DOCSLIB.ORG

Hydrogenase of the Hyperthermophile Pyrococcus Furiosus Is An

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Hydrogenase of the Hyperthermophile Pyrococcus Furiosus Is An Proc. Natl. Acad. Sci. USA Vol. 90, pp. 5341-5344, June 1993 Biochemistry Hydrogenase of the hyperthermophile Pyrococcus furiosus is an elemental sulfur reductase or sulfhydrogenase: Evidence for a sulfur-reducing hydrogenase ancestor (hydrogen activation/polysulfide reduction/geothermal biolog/evolution) KESEN MA*, RICHARD N. SCHICHOt, ROBERT M. KELLYt, AND MICHAEL W. W. ADAMS*§ *Department of Biochemistry and Center for Metalloenzyme Studies, University of Georgia, Athens, GA 30602; tDepartment of Chemical Engineering, The Johns Hopkins University, Baltimore, MD 21218; and kDepartment of Chemical Engineering, North Carolina State University, Raleigh, NC 27695-7905 Communicated by Helmut Beinert, February 22, 1993 (receivedfor review December 23, 1992) ABSTRACT Microorganisms growing near and above tion in mesophilic organisms. The activity was also shown to 100C have recently been discovered near shaflow and deep sea be membrane-associated in Wollinella succinogenes (9) and hydrothermal vents. Most are obligately dependent upon the Spirillum 5175 (10, 11), whereas it was associated with a reduction of elemental sulfur (S°) to hydrogen sulfide (H2S) for periplasmic protein in some Desulfovibrio sp. (12) and with a optimal growth, even though SO reduction readily occurs soluble protein in Chlorobium thiosulfatophilum (13). Only abioticafly at their growth temperatures. The sulfur reductase one sulfur reductase has been partially purified: that from W. activity of the anaerobic archaeon Pyrococcusfuriosus, which succinogenes, an organism that can obtain energy by S0 grows optimally at 100°C by a metabolism that produces H2S respiration (10). This membrane-bound enzyme contained an if S0 is present, was found in the cytoplasm. It was purified Fe-S cluster but no heme iron and appeared to use either SO anaerobically and was shown to be identical to the hydrogenase or polysulfide as the electron acceptor (14). that had been previously purified from this organism. Both S0 We report here on the SO-reducing activity of the hyper- and polysulfide served as substrates for H2S production, and thermophile P. furiosus, which grows optimally at 100°C (6). the SO reduction activity but not the H2-oxidation activity was This organism reduces either S0 or polysulfide to H2S during enhanced by the redox protein rubredoxin. The H2-oxidizing growth and can do so remote from the insoluble sulfur and SO-reduction activities of the enzyme also showed different particles present in the medium (15). However, the abiotic responses to pH, temperature, and inhibitors. This bifunc- reduction of S to H2S also occurs at the growth temperature tional "sulfhydrogenase" enzyme can, therefore, dispose ofthe of P. furiosus (15), and so it was far from clear whether any excess reductant generated during fermentation using either enzyme is actually involved in catalyzing S0 reduction. In protons or polysulfides as the electron acceptor. In addition, addition, assuming the reaction is enzymatic, two mecha- purified hydrogenases from both hyperthermophilic and me- nisms have been suggested by which the excess reductant sophilic representatives of the archaeal and bacterial domains produced during fermentation might be coupled to SO reduc- were shown to reduce SO to H2S. It is suggested that the function tion rather than HI reduction (H2 production) (4). These did of some form of ancestral hydrogenase was SO reduction rather not involve membrane electron transport, which might be than, or in addition to, the reduction of protons. anticipated if S0 reduction is an energy-conserving reaction (7). However, we show here that P.furiosus does have sulfur The ability of microorganisms to reduce elemental sulfur (S0) reductase activity, defined as the release of H2S from S, but is a very recent discovery (1) and is still limited in the it is located in the cytoplasmic fraction. Moreover, it is microbial world (2). The notable exceptions are the hyper- associated with and seems to be identical to the hydrogenase thermophilic archaea (formerly archaebacteria) that thrive at previously purified from this organism (16). temperatures near and even above 100°C in shallow and deep sea hydrothermal vents (3-5). All of these organisms have MATERIALS AND METHODS been shown to be dependent, to a greater or lesser extent, Growth ofBacterium. P.furiosus (DSM 3638) was routinely upon S0 reduction for optimal growth. The majority are grown at 85°C in a 500-liter fermenter, as described (16). obligately dependent upon the reduction of S0 to H2S and use Enzyme Assays. Sulfur reductase was assayed at 80°C in either H2 or organic compounds as electron donors. On the 8-ml sealed vials with H2 as the gas phase. The 2-ml assay other hand, some of these organisms can grow by fermenta- mixture routinely contained 100 mM N-(2-hydroxyethyl)- tive-type metabolisms and produce H2. In these cases addi- piperazine-N'-3-propanesulfonic acid (EPPS)/NaOH, pH tion of SO to the growth medium leads to the production of 8.4, 0.1 g of sublimed sulfur (Baker), and 0.8 mM sodium H2S and usually stimulates growth. It has been suggested that dithionite. The vials were shaken at 160 rpm in a water bath, sulfur reduction is a "detoxification" mechanism to lower the and cell-free extract or purified fractions were added to amount of H2 produced because H2 inhibits growth (6). initiate the reaction. Where indicated, P. furiosus hydrogen- However, we have recently shown that in at least one ase (5 ug) was added immediately before the addition of the fermentative hyperthermophile, Pyrococcus furiosus (6), S0 extract. At intervals, aliquots of the assay mixture were reduction appears to be an energy-conserving reaction (7). removed and assayed for hydrogen sulfide (H2S) by methyl- Little is known about the mechanism of S0 reduction in ene blue formation (17). Because the H2S is distributed hyperthermophilic organisms. Pihi et al. (8) reported that the between the gas and liquid phases, the total amount of H2S activity was membrane-bound in the autotrophic, S°- produced in this assay was calculated from a standard curve respiring Pyrodictium brockii, which grows optimally at prepared by using known amounts of sodium sulfide (Na2S) 105°C. Moreover, there have been few reports on SO reduc- under the same conditions. One unit of sulfur reductase activity is defined as the production of 1 ,umol ofH2S per min, The publication costs of this article were defrayed in part by page charge calculated from the amount of H2S produced over a 3-hr payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact. §To whom reprint requests should be addressed. 5341 Downloaded by guest on October 2, 2021 5342 Biochemistry: Ma et al. Proc. Natl. Acad. Sci. USA 90 (1993) period. Hydrogenase activity was determined by H2 evolu- that in the sulfur reductase assay used, the production ofH2S tion of H2 oxidation as described (15). One unit of activity is from SO was measured by using H2 as the electron donor. defined as 1 ,umol of H2 evolved or consumed per min. Under all conditions tested and with either cell-free extracts Polysulfide Preparation and Determination. A 0.5 M or purified fractions, H2S could not be detected unless a polysulfide solution was prepared by reaction of 12 g ofNa2S protein-containing fraction was added to the assay mixture. with 1.6 g of elemental sulfur in 100 ml of anoxic water (18). In the initial attempts to purify the sulfur reductase activity Polysulfide was measured by cold cyanolysis (19). from cell-free extracts, purified P. furiosus hydrogenase was Purification of Sulfur Reductase. All steps were performed added to the assay medium to provide a means of activating under anaerobic conditions at room temperature. All buffers H2 as a source of reductant. However, control experiments were repeatedly degassed and flushed with Ar and were showed that addition of hydrogenase, although stimulatory, maintained under a positive pressure of Ar. They all con- was not obligatory for H2S production, and it was therefore tained dithiothreitol (1 mM) to protect against trace 02 omitted for routine assays. The sulfur reductase activity was contamination. All columns were controlled by a fast protein purified -30-fold compared with the cell-free extract with an liquid chromatography system (Pharmacia LKB). P.furiosus overall yield of 10% (Table 1). Analysis of the purified cells (12 g, wet weight) were suspended in 50 mM Tris-HCl, enzyme by SDS/gel electrophoresis revealed why the addi- pH 8.1 (1 g of cells per 4-ml buffer) containing lysozyme (1 tion of hydrogenase was not required for H2 activation: the mg/ml) and DNase (0.1 mg/ml). The cells were lysed during purified sulfur reductase showed the same pattern ofsubunits incubation ofthe cell suspension at 35°C for 2 hr with stirring. as that exhibited by the hydrogenase previously purified from A cell-free extract was obtained by centrifugation at 50,000 x this organism (16). Furthermore, that the enzymes respon- g for 1 hr. The cell-free extract was applied to a column (2.8 sible for SO reduction and H2 evolution inP.furiosus were one x 13 cm) of Q Sepharose Fast Flow (Pharmacia LKB) that and the same was confirmed by the co-elution of these two was equilibrated with 50 mM Tris HCl buffer, pH 8.1, at 2.5 activities throughout the purification procedure. In addition, ml per min. The absorbed proteins were eluted with a 500-ml pure samples of P. furiosus hydrogenase previously purified lineargradient from 0 to 0.7 M NaCl in the same buffer. Sulfur in our laboratory all exhibited sulfur reductase activity. The reductase activity was located in the fractions eluting at enzymatic nature of the H2S-producing activity of what we 0.35-0.5 M NaCl.
Load more

Recommended publications
  • A Virus That Infects a Hyperthermophile Encapsidates A-Form
    RESEARCH | REPORTS we observe sets of regulatory sites that exhibit Illumina, Inc. One or more embodiments of one or more patents SUPPLEMENTARY MATERIALS patterns of coordinated regulation (e.g., LYN, and patent applications filed by Illumina may encompass the www.sciencemag.org/content/348/6237/910/suppl/DC1 encoding a tyrosine kinase involved in B cell methods, reagents, and data disclosed in this manuscript. All Materials and Methods methods for making the transposase complexes are described in signaling) (Fig. 4B), although reproducibility of Figs. S1 to S22 (18); however, Illumina will provide transposase complexes in Tables S1 and S2 these patterns across biological replicates was response to reasonable requests from the scientific community References (24–39) modest (fig. S22). Given the sparsity of the data, subject to a material transfer agreement. Some work in this study identifying pairs of coaccessible DNA elements is related to technology described in patent applications 19 March 2015; accepted 24 April 2015 WO2014142850, 2014/0194324, 2010/0120098, 2011/0287435, Published online 7 May 2015; within individual loci is statistically challenging 2013/0196860, and 2012/0208705. 10.1126/science.aab1601 and merits further development. We report chromatin accessibility maps for >15,000 single cells. Our combinatorial cellular indexing scheme could feasibly be scaled to col- VIROLOGY lect data from ~17,280 cells per experiment by using 384-by-384 barcoding and sorting 100 nu- clei per well (assuming similar cell recovery and A virus that infects a collision rates) (fig. S1) (19). Particularly as large- scale efforts to build a human cell atlas are con- templated (23), it is worth noting that because hyperthermophile encapsidates DNA is at uniform copy number, single-cell chro- matin accessibility mapping may require far fewer A-form DNA reads per single cell to define cell types, relative to single-cell RNA-seq.
    [Show full text]
  • Extremely Thermophilic Microorganisms As Metabolic Engineering Platforms for Production of Fuels and Industrial Chemicals
    REVIEW published: 05 November 2015 doi: 10.3389/fmicb.2015.01209 Extremely thermophilic microorganisms as metabolic engineering platforms for production of fuels and industrial chemicals Benjamin M. Zeldes 1, Matthew W. Keller 2, Andrew J. Loder 1, Christopher T. Straub 1, Michael W. W. Adams 2 and Robert M. Kelly 1* 1 Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, USA, 2 Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, USA Enzymes from extremely thermophilic microorganisms have been of technological interest for some time because of their ability to catalyze reactions of industrial significance at elevated temperatures. Thermophilic enzymes are now routinely produced in recombinant mesophilic hosts for use as discrete biocatalysts. Genome and metagenome sequence data for extreme thermophiles provide useful information for putative biocatalysts for a wide range of biotransformations, albeit involving at most a few enzymatic steps. However, in the past several years, unprecedented progress has been made in establishing molecular genetics tools for extreme thermophiles to the point Edited by: that the use of these microorganisms as metabolic engineering platforms has become Bettina Siebers, University of Duisburg-Essen, possible. While in its early days, complex metabolic pathways have been altered or Germany engineered into recombinant extreme thermophiles, such that the production of fuels and Reviewed by: chemicals at elevated temperatures has become possible. Not only does this expand the Haruyuki Atomi, thermal range for industrial biotechnology, it also potentially provides biodiverse options Kyoto University, Japan Phillip Craig Wright, for specific biotransformations unique to these microorganisms. The list of extreme University of Sheffield, UK thermophiles growing optimally between 70 and 100◦C with genetic toolkits currently *Correspondence: available includes archaea and bacteria, aerobes and anaerobes, coming from genera Robert M.
    [Show full text]
  • Do Ultrastable Proteins from Hyperthermophiles Have High Or Low Conformational Rigidity?
    Commentary Do ultrastable proteins from hyperthermophiles have high or low conformational rigidity? Rainer Jaenicke* Institute of Biophysics and Physical Biochemistry, University of Regensburg, D-93040 Regensburg, Germany ife on earth has an unbelievable adaptive parisons of their protein inventories with Lcapacity. Except for centers of volcanic those of suitable mesophilic counterparts, a activity, the entire surface of our planet is a wealth of data has been accumulated that biosphere. In this context, the most surpris- indicated that stabilization involves all levels ing discovery in our lifetime was the expan- of the hierarchy of protein structure, i.e., sion from the anthropocentrically defined secondary, supersecondary, tertiary, and ‘‘normal temperature’’ of mesophiles quaternary interactions. The common con- (Ͻ40°C) to the optimum temperature range clusion from model studies was that the of hyperthermophiles around and above the stability of proteins from extremophiles is boiling point of water. That in this class of optimized to maintain corresponding func- microorganisms high temperature is re- tional states under a given set of environ- quired for growth rather than tolerated im- mental conditions. For the standard state at plies that the whole repertoire of their bi- 25°C, enhanced thermal stability of hyper- omolecules must be sufficiently stable to thermophile proteins would then be the allow the cellular microcosm to work. The result of enhanced conformational rigidity Fig. 1. Three-dimensional structure of rubre- strategies nature has used to stabilize the in their folded native state (5). doxin from P. furiosus. Numbered residues mark inventory of the cell, especially proteins, Evidence from recent amide hydrogen the most slowly exchanging hydrogens, close to under extreme conditions are still enig- exchange experiments reported in this is- the two cysteine knuckles (7–9).
    [Show full text]
  • Bhattacharya.1999.Thermophiles.Pdf
    THE PHYLOGENY OF THERMOPHILES AND HYPERTHERMOPHILES AND THE THREE DOMAINS OF LIFE The Phylogeny of Thermophiles DEBASHISH BHATTACHARYA University of Iowa Department of Biological Sciences Biology Building, Iowa City, Iowa 52242-1324 United States THOMAS FRIEDL Department of Biology, General Botany University of Kaiserslautern P.O. Box 3049, D-67653 Kaiserslautern, Germany HEIKO SCHMIDT Deutsches Krebsforschungszentrum Theoretische Bioinformatik Im Neuenheimer Feld 280 , D-69120 Heidelberg, Germany 1. Introduction The nature of the first cells and the environment in which they lived are two of the most interesting problems in evolutionary biology. All living things are descendents of these primordial cells and are divided into three fundamental lineages or domains, Archaea (formerly known as Archaebacteria), Bacteria (formerly known as Eubacteria), and the Eucarya (formerly known as Eukaryotes, Woese et al. 1990). The Archaea and Bacteria are prokaryotic domains whereas the Eucarya includes all other living things that have a nucleus (i.e., the genetic material is separated from the cytoplasm by a nuclear envelope). The observation of the three primary domains, first made on the basis of small subunit (i.e., 16S, 18S) ribosomal DNA (rDNA) sequence comparisons (Woese 1987), has created a framework with which the nature of the last common ancestor (LCA) can be addressed. In this review we present phylogenies of the prokaryotic domains to understand the origin and distribution of the thermophiles (organisms able to grow in temperatures > 45°C) and the hyperthermophiles (organisms able to grow in temperatures > 80°C). Hyperthermophiles are limited to the Archaea and Bacteria. In addition, we inspect the distribution of extremophiles within the cyanobacteria.
    [Show full text]
  • ATP Synthases from Archaea: the Beauty of a Molecular Motor
    Biochimica et Biophysica Acta 1837 (2014) 940–952 Contents lists available at ScienceDirect Biochimica et Biophysica Acta journal homepage: www.elsevier.com/locate/bbabio Review ATP synthases from archaea: The beauty of a molecular motor Gerhard Grüber a,⁎, Malathy Sony Subramanian Manimekalai a, Florian Mayer b, Volker Müller b,⁎ a School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, Republic of Singapore b Molecular Microbiology & Bioenergetics, Institute of Molecular Biosciences, Johann Wolfgang Goethe University Frankfurt/Main, Max-von-Laue-Str. 9, 60438 Frankfurt, Germany article info abstract Article history: Archaea live under different environmental conditions, such as high salinity, extreme pHs and cold or hot tem- Received 13 November 2013 peratures. How energy is conserved under such harsh environmental conditions is a major question in cellular Received in revised form 7 March 2014 bioenergetics of archaea. The key enzymes in energy conservation are the archaeal A1AO ATP synthases, a class Accepted 11 March 2014 of ATP synthases distinct from the F F ATP synthase ATP synthase found in bacteria, mitochondria and chloro- Available online 17 March 2014 1 O plasts and the V1VO ATPases of eukaryotes. A1AO ATP synthases have distinct structural features such as a collar- Keywords: like structure, an extended central stalk, and two peripheral stalks possibly stabilizing the A1AO ATP synthase dur- ATPase ing rotation in ATP synthesis/hydrolysis at high temperatures as well as to provide the storage of transient elastic Na+ bioenergetics energy during ion-pumping and ATP synthesis/-hydrolysis. High resolution structures of individual subunits and Energy conservation subcomplexes have been obtained in recent years that shed new light on the function and mechanism of this Methanogenesis unique class of ATP synthases.
    [Show full text]
  • Life in Extreme Environments
    insight review articles Life in extreme environments Lynn J. Rothschild & Rocco L. Mancinelli NASA Ames Research Center, Moffett Field, California 94035-1000, USA (e-mail: [email protected]; [email protected]) Each recent report of liquid water existing elsewhere in the Solar System has reverberated through the international press and excited the imagination of humankind. Why? Because in the past few decades we have come to realize that where there is liquid water on Earth, virtually no matter what the physical conditions, there is life. What we previously thought of as insurmountable physical and chemical barriers to life, we now see as yet another niche harbouring ‘extremophiles’. This realization, coupled with new data on the survival of microbes in the space environment and modelling of the potential for transfer of life between celestial bodies, suggests that life could be more common than previously thought. Here we examine critically what it means to be an extremophile, and the implications of this for evolution, biotechnology and especially the search for life in the Universe. ormal is passé; extreme is chic. While thriving in biological extremes (for example, nutritional Aristotle cautioned “everything in extremes, and extremes of population density, parasites, moderation”, the Romans, known for their prey, and so on). excesses, coined the word ‘extremus’, the ‘Extremophile’ conjures up images of prokaryotes, yet the superlative of exter (‘being on the outside’). taxonomic range spans all three domains. Although all NBy the fifteenth century ‘extreme’ had arrived, via Middle hyperthermophiles are members of the Archaea and French, to English. At the dawning of the twenty-first Bacteria, eukaryotes are common among the psychrophiles, century we know that the Solar System, and even Earth, acidophiles, alkaliphiles, piezophiles, xerophiles and contain environmental extremes unimaginable to the halophiles (which respectively thrive at low temperatures, low ‘ancients’ of the nineteenth century.
    [Show full text]
  • 4 Metabolic and Taxonomic Diversification in Continental Magmatic Hydrothermal Systems
    Maximiliano J. Amenabar, Matthew R. Urschel, and Eric S. Boyd 4 Metabolic and taxonomic diversification in continental magmatic hydrothermal systems 4.1 Introduction Hydrothermal systems integrate geological processes from the deep crust to the Earth’s surface yielding an extensive array of spring types with an extraordinary diversity of geochemical compositions. Such geochemical diversity selects for unique metabolic properties expressed through novel enzymes and functional characteristics that are tailored to the specific conditions of their local environment. This dynamic interaction between geochemical variation and biology has played out over evolu- tionary time to engender tightly coupled and efficient biogeochemical cycles. The timescales by which these evolutionary events took place, however, are typically in- accessible for direct observation. This inaccessibility impedes experimentation aimed at understanding the causative principles of linked biological and geological change unless alternative approaches are used. A successful approach that is commonly used in geological studies involves comparative analysis of spatial variations to test ideas about temporal changes that occur over inaccessible (i.e. geological) timescales. The same approach can be used to examine the links between biology and environment with the aim of reconstructing the sequence of evolutionary events that resulted in the diversity of organisms that inhabit modern day hydrothermal environments and the mechanisms by which this sequence of events occurred. By combining molecu- lar biological and geochemical analyses with robust phylogenetic frameworks using approaches commonly referred to as phylogenetic ecology [1, 2], it is now possible to take advantage of variation within the present – the distribution of biodiversity and metabolic strategies across geochemical gradients – to recognize the extent of diversity and the reasons that it exists.
    [Show full text]
  • Hyperthermophilic Microorganisms - Karl O
    EXTREMOPHILES - Vol. I - Hyperthermophilic Microorganisms - Karl O. Stetter HYPERTHERMOPHILIC MICROORGANISMS Karl O. Stetter Universität Regensburg, Lehrstuhl für Mikrobiologie, Universitätsstraße 31, D-93053 Regensburg, Germany Keywords: Thermophilic, hyperthermophile, extremophile, prokaryote, archaea, bacteria, primitive, phylogeny, physiology, autotrophic, heterotrophic, heat stability, protein, nucleic acid, biotechnology, biotope, volcanic, hydrothermal, geothermal, solfataric, abyssal, antarctic, extraterrestrial, Mars Contents 1. Introduction 2. Biotopes of hyperthermophiles 2.1. Terrestrial biotopes 2.2. Marine biotopes 3. Phylogeny of hyperthermophiles 4. Taxonomy of hyperthermophiles 5. Sampling and isolation of hyperthermophiles 6. Strategies of life and environmental adaptations of hyperthermophiles 6.1. General metabolic potentialities 6.2. Physiological properties of the different groups of hyperthermophiles 6.2.1. Terrestrial hyperthermophiles 6.2.2. Marine hyperthermophiles 7. Distribution of species and complexity in hyperthermophilic ecosystems 8. Basis of heat stability and the upper temperature limit for life 9. Conclusions: hyperthermophiles in the history of life Acknowledgments Bibliography Biographical Sketch Summary Hyperthermophilic Archaea and Bacteria with optimal growth temperatures between 80 and 110° C have been isolated from geo- and hydrothermally heated terrestrial and submarineUNESCO environments. Small subunit – rR NAEOLSS sequence comparisons indicate great phylogenetic diversity among the 32genera
    [Show full text]
  • Energetic Limitations of Thermophilic Methanogens and Thiosulfate Reducers in the Subsurface Biosphere at Deep-Sea Hydrothermal Vents
    University of Massachusetts Amherst ScholarWorks@UMass Amherst Doctoral Dissertations Dissertations and Theses November 2015 Energetic Limitations Of Thermophilic Methanogens And Thiosulfate Reducers In The Subsurface Biosphere At Deep-Sea Hydrothermal Vents Lucy C. Stewart University of Massachusetts Amherst Follow this and additional works at: https://scholarworks.umass.edu/dissertations_2 Part of the Environmental Microbiology and Microbial Ecology Commons Recommended Citation Stewart, Lucy C., "Energetic Limitations Of Thermophilic Methanogens And Thiosulfate Reducers In The Subsurface Biosphere At Deep-Sea Hydrothermal Vents" (2015). Doctoral Dissertations. 464. https://doi.org/10.7275/7403929.0 https://scholarworks.umass.edu/dissertations_2/464 This Open Access Dissertation is brought to you for free and open access by the Dissertations and Theses at ScholarWorks@UMass Amherst. It has been accepted for inclusion in Doctoral Dissertations by an authorized administrator of ScholarWorks@UMass Amherst. For more information, please contact [email protected]. Energetic Limitations Of Thermophilic Methanogens And Thiosulfate Reducers In The Subsurface Biosphere At Deep-Sea Hydrothermal Vents A Dissertation Presented By LUCY C. STEWART Submitted to the Graduate School of the University of Massachusetts Amherst in partial fulfilment of the requirements for the degree of Doctor of Philosophy September 2015 Microbiology Department © Copyright by Lucy C. Stewart 2015 All Rights Reserved Energetic Limitations Of Thermophilic Methanogens
    [Show full text]
  • Hydrogen Stress and Syntrophy of Hyperthermophilic Heterotrophs and Methanogens
    University of Massachusetts Amherst ScholarWorks@UMass Amherst Doctoral Dissertations Dissertations and Theses July 2018 HYDROGEN STRESS AND SYNTROPHY OF HYPERTHERMOPHILIC HETEROTROPHS AND METHANOGENS Begum Topcuoglu University of Massachusetts Amherst Follow this and additional works at: https://scholarworks.umass.edu/dissertations_2 Part of the Environmental Microbiology and Microbial Ecology Commons, and the Microbial Physiology Commons Recommended Citation Topcuoglu, Begum, "HYDROGEN STRESS AND SYNTROPHY OF HYPERTHERMOPHILIC HETEROTROPHS AND METHANOGENS" (2018). Doctoral Dissertations. 1299. https://doi.org/10.7275/11912692.0 https://scholarworks.umass.edu/dissertations_2/1299 This Open Access Dissertation is brought to you for free and open access by the Dissertations and Theses at ScholarWorks@UMass Amherst. It has been accepted for inclusion in Doctoral Dissertations by an authorized administrator of ScholarWorks@UMass Amherst. For more information, please contact [email protected]. HYDROGEN STRESS AND SYNTROPHY OF HYPERTHERMOPHILIC HETEROTROPHS AND METHANOGENS A Dissertation Presented by BEGÜM D. TOPÇUOĞLU Submitted to the Graduate School of the University of Massachusetts Amherst in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY May 2018 Microbiology © Copyright by Begüm Topçuoğlu 2018 All Rights Reserved HYDROGEN STRESS AND SYNTROPHY OF HYPERTHERMOPHILIC HETEROTROPHS AND METHANOGENS A Dissertation Presented by BEGÜM D. TOPÇUOĞLU Approved as to style and content by: ____________________________________
    [Show full text]
  • Downloaded (July 2018) and Aligned Using Msaprobs V0.9.7 (16)
    bioRxiv preprint doi: https://doi.org/10.1101/524215; this version posted January 20, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Positively twisted: The complex evolutionary history of Reverse Gyrase suggests a non- hyperthermophilic Last Universal Common Ancestor Ryan Catchpole1,2 and Patrick Forterre1,2 1Institut Pasteur, Unité de Biologie Moléculaire du Gène chez les Extrêmophiles (BMGE), Département de Microbiologie F-75015 Paris, France 2Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Univ. Paris-Sud, Univ. Paris-Saclay, 91198, Gif-sur-Yvette Cedex, France 1 bioRxiv preprint doi: https://doi.org/10.1101/524215; this version posted January 20, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Abstract Reverse gyrase (RG) is the only protein found ubiquitously in hyperthermophilic organisms, but absent from mesophiles. As such, its simple presence or absence allows us to deduce information about the optimal growth temperature of long-extinct organisms, even as far as the last universal common ancestor of extant life (LUCA). The growth environment and gene content of the LUCA has long been a source of debate in which RG often features. In an attempt to settle this debate, we carried out an exhaustive search for RG proteins, generating the largest RG dataset to date.
    [Show full text]
  • Insight Into the Evolution of Microbial Metabolism from the Deep- 2 Branching Bacterium, Thermovibrio Ammonificans 3 4 5 Donato Giovannelli1,2,3,4*, Stefan M
    1 Insight into the evolution of microbial metabolism from the deep- 2 branching bacterium, Thermovibrio ammonificans 3 4 5 Donato Giovannelli1,2,3,4*, Stefan M. Sievert5, Michael Hügler6, Stephanie Markert7, Dörte Becher8, 6 Thomas Schweder 8, and Costantino Vetriani1,9* 7 8 9 1Institute of Earth, Ocean and Atmospheric Sciences, Rutgers University, New Brunswick, NJ 08901, 10 USA 11 2Institute of Marine Science, National Research Council of Italy, ISMAR-CNR, 60100, Ancona, Italy 12 3Program in Interdisciplinary Studies, Institute for Advanced Studies, Princeton, NJ 08540, USA 13 4Earth-Life Science Institute, Tokyo Institute of Technology, Tokyo 152-8551, Japan 14 5Biology Department, Woods Hole Oceanographic Institution, Woods Hole, MA 02543, USA 15 6DVGW-Technologiezentrum Wasser (TZW), Karlsruhe, Germany 16 7Pharmaceutical Biotechnology, Institute of Pharmacy, Ernst-Moritz-Arndt-University Greifswald, 17 17487 Greifswald, Germany 18 8Institute for Microbiology, Ernst-Moritz-Arndt-University Greifswald, 17487 Greifswald, Germany 19 9Department of Biochemistry and Microbiology, Rutgers University, New Brunswick, NJ 08901, USA 20 21 *Correspondence to: 22 Costantino Vetriani 23 Department of Biochemistry and Microbiology 24 and Institute of Earth, Ocean and Atmospheric Sciences 25 Rutgers University 26 71 Dudley Rd 27 New Brunswick, NJ 08901, USA 28 +1 (848) 932-3379 29 [email protected] 30 31 Donato Giovannelli 32 Institute of Earth, Ocean and Atmospheric Sciences 33 Rutgers University 34 71 Dudley Rd 35 New Brunswick, NJ 08901, USA 36 +1 (848) 932-3378 37 [email protected] 38 39 40 Abstract 41 Anaerobic thermophiles inhabit relic environments that resemble the early Earth. However, the 42 lineage of these modern organisms co-evolved with our planet.
    [Show full text]