; Veeger, DirkJan (H. E. J.); Gwinn, T

    1997-01-01

    To study the anaerobic work capacity in wheelchair athletes, 67 elite wheelchair athletes (50 male) were studied in a 30-second sprint test on a computer-controlled wheelchair ergometer during the World Championships and Games for the Disabled in Assen (1990). The experimental set-up (ergometer,

  5. Anaerobic Toxicity of Cationic Silver Nanoparticles

    Science.gov (United States)

    The microbial toxicity of silver nanoparticles (AgNPs) stabilized with different capping agents was compared to that of Ag+ under anaerobic conditions. Three AgNPs were investigated: (1) negatively charged citrate-coated AgNPs (citrate-AgNPs), (2) minimally charged p...

  6. Anaerobic Pre-treatment of Strong Sewage

    NARCIS (Netherlands)

    Halalsheh, M.M.

    2002-01-01

    The main objective of this research was to assess the feasibility of applying low cost anaerobic technology for the treatment of relatively high strength sewage of Jordan using two-stage and one-stage UASB reactors operated at ambient temperatures. The wastewater produced in Jordan is characterised

  7. Potential for anaerobic conversion of xenobiotics

    DEFF Research Database (Denmark)

    Mogensen, Anders Skibsted; Dolfing, J.; Haagensen, Frank

    2003-01-01

    regarding the treatment of xenobiotics is shown with the focus on the UASB reactor, but the applicability of other reactor designs for treatment of hazardous waste is also included. Bioaugmentation has proved to be a viable technique to enhance a specific activity in anaerobic reactors and recent research...

  8. An anaerobic mitochondrion that produces hydrogen

    NARCIS (Netherlands)

    Boxma, B.; Graaf, de R.M.; Staay, van der G.W.M.; Alen, T.A.; Richard, G.; Gabalon, T.; Hoek, van A.H.A.M.; Moon - van der Staay, S.Y.; Koopman, W.J.H.; Hellemond, van J.J.; Tielens, A.G.M.; Friedrich, T.; Veenhuis, M.; Huynen, M.A.; Hackstein, J.H.P.

    2005-01-01

    Hydrogenosomes are organelles that produce ATP and hydrogen(1), and are found in various unrelated eukaryotes, such as anaerobic flagellates, chytridiomycete fungi and ciliates(2). Although all of these organelles generate hydrogen, the hydrogenosomes from these organisms are structurally and metabo

  9. Hemicellulases from anaerobic thermophiles. Progress report

    Energy Technology Data Exchange (ETDEWEB)

    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.

  10. Anaerobic membrane bioreactors: Are membranes really necessary?

    NARCIS (Netherlands)

    Davila, M.; Kassab, G.; Klapwijk, A.; Lier, van J.B.

    2008-01-01

    Membranes themselves represent a significant cost for the full scale application of anaerobic membrane bioreactors (AnMBR). The possibility of operating an AnMBR with a self-forming dynamic membrane generated by the substances present in the reactor liquor would translate into an important saving. A

  11. Sulfate-reducing bacteria in anaerobic bioreactors.

    NARCIS (Netherlands)

    Oude Elferink, S.J.W.H.

    1998-01-01

    The treatment of industrial wastewaters containing high amounts of easily degradable organic compounds in anaerobic bioreactors is a well-established process. Similarly, wastewaters which in addition to organic compounds also contain sulfate can be treated in this way. For a long time, the occurrenc

  12. Conversion of Methanogenic Substrates in Anaerobic Reactors

    NARCIS (Netherlands)

    Gonzalez-Gil, G.

    2000-01-01

    The EGSB systems represents an attractive option to extend further the use of anaerobic technology for wastewater treatment, particularly with respect to waste streams originating from chemical industries. Frequently chemical waste streams are unbalanced with respect to nutrients and/or micronutrien

  13. Anaerobic microbial LCFA degradation in bioreactors

    NARCIS (Netherlands)

    Sousa, D.Z.; Pereira, M.A.; Alves, J.I.; Smidt, H.; Stams, A.J.M.; Alves, M.M.

    2008-01-01

    This paper reviews recent results obtained on long-chain fatty acids (LCFA) anaerobic degradation. Two LCFA were used as model substrates: oleate, a mono-unsaturated LCFA, and palmitate, a saturated LCFA, both abundant in LCFA-rich wastewaters. 16S rRNA gene analysis of sludge samples submitted to c

  14. Anaerobic work capacity in elite wheelchair athletes

    NARCIS (Netherlands)

    van der Woude, L H; Bakker, W H; Elkhuizen, J W; Veeger, DirkJan (H. E. J.); Gwinn, T

    1997-01-01

    To study the anaerobic work capacity in wheelchair athletes, 67 elite wheelchair athletes (50 male) were studied in a 30-second sprint test on a computer-controlled wheelchair ergometer during the World Championships and Games for the Disabled in Assen (1990). The experimental set-up (ergometer, pro

  15. Anaerobic digestion in sustainable biomass chains

    NARCIS (Netherlands)

    Pabon Pereira, C.P.

    2009-01-01

    This thesis evaluates the potential contribution of anaerobic digestion (AD) to the sustainability of biomass chains. Results provide insights in the technological potential to recover energy and valuable by-products from energy crops and residues, and evaluate biomass cascades involving AD technolo

  16. Essential metal depletion in an anaerobic reactor

    NARCIS (Netherlands)

    Osuna, M.B.; Iza, J.M.; Zandvoort, M.H.; Lens, P.N.L.

    2003-01-01

    The effect of the absence of trace elements on the conversion of a mixture of volatile fatty acids by a distillery anaerobic granular sludge was investigated. Two UASB reactors were operated under identical operational conditions except for the influent trace metal concentrations, during 140 days. E

  17. Anaerobic effluent disinfection using ozone: Byproducts formation

    NARCIS (Netherlands)

    Silva, G.H.R.; Daniel, L.A.; Bruning, H.; Rulkens, W.H.

    2010-01-01

    This research was aimed at studying oxidation processes, coliform inactivation effectiveness and disinfection byproducts (DBPs) associated with the disinfection of anaerobic sanitary wastewater effluent with ozone applied at doses of 5.0, 8.0 and 10.0mg O(3)L(-1) for contact times of 5, 10 and 15 mi

  18. Anaerobic Toxicity of Cationic Silver Nanoparticles

    Science.gov (United States)

    The microbial toxicity of silver nanoparticles (AgNPs) stabilized with different capping agents was compared to that of Ag+ under anaerobic conditions. Three AgNPs were investigated: (1) negatively charged citrate-coated AgNPs (citrate-AgNPs), (2) minimally charged p...

  19. Anaerobic effluent disinfection using ozone: Byproducts formation

    NARCIS (Netherlands)

    Silva, G.H.R.; Daniel, L.A.; Bruning, H.; Rulkens, W.H.

    2010-01-01

    This research was aimed at studying oxidation processes, coliform inactivation effectiveness and disinfection byproducts (DBPs) associated with the disinfection of anaerobic sanitary wastewater effluent with ozone applied at doses of 5.0, 8.0 and 10.0mg O(3)L(-1) for contact times of 5, 10 and 15 mi

  20. The fate of methanol in anaerobic bioreactors

    NARCIS (Netherlands)

    Florencio, L.

    1994-01-01

    Methanol is an important component of certain industrial wastewaters. In anaerobic environments, methanol can be utilized by methanogens and acetogens. In wastewater treatment plants, the conversion of methanol into methane is preferred because this conversion is responsible for chemical

  1. Teleosts in hypoxia : Aspects of anaerobic metabolism

    NARCIS (Netherlands)

    Van den Thillart, G.; van Waarde, Aren

    1985-01-01

    Moderate hypoxia can be tolerated by many fish species, while only some species survive severe hypoxia or anoxia. Hypoxia usually activates anaerobic glycolysis, which may be temporary when the animals are able to improve their oxygen extraction capacity. Switching over to aerobic metabolism allows

  2. Anaerobic fitness tests: what are we measuring?

    Science.gov (United States)

    Van Praagh, Emmanuel

    2007-01-01

    Anaerobic fitness, during growth and development, has not received the same attention from researchers as aerobic fitness. This is surprising given the level of anaerobic energy used daily during childhood and adolescence. During physical activity and sport, the child is spontaneously more attracted to short-burst movements than to long-term activities. It is, however, well known that in anaerobic activities such as sprint cycling, sprint running or sprint swimming, the child's performance is distinctly poorer than that of the adult. This partly reflects the child's lesser ability to generate mechanical energy from chemical energy sources during short-term high-intensity work or exercise. Direct measurements of the rate or capacity of anaerobic pathways for energy turnover presents several ethical and methodological difficulties. Therefore, rather than measure energy supply, pediatric exercise scientists have concentrated on measuring short-term power output by means of standardized protocol tests such as short-term cycling power tests, running tests or vertical jump tests. There is, however, no perfect test and, therefore, it is important to acknowledge the benefits and limitations of each testing method. Mass-related short-term power output was shown to increase dramatically during growth and development, whereas the corresponding increase in peak blood lactate was considerably lower. This suggests that the observed difference between children and adolescents during short-term power output testing may be related to neuromuscular factors, hormonal factors and improved motor coordination.

  3. Biodegradability of leathers through anaerobic pathway.

    Science.gov (United States)

    Dhayalan, K; Fathima, N Nishad; Gnanamani, A; Rao, J Raghava; Nair, B Unni; Ramasami, T

    2007-01-01

    Leather processing generates huge amounts of both solid and liquid wastes. The management of solid wastes, especially tanned leather waste, is a challenging problem faced by tanners. Hence, studies on biodegradability of leather become imperative. In this present work, biodegradability of untanned, chrome tanned and vegetable tanned leather under anaerobic conditions has been addressed. Two different sources of anaerobes have been used for this purpose. The effect of detanning as a pretreatment method before subjecting the leather to biodegradation has also been studied. It has been found that vegetable tanned leather leads to more gas production than chrome tanned leather. Mixed anaerobic isolates when employed as an inoculum are able to degrade the soluble organics of vegetable tanned material and thus exhibit an increased level of gas production during the initial days, compared to the results of the treatments that received the anaerobic sludge. With chrome tanned materials, there was not much change in the volume of the gas produced from the two different sources. It has been found that detanning tends to improve the biodegradability of both types of leathers.

  4. The fate of methanol in anaerobic bioreactors.

    NARCIS (Netherlands)

    Florencio, L.

    1994-01-01

    Methanol is an important component of certain industrial wastewaters. In anaerobic environments, methanol can be utilized by methanogens and acetogens. In wastewater treatment plants, the conversion of methanol into methane is preferred because this conversion is responsible for chemical oxygen dema

  5. Essential metal depletion in an anaerobic reactor.

    NARCIS (Netherlands)

    Osuna, M.B.; Iza, J.M.; Zandvoort, M.H.; Lens, P.N.L.

    2003-01-01

    The effect of the absence of trace elements on the conversion of a mixture of volatile fatty acids by a distillery anaerobic granular sludge was investigated. Two UASB reactors were operated under identical operational conditions except for the influent trace metal concentrations, during 140 days. E

  6. Anaerobic Digestion in a Flooded Densified Leachbed

    Science.gov (United States)

    Chynoweth, David P.; Teixeira, Arthur A.; Owens, John M.; Haley, Patrick J.

    2009-01-01

    A document discusses the adaptation of a patented biomass-digesting process, denoted sequential batch anaerobic composting (SEBAC), to recycling of wastes aboard a spacecraft. In SEBAC, high-solids-content biomass wastes are converted into methane, carbon dioxide, and compost.

  7. Intraspecific variation in aerobic and anaerobic locomotion

    DEFF Research Database (Denmark)

    Svendsen, Jon Christian; Tirsgård, Bjørn; Cordero, Gerardo A.;

    2015-01-01

    Intraspecific variation and trade-off in aerobic and anaerobic traits remain poorly understood in aquatic locomotion. Using gilthead sea bream (Sparus aurata) and Trinidadian guppy (Poecilia reticulata), both axial swimmers, this study tested four hypotheses: (1) gait transition from steady...

  8. Anaerobic degradation of linear alkylbenzene sulfonate.

    Science.gov (United States)

    Mogensen, Anders S; Haagensen, Frank; Ahring, Birgitte K

    2003-04-01

    Linear alkylbenzene sulfonate (LAS) found in wastewater is removed in the wastewater treatment facilities by sorption and aerobic biodegradation. The anaerobic digestion of sewage sludge has not been shown to contribute to the removal. The concentration of LAS based on dry matter typically increases during anaerobic stabilization due to transformation of easily degradable organic matter. Hence, LAS is regarded as resistant to biodegradation under anaerobic conditions. We present data from a lab-scale semi-continuously stirred tank reactor (CSTR) spiked with linear dodecylbenzene sulfonate (C12 LAS), which show that C12 LAS was biodegradable under methanogenic conditions. Sorption of C12 LAS on sewage sludge was described with a Freundlich isotherm. The C12 LAS sorption was determined with different concentrations of total solids (TS). In the semi-continuously stirred tank reactor, 18% of the added C12 LAS was bioavailable and 20% was biotransformed when spiking with 100 mg/L of C12 LAS and a TS concentration of 14.2 mg/L. Enhanced bioavailability of C12 LAS was obtained in an upflow anaerobic sludge blanket (UASB) reactor inoculated with granular sludge and sewage sludge. Biodegradation under thermophilic conditions was 37% with LAS as sole carbon source. Benzaldehyde was produced in the UASB reactor during LAS transformation.

  9. Anaerobic hydrolysis during digestion of complex substrates

    NARCIS (Netherlands)

    Sanders, W.T.M.

    2001-01-01

    Complex waste(water) such as, raw sewage, dairy wastewater, slaughterhouse wastewater, fish processing wastewater, primary sludge and the organic fraction of municipal solid waste have been proven to be degradable under anaerobic conditions. However, during the digestion process the conversion of th

  10. EFFECTS OF ECCENTRIC EXERCISE ON ANAEROBIC POWER, STARTING SPEED AND ANAEROBIC ENDURANCE

    OpenAIRE

    Maciejczyk, Marcin; Szymura, Jadwiga; Wiecek, Magdalena; Szygula, Zbigniew; Kepinska, Magdalena; Ochalek, Katarzyna; Pokrywka, Andrzej

    2015-01-01

    The aim of this study was to evaluate the effects of eccentric exercise on anaerobic power, starting speed and anaerobic endurance. The participants performed the maximum cycling sprint test (MCST) prior to eccentric exercise (ECC), 10 minutes after, as well as one hour, 24 hours, 48 hours, and one week after ECC. The peak and mean power, time to attain peak power, time of maintaining peak power and power decrease were measured in the MCST. Before and after ECC, the myoglobin concentration...

  11. Anaerobic benzene oxidation by Geobacter species.

    Science.gov (United States)

    Zhang, Tian; Bain, Timothy S; Nevin, Kelly P; Barlett, Melissa A; Lovley, Derek R

    2012-12-01

    The abundance of Geobacter species in contaminated aquifers in which benzene is anaerobically degraded has led to the suggestion that some Geobacter species might be capable of anaerobic benzene degradation, but this has never been documented. A strain of Geobacter, designated strain Ben, was isolated from sediments from the Fe(III)-reducing zone of a petroleum-contaminated aquifer in which there was significant capacity for anaerobic benzene oxidation. Strain Ben grew in a medium with benzene as the sole electron donor and Fe(III) oxide as the sole electron acceptor. Furthermore, additional evaluation of Geobacter metallireducens demonstrated that it could also grow in benzene-Fe(III) medium. In both strain Ben and G. metallireducens the stoichiometry of benzene metabolism and Fe(III) reduction was consistent with the oxidation of benzene to carbon dioxide with Fe(III) serving as the sole electron acceptor. With benzene as the electron donor, and Fe(III) oxide (strain Ben) or Fe(III) citrate (G. metallireducens) as the electron acceptor, the cell yields of strain Ben and G. metallireducens were 3.2 × 10(9) and 8.4 × 10(9) cells/mmol of Fe(III) reduced, respectively. Strain Ben also oxidized benzene with anthraquinone-2,6-disulfonate (AQDS) as the sole electron acceptor with cell yields of 5.9 × 10(9) cells/mmol of AQDS reduced. Strain Ben serves as model organism for the study of anaerobic benzene metabolism in petroleum-contaminated aquifers, and G. metallireducens is the first anaerobic benzene-degrading organism that can be genetically manipulated.

  12. Winery and distillery wastewater treatment by anaerobic digestion.

    Science.gov (United States)

    Moletta, R

    2005-01-01

    Anaerobic digestion is widely used for wastewater treatment, especially in the food industries. Generally after the anaerobic treatment there is an aerobic post-treatment in order to return the treated water to nature. Several technologies are applied for winery wastewater treatment. They are using free cells or flocs (anaerobic contact digesters, anaerobic sequencing batch reactors and anaerobic lagoons), anaerobic granules (Upflow Anaerobic Sludge Blanket--UASB), or biofilms on fixed support (anaerobic filter) or on mobile support as with the fluidised bed. Some technologies include two strategies, e.g. a sludge bed with anaerobic filter as in the hybrid digester. With winery wastewaters (as for vinasses from distilleries) the removal yield for anaerobic digestion is very high, up to 90-95% COD removal. The organic loads are between 5 and 15 kgCOD/m3 of digester/day. The biogas production is between 400 and 600 L per kg COD removed with 60 to 70% methane content. For anaerobic and aerobic post-treatment of vinasses in the Cognac region, REVICO company has 99.7% COD removal and the cost is 0.52 Euro/m3 of vinasses.

  13. Anaerobic protozoa and their growth in biomethanation systems.

    Science.gov (United States)

    Priya, M; Haridas, Ajit; Manilal, V B

    2008-04-01

    This study was to investigate growth of protozoa and its influence on biodegradation in anaerobic treatment systems. It was done by specifically controlling and monitoring growth of protozoa versus degradation in continuous stirred anaerobic reactors and batch anaerobic reactors. Occurrence of a diverse protozoa population such as the ciliates, Prorodon, Vorticella, Cyclidium, Spathidium, Loxodes, Metopus were observed in stable anaerobic systems and the flagellates, Rhynchomonas, Naeglaria, Amoeboflagellates, Tetramitus, Trepomonas and Bodo during increased VFA concentration and affected periods of biomethanation. The abundance of ciliates in the anaerobic system had significant correlation with the reduction of MLSS, increased rate of COD removal and higher methane production. The results of this study thus tend to relate increased anaerobic degradation with the abundance of protozoa, mainly ciliates, which indicate their possible involvement in the process. Present study also reveals that performance of anaerobic process can be assessed by monitoring the protozoa population in the system.

  14. Affinity of ribosomal protein S8 from mesophilic and (hyper)thermophilic archaea and bacteria for 16S rRNA correlates with the growth temperatures of the organisms.

    Science.gov (United States)

    Gruber, Thomas; Köhrer, Caroline; Lung, Birgit; Shcherbakov, Dmitri; Piendl, Wolfgang

    2003-08-14

    The ribosomal protein S8 plays a pivotal role in the assembly of the 30S ribosomal subunit. Using filter binding assays, S8 proteins from mesophilic, and (hyper)thermophilic species of the archaeal genus Methanococcus and from the bacteria Escherichia coli and Thermus thermophilus were tested for their affinity to their specific 16S rRNA target site. S8 proteins from hyperthermophiles exhibit a 100-fold and S8 from thermophiles exhibit a 10-fold higher affinity than their mesophilic counterparts. Thus, there is a striking correlation of affinity of S8 proteins for their specific RNA binding site and the optimal growth temperatures of the respective organisms. The stability of individual rRNA-protein complexes might modulate the stability of the ribosome, providing a maximum of thermostability and flexibility at the growth temperature of the organism.

  15. ISOLATION OF ANAEROBES IN DEEP SEATED PRESSURE ULCERS USING A NOVEL INNOVATIVE TECHNIQUE OF ANAEROBE ISOLATION

    Directory of Open Access Journals (Sweden)

    Lalbiaktluangi

    2015-12-01

    Full Text Available BACKGROUND Isolation of an anaerobe is usually neglected in hospitals with limited resources due to the expensive and complicated technique of anaerobic isolation methods, which is difficult to arrange in such resource poor settings. Conventionally adopted anaerobic culture methods such as Anaerobic jar, Gas-Pak, Anoxomat or Automated glove-box systems are extremely costly and cumbersome for single unit testing, but not suitable for small scale laboratories. However, anaerobic bacteria are not to be overlooked as they have made a comeback in clinical settings and are even showing resistance to Metronidazole, once thought to be the gold standard bullet against anaerobes. Deep seated pressure ulcers are usually the site where anaerobe causes an infection in synergy with aerobes. AIMS AND OBJECTIVES Isolation of anaerobes in deep seated pressure ulcers using a novel innovative technique and to study their antibiogram profile. MATERIALS AND METHODS Swabs taken from depth of deep seated pressure ulcers were immediately inoculated in Brucella blood agar at bedside and placed in polycarbonate airtight jar for anaerobic incubation using a novel innovative Modified Candle Jar technique. In this technique five grams of grease-free grade zero steel wool were dipped in 50ml freshly prepared acidified copper sulphate solution until the copper colour appeared. Excess solution was drained and the steel wool was moulded into a loose pad to fit on an open Petri plate placed on top of the inoculated Brucella blood agar plates. A white-wax candle was placed at the centre of this plate. A small test tube containing mixture of 0.5g sodium-bicarbonate and 0.5g magnesium carbonate was kept ready to be placed inside the jar, just after placing the inoculated plate and incubated for 48 hours. RESULTS Peptostreptococcus anaerobius and Bacteroides fragilis were successfully isolated from deep seated pressure ulcers by this method. Antibiogram studies were done using the

  16. Viruses of hyperthermophilic Crenarchaea

    DEFF Research Database (Denmark)

    Prangishvili, D.; Garrett, R. A.

    2005-01-01

    Since the discovery of the Archaea - the third domain of life - by Woese and colleagues in 1977, the subsequent developments in molecular and cell biology, and also genomics, have strongly reinforced the view that archaea and eukarya co-evolved, separately from bacteria, over a long time. However......, when one examines the archaeal viruses, the picture appears complex. Most viruses that are known to infect members of the kingdom Euryarchaeota resemble bacterial viruses, whereas those associated with the kingdom Crenarchaeota show little resemblance to either bacterial or eukaryal viruses...

  17. In situ detection of anaerobic alkane metabolites in subsurface environments

    Directory of Open Access Journals (Sweden)

    Lisa eGieg

    2013-06-01

    Full Text Available Alkanes comprise a substantial fraction of crude oil and refined fuels. As such, they are prevalent within deep subsurface fossil fuel deposits and in shallow subsurface environments such as aquifers that are contaminated with hydrocarbons. These environments are typically anaerobic, and host diverse microbial communities that can potentially use alkanes as substrates. Anaerobic alkane biodegradation has been reported to occur under nitrate-reducing, sulfate-reducing, and methanogenic conditions. Elucidating the pathways of anaerobic alkane metabolism has been of interest in order to understand how microbes can be used to remediate contaminated sites. Alkane activation primarily occurs by addition to fumarate, yielding alkylsuccinates, unique anaerobic metabolites that can be used to indicate in situ anaerobic alkane metabolism. These metabolites have been detected in hydrocarbon-contaminated shallow aquifers, offering strong evidence for intrinsic anaerobic bioremediation. Recently, studies have also revealed that alkylsuccinates are present in oil and coal seam production waters, indicating that anaerobic microbial communities can utilize alkanes in these deeper subsurface environments. In many crude oil reservoirs, the in situ anaerobic metabolism of hydrocarbons such as alkanes may be contibuting to modern-day detrimental effects such as oilfield souring, or may lead to more benefical technologies such as enhanced energy recovery from mature oilfields. In this review, we briefly describe the key metabolic pathways for anaerobic alkane (including n-alkanes, isoalkanes, and cyclic alkanes metabolism and highlight several field reports wherein alkylsuccinates have provided evidence for anaerobic in situ alkane metabolism in shallow and deep subsurface environments.

  18. In situ detection of anaerobic alkane metabolites in subsurface environments.

    Science.gov (United States)

    Agrawal, Akhil; Gieg, Lisa M

    2013-01-01

    Alkanes comprise a substantial fraction of crude oil and refined fuels. As such, they are prevalent within deep subsurface fossil fuel deposits and in shallow subsurface environments such as aquifers that are contaminated with hydrocarbons. These environments are typically anaerobic, and host diverse microbial communities that can potentially use alkanes as substrates. Anaerobic alkane biodegradation has been reported to occur under nitrate-reducing, sulfate-reducing, and methanogenic conditions. Elucidating the pathways of anaerobic alkane metabolism has been of interest in order to understand how microbes can be used to remediate contaminated sites. Alkane activation primarily occurs by addition to fumarate, yielding alkylsuccinates, unique anaerobic metabolites that can be used to indicate in situ anaerobic alkane metabolism. These metabolites have been detected in hydrocarbon-contaminated shallow aquifers, offering strong evidence for intrinsic anaerobic bioremediation. Recently, studies have also revealed that alkylsuccinates are present in oil and coal seam production waters, indicating that anaerobic microbial communities can utilize alkanes in these deeper subsurface environments. In many crude oil reservoirs, the in situ anaerobic metabolism of hydrocarbons such as alkanes may be contributing to modern-day detrimental effects such as oilfield souring, or may lead to more beneficial technologies such as enhanced energy recovery from mature oilfields. In this review, we briefly describe the key metabolic pathways for anaerobic alkane (including n-alkanes, isoalkanes, and cyclic alkanes) metabolism and highlight several field reports wherein alkylsuccinates have provided evidence for anaerobic in situ alkane metabolism in shallow and deep subsurface environments.

  19. Atrazine removal in Danish anaerobic aquifers

    DEFF Research Database (Denmark)

    Pedersen, Philip Grinder; Arildskov, N.P.; Albrechtsen, Hans-Jørgen

    2002-01-01

    The pesticide atrazine (6-chloro-N-2-ethyl-N-4-isopropyl-1,3,5-triazine -2,4-diamine) was removed from the water phase in anaerobic laboratory batch incubations with sediment and groundwater from a number of Danish anaerobic aquifers, but not in incubations from aerobic aquifers. The removal...... process was abiotic since atrazine was also removed from microbially inhibited autoclaved and chloroform amended controls, although in controls amended with mercury, atrazine removal was slowed down. (ring-U-C-14)- atrazine amended samples showed no mineralization to (CO2)-C-14 or transformation...... to soluble degradation products, indicating that a slow sorption process was responsible for the atrazine removal. Approximately 20% of the applied C-14-atrazine was present in a non-extractable residual sediment bound fraction, indicating the slow sorption process to be in part irreversible...

  20. Instrumentation and Control in Anaerobic Digestion

    DEFF Research Database (Denmark)

    Anaerobic digestion is a multistep process, and is most applied to solids destruction and wastewater treatment for energy production. Despite wide application, and long-term industrial proof of application, some industries are still reluctant to apply this technology. One of the classical reasons...... benchmark. There has therefore been, overall, a quantum advance in application and sophistication of instrumentation and control in anaerobic digestion, and it is an effective option for improved process loading rate and conversion efficiency....... are still a limitation, but this is being partly addressed by the increased complexity of digestion processes. Methods for control benchmarking have also been improved, as there is now an industry standard model (the IWA ADM1), and this is being applied in an improved whole wastewater treatment plant...

  1. Anaerobic treatment of wastewater containing methanol in upflow anaerobic sludge bed (UASB) reactor

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The direct conversion of methanol into methane is the main process in anaerobic treatment of methanol containing wastewater.However,acetic acid can also be produced from methanol theoretically,which may probably result in an abrupt pH drop and deteriorate the anaerobic process.Therefore,it is interesting to know what would really happen in an anaerobic reactor treating methanol wastewater.In this study,an up-flow anaerobic sludge bed (UASB) reactor treating methanol wastewater was operated.The chemical oxygen demand (COD),acetic acid and pH of the effluent were monitored at different loadings and influent alkalinity.The results showed that the anaerobic reactor could be operated steadily at as low as 119 mg/L of influent alkalinity and high organic loading rate with no obvious pH drops.Volatile fatty acids accumulation was not observed even at strong shock loadings.The microorganisms in the sludge at the end of the test became homogeneous in morphology,which were mainly spherical or spheroidal in shape.

  2. Cultivation of anaerobic and facultatively anaerobic bacteria from spacecraft-associated clean rooms.

    Science.gov (United States)

    Stieglmeier, Michaela; Wirth, Reinhard; Kminek, Gerhard; Moissl-Eichinger, Christine

    2009-06-01

    In the course of this biodiversity study, the cultivable microbial community of European spacecraft-associated clean rooms and the Herschel Space Observatory located therein were analyzed during routine assembly operations. Here, we focused on microorganisms capable of growing without oxygen. Anaerobes play a significant role in planetary protection considerations since extraterrestrial environments like Mars probably do not provide enough oxygen for fully aerobic microbial growth. A broad assortment of anaerobic media was used in our cultivation strategies, which focused on microorganisms with special metabolic skills. The majority of the isolated strains grew on anaerobic, complex, nutrient-rich media. Autotrophic microorganisms or microbes capable of fixing nitrogen were also cultivated. A broad range of facultatively anaerobic bacteria was detected during this study and also, for the first time, some strictly anaerobic bacteria (Clostridium and Propionibacterium) were isolated from spacecraft-associated clean rooms. The multiassay cultivation approach was the basis for the detection of several bacteria that had not been cultivated from these special environments before and also led to the discovery of two novel microbial species of Pseudomonas and Paenibacillus.

  3. Enhancing post anaerobic digestion of full-scale anaerobically digested sludge using free nitrous acid treatment.

    Science.gov (United States)

    Zhang, Tingting; Wang, Qilin; Ye, Liu; Yuan, Zhiguo

    2016-05-01

    In some wastewater treatment plants (WWTPs), the ever increasing production of sludge with the expanding population overloaded the anaerobic digestion which compromises the sludge reduction efficiency. Post anaerobic digestion of anaerobically digested sludge (ADS) has been applied to enhance sludge reduction, however, to a very limited extent. This study verified the effectiveness of free nitrous acid (FNA i.e. HNO2) pre-treatment on enhancing full-scale ADS degradation in post anaerobic digestion. The ADS collected from a full-scale WWTP was subject to FNA treatment at concentrations of 0.77, 1.54, 2.31, 3.08, and 3.85 mg N/L for 24 h followed by biochemical methane potential tests. The FNA treatment at all concentrations resulted in an increase (from 1.5-3.1 % compared to the control) in sludge reduction with the highest improvement achieved at 0.77 mg HNO2-N/L. The FNA treatment at this concentration also resulted in the highest increase in methane production (40 %) compared to the control. The economic analysis indicates that FNA treatment is economically attractive for enhancing post anaerobic digestion of full-scale ADS.

  4. Monitoring and control of anaerobic reactors

    DEFF Research Database (Denmark)

    Pind, Peter Frode; Angelidaki, Irini; Ahring, Birgitte Kiær;

    2003-01-01

    control approaches that have been used are comprehensively described. These include simple and adaptive controllers, as well as more recent developments such as fuzzy controllers, knowledge-based controllers and controllers based on neural networks.......The current status in monitoring and control of anaerobic reactors is reviewed. The influence of reactor design and waste composition on the possible monitoring and control schemes is examined. After defining the overall control structure, and possible control objectives, the possible process...

  5. Anaerobic co-digestion of organic wastes

    OpenAIRE

    2009-01-01

    Tese de doutoramento em Engenharia Química e Biológica Anaerobic digestion is an already established process but the increasing need of bio‐waste recovery has determined the emergence of new substrates, revamping the research in this field. Contrary to some other European countries, in Portugal this technology is still scarcely in use. Nonetheless, the current legislation endorses this application as a waste management and as an energy recovery process. The rapid growth of the ...

  6. Conversion of Methanogenic Substrates in Anaerobic Reactors

    OpenAIRE

    Gonzalez-Gil, G.

    2000-01-01

    The EGSB systems represents an attractive option to extend further the use of anaerobic technology for wastewater treatment, particularly with respect to waste streams originating from chemical industries. Frequently chemical waste streams are unbalanced with respect to nutrients and/or micronutrients and furthermore these streams may contain toxic-biodegradable compounds. To reduce toxicity high recycle ratios may be applied as in the case of EGSB reactors however, this at the same time may ...

  7. Vinasses treatment in anaerobic fludized bed reactor.

    Directory of Open Access Journals (Sweden)

    Francisco J. C. Terán

    2009-03-01

    Full Text Available The agricultural use of vinasse produced by the sugar industry has gone through many changes over the years. Coupled with concern over the increased agronomic efficiency and optimizing the management of the use of such waste, you can highlight the major global ecological awareness, developed after 90s. This study aims at the construction and operation of a reactor anaerobic cracker (RALF on pilot scale to verify the burden of chemical demand of oxygen (DQO of vinasse, under mesophilic. The stillage used for feeding the reactor was from a sugar cane processing plant, located in the city of Regente Feijó, São Paulo State. The inoculum was anaerobic sludge from a reactor and upward flow anaerobic sludge blanket (UASB treating wastewater from a factory of soda. The concentrations of vinasse to be treated ranged 17,239 mg DQO L-1 up to 28,174 mg DQO L-1. The effluent pH was maintained between 6.4 and 8.6 during the research. The productivity of biogas in the reactor has not achieved the expected rates, reaching only 46 mL day-1. Maximum efficiency attained during operation was 51.1 %, corresponding to a 14-day operation time, vinasses organic loading of 19.5 kg DQO m-3 dia-1 and to an hydraulic detention time of one day.

  8. Anaerobic Capacity of Sailors with Disabilities

    Directory of Open Access Journals (Sweden)

    Prokopowicz Grzegorz

    2017-03-01

    Full Text Available Introduction. A review of Polish and international literature does not give a clear indication of the level of anaerobic capacity that sailors with disabilities demonstrate with regard to their functional capacities. This study sought to determine differences in functional capacity levels between sailors from three medical and functional groups. Material and methods. The research was carried out during a sports camp at the National Sailing Centre in Górki Zachodnie in 2014. Eighteen males with locomotor disabilities were included in the study. The athletes were members of the National Team of Sailors with Disabilities of the Polish Yachting Association. The sportsmen competed in the Skud 18 and 2.4mR Paralympic classes. A 30-second Wingate test for upper limbs was employed in the study. Results. Significant differences in mean power (MP values were noted between the groups under investigation. The group of wheelchair sailors with improper core stability (A and the group of wheelchair sailors with proper core stability (B had significantly lower scores than the group of study participants who were able to move freely, that is to walk (C. Conclusions. The study revealed that a 30-second anaerobic capacity test performed on an arm ergometer differentiated disabled sailors from selected groups in terms of mean power. Research on anaerobic capacity may be used to verify the current classification in Paralympic sailing and will make it possible to differentiate present competition categories.

  9. Microbial Aspects of Anaerobic BTEX Degradation

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Combined with conventional methods, developments in both geochemical (delineation of redox processes) and molecular microbial methods (analysis of 16S rDNA genes and functional genes) have allowed us to study in details microorganisms and genes involved in the anaerobic degradation of benzene, toluene, ethylbenzene and xylene (BTEX) under specific redox conditions. This review summarizes recent research in this field. The potential for anaerobic BTEX degradation is widely spread. Specific groups of microorganisms appear to be involved in degradation under different redox conditions. Members of the Azoarcus/Thauera cluster perform BTEX degradation under denitrifying conditions, Geobacteraceae under Fe (III) reducing conditions and Desulfobacteriaceae under sulfate reducing conditions. The information so far obtained on biochemistry and molecular genetics of BTEX degradation indicates that each BTEX compound is funneled into the central benzyol-CoA pathway by a different peripheral pathway. The peripheral pathways of per BTEX compound show similarities among different physiological groups of microorganisms. We also describe how knowledge obtained on the microbial aspects of BTEX degradation can be used to enhance and monitor anaerobic BTEX degradation.

  10. Alternating Current Influences Anaerobic Electroactive Biofilm Activity.

    Science.gov (United States)

    Wang, Xin; Zhou, Lean; Lu, Lu; Lobo, Fernanda Leite; Li, Nan; Wang, Heming; Park, Jaedo; Ren, Zhiyong Jason

    2016-09-06

    Alternating current (AC) is known to inactivate microbial growth in suspension, but how AC influences anaerobic biofilm activities has not been systematically investigated. Using a Geobacter dominated anaerobic biofilm growing on the electrodes of microbial electrochemical reactors, we found that high frequency AC ranging from 1 MHz to 1 kHz (amplitude of 5 V, 30 min) showed only temporary inhibition to the biofilm activity. However, lower frequency (100 Hz, 1.2 or 5 V) treatment led to 47 ± 19% permanent decrease in limiting current on the same biofilm, which is attributed to the action of electrohydrodynamic force that caused biofilm damage and loss of intercellular electron transfer network. Confocal microscopy images show such inactivation mainly occurred at the interface between the biofilm and the electrode. Reducing the frequency further to 1 Hz led to water electrolysis, which generated gas bubbles that flushed all attached cells out of the electrode. These findings provide new references on understanding and regulating biofilm growth, which has broader implications in biofouling control, anaerobic waste treatment, energy and product recovery, and general understanding of microbial ecology and physiology.

  11. Anaerobic Treatment of Palm Oil Mill Effluent in Pilot-Scale Anaerobic EGSB Reactor.

    Science.gov (United States)

    Wang, Jin; Mahmood, Qaisar; Qiu, Jiang-Ping; Li, Yin-Sheng; Chang, Yoon-Seong; Li, Xu-Dong

    2015-01-01

    Large volumes of untreated palm oil mill effluent (POME) pose threat to aquatic environment due to the presence of very high organic content. The present investigation involved two pilot-scale anaerobic expanded granular sludge bed (EGSB) reactors, continuously operated for 1 year to treat POME. Setting HRT at 9.8 d, the anaerobic EGSB reactors reduced COD from 71179 mg/L to 12341 mg/L and recycled half of sludge by a dissolved air flotation (DAF). The average effluent COD was 3587 mg/L with the consistent COD removal efficiency of 94.89%. Adding cationic polymer (PAM) dose of 30 mg/L to DAF unit and recycling its half of sludge caused granulation of anaerobic sludge. Bacilli and small coccid bacteria were the dominant microbial species of the reactor. The reactor produced 27.65 m(3) of biogas per m(3) of POME which was utilized for electricity generation.

  12. Anaerobic filters for the treatment of coal gasification wastewater.

    Science.gov (United States)

    Suidan, M T; Siekerka, G L; Kao, S W; Pfeffer, J T

    1983-06-01

    A process train consisting of the following sequence of unit processes, a berl-saddle-packed anaerobic filter, an expanded bed, granular activated carbon anaerobic filter, and an activated sludge nitrification system was evaluated for the treatment of a synthetically prepared coal gasification wastewater. The first-stage anaerobic filter resulted in very little removal of organic matter and no methane production. Excellent reduction in organic matter occurred in the granular activated carbon anaerobic filter. The removal mechanism was initially adsorptive and near the end of the study, removal of organic matter was primarily through conversion to methane gas. It is felt that the success of the activated carbon anaerobic filter was due to the ability of the activated carbon to sequester some components of the wastewater that were toxic to the mixed culture of anaerobic microorganisms. The activated sludge nitrification system resulted in complete ammonia oxidation and was very efficient in final effluent polishing.

  13. METRONIDAZOLE RESISTANCE IN ANAEROBES ISOLATED FROM CHRONIC PERIODONTITIS CASES

    Directory of Open Access Journals (Sweden)

    Sushma Narayan

    2016-01-01

    Full Text Available Periodontitis is the most frequent oral health problem in the world. The infection is primarily caused by anaerobic microorganisms. Metronidazole is the most commonly used drug to treat the infection but recently the anaerobes have shown the resistance to this drug. Therefore, the present study was undertaken to isolate and identify the anaerobes associated with periodontitis and study their susceptibility pattern to the Metronidazole. Total 90 samples were collected from chronic periodontitis cases. Anaerobes were isolated in 71% of periodontitis cases. Gram positive organisms were more predominantly isolated than Gram negative organisms. The Gram negative anaerobes were found to be 100% sensitive to Metronidazole while Gram positive anaerobes showed 8% resistance to Metronidazole.

  14. Adenosine monophosphate-activated protein kinase and myofibrillar protein degradation%腺苷酸活化蛋白激酶与骨骼肌蛋白质降解

    Institute of Scientific and Technical Information of China (English)

    马延超; 朱荣; 李俊平

    2012-01-01

    BACKGROUND: Adenosine monophosphate-activated protein kinase (AMPK) is an intracellular energy sensor in skeletal muscle, which can be activated by exercise. AMPK, widely existing in eucaryotic cells, is the serine/threonine protein kinase. OBJECTIVE: To review the structure and the function of AMPK, changes of AMPK activity and the influence of AMPK activity on skeletal muscle protein degeneration during exercise. METHODS: A computer-based online retrieval of China National Knowledge Infrastructure (CNKI), Vip database, http://highwire.stanford.edu and www.ncbi.nlm.nih.gov/pubmed was performed to search papers regarding AMPK and myofibrillar protein degradation. The structure and the function of AMPK, the change of AMPK activity in exercise, and the effect of AMPK activation on myofibrillar protein degradation were retrieved. RESULTS AND CONCLUSION: A total of 35 papers were retrieved. This study summarized the structure and the function of AMPK. In the resistance exercise and in the moderate and high intensity cycle exercise, AMPK activity may be increased, and in the low intensity cycle exercise, AMPK activity may not be increased. Activated AMPK may promote the protein degradation.%背景:机体运动时骨骼肌收缩,ATP被大量消耗,产生大量腺苷一磷酸,导致腺苷酸活化蛋白激酶的激活.目的:综述不同运动过程中腺苷酸活化蛋白激酶活性的变化,以及腺苷酸活化蛋白激酶对骨骼肌蛋白质降解的研究成果.方法:检索中国期刊网、维普期刊数据库、www.ncbi.nlm.nih.gov/pubmed和http://highwire.stanford.edu/网站与腺苷酸活化蛋白激酶、运动、蛋白质降解研究相关的文章.并对腺苷酸活化蛋白激酶的结构与作用,不同运动过程中腺苷酸活化蛋白激酶活性的变化,以及腺苷酸活化蛋白激酶升高对骨骼肌蛋白质降解的内容进行分析综述.结果与结论:共纳入相关文献35篇.本文综述了腺苷酸活化蛋白激酶的结构、作用

  15. A bio-electrochemical system for removing inhibitors of anaerobic digestion processes from anaerobic reactors

    DEFF Research Database (Denmark)

    2014-01-01

    Inhibition of anaerobic digestion process by high level of ammonia (NH4 +/I\\IH3) is the most serious problem existing in biogas plants. No viable/applicable method to overcome this problem has been found up to now. This invention proposes an innovative submersible bio-electrochemical membrane...... reactor to recover ammonia from anaerobic digestion reactor, and thereby alleviate or counteract ammonia inhibition and enhance the conversion of ammonia-rich wastes to biogas. The invention may further reduce overall cost, giving synergistic advantages for both ammonia recycling and biogas plants...

  16. Inhibition of the anaerobic digestion process by linear alkylbenzene sulfonates

    DEFF Research Database (Denmark)

    Gavala, Hariklia N.; Ahring, Birgitte Kiær

    2002-01-01

    of the anaerobic digestion process should be seriously taken into consideration when wastewater from a surfactant producing industry is to be treated biologically or enter a municipal wastewater treatment plant that employs anaerobic technology. The upper allowable biomass specific LAS concentration should be 14......Linear Alkylbenzene Sulfonates (LAS) are the most widely used synthetic anionic surfactants. They are anthropogenic, toxic compounds and are found in the primary sludge generated in municipal wastewater treatment plants. Primary sludge is usually stabilized anaerobically and therefore...

  17. Transforming growth factor-β3 regulates cell junction restructuring via MAPK-mediated mRNA destabilization and Smad-dependent protein degradation of junctional adhesion molecule B (JAM-B).

    Science.gov (United States)

    Zhang, Xu; Lui, Wing-Yee

    2015-06-01

    Junctional adhesion molecule-B (JAM-B) is found between Sertoli cells at the blood-testis barrier (BTB) as well as between Sertoli and germ cells at the apical ectoplasmic specializations (ES) in the testis. The expression of JAM-B is tightly regulated to modulate the passage of spermatocytes across the BTB as well as the release of mature spermatozoa from the seminiferous epithelium. Transforming growth factor beta (TGF-β) family is implicated in the regulation of testicular cell junction dynamics during spermatogenesis. This study aims to investigate the effects of TGF-β3 on the expression of JAM-B as well as the underlying mechanisms on how TGF-β3 regulates JAM-B expression to facilitate the disassembly of the BTB and apical ES. Our results revealed that TGF-β3 suppresses JAM-B at post-transcriptional and post-translational levels. Inhibitor, siRNA knockdown and co-immunoprecipitation have shown that TGF-β3 induces JAM-B protein degradation via ubiquitin-proteasome pathway. Immunofluorescence staining further confirmed that blockage of ubiquitin-proteasome pathway could abrogate TGF-β3-induced loss of JAM-B at the cell-cell interface. siRNA knockdown and immunofluorescence staining also demonstrated that activation of Smad signaling is required for TGF-β3-induced JAM-B protein degradation. In addition, TGF-β3 reduces JAM-B mRNA levels, at least in part, via post-transcriptional regulation. mRNA stability assay has confirmed that TGF-β3 promotes the degradation of JAM-B transcript and TGF-β3-mediated mRNA destabilization requires the activation of ERK1/2 and p54 JNK signal cascades. Taken together, TGF-β3 significantly downregulates JAM-B expression via post-transcriptional and post-translational modulation and results in the disruption of BTB and apical ES.

  18. Anaerobic Metabolism and Bioremediation of Explosives-Contaminated Soil

    Science.gov (United States)

    Boopathy, Raj

    Nitroaromatic compounds pollute soil, water, and food via use of pesticides, plastics, pharmaceuticals, landfill dumping of industrial wastes, and the military use of explosives. Biotransformation of trinitrotoluene and other nitroaromatics by aerobic bacteria in the laboratory has been frequently reported, but the anaerobic bacterial metabolism of nitroaromatics has not been studied as extensively perhaps due to the difficulty in working with anaerobic cultures and the slow growth of anaerobes. Sulfate-reducing and methanogenic bacteria can metabolize nitroaromatic compounds under anaerobic conditions if appropriate electron donors and electron acceptors are present in the environment.

  19. Inhibition of the anaerobic digestion process by linear alkylbenzene sulfonates

    DEFF Research Database (Denmark)

    Gavala, Hariklia N.; Ahring, Birgitte Kiær

    2002-01-01

    it is important to investigate the effect of these xenobiotic compounds on an anaerobic environment. The inhibitory effect of Linear Alkylbenzene Sulfonates (LAS) on the acetogenic and methanogenic step of the anaerobic digestion process was studied. LAS inhibit both acetogenesis from propionate...... of the anaerobic digestion process should be seriously taken into consideration when wastewater from a surfactant producing industry is to be treated biologically or enter a municipal wastewater treatment plant that employs anaerobic technology. The upper allowable biomass specific LAS concentration should be 14...

  20. Energy conservation by oxidation of formate to carbon dioxide and hydrogen via a sodium ion current in a hyperthermophilic archaeon.

    Science.gov (United States)

    Lim, Jae Kyu; Mayer, Florian; Kang, Sung Gyun; Müller, Volker

    2014-08-05

    Thermococcus onnurineus NA1 is known to grow by the anaerobic oxidation of formate to CO2 and H2, a reaction that operates near thermodynamic equilibrium. Here we demonstrate that this reaction is coupled to ATP synthesis by a transmembrane ion current. Formate oxidation leads to H(+) translocation across the cytoplasmic membrane that then drives Na(+) translocation. The ion-translocating electron transfer system is rather simple, consisting of only a formate dehydrogenase module, a membrane-bound hydrogenase module, and a multisubunit Na(+)/H(+) antiporter module. The electrochemical Na(+) gradient established then drives ATP synthesis. These data give a mechanistic explanation for chemiosmotic energy conservation coupled to formate oxidation to CO2 and H2. Because it is discussed that the membrane-bound hydrogenase with the Na(+)/H(+) antiporter module are ancestors of complex I of mitochondrial and bacterial electron transport these data also shed light on the evolution of ion transport in complex I-like electron transport chains.

  1. ANAEROBIC EXERCISE TESTING IN REHABILITATION : A SYSTEMATIC REVIEW OF AVAILABLE TESTS AND PROTOCOLS

    NARCIS (Netherlands)

    Krops, Leonie A.; Albada, Trijntje; van der Woude, Lucas H. V.; Hijmans, Juha M.; Dekker, Rienk

    Objective: Anaerobic capacity assessment in rehabilitation has received increasing scientific attention in recent years. However, anaerobic capacity is not tested consistently in clinical rehabilitation practice. This study reviews tests and protocols for anaerobic capacity in adults with various

  2. [Distribution and removal of anaerobic antibiotic resistant bacteria during mesophilic anaerobic digestion of sewage sludge].

    Science.gov (United States)

    Tong, Juan; Wang, Yuan-Yue; Wei Yuan, Song

    2014-10-01

    Sewage sludge is one of the major sources that releasing antibiotic resistant bacteria (ARB) and antibiotic resistant genes (ARG) into the environment since it contains large amount of ARB, but there is little information about the fate of the anaerobic ARB in the anaerobic digestion of sewage sludge. Therefore, the distribution, removal and seasonal changes of tetracycline and β-lactam antibiotics resistant bacteria in the mesophilic egg-shaped digesters of a municipal wastewater treatment plant were investigated for one year in this study. Results showed that there were higher amounts of ARB and higher resistance rate of β-lactam antibiotics than that of tetracycline antibiotics in the sewage sludge. All ARB could be significantly reduced during the mesophilic anaerobic digestion process by 1.48-1.64 log unit (P resistance rates were significantly increased after anaerobic digestion by 12.0% and 14.3%, respectively (P change characteristics. Except for chlorotetracycline resistant bacteria, there were more ARB in the sewage sludge in cold season than in warm season (P < 0.05).

  3. Application of Anaerobic Digestion Model No. 1 for simulating anaerobic mesophilic sludge digestion

    Energy Technology Data Exchange (ETDEWEB)

    Mendes, Carlos, E-mail: carllosmendez@gmail.com; Esquerre, Karla, E-mail: karlaesquerre@ufba.br; Matos Queiroz, Luciano, E-mail: lmqueiroz@ufba.br

    2015-01-15

    Highlights: • The behavior of a anaerobic reactor was evaluated through modeling. • Parametric sensitivity analysis was used to select most sensitive of the ADM1. • The results indicate that the ADM1 was able to predict the experimental results. • Organic load rate above of 35 kg/m{sup 3} day affects the performance of the process. - Abstract: Improving anaerobic digestion of sewage sludge by monitoring common indicators such as volatile fatty acids (VFAs), gas composition and pH is a suitable solution for better sludge management. Modeling is an important tool to assess and to predict process performance. The present study focuses on the application of the Anaerobic Digestion Model No. 1 (ADM1) to simulate the dynamic behavior of a reactor fed with sewage sludge under mesophilic conditions. Parametric sensitivity analysis is used to select the most sensitive ADM1 parameters for estimation using a numerical procedure while other parameters are applied without any modification to the original values presented in the ADM1 report. The results indicate that the ADM1 model after parameter estimation was able to predict the experimental results of effluent acetate, propionate, composites and biogas flows and pH with reasonable accuracy. The simulation of the effect of organic shock loading clearly showed that an organic shock loading rate above of 35 kg/m{sup 3} day affects the performance of the reactor. The results demonstrate that simulations can be helpful to support decisions on predicting the anaerobic digestion process of sewage sludge.

  4. Oxygen tolerance capacity of upflow anaerobic solid-state (UASS) with anaerobic filter (AF) system.

    Science.gov (United States)

    Meng, Yao; Jost, Carsten; Mumme, Jan; Wang, Kaijun; Linke, Bernd

    2016-07-01

    In order to investigate the oxygen tolerance capacity of upflow anaerobic solid-state (UASS) with anaerobic filter (AF) system, the effect of microaeration on thermophilic anaerobic digestion of maize straw was investigated under batch conditions and in the UASS with AF system. Aeration intensities of 0-431mL O2/gvs were conducted as pretreatment under batch conditions. Aeration pretreatment obviously enhanced anaerobic digestion and an aeration intensity of 431mL O2/gvs increased the methane yield by 82.2%. Aeration intensities of 0-355mL O2/gvs were conducted in the process liquor circulation of the UASS with AF system. Dissolved oxygen (DO) of UASS and AF reactors kept around 1.39±0.27 and 0.99±0.38mg/L, respectively. pH was relatively stable around 7.11±0.04. Volatile fatty acids and soluble chemical oxygen demand concentration in UASS reactor were higher than those in AF reactor. Methane yield of the whole system was almost stable at 85±7mL/gvs as aeration intensity increased step by step. The UASS with AF system showed good oxygen tolerance capacity.

  5. Wingate Anaerobic Test Peak Power and Anaerobic Capacity Classification for Men and Women Intercollegiate Athletes

    Science.gov (United States)

    2009-12-01

    Ponorac et al. (1 7) Sbriccol i et al. Starling et al. (20) Watson and Sargeant (22) Weber et al. (23) Wiegman et al. (24) 2008 2001 2004 2002 2007 1 998... Wiegman , JE Burton, RR, and Forster, EM. The role of anaerobic power in human tolerance to simulated aerial combat maneuvers. Aotat Span Emimn Med 66

  6. The challenges of anaerobic digestion and the role of biochar in optimizing anaerobic digestion.

    Science.gov (United States)

    Fagbohungbe, Michael O; Herbert, Ben M J; Hurst, Lois; Ibeto, Cynthia N; Li, Hong; Usmani, Shams Q; Semple, Kirk T

    2017-03-01

    Biochar, like most other adsorbents, is a carbonaceous material, which is formed from the combustion of plant materials, in low-zero oxygen conditions and results in a material, which has the capacity to sorb chemicals onto its surfaces. Currently, research is being carried out to investigate the relevance of biochar in improving the soil ecosystem, digestate quality and most recently the anaerobic digestion process. Anaerobic digestion (AD) of organic substrates provides both a sustainable source of energy and a digestate with the potential to enhance plant growth and soil health. In order to ensure that these benefits are realised, the anaerobic digestion system must be optimized for process stability and high nutrient retention capacity in the digestate produced. Substrate-induced inhibition is a major issue, which can disrupt the stable functioning of the AD system reducing microbial breakdown of the organic waste and formation of methane, which in turn reduces energy output. Likewise, the spreading of digestate on land can often result in nutrient loss, surface runoff and leaching. This review will examine substrate inhibition and their impact on anaerobic digestion, nutrient leaching and their environmental implications, the properties and functionality of biochar material in counteracting these challenges. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Horse manure as feedstock for anaerobic digestion.

    Science.gov (United States)

    Hadin, Sa; Eriksson, Ola

    2016-10-01

    Horse keeping is of great economic, social and environmental benefit for society, but causes environmental impacts throughout the whole chain from feed production to manure treatment. According to national statistics, the number of horses in Sweden is continually increasing and is currently approximately 360,000. This in turn leads to increasing amounts of horse manure that have to be managed and treated. Current practices could cause local and global environmental impacts due to poor performance or lack of proper management. Horse manure with its content of nutrients and organic material can however contribute to fertilisation of arable land and recovery of renewable energy following anaerobic digestion. At present anaerobic digestion of horse manure is not a common treatment. In this paper the potential for producing biogas and biofertiliser from horse manure is analysed based on a thorough literature review in combination with mathematical modelling and simulations. Anaerobic digestion was chosen as it has a high degree of resource conservation, both in terms of energy (biogas) and nutrients (digestate). Important factors regarding manure characteristics and operating factors in the biogas plant are identified. Two crucial factors are the type and amount of bedding material used, which has strong implications for feedstock characteristics, and the type of digestion method applied (dry or wet process). Straw and waste paper are identified as the best materials in an energy point of view. While the specific methane yield decreases with a high amount of bedding, the bedding material still makes a positive contribution to the energy balance. Thermophilic digestion increases the methane generation rate and yield, compared with mesophilic digestion, but the total effect is negligible.

  8. Crystal structure of 1-deoxy-d-xylulose 5-phosphate reductoisomerase from the hyperthermophile Thermotoga maritima for insights into the coordination of conformational changes and an inhibitor binding.

    Science.gov (United States)

    Takenoya, Mihoko; Ohtaki, Akashi; Noguchi, Keiichi; Endo, Kiwamu; Sasaki, Yasuyuki; Ohsawa, Kanju; Yajima, Shunsuke; Yohda, Masafumi

    2010-06-01

    Isopentenyl diphosphate is a precursor of various isoprenoids and is produced by the 2-C-methyl-d-erythritol 4-phosphate (MEP) pathway in plastids of plants, protozoa and many eubacteria. A key enzyme in the MEP pathway, 1-deoxy-d-xylulose 5-phosphate reductoisomerase (DXR), has been shown to be the target of fosmidomycin, which works as an antimalarial, antibacterial and herbicidal compound. In this paper, we report studies of kinetics and the crystal structures of the thermostable DXR from the hyperthermophile Thermotoga maritima. Unlike the mesophilic DXRs, Thermotoga DXR (tDXR) showed activity only with Mg(2+) at its growth temperature. We solved the crystal structures of tDXR with and without fosmidomycin. The structure without fosmidomycin but unexpectedly bound with 2-methyl-2,4-pentanediol (MPD), revealing a new extra space available for potential drug design. This structure adopted the closed form by rigid domain rotation but without the flexible loop over the active site, which was considered as a novel conformation. Further, the conserved Asp residue responsible for cation binding seemed to play an important role in adjusting the position of fosmidomycin. Taken together, our kinetic and the crystal structures illustrate the binding mode of fosmidomycin that leads to its slow, tight binding according to the conformational changes of DXR.

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

    Directory of Open Access Journals (Sweden)

    Ron S. Ronimus

    2003-01-01

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

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

    Science.gov (United States)

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

    2015-08-01

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

  11. Co-expression with RadA and the characterization of stRad55B, a RadA paralog from the hyperthermophilic crenarchaea Sulfolobus tokodaii

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    ST0838 (designed stRad55B) is one of the four RadA paralogs (or Rad55 homologues) in the genome of the hyperthermophilic crenarchaeon Sulfolobus tokodaii. The gene is induced by UV irradiation, sug-gesting that it is involved in DNA recombinational repair in this organism. However, this protein could not be expressed normally in vitro. In this study, thermostable and soluble stRad55B was obtained by co-expression with S. tokodaii RadA (stRadA) in E. coli, and the enzymatic properties were examined. It was found that stRad55B bound ssDNA preferentially and had a very weak ATPase activity that was not stimulated by DNA. The recombinant protein inhibited the strand exchange activity promoted by stRadA, indicating that stRad55B might be an inhibitor to the homologous recombination in this ar-chaeon. The results will be helpful for further functional and interaction analysis of RadA paralogs and for the understanding of the mechanism of recombinational repair in archaea.

  12. Co-expression with RadA and the characterization of stRad55B, a RadA paralog from the hyperthermophilic crenarchaea Sulfolobus tokodaii

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    ST0838 (designed stRad55B) is one of the four RadA paralogs (or Rad55 homologues) in the genome of the hyperthermophilic crenarchaeon Sulfolobus tokodaii. The gene is induced by UV irradiation, suggesting that it is involved in DNA recombinational repair in this organism. However, this protein could not be expressed normally in vitro. In this study, thermostable and soluble stRad55B was obtained by co-expression with S. tokodaii RadA (stRadA) in E. coli, and the enzymatic properties were examined. It was found that stRad55B bound ssDNA preferentially and had a very weak ATPase activity that was not stimulated by DNA. The recombinant protein inhibited the strand exchange activity promoted by stRadA, indicating that stRad55B might be an inhibitor to the homologous recombination in this archaeon. The results will be helpful for further functional and interaction analysis of RadA paralogs and for the understanding of the mechanism of recombinational repair in archaea.

  13. Crystal structure, SAXS and kinetic mechanism of hyperthermophilic ADP-dependent glucokinase from Thermococcus litoralis reveal a conserved mechanism for catalysis.

    Directory of Open Access Journals (Sweden)

    Jaime Andrés Rivas-Pardo

    Full Text Available ADP-dependent glucokinases represent a unique family of kinases that belong to the ribokinase superfamily, being present mainly in hyperthermophilic archaea. For these enzymes there is no agreement about the magnitude of the structural transitions associated with ligand binding and whether they are meaningful to the function of the enzyme. We used the ADP-dependent glucokinase from Thermococcus litoralis as a model to investigate the conformational changes observed in X-ray crystallographic structures upon substrate binding and to compare them with those determined in solution in order to understand their interplay with the glucokinase function. Initial velocity studies indicate that catalysis follows a sequential ordered mechanism that correlates with the structural transitions experienced by the enzyme in solution and in the crystal state. The combined data allowed us to resolve the open-closed conformational transition that accounts for the complete reaction cycle and to identify the corresponding clusters of aminoacids residues responsible for it. These results provide molecular bases for a general mechanism conserved across the ADP-dependent kinase family.

  14. DNA Polymerases BI and D from the Hyperthermophilic Archaeon Pyrococcus furiosus Both Bind to Proliferating Cell Nuclear Antigen with Their C-Terminal PIP-Box Motifs▿

    Science.gov (United States)

    Tori, Kazuo; Kimizu, Megumi; Ishino, Sonoko; Ishino, Yoshizumi

    2007-01-01

    Proliferating cell nuclear antigen (PCNA) is the sliding clamp that is essential for the high processivity of DNA synthesis during DNA replication. Pyrococcus furiosus, a hyperthermophilic archaeon, has at least two DNA polymerases, polymerase BI (PolBI) and PolD. Both of the two DNA polymerases interact with the archaeal P. furiosus PCNA (PfuPCNA) and perform processive DNA synthesis in vitro. This phenomenon, in addition to the fact that both enzymes display 3′-5′ exonuclease activity, suggests that both DNA polymerases work in replication fork progression. We demonstrated here that both PolBI and PolD functionally interact with PfuPCNA at their C-terminal PIP boxes. The mutant PolBI and PolD enzymes lacking the PIP-box sequence do not respond to the PfuPCNA at all in an in vitro primer extension reaction. This is the first experimental evidence that the PIP-box motif, located at the C termini of the archaeal DNA polymerases, is actually critical for PCNA binding to form a processive DNA-synthesizing complex. PMID:17496095

  15. DNA polymerases BI and D from the hyperthermophilic archaeon Pyrococcus furiosus both bind to proliferating cell nuclear antigen with their C-terminal PIP-box motifs.

    Science.gov (United States)

    Tori, Kazuo; Kimizu, Megumi; Ishino, Sonoko; Ishino, Yoshizumi

    2007-08-01

    Proliferating cell nuclear antigen (PCNA) is the sliding clamp that is essential for the high processivity of DNA synthesis during DNA replication. Pyrococcus furiosus, a hyperthermophilic archaeon, has at least two DNA polymerases, polymerase BI (PolBI) and PolD. Both of the two DNA polymerases interact with the archaeal P. furiosus PCNA (PfuPCNA) and perform processive DNA synthesis in vitro. This phenomenon, in addition to the fact that both enzymes display 3'-5' exonuclease activity, suggests that both DNA polymerases work in replication fork progression. We demonstrated here that both PolBI and PolD functionally interact with PfuPCNA at their C-terminal PIP boxes. The mutant PolBI and PolD enzymes lacking the PIP-box sequence do not respond to the PfuPCNA at all in an in vitro primer extension reaction. This is the first experimental evidence that the PIP-box motif, located at the C termini of the archaeal DNA polymerases, is actually critical for PCNA binding to form a processive DNA-synthesizing complex.

  16. Anaerobic wastewater treatment of concentrated sewage using a two-stage upflow anaerobic sludge blanket- anaerobic filter system.

    Science.gov (United States)

    Halalsheh, Maha M; Abu Rumman, Zainab M; Field, Jim A

    2010-01-01

    A two-stage pilot-scale upflow anaerobic sludge blanket - anaerobic filter (UASB-AF) reactors system treating concentrated domestic sewage was operated at 23 degrees C and at hydraulic retention times (HRT) of 15 and 4 h, respectively. Excess sludge from the downstream AF stage was returned to the upstream UASB reactor. The aim was to obtain higher sludge retention time (SRT) in the UASB reactor for better methanization of suspended COD. The UASB-AF system removed 55% and 65% of the total COD (COD(tot)) and suspended COD (COD(ss)), respectively. The calculated SRT in the UASB reactor ranged from 20-35 days. The AF reactor removed the washed out sludge from the first stage reactor with average COD(ss) removal efficiency of 55%. The volatile fatty acids concentration in the effluent of the AF was 39 mg COD/L compared with 78 mg COD/L measured for the influent. The slightly higher COD(tot) removal efficiency obtained in this study compared with a single stage UASB reactor was achieved at 17% reduction in the total volume.

  17. Improvement of anaerobic digestion of sludge

    Energy Technology Data Exchange (ETDEWEB)

    Dohanyos, Michael; Zabranska, Jana; Kutil, Josef; Jenicek, Pavel

    2003-07-01

    Anaerobic digestion improvement can be accomplished by different methods. Besides optimization of process conditions is frequently used pretreatment of input sludge and increase of process temperature. Thermophilic process brings a higher solids reduction and biogas production, the high resistance to foaming, no problems with odour, the higher effect of destroying pathogens and the improvement of the energy balance of the whole treatment plant. Disintegration of excess activated sludge in lysate centrifuge was proved in full-scale conditions causing increase of biogas production. The rapid thermal conditioning of digested sludge is acceptable method of particulate matter disintegration and solubilization. (author)

  18. Monitoring and control of anaerobic reactors

    DEFF Research Database (Denmark)

    Pind, Peter Frode; Angelidaki, Irini; Ahring, Birgitte Kiær;

    2003-01-01

    measurements are reviewed in detail. In the sequel, possible manipulated variables, such as the hydraulic retention time, the organic loading rate, the sludge retention time, temperature, pH and alkalinity are evaluated with respect to the two main reactor types: high-rate and low-rate. Finally, the different......The current status in monitoring and control of anaerobic reactors is reviewed. The influence of reactor design and waste composition on the possible monitoring and control schemes is examined. After defining the overall control structure, and possible control objectives, the possible process...

  19. Hybrid modelling of anaerobic wastewater treatment processes.

    Science.gov (United States)

    Karama, A; Bernard, O; Genovesi, A; Dochain, D; Benhammou, A; Steyer, J P

    2001-01-01

    This paper presents a hybrid approach for the modelling of an anaerobic digestion process. The hybrid model combines a feed-forward network, describing the bacterial kinetics, and the a priori knowledge based on the mass balances of the process components. We have considered an architecture which incorporates the neural network as a static model of unmeasured process parameters (kinetic growth rate) and an integrator for the dynamic representation of the process using a set of dynamic differential equations. The paper contains a description of the neural network component training procedure. The performance of this approach is illustrated with experimental data.

  20. Anaerobic digestion of slaughterhouse by-products

    DEFF Research Database (Denmark)

    Hejnfelt, Anette; Angelidaki, Irini

    2009-01-01

    flour, fat, blood, hair, meat, ribs, raw waste were: 225, 497,487, 561, 582, 575, 359, 619 dm(3) kg(-1) respectively, corresponding to 50-100% of the calculated theoretical methane potential. Dilution of the by-products had a positive effect on the specific methane yield with the highest dilutions......Anaerobic digestion of animal by-products was investigated in batch and semi-continuously fed, reactor experiments at 55 degrees C and for some experiments also at 37 degrees C. Separate or mixed by-products from pigs were tested. The methane potential measured by batch assays for meat- and bone...

  1. Electrochemical monitoring of ammonia during anaerobic digestion

    DEFF Research Database (Denmark)

    Zhao, Nannan; Angelidaki, Irini; Zhang, Yifeng

    Ammonia is known as key inhibitor to methanogens in anaerobic digestion (AD) process. It’s of importance to develop efficient tool for ammonia monitoring. In this study, an electrolysis cell (EC) coupled with a complete nitrification reactor was developed as sensor for real time and online......-rich digesters. It was observed that the initial transient currents (0 min) were linearly corresponding to the ammonia levels (from 0 to 95.75 mg/L NH4+-N, R2 = 0.9673). Finally, this new sensor was tested with real AD effluent and the results showed no significant difference with that measured by conventional...

  2. Anaerobic xylose fermentation by Spathaspora passalidarum

    DEFF Research Database (Denmark)

    Hou, Xiaoru

    2012-01-01

    A cost-effective conversion of lignocellulosic biomass into bioethanol requires that the xylose released from the hemicellulose fraction (20–40% of biomass) can be fermented. Baker’s yeast, Saccharomyces cerevisiae, efficiently ferments glucose but it lacks the ability to ferment xylose. Xylose-fermenting...... yeast such as Pichia stipitis requires accurately controlled microaerophilic conditions during the xylose fermentation, rendering the process technically difficult and expensive. In this study, it is demonstrated that under anaerobic conditions Spathaspora passalidarum showed high ethanol production...

  3. Methanol conversion in high-rate anaerobic reactors

    NARCIS (Netherlands)

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

    2001-01-01

    An overview on methanol conversion in high-rate anaerobic reactors is presented, with the focus on technological as well as microbiological aspects. The simple C1-compound methanol can be degraded anaerobically in a complex way, in which methanogens, sulfate reducing bacteria and homoacetogens

  4. Balancing hygienization and anaerobic digestion of raw sewage sludge.

    Science.gov (United States)

    Astals, S; Venegas, C; Peces, M; Jofre, J; Lucena, F; Mata-Alvarez, J

    2012-12-01

    The anaerobic digestion of raw sewage sludge was evaluated in terms of process efficiency and sludge hygienization. Four different scenarios were analyzed, i.e. mesophilic anaerobic digestion, thermophilic anaerobic digestion and mesophilic anaerobic digestion followed by a 60 °C or by an 80 °C hygienization treatment. Digester performance (organic matter removal, process stability and biogas yield) and the hygienization efficiency (reduction of Escherichia coli, somatic coliphages and F-specific RNA phages) were the main examined factors. Moreover, a preliminary economical feasibility study of each option was carried out throughout an energy balance (heat and electricity). The obtained results showed that both thermophilic anaerobic digestion and mesophilic anaerobic digestion followed by a hygienization step were able to produce an effluent sludge that fulfills the American and the European legislation for land application. However, higher removal efficiencies of indicators were obtained when a hygienization post-treatment was present. Regarding the energy balance, it should be noted that all scenarios have a significant energy surplus. Particularly, positive heat balances will be obtained for the thermophilic anaerobic digestion and for the mesophilic anaerobic digestion followed by 60 °C hygienization post-treatment if an additional fresh-sludge/digested sludge heat exchanger is installed for energy recovery.

  5. A fuzzy logic approach to control anaerobic digestion

    NARCIS (Netherlands)

    Domnanovich, A.M.; Strik, D.P.B.T.B.; Pfeiffer, B.; Karlovits, M.; Zani, L.; Braun, R.; Holubar, P.

    2003-01-01

    One of the goals of the EU-Project AMONCO (Advanced Prediction, Monitoring and Controlling of Anaerobic Digestion Process Behaviour towards Biogas Usage in Fuel Cells) is to create a control tool for the anaerobic digestion process, which predicts the volumetric organic loading rate (Bv) for the nex

  6. A fuzzy logic approach to control anaerobic digestion

    NARCIS (Netherlands)

    Domnanovich, A.M.; Strik, D.P.B.T.B.; Pfeiffer, B.; Karlovits, M.; Zani, L.; Braun, R.; Holubar, P.

    2003-01-01

    One of the goals of the EU-Project AMONCO (Advanced Prediction, Monitoring and Controlling of Anaerobic Digestion Process Behaviour towards Biogas Usage in Fuel Cells) is to create a control tool for the anaerobic digestion process, which predicts the volumetric organic loading rate (Bv) for the nex

  7. TEST RESULTS FOR FUEL CELL OPERATION ON ANAEROBIC DIGESTER GAS

    Science.gov (United States)

    EPA, in conjunction with ONSI Corp., embarked on a project to define, design, test, and assess a fuel cell energy recovery system for application at anaerobic digester waste water (sewage) treatment plants. Anaerobic digester gas (ADG) is produced at these plants during the proce...

  8. Anaerobic catabolism of aromatic compounds: a genetic and genomic view.

    Science.gov (United States)

    Carmona, Manuel; Zamarro, María Teresa; Blázquez, Blas; Durante-Rodríguez, Gonzalo; Juárez, Javier F; Valderrama, J Andrés; Barragán, María J L; García, José Luis; Díaz, Eduardo

    2009-03-01

    Aromatic compounds belong to one of the most widely distributed classes of organic compounds in nature, and a significant number of xenobiotics belong to this family of compounds. Since many habitats containing large amounts of aromatic compounds are often anoxic, the anaerobic catabolism of aromatic compounds by microorganisms becomes crucial in biogeochemical cycles and in the sustainable development of the biosphere. The mineralization of aromatic compounds by facultative or obligate anaerobic bacteria can be coupled to anaerobic respiration with a variety of electron acceptors as well as to fermentation and anoxygenic photosynthesis. Since the redox potential of the electron-accepting system dictates the degradative strategy, there is wide biochemical diversity among anaerobic aromatic degraders. However, the genetic determinants of all these processes and the mechanisms involved in their regulation are much less studied. This review focuses on the recent findings that standard molecular biology approaches together with new high-throughput technologies (e.g., genome sequencing, transcriptomics, proteomics, and metagenomics) have provided regarding the genetics, regulation, ecophysiology, and evolution of anaerobic aromatic degradation pathways. These studies revealed that the anaerobic catabolism of aromatic compounds is more diverse and widespread than previously thought, and the complex metabolic and stress programs associated with the use of aromatic compounds under anaerobic conditions are starting to be unraveled. Anaerobic biotransformation processes based on unprecedented enzymes and pathways with novel metabolic capabilities, as well as the design of novel regulatory circuits and catabolic networks of great biotechnological potential in synthetic biology, are now feasible to approach.

  9. Molecular AND logic gate based on bacterial anaerobic respiration.

    Science.gov (United States)

    Arugula, Mary Anitha; Shroff, Namita; Katz, Evgeny; He, Zhen

    2012-10-21

    Enzyme coding genes that integrate information for anaerobic respiration in Shewanella oneidensis MR-1 were used as input for constructing an AND logic gate. The absence of one or both genes inhibited electrochemically-controlled anaerobic respiration, while wild type bacteria were capable of accepting electrons from an electrode for DMSO reduction.

  10. Xylan degradation by the anaerobic bacterium Bacteroides xylanolyticus

    NARCIS (Netherlands)

    Schyns, P.J.Y.M.L.

    1997-01-01

    Plant cell walls are the major reservoir of fixed carbon in nature. The mineralization of the fiber material, the so called lignocellulosic complex, proceeds almost exclusively by microbial processes in both aerobic and anaerobic environments. In anaerobic microbial processes the energy of

  11. Microbiology of anaerobic digestion; Microbiologia da digestao anaerobica

    Energy Technology Data Exchange (ETDEWEB)

    Novaes, Rosana Filomena Vazoller [CETESB, Sao Paulo, SP (Brazil)

    1988-12-31

    Considerations was made about the microorganisms involved in the anaerobic digestion of wastes. Are also presented, the main results on this subject obtained, until now, in the studies carried on the group of anaerobic microbiology researchers from the Sanitary Company of Sao Paulo State, Brazil. (author) 23 refs., 8 figs., 3 tabs.

  12. A fuzzy logic approach to control anaerobic digestion

    NARCIS (Netherlands)

    Domnanovich, A.M.; Strik, D.P.B.T.B.; Pfeiffer, B.; Karlovits, M.; Zani, L.; Braun, R.; Holubar, P.

    2003-01-01

    One of the goals of the EU-Project AMONCO (Advanced Prediction, Monitoring and Controlling of Anaerobic Digestion Process Behaviour towards Biogas Usage in Fuel Cells) is to create a control tool for the anaerobic digestion process, which predicts the volumetric organic loading rate (Bv) for the

  13. Anaerobic digestion of pot-ale

    Energy Technology Data Exchange (ETDEWEB)

    Mosey, F.E.

    1990-12-01

    In the production of whisky, the fermented wash is distilled twice and each bushel of grain yields about 15.5 gallons of pot-ale, 6.0 gallons of spent lees and 2.7 gallons of proof spirit. Disposal of pot-ale, the strong residue from the first distillation, containing all the non-volatile and unfermented components of the wash, will always be difficult. Anaerobic digestion provides a possible option. By destroying most of the biodegradable solids and converting them to biogas, it provides an intermediate effluent which conventional treatment technology can purify to river discharge standards. Pilot-scale trials confirm that pot-ale can be treated by anaerobic digestion. The most severe problems are the high purification efficiencies required to achieve UK river discharge standards and the quality and settling properties of the biological sludges produced. To achieved these standards, the design and operation of the entire treatment chain is dominated by the need to capture and concentrate suspended solids (SS) produced by the biological fermentations. Overall performance targets are 99.95% removal of biological oxygen demand (BOD), 99% removal of ammonia and a surplus sludge production of less than 20% of the incoming flow. (author).

  14. Occurrence of ethylene in anaerobic soil

    Energy Technology Data Exchange (ETDEWEB)

    Smith, K.A.; Restall, S.W.F.

    1971-01-01

    The production of ethylene and other hydrocarbon gases by soils under anaerobic conditions was measured by gas chromatography. Ethylene was the only hydrocarbon gas which occurred in physiologically significant concentrations; more than 20 ppm was found in several soils after 10 days at 20/sup 0/C. These concentrations were considerably higher than those which were known to cause severe reductions in the extension of root axes of some plant species. Experiments with sterilized and unsterilized soil indicated that ethylene was produced by enzyme activity and not by chemical action. The gas was found in soil when the oxygen concentration fell below 2%; total evolution was correlated with organic matter content, and was affected by drying and rewetting and by the growth of plant roots. The rate of production was increased by raising the temperature and by addition of glucose or peptone; high concentrations of nitrate depressed the rate, but sulfate and phosphate had little effect. It is concluded that ethylene may be a significant factor in causing injury to crop plants under waterlogged conditions and also in situations where anaerobic pockets occur within a mainly aerobic soil structure, provided that escape of the gas from the soil is impeded sufficiently to allow inhibitory concentrations to build up in the vicinity of plant roots. 31 references, 7 figures, 3 tables.

  15. Anaerobic and aerobic transformation of TNT

    Energy Technology Data Exchange (ETDEWEB)

    Kulpa, C.F. [Univ. of Notre Dame, IN (United States). Dept. of Biological Sciences; Boopathy, R.; Manning, J. [Argonne National Lab., IL (United States). Environmental Research Div.

    1996-12-31

    Most studies on the microbial metabolism of nitroaromatic compounds have used pure cultures of aerobic microorganisms. In many cases, attempts to degrade nitroaromatics under aerobic conditions by pure cultures result in no mineralization and only superficial modifications of the structure. However, mixed culture systems properly operated result in the transformation of 2,4,6-trinitrotoluene (TNT) and in some cases mineralization of TNT occurs. In this paper, the mixed culture system is described with emphasis on intermediates and the characteristics of the aerobic microbial process including the necessity for a co-substrate. The possibility of removing TNT under aerobic/anoxic conditions is described in detail. Another option for the biodegradation of TNT and nitroaromatics is under anaerobic, sulfate reducing conditions. In this instance, the nitroaromatic compounds undergo a series of reductions with the formation of amino compounds. TNT under sulfate reducing conditions is reduced to triaminotoluene presumably by the enzyme nitrite reductase, which is commonly found in many Desulfovibrio spp. The removal of nitro groups from TNT is achieved by a series of reductive reactions with the formation of ammonia and toluene by Desulfovibrio sp. (B strain). These metabolic processes could be applied to other nitroaromatic compounds like nitrobenzene, nitrobenzoic acids, nitrophenols, and aniline. The data supporting the anaerobic transformation of TNT under different growth condition are reviewed in this report.

  16. Anaerobic performances of sedentary and trained subjects.

    Science.gov (United States)

    Serresse, O; Ama, P F; Simoneau, J A; Lortie, G; Bouchard, C; Boulay, M R

    1989-03-01

    The objective of this report was to compare the performance of sedentary individuals, physical education students, and athletes of various disciplines in 10 s and 90 s maximal cycle ergometer tests. The 10 s power was the highest power output in one second from the 10 s test, while capacities were defined as the total work output during the best 10 s trial and the 90 s test. ANOVA and Duncan multiple range test indicated that the mean values of the 10 S power and capacity and the 90 S capacity tests were significantly higher in sprinter than in sedentary groups. Sprinters performed significantly better than marathon runners only in the 10 s capacity and power. Bodybuilders and sedentary subjects had similar results in the 90 s capacity test. Mean performance values per kilogram of body weight in sedentary females reached about 60% of sedentary males while marathon runners, physical education students and sprinter females reached about 80% of the male performances for the three indicators. When expressed per kilogram of fat-free mass, females reached a higher proportion of the male values for all performances. These results indicate that: a) there are differences for the power and capacity measured in predominantly anaerobic tests between athletes from different disciplines and sedentary individuals, and b) gender differences exist for these anaerobic performance indicators, but they appear attenuated in trained subjects.

  17. Electrolysis-enhanced anaerobic digestion of wastewater.

    Science.gov (United States)

    Tartakovsky, B; Mehta, P; Bourque, J-S; Guiot, S R

    2011-05-01

    This study demonstrates enhanced methane production from wastewater in laboratory-scale anaerobic reactors equipped with electrodes for water electrolysis. The electrodes were installed in the reactor sludge bed and a voltage of 2.8-3.5 V was applied resulting in a continuous supply of oxygen and hydrogen. The oxygen created micro-aerobic conditions, which facilitated hydrolysis of synthetic wastewater and reduced the release of hydrogen sulfide to the biogas. A portion of the hydrogen produced electrolytically escaped to the biogas improving its combustion properties, while another part was converted to methane by hydrogenotrophic methanogens, increasing the net methane production. The presence of oxygen in the biogas was minimized by limiting the applied voltage. At a volumetric energy consumption of 0.2-0.3 Wh/L(R), successful treatment of both low and high strength synthetic wastewaters was demonstrated. Methane production was increased by 10-25% and reactor stability was improved in comparison to a conventional anaerobic reactor.

  18. Anaerobic microbial degradation of organochlorine insecticides Aldrin

    Energy Technology Data Exchange (ETDEWEB)

    Chiu, T.C.; Yen, J.H.; Wang, Y.S. [National Taiwan Univ. (Taiwan)

    2004-09-15

    Aldrin (1,2,3,4,10,10-hexachloro-1,4,4a,5,8,8a-hexahydro-1,4-endo-exo-5,8-dimethanonnaphthalene), a cyclodiene organochlorine insecticide, was banned by nations and classified as B2 carcinogen by United States Environmental Protection Agency (EPA). Because of its chemical stability and lipophilicity, aldrin is regarded as a persistent and recalcitrant compound. Aldrin is easily adsorbed to soil and sediment after spreading to the environments, furthermore, it may be accumulated in animal's tissue or milk and then cause adverse effects by food-chain. The dissipation process of aldrin in environments has continuously been paid much attention by researchers. In general, the dissipation of aldrin has been thought as relating to three mechanisms: photo-degradation, chemical hydrolysis, and microbial degradation. And it has been well known that microbial degradation is the most important agent for breakdown of organochlorine pesticides. There has been shown that aldrin could be transformed to its metabolites, such as dieldrin or photo-dieldrin, by microorganisms under aerobic conditions, however, limited information has been shown under anaerobic conditions. For this reason, the degradation potential of aldrin by anaerobic microorganisms obtained from indigenous river sediment was evaluated, and the effect of environmental factors such as temperatures and nutrients on the aldrin degradation was also investigated in this study.

  19. Performance of sulfate-dependent anaerobic ammo-nium oxidation

    Institute of Scientific and Technical Information of China (English)

    ZHANG Lei; ZHENG Ping; HE YuHui; JIN RenCun

    2009-01-01

    The performance of sulfate-dependent anaerobic ammonium oxidation was studied. The results showed that both SO42- and NH4+ were chemically stable under anaerobic conditions. They did not react with each other in the absence of biological catalyst (sludge). The anaerobic digested sludge cultivated in an anaerobic reactor for three years took on the ability of oxidizing ammonium with sulfate anaero-bically. The average reduction of sulfate and ammonium was 71.67 mg.L-1 and 56.82 mg.L-1 at high concentrations.The reaction between SO42- and NH4+ was difficult, though feasible, due to its low standard Gibbs free energy change. The experiment demonstrated that high substrate concentrations and low oxidation-reduction potential (ORP) may be favourable for the biological reaction.

  20. Mathematical modelling of anaerobic digestion processes: applications and future needs

    DEFF Research Database (Denmark)

    Batstone, Damien J.; Puyol, Daniel; Flores Alsina, Xavier

    2015-01-01

    of the role of the central carbon catabolic metabolism in anaerobic digestion, with an increased importance of phosphorous, sulfur, and metals as electron source and sink, and consideration of hydrogen and methane as potential electron sources. The paradigm of anaerobic digestion is challenged by anoxygenic...... phototrophism, where energy is relatively cheap, but electron transfer is expensive. These new processes are commonly not compatible with the existing structure of anaerobic digestion models. These core issues extend to application of anaerobic digestion in domestic plant-wide modelling, with the need......Anaerobic process modelling is a mature and well-established field, largely guided by a mechanistic model structure that is defined by our understanding of underlying processes. This led to publication of the IWA ADM1, and strong supporting, analytical, and extension research in the 15 years since...

  1. Biochemistry and Evolution of Anaerobic Energy Metabolism in Eukaryotes

    Science.gov (United States)

    Müller, Miklós; Mentel, Marek; van Hellemond, Jaap J.; Henze, Katrin; Woehle, Christian; Gould, Sven B.; Yu, Re-Young; van der Giezen, Mark

    2012-01-01

    Summary: Major insights into the phylogenetic distribution, biochemistry, and evolutionary significance of organelles involved in ATP synthesis (energy metabolism) in eukaryotes that thrive in anaerobic environments for all or part of their life cycles have accrued in recent years. All known eukaryotic groups possess an organelle of mitochondrial origin, mapping the origin of mitochondria to the eukaryotic common ancestor, and genome sequence data are rapidly accumulating for eukaryotes that possess anaerobic mitochondria, hydrogenosomes, or mitosomes. Here we review the available biochemical data on the enzymes and pathways that eukaryotes use in anaerobic energy metabolism and summarize the metabolic end products that they generate in their anaerobic habitats, focusing on the biochemical roles that their mitochondria play in anaerobic ATP synthesis. We present metabolic maps of compartmentalized energy metabolism for 16 well-studied species. There are currently no enzymes of core anaerobic energy metabolism that are specific to any of the six eukaryotic supergroup lineages; genes present in one supergroup are also found in at least one other supergroup. The gene distribution across lineages thus reflects the presence of anaerobic energy metabolism in the eukaryote common ancestor and differential loss during the specialization of some lineages to oxic niches, just as oxphos capabilities have been differentially lost in specialization to anoxic niches and the parasitic life-style. Some facultative anaerobes have retained both aerobic and anaerobic pathways. Diversified eukaryotic lineages have retained the same enzymes of anaerobic ATP synthesis, in line with geochemical data indicating low environmental oxygen levels while eukaryotes arose and diversified. PMID:22688819

  2. When RNA and protein degradation pathways meet

    Directory of Open Access Journals (Sweden)

    Pascal eGENSCHIK

    2014-04-01

    Full Text Available RNA silencing has become a major focus of molecular and biomedical research in the last decade. This mechanism, which is conserved in most eukaryotes, has been extensively studied and is associated to various pathways implicated in the regulation of development, in the control of transposition events, heterochromatin maintenance and also playing a role in defense against viruses. Despite of its importance, the regulation of the RNA silencing machinery itself remains still poorly explored. Recently several reports in both plants and metazoans revealed that key components of RNA silencing, such as RNA-induced silencing complex (RISC component ARGONAUTE proteins, but also the endonuclease Dicer are subjected to proteasomal and autophagic pathways. Here we will review these post-translational proteolytic regulations with a special emphasis on plant research and also discuss their functional relevance.

  3. Validity of the Pediatric Running-Based Anaerobic Sprint Test to Determine Anaerobic Performance in Healthy Children

    NARCIS (Netherlands)

    Bongers, Bart C.; Werkman, Maarten S.; Blokland, Donna; Eijsermans, Maria J. C.; van der Torre, Patrick; Bartels, Bart; Verschuren, Olaf; Takken, Tim

    2015-01-01

    Purpose: To determine criterion validity of the pediatric running-based anaerobic sprint test (RAST) as a nonsophisticated field test for evaluating anaerobic performance in healthy children and adolescents. Methods: Data from 65. healthy children (28 boys and 37 girls between 6 and 18 years of age,

  4. The effect of biological sulfate reduction on anaerobic color removal in anaerobic-aerobic sequencing batch reactors.

    Science.gov (United States)

    Cirik, Kevser; Kitis, Mehmet; Cinar, Ozer

    2013-05-01

    Combination of anaerobic-aerobic sequencing processes result in both anaerobic color removal and aerobic aromatic amine removal during the treatment of dye-containing wastewaters. The aim of the present study was to gain more insight into the competitive biochemical reactions between sulfate and azo dye in the presence of glucose as electron donor source. For this aim, anaerobic-aerobic sequencing batch reactor fed with a simulated textile effluent including Remazol Brilliant Violet 5R (RBV 5R) azo dye was operated with a total cycle time of 12 h including anaerobic (6 h) and aerobic cycles (6 h). Microorganism grown under anaerobic phase of the reactor was exposed to different amounts of competitive electron acceptor (sulfate). Performance of the anaerobic phase was determined by monitoring color removal efficiency, oxidation reduction potential, color removal rate, chemical oxygen demand (COD), color, specific anaerobic enzyme (azo reductase) and aerobic enzyme (catechol 1,2-dioxygenase), and formation of aromatic amines. The presence of sulfate was not found to significantly affect dye decolorization. Sulfate and azo dye reductions took place simultaneously in all operational conditions and increase in the sulfate concentration generally stimulated the reduction of RBV 5R. However, sulfate accumulation under anaerobic conditions was observed proportional to increasing sulfate concentration.

  5. Anaerobic methanethiol degradation in upflow anaerobic sludge bed reactors at high salinity (> 0.5 M Na+)

    NARCIS (Netherlands)

    Leerdam, van R.C.; Bok, de F.A.M.; Lens, P.N.L.; Stams, A.J.M.; Janssen, A.J.H.

    2007-01-01

    The feasibility of anaerobic methanethiol (MT) degradation at elevated sodium concentrations was investigated in a mesophilic (30°C) lab-scale upflow anaerobic sludge bed (UASB) reactor, inoculated with estuarine sediment originating from the Wadden Sea (The Netherlands). MT was almost completely de

  6. Thermophilic anaerobic digestion of coffee grounds with and without waste activated sludge as co-substrate using a submerged AnMBR: system amendments and membrane performance.

    Science.gov (United States)

    Qiao, Wei; Takayanagi, Kazuyuki; Shofie, Mohammad; Niu, Qigui; Yu, Han Qing; Li, Yu-You

    2013-12-01

    Coffee grounds are deemed to be difficult for degradation by thermophilic anaerobic process. In this research, a 7 L AnMBR accepting coffee grounds was operated for 82 days and failed with pH dropping to 6.6. The deficiency of micronutrients in the reactor was identified. The system was recovered by supplying micronutrient, pH adjustment and influent ceasing for 22 days. In the subsequent 160 days of co-digestion experiment, waste activated sludge (15% in the mixture) was mixed into coffee grounds. The COD conversion efficiency of 67.4% was achieved under OLR of 11.1 kg-COD/m(3) d and HRT of 20 days. Tannins was identified affecting protein degradation by a batch experiment. Quantitative supplements of NH4HCO3 (0.12 g-N/g-TSin) were effective to maintain alkalinity and pH. The solid concentration in the AnMBR reached 75 g/L, but it did not significantly affect membrane filtration under a flux of 5.1 L/m(2) h. Soluble carbohydrate, lipid and protein were partially retained by the membrane. Copyright © 2013 Elsevier Ltd. All rights reserved.

  7. Anaerobic toxicity of cationic silver nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Gitipour, Alireza; Thiel, Stephen W. [Biomedical, Chemical, and Environmental Engineering, University of Cincinnati, Cincinnati, OH (United States); Scheckel, Kirk G. [USEPA, Office of Research and Development, Cincinnati, OH (United States); Tolaymat, Thabet, E-mail: tolaymat.thabet@epa.gov [USEPA, Office of Research and Development, Cincinnati, OH (United States)

    2016-07-01

    The microbial toxicity of silver nanoparticles (AgNPs) stabilized with different capping agents was compared to that of Ag{sup +} under anaerobic conditions. Three AgNPs were investigated: (1) negatively charged citrate-coated AgNPs (citrate-AgNPs), (2) minimally charged polyvinylpyrrolidone coated AgNPs (PVP-AgNPs) and (3) positively charged branched polyethyleneimine coated AgNPs (BPEI-AgNPs). The AgNPs investigated in this experiment were similar in size (10–15 nm), spherical in shape, but varied in surface charge which ranged from highly negative to highly positive. While, at AgNPs concentrations lower than 5 mg L{sup −1}, the anaerobic decomposition process was not influenced by the presence of the nanoparticles, there was an observed impact on the diversity of the microbial community. At elevated concentrations (100 mg L{sup −1} as silver), only the cationic BPEI-AgNPs demonstrated toxicity similar in magnitude to that of Ag{sup +}. Both citrate and PVP-AgNPs did not exhibit toxicity at the 100 mg L{sup −1} as measured by biogas evolution. These findings further indicate the varying modes of action for nanoparticle toxicity and represent one of the few studies that evaluate end-of-life management concerns with regards to the increasing use of nanomaterials in our everyday life. These findings also highlight some of the concerns with a one size fits all approach to the evaluation of environmental health and safety concerns associated with the use of nanoparticles. - Highlights: • At concentrations -1 the anaerobic decomposition process was not impacted. • An impact on the microbial community at concentrations -1 were observed. • At high concentrations (100 mg L{sup −1}), the cationic BPEI-AgNPs demonstrated toxicity. • Toxicity was demonstrated without the presence of oxidative dissolution of silver. • A one size fits all approach for the evaluation of NPs may not be accurate.

  8. Unique cellular events occurring during the initial interaction of macrophages with matrix-retained or methylated aggregated low density lipoprotein (LDL). Prolonged cell-surface contact during which ldl-cholesteryl ester hydrolysis exceeds ldl protein degradation.

    Science.gov (United States)

    Buton, X; Mamdouh, Z; Ghosh, R; Du, H; Kuriakose, G; Beatini, N; Grabowski, G A; Maxfield, F R; Tabas, I

    1999-11-05

    A critical event in atherogenesis is the interaction of arterial wall macrophages with subendothelial lipoproteins. Although most studies have investigated this interaction by incubating cultured macrophages with monomeric lipoproteins dissolved in media, arterial wall macrophages encounter lipoproteins that are mostly bound to subendothelial extracellular matrix, and these lipoproteins are often aggregated or fused. Herein, we utilize a specialized cell-culture system to study the initial interaction of macrophages with aggregated low density lipoprotein (LDL) bound to extracellular matrix. The aggregated LDL remains extracellular for a relatively prolonged period of time and becomes lodged in invaginations in the surface of the macrophages. As expected, the degradation of the protein moiety of the LDL was very slow. Remarkably, however, hydrolysis of the cholesteryl ester (CE) moiety of the LDL was 3-7-fold higher than that of the protein moiety, in stark contrast to the situation with receptor-mediated endocytosis of acetyl-LDL. Similar results were obtained using another experimental system in which the degradation of aggregated LDL protein was delayed by LDL methylation rather than by retention on matrix. Additional experiments indicated the following properties of this interaction: (a) LDL-CE hydrolysis is catalyzed by lysosomal acid lipase; (b) neither scavenger receptors nor the LDL receptor appear necessary for the excess LDL-CE hydrolysis; and (c) LDL-CE hydrolysis in this system is resistant to cellular potassium depletion, which further distinguishes this process from receptor-mediated endocytosis. In summary, experimental systems specifically designed to mimic the in vivo interaction of arterial wall macrophages with subendothelial lipoproteins have demonstrated an initial period of prolonged cell-surface contact in which CE hydrolysis exceeds protein degradation.

  9. Anaerobic Digestion Modeling: from One to Several Bacterial Populations

    Directory of Open Access Journals (Sweden)

    Iván D. Ramírez-Rivas

    2013-11-01

    Full Text Available Anaerobic digestion systems are complex processes that unfortunately often suffer from instability causing digester failure. In order to be able to design, optimizing and operate efficiently anaerobic digestion systems, appropriate control strategies need to be designed. Such strategies require, in general, the development of mathematical models. The anaerobic digestion process comprises a complex network of sequential and parallel reactions of biochemical and physicochemical nature. Usually, such reactions contain a particular step, the so called rate-limiting step which, being the slowest, limits the reaction rate of the overall process. The first attempts for modeling anaerobic digestion led to models describing only the limiting step. However, over a wide range of operating conditions, the limiting step is not always the same. It may depend on wastewater characteristics, hydraulic loading, temperature, etc. It is apparent that the "limiting step hypothesis" leads to simple and readily usable models. Such models, however, do not describe very well the digester behavior, especially under transient operating conditions. This work reviews the current state-of-the-art in anaerobic digestion modeling. We give a brief description of the key anaerobic digestion models that have been developed so far for describing biomass growth systems, including the International Water Association’s Anaerobic Digestion Model 1 (ADM1 and we identify the areas that require further research endeavors.

  10. Trace metal speciation and bioavailability in anaerobic digestion: A review.

    Science.gov (United States)

    Thanh, Pham Minh; Ketheesan, Balachandran; Yan, Zhou; Stuckey, David

    2016-01-01

    Trace metals are essential for the growth of anaerobic microorganisms, however, in practice they are often added to anaerobic digesters in excessive amounts, which can lead to inhibition. The concept of bioavailability of metals in anaerobic digestion has been poorly understood in the past, and a lack of deep understanding of the relationship between trace metal speciation and bioavailability can result in ineffective metal dosing strategies for anaerobic digesters. Sequential extraction schemes are useful for fractionating trace metals into their different forms, and metal sulfides can serve as a store and source for trace metals during anaerobic digestion, while natural/synthetic chelating agents (soluble microbial products-SMPs, extracellular polysaccharides-EPS, and EDTA/NTA) are capable of controlling trace metal bioavailability. Nevertheless, more work is needed to: investigate the speciation and bioavailability of Ca, Mg, Mn, W, and Se; compare the bioavailability of different forms of trace metals e.g. carbonates, sulfides, phosphates to different anaerobic trophic groups; determine what factors influence metal sulfide dissolution; investigate whether chelating agents can increase trace metal bioavailability; develop and adapt specialized analytical techniques, and; determine how trace metal dynamics change in an anaerobic membrane bioreactor (AnMBR).

  11. Anaerobic Treatment of Palm Oil Mill Effluent in Pilot-Scale Anaerobic EGSB Reactor

    Science.gov (United States)

    Wang, Jin; Mahmood, Qaisar; Qiu, Jiang-Ping; Li, Yin-Sheng; Chang, Yoon-Seong; Li, Xu-Dong

    2015-01-01

    Large volumes of untreated palm oil mill effluent (POME) pose threat to aquatic environment due to the presence of very high organic content. The present investigation involved two pilot-scale anaerobic expanded granular sludge bed (EGSB) reactors, continuously operated for 1 year to treat POME. Setting HRT at 9.8 d, the anaerobic EGSB reactors reduced COD from 71179 mg/L to 12341 mg/L and recycled half of sludge by a dissolved air flotation (DAF). The average effluent COD was 3587 mg/L with the consistent COD removal efficiency of 94.89%. Adding cationic polymer (PAM) dose of 30 mg/L to DAF unit and recycling its half of sludge caused granulation of anaerobic sludge. Bacilli and small coccid bacteria were the dominant microbial species of the reactor. The reactor produced 27.65 m3 of biogas per m3 of POME which was utilized for electricity generation. PMID:26167485

  12. Anaerobic digestion of tannery waste: semi-continuous and anaerobic sequencing batch reactor processes.

    Science.gov (United States)

    Zupancic, Gregor D; Jemec, A

    2010-01-01

    Disposal of the vast amounts of tannery waste that are currently generated is a significant problem. Anaerobic treatment of different types of tannery waste (fleshings, skin trimmings and wastewater sludge) was investigated. The biochemical methane potential is the same at 37 degrees C or 55 degrees C and an assay of this was shown to be an appropriate screening tool with which to estimate the susceptibility of a substrate to anaerobic digestion. The start-up procedure of a tannery waste thermophilic anaerobic digestion in 100 days using seed from mesophilic digester processing municipal sludge is presented. The specific methane production potential at 55 degrees C is estimated to be 0.617 m(3)kg(-1) of volatile suspended solids for tannery waste sludge, 0.377 m(3)kg(-1) for tannery waste trimmings and 0.649 m(3)kg(-1) for tannery waste fleshings. Additional concerns such as chromium content, salinity and temperature fluctuations were also addressed. Chromium content and salinity showed no adverse effects; however a reactor temperature reduction of 4.4 degrees C led to a drop in biogas production of 25%, indicating a requirement to keep the temperature constant at 55 degrees C.

  13. Anaerobic biodegradation of linear alkylbenzene sulfonate (LAS) in upflow anaerobic sludge blanket (UASB) reactors.

    Science.gov (United States)

    Sanz, José L; Culubret, Elayne; de Ferrer, Juan; Moreno, Alfonso; Berna, José L

    2003-01-01

    The anaerobic biodegradation of Linear Alkylbenzene Sulfonate (LAS) was studied in Upflow Anaerobic Sludge Blanket Reactors (UASB). One reactor was fed with easily degradable substrates and commercial LAS solution during a period of 3 months (Reactor 1), meanwhile a second reactor was fed with a commercial LAS solution without co-substrate (Reactor 2) during 4 months. Both reactors were operated with an organic loading rate of 4-5 mg-LAS/l x day and a hydraulic retention time of one day. The LAS biodegradation was determined by full mass balance. LAS was analysed by HPLC in the liquid phase (influent and effluent streams of the reactors) as well as in the solid phase (granular sludge used as biomass). The results indicate a high level of removal (primary biodegradation: 64-85%). Biodegradation was higher in the absence of external co-substrates than in the presence of additional sources of carbon. This indicates that the surfactant can be partially used as carbon and energy source by anaerobic bacteria. Under the operating conditions used, inhibition of the methanogenic activity or any other negative effects on the biomass due to the presence of LAS were not observed. The methanogenic activity remained high and stable throughout the experiment.

  14. Growth media in anaerobic fermentative processes: The underestimated potential of thermophilic fermentation and anaerobic digestion.

    Science.gov (United States)

    Hendriks, A T W M; van Lier, J B; de Kreuk, M K

    2017-09-01

    Fermentation and anaerobic digestion of organic waste and wastewater is broadly studied and applied. Despite widely available results and data for these processes, comparison of the generated results in literature is difficult. Not only due to the used variety of process conditions, but also because of the many different growth media that are used. Composition of growth media can influence biogas production (rates) and lead to process instability during anaerobic digestion. To be able to compare results of the different studies reported, and to ensure nutrient limitation is not influencing observations ascribed to process dynamics and/or reaction kinetics, a standard protocol for creating a defined growth medium for anaerobic digestion and mixed culture fermentation is proposed. This paper explains the role(s) of the different macro- and micronutrients, as well as the choices for a growth medium formulation strategy. In addition, the differences in nutrient requirements between mesophilic and thermophilic systems are discussed as well as the importance of specific trace metals regarding specific conversion routes and the possible supplementary requirement of vitamins. The paper will also give some insight into the bio-availability and toxicity of trace metals. A remarkable finding is that mesophilic and thermophilic enzymes are quite comparable at their optimum temperatures. This has consequences for the trace metal requirements of thermophiles under certain conditions. Under non-limiting conditions, the trace metal requirement of thermophilic systems is about 3 times higher than for mesophilic systems. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. Photochemistry of hypocrellin derivatives under anaerobic conditions

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    To improve the red absorption and solubility of hypocrellin, we have synthesized a series of hypocrellin B derivatives. The photochemistry of these new compounds in anaerobic media has been investigated by using electronic paramagnetic resonance (EPR) and spectrophotometric methods. The semiquinone anion radicals can be produced by self-electron transfer on irradiation, with the formation efficiency and EPR hyperfine structures of the semiquinone anion radicals dependent on the structures of the derivatives. When an electron donor is present, the electron transfer from electron donor to hypocrellin B derivatives enhanced the production of the corresponding semiquinone anion radical; in addition, the semiquinone anion radical and hydroquinone can be detected spectrophotometrically. Structural modifications exert little effect on the absorption position of semiquinone anion radical and hydroquinone, but influence their formation efficiency significantly.

  16. [Antimicrobial susceptibility testing of anaerobic bacteria].

    Science.gov (United States)

    García-Sánchez, José E; García-Sánchez, Enrique; García-García, María Inmaculada

    2014-02-01

    The anaerobic bacteria resistance to antibiotics is increasing, and even has appeared against the most active of those, like metronidazol and carbapenems. This fact forces to make and periodical sensibility tests -at least in the most aggressive and virulent species, in cases that they are isolated from life locations and in the absence of therapeutic response- to check the local sensibility and to establish suitable empiric therapies, all based on multicentric studies carried out in order to this or well to check the activity of new antibiotics. For the laboratory routine, the easiest sensibility method is the E-test/MIC evaluator. Another alternative is microdilution, that's only normalized for Bacteroides. There are preliminary facts that allow the use of disc diffusion method in some species of Bacteroides and Clostridium. For the temporal and multicentric studies, the procedure is dilution in agar plate, the reference method. Copyright © 2014 Elsevier España, S.L. All rights reserved.

  17. Hydrogen production from glucose by anaerobes.

    Science.gov (United States)

    Ogino, Hiroyasu; Miura, Takashi; Ishimi, Kosaku; Seki, Minoru; Yoshida, Hiroyuki

    2005-01-01

    Various anaerobes were cultivated in media containing glucose. When 100 mL of thioglycollate medium containing 2.0% (w/v) glucose was used, Clostridium butyricum ATCC 859, NBRC 3315, and NBRC 13949 evolved 227-243 mL of biogas containing about 180 mL of hydrogen in 1 day. Although some strains had some resistance against oxygen, C. butyricum ATCC 859 and 860 did not have it. C. butyricum NBRC 3315 and Enterobacter aerogenes NBRC 13534 produced hydrogen in the presence of glucose or pyruvic acid, and E. aerogenes NBRC 13534 produced hydrogen by not only glucose and pyruvic acid but also dextrin, sucrose, maltose, galactose, fructose, mannose, and mannitol. When a medium containing 0.5% (w/v) yeast extract and 2.0% (w/v) glucose was used, E. aerogenes NBRC 13534 evolved more biogas and hydrogen than C. butyricum NBRC 3315 in the absence of reducing agent.

  18. Applications of the anaerobic digestion process

    DEFF Research Database (Denmark)

    Angelidaki, Irini; Ellegaard, L.; Ahring, Birgitte Kiær

    2003-01-01

    and incineration of organic waste has become less desirable, and legislation, both in Europe and elsewhere, tends to favor biological treatment as a way of recycling minerals and nutrients of organic wastes from society back to the food production and supply chain. Removing the relatively wet organic waste from...... and resource/energy recovery have been developed. Treatment of biowastes by anaerobic digestion processes is in many cases the optimal way to convert organic waste into useful products such as energy (in the form of biogas) and a fertilizer product. Other waste management options, such as land filling......At the start of the new millennium waste management has become a political priority in many countries. One of the main problems today is to cope with an increasing amount of primary waste in an environmentally acceptable way. Biowastes, i.e., municipal, agricultural or industrial organic waste...

  19. Peritoneal dialysis peritonitis by anaerobic pathogens: a retrospective case series

    Science.gov (United States)

    2013-01-01

    Background Bacterial infections account for most peritoneal dialysis (PD)-associated peritonitis episodes. However, anaerobic PD peritonitis is extremely rare and intuitively associated with intra-abdominal lesions. In this study, we examined the clinical characteristics of PD patients who developed anaerobic peritonitis. Methods We retrospectively identified all anaerobic PD peritonitis episodes from a prospectively collected PD registry at a single center between 1990 and 2010. Only patients receiving more than 3 months of PD were enrolled. We analyzed clinical features as well as outcomes of anaerobic PD peritonitis patients. Results Among 6 patients, 10 episodes of PD-associated peritonitis were caused by anaerobic pathogens (1.59% of all peritonitis episodes during study the period), in which the cultures from 5 episodes had mixed growth. Bacteroides fragilis was the most common species identified (4 isolates). Only 3 episodes were associated with gastrointestinal lesions, and 4 episodes were related to a break in sterility during exchange procedures. All anaerobic pathogens were susceptible to clindamycin and metronidazole, but penicillin resistance was noted in 4 isolates. Ampicillin/sulbactam resistance was found in 2 isolates. In 5 episodes, a primary response was achieved using the first-generation cephalosporin and ceftazidime or aminoglycoside. In 3 episodes, the first-generation cephalosporin was replaced with aminoglycosides. Tenckhoff catheter removal was necessary in 2 episodes. Only one episode ended with mortality (due to a perforated bowel). Conclusion Anaerobic PD-associated peritonitis might be predominantly caused by contamination, rather than intra-abdominal events. Half of anaerobic PD-associated peritonitis episodes had polymicrobial growth. The overall outcome of anaerobic peritonitis is fair, with a high catheter survival rate. PMID:23705895

  20. Anaerobic digestion for sustainable development: a natural approach

    Energy Technology Data Exchange (ETDEWEB)

    Gijzen, H.J.

    2002-07-01

    After the discovery of methane gas by Alessandro Volta in 1776, it took about 100 years before anaerobic processes for the treatment of wastewater and sludges were introduced. The development of high rate anaerobic digesters for the treatment of sewage and industrial wastewater took until the nineteen-seventies and for solid waste even till the nineteen-eighties. All digesters have in common that they apply natural anaerobic consortia of microorganisms for degradation and transformation processes. In view of this, it could be rewarding to evaluate the efficiency of natural ecosystems for their possible application. Examples of high rate anaerobic natural systems include the forestomach of ruminants and the hindgut of certain insects, such as termites and cockroaches. These ''natural reactors'' exhibit volumetric methane production rates as high as 35 l/l.d. The development of anaerobic reactors based on such natural anaerobic systems could produce eco-technologies for the effective management of a wide variety of solid wastes and industrial wastewater. Important limitations of anaerobic treatment of domestic sewage relate to the absence of nutrient and pathogen removal. A combination of anaerobic pre-treatment followed by photosynthetic post-treatment is proposed for the effective recovery of energy and nutrients from sewage. This eco-technology approach is based on the recognition that the main nutrient assimilating capacity is housed in photosynthetic plants. The proposed anaerobic-photosynthetic process is energy efficient, cost effective and applicable under a wide variety of rural and urban conditions. In conclusion: a natural systems approach towards waste management could generate affordable eco-technologies for effective treatment and resource recovery. (author)

  1. Invited review: anaerobic fermentation of dairy food wastewater.

    Science.gov (United States)

    Hassan, A N; Nelson, B K

    2012-11-01

    Dairy food wastewater disposal represents a major environmental problem. This review discusses microorganisms associated with anaerobic digestion of dairy food wastewater, biochemistry of the process, factors affecting anaerobic digestion, and efforts to develop defined cultures. Anaerobic digestion of dairy food wastewater offers many advantages over other treatments in that a high level of waste stabilization is achieved with much lower levels of sludge. In addition, the process produces readily usable methane with low nutrient requirements and no oxygen. Anaerobic digestion is a series of complex reactions that broadly involve 2 groups of anaerobic or facultative anaerobic microorganisms: acidogens and methanogens. The first group of microorganisms breaks down organic compounds into CO(2) and volatile fatty acids. Some of these organisms are acetogenic, which convert long-chain fatty acids to acetate, CO(2), and hydrogen. Methanogens convert the acidogens' products to methane. The imbalance among the different microbial groups can lead not only to less methane production, but also to process failure. This is due to accumulation of intermediate compounds, such as volatile fatty acids, that inhibit methanogens. The criteria used for evaluation of the anaerobic digestion include levels of hydrogen and volatile fatty acids, methane:carbon ratio, and the gas production rate. A steady state is achieved in an anaerobic digester when the pH, chemical oxygen demand of the effluent, the suspended solids of the effluent, and the daily gas production remain constant. Factors affecting efficiency and stability of the process are types of microorganisms, feed C:N ratio, hydraulic retention time, reactor design, temperature, pH control, hydrogen pressure, and additives such as manure and surfactants. As anaerobic digesters become increasingly used in dairy plants, more research should be directed toward selecting the best cultures that maximize methane production from dairy

  2. Disruption of a sugar transporter gene cluster in a hyperthermophilic archaeon using a host-marker system based on antibiotic resistance.

    Science.gov (United States)

    Matsumi, Rie; Manabe, Kenji; Fukui, Toshiaki; Atomi, Haruyuki; Imanaka, Tadayuki

    2007-04-01

    We have developed a gene disruption system in the hyperthermophilic archaeon Thermococcus kodakaraensis using the antibiotic simvastatin and a fusion gene designed to overexpress the 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase gene (hmg(Tk)) with the glutamate dehydrogenase promoter. With this system, we disrupted the T. kodakaraensis amylopullulanase gene (apu(Tk)) or a gene cluster which includes apu(Tk) and genes encoding components of a putative sugar transporter. Disruption plasmids were introduced into wild-type T. kodakaraensis KOD1 cells, and transformants exhibiting resistance to 4 microM simvastatin were isolated. The transformants exhibited growth in the presence of 20 microM simvastatin, and we observed a 30-fold increase in intracellular HMG-CoA reductase activity. The expected gene disruption via double-crossover recombination occurred at the target locus, but we also observed recombination events at the hmg(Tk) locus when the endogenous hmg(Tk) gene was used. This could be avoided by using the corresponding gene from Pyrococcus furiosus (hmg(Pf)) or by linearizing the plasmid prior to transformation. While both gene disruption strains displayed normal growth on amino acids or pyruvate, cells without the sugar transporter genes could not grow on maltooligosaccharides or polysaccharides, indicating that the gene cluster encodes the only sugar transporter involved in the uptake of these compounds. The Deltaapu(Tk) strain could not grow on pullulan and displayed only low levels of growth on amylose, suggesting that Apu(Tk) is a major polysaccharide-degrading enzyme in T. kodakaraensis.

  3. Lesion-Induced Mutation in the Hyperthermophilic Archaeon Sulfolobus acidocaldarius and Its Avoidance by the Y-Family DNA Polymerase Dbh.

    Science.gov (United States)

    Sakofsky, Cynthia J; Grogan, Dennis W

    2015-10-01

    Hyperthermophilic archaea offer certain advantages as models of genome replication, and Sulfolobus Y-family polymerases Dpo4 (S. solfataricus) and Dbh (S. acidocaldarius) have been studied intensively in vitro as biochemical and structural models of trans-lesion DNA synthesis (TLS). However, the genetic functions of these enzymes have not been determined in the native context of living cells. We developed the first quantitative genetic assays of replication past defined DNA lesions and error-prone motifs in Sulfolobus chromosomes and used them to measure the efficiency and accuracy of bypass in normal and dbh(-) strains of Sulfolobus acidocaldarius. Oligonucleotide-mediated transformation allowed low levels of abasic-site bypass to be observed in S. acidocaldarius and demonstrated that the local sequence context affected bypass specificity; in addition, most erroneous TLS did not require Dbh function. Applying the technique to another common lesion, 7,8-dihydro-8-oxo-deoxyguanosine (8-oxo-dG), revealed an antimutagenic role of Dbh. The efficiency and accuracy of replication past 8-oxo-dG was higher in the presence of Dbh, and up to 90% of the Dbh-dependent events inserted dC. A third set of assays, based on phenotypic reversion, showed no effect of Dbh function on spontaneous -1 frameshifts in mononucleotide tracts in vivo, despite the extremely frequent slippage at these motifs documented in vitro. Taken together, the results indicate that a primary genetic role of Dbh is to avoid mutations at 8-oxo-dG that occur when other Sulfolobus enzymes replicate past this lesion. The genetic evidence that Dbh is recruited to 8-oxo-dG raises questions regarding the mechanism of recruitment, since Sulfolobus spp. have eukaryotic-like replisomes but no ubiquitin.

  4. Studies of the hyperthermophile Thermotoga maritima by random sequencing of cDNA and genomic libraries. Identification and sequencing of the trpEG (D) operon.

    Science.gov (United States)

    Kim, C W; Markiewicz, P; Lee, J J; Schierle, C F; Miller, J H

    1993-06-20

    Random sequencing of cDNA and genomic libraries has been used to study the genome of the hyperthermophile Thermotoga maritima. To date, 175 unique clones have been analyzed by comparing short sequence tags with known proteins in the PIR and GenBank databases. We find that a significant proportion of sequences can be matched to previously identified protein from non-Thermotoga sources. A high match rate was obtained from an oligo(dT)-primed cDNA library, where one-third of all unique sequences analyzed (21/65) shared high amino acid sequence similarity with proteins in the PIR and GenBank databases. Also, approximately one-third of the unique sequences from a second cDNA library (28/89), constructed with random oligo primers, could be matched to sequences in PIR and GenBank. Identification of genes from the oligo(dT)-primed cDNA library indicates that some Thermotoga mRNAs are polyadenylated. Genes have also been identified from a 1 to 2 kb genomic DNA library. Here, (3/21) of genomic sequences analyzed could be matched to protein in PIR and GenBank. One of the genomic clones had high sequence similarity to the tryptophan synthesis gene anthranilate synthase component I (trpE). Using this sequence tag, the Thermotoga trp operon was isolated and sequenced. The Thermotoga maritima trp operon is arranged with trpE forming an overlapping transcript with a second protein consisting of a fusion of anthranilate synthase component II (trpG) and anthranilate phosphoribosyltransferse (trpD). With regard to the fusion, the operon organization is similar to Escherichia coli and Salmonella typhimurium, but lacks the classic attenuation system of enteric bacteria. Amino acid sequence comparison with 19 trpE, 18 trpG and 14 trpD genes from other organisms suggest that the Thermotoga trp genes resemble corresponding genes from other thermophiles more closely than expected.

  5. Electron beam/biological processing of anaerobic and aerobic sludge

    Science.gov (United States)

    Čuba, V.; Pospíšil, M.; Múčka, V.; Jeníček, P.; Dohányos, M.; Zábranská, J.

    2003-01-01

    Besides common chemical and biological methods, the radiation technology is a promising way of sludge treatment. The paper describes possibilities of combined accelerated electrons/biological processing of both anaerobic and aerobic sludge. Besides one-shot experiments, experimental reactors for the simulation of anaerobic processes have been used. Main effort has been aimed to decrease organic compounds concentration and overall volume of solids, to improve some physico-chemical parameters of sludge, to validate hygienisation effects of the ionising radiation, and in the case of anaerobic sludge, to increase the volume of the produced biogas. Positive effects of the electron beam processing have been observed on all previously named parameters.

  6. ANAEROBIC MEMBRANE BIOREACTORS FOR DOMESTIC WASTEWATER TREATMENT. PRELIMINARY STUDY

    Directory of Open Access Journals (Sweden)

    Luisa Vera

    2014-12-01

    Full Text Available The operation of submerged anaerobic membrane bioreactors (SAnMBRs for domestic wastewaters treatment was studied in laboratory scale, with the objective to define sustainable filtration conditions of the suspensions along the process. During continuous experiments, the organic matter degradation by anaerobic way showed an average DQOT removal of 85% and 93%. Indeed, the degradation generated biogas after 12 days of operation and its relative methane composition was of 60% after 25 days of operation. Additionally, the comparison between membrane bioreactors (MBRs performance in aerobic and anaerobic conditions in filterability terms, reported that both systems behave similarly once reached the stationary state.

  7. Methane and hydrogen production by human intestinal anaerobic bacteria.

    Science.gov (United States)

    McKay, L F; Holbrook, W P; Eastwood, M A

    1982-06-01

    The gas above liquid cultures of a variety of human intestinal anaerobic bacteria was sampled and analysed by headspace gas chromatography. Hydrogen production was greatest with strains of the genus Clostridium, intermediate with anaerobic cocci and least with Bacteroides sp. Very few strains produced methane although small amounts were detected with one strain of B. thetaiotaomicron, C. perfringens and C. histolyticum. There may be a relationship between these anaerobic bacteria and several gastrointestinal disorders in which there is a build up of hydrogen or methane in the intestines.

  8. Decolorization of azo dyes under batch anaerobic and sequential anaerobic/aerobic conditions.

    Science.gov (United States)

    Işik, Mustafa; Sponza, Delia Teresa

    2004-01-01

    Batch anaerobic and sequential anaerobic upflow anaerobic sludge blanket (UASB)/aerobic continuous stirred tank reactor (CSTR) were used to determine the color and COD removals under anaerobic/aerobic conditions. Two azo dyes namely "Reactive Black 5 (RB 5)," "Congo Red (CR)," and glucose as a carbon source were used for synthetic wastewater. The course of the decolorization process approximates to first order and zero order kinetics with respect to dye concentration for RB 5 and Congo Red azo dyes, respectively, in batch conditions. The decolorization kinetic constant (K0) values increased from 3.6 to 11.8 mg(L h)(-1) as increases in dye concentrations from 200 to 3200 mg L(-1) for CR. Increases in dye concentrations from 0 to 3200 mg L(-1) reduce the decolorization rate constant (k1) values from 0.0141 to 0.0019 h(-1) in batch studies performed with RB 5. Decolorization was achieved effectively under test conditions but ultimate decolorization of azo dyes was not observed at all dye concentrations in batch assay conditions. Dye concentrations of 100 mg L(-1) and 3000 mg L(-1) of glucose-COD containing basal medium were used for continuous studies. The effect of organic loadings and HRT, on the color removal efficiencies and methane gas productions were monitored. 94.1-45.4% COD and 79-73% color removal efficiencies were obtained at an organic system during decolorization of Reactive Black 5. 92.3-77.0% COD and 95.3-92.2% decolorization efficiencies were achieved at a organic loading rate of 1.03-6.65 kg (m3 day)(-1) and a HRT of 3.54-0.49 for Congo Red treatment. The results of this study showed that, although decolorization continued, COD removal efficiencies and methane gas production were depressed at high organic loadings under anaerobic conditions. Furthermore, VFA accumulation, alkalinity consumption, and methane gas percentage were monitored at organic loading as high as 2.49-4.74 kg (m3 day)(-1) and 24.60-30.62 kg (m3 day)(-1), respectively, through the

  9. Atmospheric vs. anaerobic processing of metabolome samples for the metabolite profiling of a strict anaerobic bacterium, Clostridium acetobutylicum.

    Science.gov (United States)

    Lee, Sang-Hyun; Kim, Sooah; Kwon, Min-A; Jung, Young Hoon; Shin, Yong-An; Kim, Kyoung Heon

    2014-12-01

    Well-established metabolome sample preparation is a prerequisite for reliable metabolomic data. For metabolome sampling of a Gram-positive strict anaerobe, Clostridium acetobutylicum, fast filtration and metabolite extraction with acetonitrile/methanol/water (2:2:1, v/v) at -20°C under anaerobic conditions has been commonly used. This anaerobic metabolite processing method is laborious and time-consuming since it is conducted in an anaerobic chamber. Also, there have not been any systematic method evaluation and development of metabolome sample preparation for strict anaerobes and Gram-positive bacteria. In this study, metabolome sampling and extraction methods were rigorously evaluated and optimized for C. acetobutylicum by using gas chromatography/time-of-flight mass spectrometry-based metabolomics, in which a total of 116 metabolites were identified. When comparing the atmospheric (i.e., in air) and anaerobic (i.e., in an anaerobic chamber) processing of metabolome sample preparation, there was no significant difference in the quality and quantity of the metabolomic data. For metabolite extraction, pure methanol at -20°C was a better solvent than acetonitrile/methanol/water (2:2:1, v/v/v) at -20°C that is frequently used for C. acetobutylicum, and metabolite profiles were significantly different depending on extraction solvents. This is the first evaluation of metabolite sample preparation under aerobic processing conditions for an anaerobe. This method could be applied conveniently, efficiently, and reliably to metabolome analysis for strict anaerobes in air. © 2014 Wiley Periodicals, Inc.

  10. Bacterial ecology of abattoir wastewater treated by an anaerobic digestor

    Directory of Open Access Journals (Sweden)

    Linda Jabari

    2016-03-01

    Full Text Available Abstract Wastewater from an anaerobic treatment plant at a slaughterhouse was analysed to determine the bacterial biodiversity present. Molecular analysis of the anaerobic sludge obtained from the treatment plant showed significant diversity, as 27 different phyla were identified. Firmicutes, Proteobacteria, Bacteroidetes, Thermotogae, Euryarchaeota (methanogens, and msbl6 (candidate division were the dominant phyla of the anaerobic treatment plant and represented 21.7%, 18.5%, 11.5%, 9.4%, 8.9%, and 8.8% of the total bacteria identified, respectively. The dominant bacteria isolated were Clostridium, Bacteroides, Desulfobulbus, Desulfomicrobium, Desulfovibrio and Desulfotomaculum. Our results revealed the presence of new species, genera and families of microorganisms. The most interesting strains were characterised. Three new bacteria involved in anaerobic digestion of abattoir wastewater were published.

  11. POLISHING THE EFFLUENT FROM AN ANAEROBIC BIOLOGICAL PERCHLORATE TREATMENT PROCESS

    Science.gov (United States)

    Anaerobic biological processes effectively reduce perchlorate to chloride. However, the effluent can be biologically unstable, high in particulates and high in disinfection by-product precursor compounds. Such an effluent would be unsuitable for transmission into a drinking water...

  12. Sequential anaerobic-aerobic treatment for domestic wastewater - A review

    NARCIS (Netherlands)

    Kassab, G.; Halalsheh, M.; Klapwijk, A.; Fayyad, M.; Lier, van J.B.

    2010-01-01

    Introduction, consolidation and even standardization of expensive conventional aerobic systems for domestic wastewater treatment imposed significant financial constraints on the expansion of sanitary services including treatment in developing countries. A viable alternative is the sequential anaerob

  13. Cobalt toxicity in anaerobic granular sludge: influence of chemical speciation

    NARCIS (Netherlands)

    Bartacek, J.; Fermoso, F.G.; Baldo-Urrutia, A.M.; Hullebusch, van E.D.; Lens, P.N.L.

    2008-01-01

    The influence of cobalt speciation on the toxicity of cobalt to methylotrophic methanogenesis in anaerobic granular sludge was investigated. The cobalt speciation was studied with three different media that contained varying concentrations of complexing ligands [carbonates, phosphates and ethylenedi

  14. Performance of sulfate-dependent anaerobic ammonium oxidation

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    The performance of sulfate-dependent anaerobic ammonium oxidation was studied.The results showed that both SO42-and NH4+ were chemically stable under anaerobic conditions.They did not react with each other in the absence of biological catalyst(sludge).The anaerobic digested sludge cultivated in an anaerobic reactor for three years took on the ability of oxidizing ammonium with sulfate anaero-bically.The average reduction of sulfate and ammonium was 71.67 mg.L-1 and 56.82 mg.L-1 at high concentrations.The reaction between SO42-and NH4+ was difficult,though feasible,due to its low standard Gibbs free energy change.The experiment demonstrated that high substrate concentrations and low oxidation-reduction potential(ORP) may be favourable for the biological reaction.

  15. Tar water digestion in an upflow anaerobic sludge blanket reactor

    Energy Technology Data Exchange (ETDEWEB)

    Skibsted Mogensen, A.; Angelidaki, I.; Schmidt, J.E.; Ahring, B.K. [Technical Univ., Dept. of Environmental Science and Engineering, Lyngby (Denmark)

    1998-08-01

    The water from the gasification and wet oxidised tar water has been digested anaerobically in UASB reactors and were digested in respectively 10 and 50% in batches. Though the tar water show inhibition at very low concentrations to aerobic microorganisms, the granular sludge used in UASB reactors degrades tar water in concentrations that reveal total inhibition of e.g. bacteria conducting the nitrification process. The value of waste waters are determined, showing that the tar water produces more biogas in the anaerobic digestion. A wide range of xenobiotics, especially phenolic compounds can be transformed in the anaerobic digestion process. Seven phenolic are followed in batch experiments and UASB reactor experiments, and their particular fate in the anaerobic systems embody large differences in the transformation pattern. (au) 24 refs.

  16. Moroccan rock phosphate solubilization during a thermo-anaerobic ...

    African Journals Online (AJOL)

    SWEET

    2013-12-04

    Dec 4, 2013 ... thermo-anaerobic grassland waste biodegradation process. Moussa S. .... plating the nutrient agar isolate on the solid NBRIP medium. (Nautiyal, 1999) ..... solubilizing fungi isolated from phosphate mines. Ecol. Eng. 33:187-.

  17. Sequential anaerobic-aerobic treatment for domestic wastewater - A review

    NARCIS (Netherlands)

    Kassab, G.; Halalsheh, M.; Klapwijk, A.; Fayyad, M.; Lier, van J.B.

    2010-01-01

    Introduction, consolidation and even standardization of expensive conventional aerobic systems for domestic wastewater treatment imposed significant financial constraints on the expansion of sanitary services including treatment in developing countries. A viable alternative is the sequential anaerob

  18. Hydrogen as clean fuel via continuous fermentation by anaerobic ...

    African Journals Online (AJOL)

    Administrator

    anaerobic photosynthetic bacterium catalyzed water gas shift reaction which was used in this research. The synthesis gas ... commercial technology for syngas was steam methane reforming, in ..... Innovations (MOSTI). The authors wish to ...

  19. Characteristics, Process Parameters, and Inner Components of Anaerobic Bioreactors

    Science.gov (United States)

    Abdelgadir, Awad; Chen, Xiaoguang; Liu, Jianshe; Xie, Xuehui; Zhang, Jian; Zhang, Kai; Wang, Heng; Liu, Na

    2014-01-01

    The anaerobic bioreactor applies the principles of biotechnology and microbiology, and nowadays it has been used widely in the wastewater treatment plants due to their high efficiency, low energy use, and green energy generation. Advantages and disadvantages of anaerobic process were shown, and three main characteristics of anaerobic bioreactor (AB), namely, inhomogeneous system, time instability, and space instability were also discussed in this work. For high efficiency of wastewater treatment, the process parameters of anaerobic digestion, such as temperature, pH, Hydraulic retention time (HRT), Organic Loading Rate (OLR), and sludge retention time (SRT) were introduced to take into account the optimum conditions for living, growth, and multiplication of bacteria. The inner components, which can improve SRT, and even enhance mass transfer, were also explained and have been divided into transverse inner components, longitudinal inner components, and biofilm-packing material. At last, the newly developed special inner components were discussed and found more efficient and productive. PMID:24672798

  20. Decolourisation of textile wastewater in a submerged anaerobic membrane bioreactor.

    Science.gov (United States)

    Spagni, Alessandro; Casu, Stefania; Grilli, Selene

    2012-08-01

    Azo dye decolourisation can be easily achieved by biological reduction under anaerobic conditions. The aim of this study was to evaluate the applicability of submerged anaerobic membrane bioreactors (SAMBRs) for the decolourisation of dyeing wastewater containing azo dyes. The reactive orange 16 was used as model of an azo dye. The results demonstrated that very high decolourisation (higher than 99%) can be achieved by SAMBRs. Although decolourisation was not significantly influenced by the azo dye concentrations up to 3.2 g L(-1), methane production was greatly inhibited (up to 80-85%). Since volatile fatty acids accumulated in the treatment system with the azo dye concentration increase, methanogenes seem to be the most sensitive microbial populations of the anaerobic ecological community. The results demonstrated that anaerobic process combined with membrane filtration can deal with highly concentrated wastewaters that result from stream separation of industrial discharges.

  1. Anaerobic ammonium oxidation in the old trickling filters at Daspoort ...

    African Journals Online (AJOL)

    An aerobic and anaerobic bench top reactor, both inoculated with ... stoichiometric analyses of chemical oxygen demand (COD), NH4+-N, NO2. ... In the aerobic reactor, the COD concentration decreased over the 56 h batch reaction period ...

  2. Anaerobic composting of pyrethrum waste with and without effective ...

    African Journals Online (AJOL)

    user

    laboratory scale experiment involved composting of the waste as substrate mixed with EM at different .... The arrangement consists of 5 L plastic cane for anaerobic .... and Documentation Project on Recycling of Domestic Solid Waste.

  3. A simple anaerobic system for onsite treatment of domestic wastewater

    African Journals Online (AJOL)

    user

    The use of anaerobic process for domestic wastewater treatment would achieve lower carbon footprint ... However, its application is still limited to industrial wastewater treatment. ...... Department of Biotechnology, Lund University, Sweden.

  4. Estimation of methane generation based on anaerobic digestion ...

    African Journals Online (AJOL)

    Drake

    generation of methane from waste at Kiteezi landfill was measured using laboratory-scale anaerobic ... environmental and health challenges (Komakech, 2014). ..... Z (2007). Climate Change 2007. The Physical Science Basis. Contribution of ...

  5. Sequential anaerobic-adsorption treatment of chemical industry wastewater.

    Science.gov (United States)

    Daga, Kailash; Pallavi, V; Patel, Dharmendra

    2011-10-01

    Treatment technologies needed to reduce the pollutant load of chemical industry effluent have been found to involve exorbitantly high costs. The present investigation aimed to treat the wastewater from chemical industry by cost effective sequential anaerobic-adsorption treatment. Wastewaters from chemical industry that are rich in biodegradable organics are tested for anaerobic treatability. The efficiency of anaerobic reactor is relatively lower 79.3%, and therefore post treatment of effluent was done by adsorption using Poly vinyl alcohol coated Datura stramonium (PVAC-DS) as an adsorbent. An overall COD removal of 93.8 % was achieved after sequential Anaerobic-Adsorption treatment, which lead to a better final effluent and a more economical treatment system.

  6. Cobalt toxicity in anaerobic granular sludge: influence of chemical speciation

    NARCIS (Netherlands)

    Bartacek, J.; Fermoso, F.G.; Baldo-Urrutia, A.M.; Hullebusch, van E.D.; Lens, P.N.L.

    2008-01-01

    The influence of cobalt speciation on the toxicity of cobalt to methylotrophic methanogenesis in anaerobic granular sludge was investigated. The cobalt speciation was studied with three different media that contained varying concentrations of complexing ligands [carbonates, phosphates and

  7. Anaerobic desulphurisation of thiophenes by mixed microbial communities from oilfields

    NARCIS (Netherlands)

    Marcelis, C.L.M.; Ivanova, A.E.; Janssen, A.J.H.; Stams, A.J.M.

    2003-01-01

    Anaerobic enrichment cultures obtained from oil fields degraded various thiophenic compounds i.e. thiophene, benzothiophene and dibenzothiophene, with the concomitant formation of sulphide using hydrogen, lactate and ethanol as possible electron donors. It was demonstrated that dibenzothiophene was

  8. Anaerobic digestion as final step of a cellulosic ethanol biorefinery:

    DEFF Research Database (Denmark)

    Uellendahl, Hinrich; Ahring, Birgitte Kiær

    2010-01-01

    In order to lower the costs for second generation bioethanol from lignocellulosic biomass anaerobic digestion of the effluent from ethanol fermentation was implemented using an upflow anaerobic sludge blanket (UASB) reactor system in a pilot-scale biorefinery plant. Both thermophilic (538C...... were, however, higher under mesophilic conditions compared to thermophilic conditions. The conversion of dissolved organic matter (VSdiss) was between 68% and 91%. The effluent from the ethanol fermentation showed no signs of toxicity to the anaerobic microorganisms. However, a high content...... of suspended matter reduced the degradation efficiency. The retention time of the anaerobic system could be reduced from 70 to 7 h by additional removal of suspended matter by clarification. Implementation of the biogas production from the fermentation effluent accounted for about 30% higher carbon utilization...

  9. Cobalt toxicity in anaerobic granular sludge: influence of chemical speciation

    NARCIS (Netherlands)

    Bartacek, J.; Fermoso, F.G.; Baldo-Urrutia, A.M.; Hullebusch, van E.D.; Lens, P.N.L.

    2008-01-01

    The influence of cobalt speciation on the toxicity of cobalt to methylotrophic methanogenesis in anaerobic granular sludge was investigated. The cobalt speciation was studied with three different media that contained varying concentrations of complexing ligands [carbonates, phosphates and ethylenedi

  10. Ladderanes as tracers for present and past anaerobic ammonium oxidation

    NARCIS (Netherlands)

    Rush, D.J.

    2012-01-01

    Ladderane lipids are membrane lipids produced exclusively by anaerobic ammonium oxidising (anammox) bacteria. Anammox bacteria are key players in the marine nitrogen cycle, performing the anammox reaction, converting equal parts of ammonium and nitrite into dinitrogen gas. This process is

  11. Skeletal muscle protein degradation mechanism in chronic obstructive pulmonary disease rats%慢性阻塞性肺疾病大鼠模型中骨骼肌蛋白降解机制的研究

    Institute of Scientific and Technical Information of China (English)

    曾祥富; 刘朝晖; 梁志科; 范伟

    2011-01-01

    目的 研究慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)大鼠模型的骨骼肌蛋白降解机制,为有效防治COPD患者骨骼肌蛋白高分解,功能障碍等难题提供科学理论和依据.方法 成年雄性SD大鼠30只,分为模型组18只,对照组12只,采用反复熏香烟和气管内注入脂多糖复制COPD大鼠模型.酶联免疫吸附法(ELISA)测定血清和膈肌组织中的白介素6(IL-6),肿瘤坏死因子α(TNF-α),γ干扰素(IFN-γ)等含量.蛋白免疫印迹法测定大鼠膈肌和腓肠肌内的E2-14K、MAFbx、MuRF-1的蛋白表达.结果 在模型组中,血清和膈肌内的IL-6、TNF-α均较对照组显著升高,IFN-γ则较对照组显著降低.Western blot的结果显示:膈肌组织中E2-14K、MAFbx、MuRF-1均较对照组表达明显增强(各蛋白的相对表达量分别为0.96±0.12 vs 0.53±0.09、0.99±0.10 vs 0.53士0.08、0.95±0.08 vs 0.51±0.16,P<0.0l).腓肠肌中E2-14K、MAFbx、MuRF-1亦较对照组表达增强(各蛋白的相对表达量分别为0.98±0.06 vs 0.83士0.05、1.00±0.04 vs 0.81±0.05、0.92±0.05 vs0.71±0.05,P<0.05).体质量与TNF-α呈显著负相关(血清r=-0.84,膈肌r=-0.75,P值均<0.01),与IFN-γ呈正相关(血清r=-0.71,膈肌r=-0.69,P值均<0.01);中性粒细胞百分比与1L-6呈正相关(血清r =0.82,膈肌r=0.91,P值均<0.01);膈肌组织中,IL-6、TNF-α的水平与膈肌的E2-14K、MAFbx、MuRF-1灰度值呈显著正相关,相关系数分别为r=0.86、r=0.94、r=0.89,P<0.01和r=0.84、r=0.83、r=0.84,P<0.01;IFN-y与E2-14K、MAFbx、MuRF-1灰度值呈显著负相关,相关系数为r =0.81、r=0.80、r=0.78,P<0.01.结论 在COPD大鼠模型中,炎症因子、免疫调节因子的表达失衡和骨骼肌内泛素及泛素化蛋白的过度表达可能是引起骨骼肌蛋白降解加快的重要原因.%Objective To investgate protein degradation in skeletal muscle of chronic obstructive pulmonary disease (COPD) rats,in order to prevent

  12. Applications for the pressurized anaerobic bioconversion; Paineistetun anaerobisen biokonversion sovellusmahdollisuudet

    Energy Technology Data Exchange (ETDEWEB)

    Kantero, M. [Preseco Oy, Espoo (Finland)

    2004-07-01

    The aim of the project is to develop more practical applications for the hygienization and gasification of biowaste based on the pressurized anaerobic bioconversion. Previous studies on the subject will be used as a basis for the project. The main research objectives are the hygienization effect of the pressurized anaerobic bioconversion, the maximizing of the biogas production and the methane ratio, and the development of the automation programs related to the previous objects. (orig.)

  13. Anaerobic depuration of waste waters; Depuracion anaerobia de aguas residuales

    Energy Technology Data Exchange (ETDEWEB)

    Mejias Sanchez, G.; Vazquez Berger, E.; Magana Pietra, A.H. [Facultad de Ingenieria, Universidad Autonoma de yucatan, Merida (Mexico)

    1996-08-01

    Trials were carried out at a 500 l semi-experimental plant using there reactor models-anaerobic filter, fixed film and UASB type-for the anaerobic treatment of waste from different sources. The results after 24 and 48 hours were compared. The greatest efficiency was obtained after 48 hours the aerobic filter reactor (66% displacement), followed by the fixed film reactor (50%) and the UASB model (41%). (Author) 16 refs.

  14. Modeling for Anaerobic Fixed-Bed Biofilm Reactors

    Energy Technology Data Exchange (ETDEWEB)

    Liu, B. Y. M.; Pfeffer, J. T.

    1989-06-01

    The specific objectives of this research were: 1. to develop an equilibrium model for chemical aspects of anaerobic reactors; 2. to modify the equilibrium model for non-equilibrium conditions; 3. to incorporate the existing biofilm models into the models above to study the biological and chemical behavior of the fixed-film anaerobic reactors; 4. to experimentally verify the validity of these models; 5. to investigate the biomass-holding ability of difference packing materials for establishing reactor design criteria.

  15. MODELLING OF BACTERIAL SULPHATE REDUCTION IN ANAEROBIC PONDS : KINETIC INVESTIGATIONS

    OpenAIRE

    Harerimana, Casimir; Vasel, Jean-Luc; Jupsin, Hugues; Ouali, Amira

    2011-01-01

    The aim of the study was first to develop a simple and practical model of anaerobic digestion including sulphate-reduction in anaerobic ponds. The basic microbiology of our model consists of three steps, namely, acidogenesis, methanogenesis, and sulphate reduction. This model includes multiple reaction stoichiometry and substrate utilization kinetics. The second aim was to determine some kinetic parameters associated with this model. The values of these parameters for sulfidogenic bacteria ar...

  16. Modeling flow inside an anaerobic digester by CFD techniques

    OpenAIRE

    2011-01-01

    Anaerobic processes are used to treat high strength organic wastewater as well as for the treatment of primary and secondary sludge from conventional wastewater treatment plants. In these processes, heterotrophic microorganisms convert biodegradable organic matter to methane and carbon dioxide in the absence of dissolved oxygen and nitrate. Some of the most important aspects of the design of anaerobic digesters are related to hydraulic considerations. In spite of its impor...

  17. Microbiological Hydrogen Production by Anaerobic Fermentation and Photosynthetic Process

    Energy Technology Data Exchange (ETDEWEB)

    Asada, Y.; Ohsawa, M.; Nagai, Y.; Fukatsu, M.; Ishimi, K.; Ichi-ishi, S.

    2009-07-01

    Hydrogen gas is a clean and renewable energy carrier. Microbiological hydrogen production from glucose or starch by combination used of an anaerobic fermenter and a photosynthetic bacterium, Rhodobacter spheroides RV was studied. In 1984, the co-culture of Clostridium butyricum and RV strain to convert glucose to hydrogen was demonstrated by Miyake et al. Recently, we studied anaerobic fermentation of starch by a thermophilic archaea. (Author)

  18. In situ detection of anaerobic alkane metabolites in subsurface environments

    OpenAIRE

    Lisa eGieg; Akhil eAgrawal

    2013-01-01

    Alkanes comprise a substantial fraction of crude oil and refined fuels. As such, they are prevalent within deep subsurface fossil fuel deposits and in shallow subsurface environments such as aquifers that are contaminated with hydrocarbons. These environments are typically anaerobic, and host diverse microbial communities that can potentially use alkanes as substrates. Anaerobic alkane biodegradation has been reported to occur under nitrate-reducing, sulfate-reducing, and methanogenic cond...

  19. In situ detection of anaerobic alkane metabolites in subsurface environments

    OpenAIRE

    Agrawal, Akhil; Gieg, Lisa M.

    2013-01-01

    Alkanes comprise a substantial fraction of crude oil and refined fuels. As such, they are prevalent within deep subsurface fossil fuel deposits and in shallow subsurface environments such as aquifers that are contaminated with hydrocarbons. These environments are typically anaerobic, and host diverse microbial communities that can potentially use alkanes as substrates. Anaerobic alkane biodegradation has been reported to occur under nitrate-reducing, sulfate-reducing, and methanogenic conditi...

  20. Contribution of anaerobic energy expenditure to whole body thermogenesis

    Directory of Open Access Journals (Sweden)

    Scott Christopher B

    2005-06-01

    Full Text Available Abstract Heat production serves as the standard measurement for the determination of energy expenditure and efficiency in animals. Estimations of metabolic heat production have traditionally focused on gas exchange (oxygen uptake and carbon dioxide production although direct heat measurements may include an anaerobic component particularly when carbohydrate is oxidized. Stoichiometric interpretations of the ratio of carbon dioxide production to oxygen uptake suggest that both anaerobic and aerobic heat production and, by inference, all energy expenditure – can be accounted for with a measurement of oxygen uptake as 21.1 kJ per liter of oxygen. This manuscript incorporates contemporary bioenergetic interpretations of anaerobic and aerobic ATP turnover to promote the independence of these disparate types of metabolic energy transfer: each has different reactants and products, uses dissimilar enzymes, involves different types of biochemical reactions, takes place in separate cellular compartments, exploits different types of gradients and ultimately each operates with distinct efficiency. The 21.1 kJ per liter of oxygen for carbohydrate oxidation includes a small anaerobic heat component as part of anaerobic energy transfer. Faster rates of ATP turnover that exceed mitochondrial respiration and that are supported by rapid glycolytic phosphorylation with lactate production result in heat production that is independent of oxygen uptake. Simultaneous direct and indirect calorimetry has revealed that this anaerobic heat does not disappear when lactate is later oxidized and so oxygen uptake does not adequately measure anaerobic efficiency or energy expenditure (as was suggested by the "oxygen debt" hypothesis. An estimate of anaerobic energy transfer supplements the measurement of oxygen uptake and may improve the interpretation of whole-body energy expenditure.