DOCSLIB.ORG

Photosynthetic Bacterium Chloroflexus Aurantiacus BEVERLY K

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Photosynthetic Bacterium Chloroflexus Aurantiacus BEVERLY K JOURNAL OF BACTERIOLOGY, JUIY 1984, P. 222-227 Vol. 159, No. 1 0021-9193/84/070222-06$02.00/0 Copyright © 1984, American Society for Microbiology Isolation of Pigmentation Mutants of the Green Filamentous Photosynthetic Bacterium Chloroflexus aurantiacus BEVERLY K. PIERSON,* LYNN M. KEITH, AND JOANNE G. LEOVY Biology Department, University of Pluget Solund, Tacoma, Washington 98416 Received 19 December 1983/Accepted 16 April 1984 Mutants deficient in the production of bacteriochlorophyll c (Bchl c) and one mutant lacking colored carotenoids were isolated from the filamentous gliding bacterium Chloroflexus aurantiacus. Mutagenesis was achieved by using UV radiation or N-methyl-N'-nitro-N-nitrosoguanidine. Several clones were isolated that were deficient in Bchl c synthesis. All reverted. One double mutant deficient both in Bchl c synthesis and in the synthesis of colored carotenoids under anaerobic conditions was isolated. Isolation of a revertant in Bchl c synthesis from this double mutant produced a mutant strain of Chloroflexus that grew photosynthetically under anaerobic conditions and lacked colored carotenoids. Analysis of pigment contents and growth rates of the mutants revealed a positive association between growth rate and content of Bchl c under light-limiting conditions. Techniques for mutagenesis and the selection of photosyn- short pieces (less than 20 ,um in length) by brief (10 to 20 s) thetic and pigmentation mutants have been developed andI ultrasonic disruption of 5- or 10-ml samples, using a Branson applied successfully to the purple bacteria (6). Much has Sonifier (model 200) with standard tip sterilized by immer- been learned about the genetics, development, and regula- sion in 95% ethanol. Optical density changes to monitor tion of the photosynthetic pigment systems of the purple disruption were measured with a Turner model 330 spectro- bacteria through the use of mutants (7). Little has been photometer at 650 nm. Lengths offragments were measured reported on the mutagenesis or selection of pigmentationI by using a calibrated ocular micrometer with a Zeiss 16 mutants in the green photosynthetic bacteria (3, 5), andl standard microscope equipped with 40 and 63 x Neofluar nothing is known of the genetics of these organisms. Chloro- phase-contrast objectives. flexus aurantiacius, a green nonsulfur bacterium containing Plating. Fragmented cells were plated by the agar overlay bacteriochlorophyll (Bchls) a and c, is capable of both method, using 3 to 5 ml of top agar (0.7% [wt/vol]) overlaid phototrophic and chemotrophic growth, and hence is a on thick (two-thirds full) agar plates (1.5% [wt/vol]). Colo- suitable subject for the generation of pigmentation andI nies could also be obtained by using the spread plate photosynthetic mutants (8). Since Chloroflexus is a filamen- technique, but the gliding of the filaments (even on dry tous, gliding thermophile, standard techniques for the gener- plates) made such colonies too diffuse to allow for accurate ation and isolation of mutants from unicellular mesophilic determination of color. Colonies embedded in 0.7% agar did bacteria had to be modified. not spread, except when they broke to the surface, and This paper describes methods to mutagenize Chloroflexius differences in color could readily be determined among the and to isolate mutants deficient in Bchl c synthesis and in the compact colonies. Because filaments could move to a limited synthesis of some carotenoids. extent in the soft agar, it was not likely that color variations within a single colony would be detected. None were ob- MATERIALS AND METHODS served during the course of this study. Cultures and growth. C. aurantiacus J-10-fl was main- Plates were incubated at 50 to 55°C anaerobically (H, and tained in medium YEDM: D medium (4) containing yeast CO) by using the GasPak System (BBL Microbiology extract (2.0 g liter-1; Difco Laboratories) and glycylglycine Systems), and aerobically by stacking plates in unsealed (free base, 1.0 g liter- l; Sigma Chemical Co.). The pH of the GasPak jars. Illumination was provided to each jar by two or medium was 8.2. The medium was solidified with 1.5% agar. three 40-W, frosted, 10.5-in. (26.67-cm) tungsten tubular Cultures were maintained at 50°C in screw cap tubes 66 to lamps (General Electric Co.). Since pigment synthesis is 75% full, 6 in. (ca. 15 cm) from a 40-W tungsten lamp. regulated by both light intensity and oxygen tension, all Exponentially growing cultures were used for all experi- plates were incubated with the same light intensity. Water ments. The term "semianaerobic" refers to cultures main- filters (approximately 5 cm) were inserted between the lamps tained in this way. "Anaerobic cultures" were grown in and cultures. completely full screw cap tubes. "Strictly anaerobic" cul- Antibiotic sensitivities. Penicillin G, ampicillin, and carben- tures were grown in tubes completely filled with medium icillin (Sigma) were tested for inhibitory effects on cultures containing Na,S * 9H2O (0.1 mg ml-'). "Aerobic" cultures of Chloroflexus grown anaerobically at 50°C in the light. All were grown without shaking in 25-ml Erlenmeyer flasks antibiotics were added as freshly prepared, filter-sterilized containing 5 ml of medium. Cell density was estimated from solutions to exponentially growing cultures at an initial optical density at 650 nm. optical density at 650 nm (OD650) near 0.05. Growth after 20 Fragmentation of filaments. To obtain single cells for to 24 h was determined by measuring the increase in OD650. isolating mutant clones, filaments were fragmented into Selection with 740-nm filter. To select against wild-type cells, advantage was taken of the fact that Bchl c (but not * Corresponding author. Bchl a) absorbs in the region 735 to 745 nm. Mutagenized 222 VOL. 159, 1984 PIGMENTATION MUTANTS OF C. AURANTIACUS 223 TABLE 1. Effects of brief sonication on filaments of Chloroflexus Screening, isolation, and characterization of mutants. Colo- Time OD650, Morphology" CFU/ml nies that were obviously different in color (orange instead of olive green) were picked from anaerobic plates and cultured Before sonication 1.0 All long filaments 3.2 x 107 under semianaerobic or anaerobic conditions for analysis of After sonication 0.56 All short fragments less 3.0 x 106 for 18 s than 20 pLm: most pigments. Since screening in this way would miss photosyn- less than 10 pLm thetically incompetent mutants, random colonies from aero- bic plates that did not show obvious color differences were " Observed microscopically. subcultured aerobically in broth and then transferred to fresh broth for semianaerobic growth to measure synthesis of pigments. Because of the gliding of filaments on the surface of agar and subsequent spreading of colonies, it was not cultures were incubated anaerobically (full screw cap tubes) possible to do replica plating. in 740-nm light in the presence of ampicillin (50 pg ml-t) for For detecting the presence of Bchl ( and Bchl a, cultures 24 h to kill the majority of the wild-type cells. To incubate were centrifuged and extracted in absolute methanol until cultures in light absorbed by Bchl ( only, interference filters pellets were white. The absorption spectra of extracts were were used that transmitted light only near 740 nm (Baird- recorded on a Varian UV-VIS spectrophotometer (model Atomic and PTR Optics). The filters were 5 cm square, had 635). approximately 50% peak transmittance at 740 (PTR) and 738 Because of obvious reversion of some of the mutant nm (Baird-Atomic). and had bandwidths of approximately 10 strains, some were replated and reisolated. nm at 50% maximum transmittance. Cultures were incubat- ed directly behind the filter. The light from a 40-W tungsten RESULTS lamp was filtered through 5 cm of water before passing Fragmentation of filaments and plating. Since Chloiroflexis through the interference filter. Preliminary experiments is a gliding filamentous organism of indeterminate length, it without ampicillin showed that the wild type grew behind the was necessary to develop a method for fragmenting the filter with a doubling time of 12.5 h. compared with 8.5 h in filaments that would permit isolation of colonies arising from white light. single cells. Very short exposure to ultrasound (10 to 20 s) A preselection incubation step under aerobic conditions fragmented filaments into short pieces suitable for plating. was used to repress Bchl c synthesis in all cells, since The length of a single cell of strain J-10-fl ranges from 0.5 to oxygen inhibits synthesis of Bchl c (11). This was necessary 13 p.m, with a mean cell length of 3.5 p.m as determined on to ensure that mutants contained little or no residual Bchl c negatively stained, unfixed cells with transmission electron and therefore would not grow in the selection step. To microscopy (11). Sonication time was varied according to increase the titer of cells before plating after the ampicillin the optical density of the culture and continued until no selection step. cultures were incubated aerobically in the fragments remained greater than 15 to 20 pum in length in light since it seemed likely that Bchl c-minus mutants would several fields. The majority of the fragments were shorter be photosynthetically incompetent or at least would grow at than 10 p.m. This degree of fragmentation resulted in a reduced rates under photosynthetic conditions. decrease in optical density of approximately 50%. The UV radiation mutagenesis. Exponentially growing cells mortality during ultrasonic fragmentation was high, howev- were harvested and suspended in 0.1 M MgSO4 to an OD650 er. There was also variation from experiment to experiment of 0.79. The filaments were fragmented by sonication (final since it was impossible to duplicate the fragmentation tech- OD650,, 0.33; ca. 3.5 x 105 CFU ml -t), and 5-ml aliquots nique from one batch of cells to another.
Load more

Recommended publications
  • Evolution of the 3-Hydroxypropionate Bicycle and Recent Transfer of Anoxygenic Photosynthesis Into the Chloroflexi
    Evolution of the 3-hydroxypropionate bicycle and recent transfer of anoxygenic photosynthesis into the Chloroflexi Patrick M. Shiha,b,1, Lewis M. Wardc, and Woodward W. Fischerc,1 aFeedstocks Division, Joint BioEnergy Institute, Emeryville, CA 94608; bEnvironmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720; and cDivision of Geological and Planetary Sciences, California Institute of Technology, Pasadena, CA 91125 Edited by Bob B. Buchanan, University of California, Berkeley, CA, and approved August 21, 2017 (received for review June 14, 2017) Various lines of evidence from both comparative biology and the provide a hard geological constraint on these analyses, the timing geologic record make it clear that the biochemical machinery for of these evolutionary events remains relative, thus highlighting anoxygenic photosynthesis was present on early Earth and provided the uncertainty in our understanding of when and how anoxy- the evolutionary stock from which oxygenic photosynthesis evolved genic photosynthesis may have originated. ca. 2.3 billion years ago. However, the taxonomic identity of these A less recognized alternative is that anoxygenic photosynthesis early anoxygenic phototrophs is uncertain, including whether or not might have been acquired in modern bacterial clades relatively they remain extant. Several phototrophic bacterial clades are thought recently. This possibility is supported by the observation that to have evolved before oxygenic photosynthesis emerged, including anoxygenic photosynthesis often sits within a derived position in the Chloroflexi, a phylum common across a wide range of modern the phyla in which it is found (3). Moreover, it is increasingly environments. Although Chloroflexi have traditionally been thought being recognized that horizontal gene transfer (HGT) has likely to be an ancient phototrophic lineage, genomics has revealed a much played a major role in the distribution of phototrophy (8–10).
    [Show full text]
  • Photosynthetic Reaction Centre of Chloroflexus Aurantiacus
    Volume 232, number 2, 364-368 FEB 05888 May 1988 Photosynthetic reaction centre of Chloroflexus aurantiacus Primary structure of M-subunit Yu.A. Ovchinnikovt, N.G. Abdulaev, B.E. Shmuckler, A.A. Zargarov, M.A. Kutuzov, I.N. Telezhinskaya, N.B. Levina and A.S. Zolotarev Shemyakin Institute of Bioorganic Chemistry, USSR Academy of Sciences, ul. Miklukho-Maklaya 16110, 117871 GSP Moscow V-437, USSR Received 24 March 1988 The M-subunit primary structure of the reaction centre (RC) from ChloroJexus auruntiacus composed of 306 amino acid residues has been determined by parallel analysis of the protein and corresponding DNA. The blocked N-terminus as well as replacement of the essential histidine liganding Mg of an accessory bacteriochlorophyll in purple bacteria by leucine distinguishes the M-subunit of Chloroflexus RC from that of purple bacteria. Photosynthesis; Reaction center; Subunit structure; Amino acid sequence; Nucleotide sequence; (Chloroflexus aurantiacus) 1. INTRODUCTION Chloroflexus RC to be homologous to the M- subunit of the purple bacteria. This paper presents The reaction centre (RC) is a protein-pigment the primary structure and tentative folding of the complex where the primary events of light energy polypeptide chain of the Chloroflexus RC M- conversion into chemical energy occur. The reac- subunit. tion centre of filamentous green bacterium Chloroflexus aurantiacus, the smallest RC known 2. MATERIALS AND METHODS today, consists of two protein subunits [l]. The primary structure of one subunit has been recently Experimental procedures applied for separation of highly determined in this laboratory by parallel analysis purified preparation of Chlorofexus RC, cleavage of the pro- of the protein and corresponding DNA [2].
    [Show full text]
  • A Novel Microbialite-Associated Phototrophic Chloroflexi Lineage
    A Novel Microbialite-Associated Phototrophic Chloroflexi Lineage Exhibiting a Quasi-Clonal Pattern along Depth Aurélien Saghaï, Yvan Zivanovic, David Moreira, Rosaluz Tavera, Purificación López-García To cite this version: Aurélien Saghaï, Yvan Zivanovic, David Moreira, Rosaluz Tavera, Purificación López-García. A Novel Microbialite-Associated Phototrophic Chloroflexi Lineage Exhibiting a Quasi-Clonal Pattern along Depth. Genome Biology and Evolution, Society for Molecular Biology and Evolution, 2020, 12 (7), pp.1207 - 1216. 10.1093/gbe/evaa122. hal-03003320 HAL Id: hal-03003320 https://hal.archives-ouvertes.fr/hal-03003320 Submitted on 13 Nov 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. GBE ANovelMicrobialite-AssociatedPhototrophicChloroflexi Lineage Exhibiting a Quasi-Clonal Pattern along Depth Aurelien Saghaı¨1,2,*, Yvan Zivanovic3,DavidMoreira1,RosaluzTavera4,and Purificacion Lopez-Garc ıa1 1Ecologie Systematique Evolution, CNRS, AgroParisTech, UniversiteParis-Saclay,Orsay,France 2Department of Forest Mycology and Plant Pathology, Swedish University of Agricultural Sciences, Uppsala, Sweden 3Institut de Biologie Integrative de la Cellule, CNRS, UniversiteParis-Saclay,Orsay,France 4Departamento de Ecologıa y Recursos Naturales, Universidad Nacional Autonoma de Mexico, Mexico City, Mexico *Corresponding author: E-mail: [email protected]. Accepted: 10 June 2020 Abstract Chloroflexales (Chloroflexi) are typical members of the anoxygenic photosynthesizing component of microbial mats and have mostly been characterized from communities associated to hot springs.
    [Show full text]
  • Comparative Genomics Provides Evidence for the 3-Hydroxypropionate Autotrophic Pathway in filamentous Anoxygenic Phototrophic Bacteria and in Hot Spring Microbial Mats
    Environmental Microbiology (2007) doi:10.1111/j.1462-2920.2007.01323.x Comparative genomics provides evidence for the 3-hydroxypropionate autotrophic pathway in filamentous anoxygenic phototrophic bacteria and in hot spring microbial mats Christian G. Klatt,1* Donald A. Bryant2 and rial photosynthate produced by the reductive pentose David M. Ward1 phosphate cycle. 1Department of Land Resources and Environmental Sciences, Montana State University, Bozeman, MT, Introduction USA. 2Department of Biochemistry and Molecular Biology, The The microbial mats that develop in the effluent channels Pennsylvania State University, PA, USA. of alkaline siliceous hot springs of Yellowstone National Park are model systems for the study of microbial com- munity ecology, and they are valuable modern analogues Summary to ancient stromatolite formations (Ward et al., 1998; Stable carbon isotope signatures of diagnostic lipid 2006; van der Meer et al., 2000). Based on our molecular biomarkers have suggested that Roseiflexus spp., the and microscopic studies of Octopus and Mushroom dominant filamentous anoxygenic phototrophic bac- Springs, these mat communities are dominated by two teria inhabiting microbial mats of alkaline siliceous groups of phototrophs at 60 and 65°C: unicellular cyano- hot springs, may be capable of fixing bicarbonate via bacteria (Synechococcus spp.) and filamentous anoxy- the 3-hydroxypropionate pathway, which has been genic phototrophs (FAPs) related to Chloroflexus and characterized in their distant relative, Chloroflexus Roseiflexus spp.
    [Show full text]
  • Chloroflexus Aurantiacus 3581
    JOURNAL OF BACTERIOLOGY, June 2009, p. 3580–3587 Vol. 191, No. 11 0021-9193/09/$08.00ϩ0 doi:10.1128/JB.00110-09 Copyright © 2009, American Society for Microbiology. All Rights Reserved. Role of the AcsF Protein in Chloroflexus aurantiacusᰔ† Kuo-Hsiang Tang, Jianzhong Wen, Xianglu Li, and Robert E. Blankenship* Departments of Biology and Chemistry, Campus Box 1137, Washington University, St. Louis, Missouri 63130 Received 26 January 2009/Accepted 27 March 2009 The green phototrophic bacteria contain a unique complement of chlorophyll pigments, which self-assemble efficiently into antenna structures known as chlorosomes with little involvement of protein. The few proteins found in chlorosomes have previously been thought to have a primarily structural function. The biosynthetic pathway of the chlorosome pigments, bacteriochlorophylls c, d, and e, is not well understood. In this report, we used spectroscopic, proteomic, and gene expression approaches to investigate the chlorosome proteins of the green filamentous anoxygenic phototrophic bacterium Chloroflexus aurantiacus. Surprisingly, Mg-protoporphy- rin IX monomethyl ester (oxidative) cyclase, AcsF, was identified under anaerobic growth conditions. The AcsF protein was found in the isolated chlorosome fractions, and the proteomics analysis suggested that significant Downloaded from portions of the AcsF proteins are not accessible to protease digestion. Additionally, quantitative real-time PCR studies showed that the transcript level of the acsF gene is not lower in anaerobic growth than in semiaerobic growth. Since the proposed enzymatic activity of AcsF requires molecular oxygen, our studies suggest that the roles of AcsF in C. aurantiacus need to be investigated further. jb.asm.org The unique chlorosome antenna complexes found in green monomethyl ester anaerobic cyclase (BchE).
    [Show full text]
  • Primary Photochemistry in the Facultatively Aerobic Green
    Proc. Natl Acad. Sci. USA Vol. 79, pp. 6532-6536, November 1982 Biophysics Primary photochemistry in the facultatively aerobic green photosynthetic bacterium Chloroflexus aurantiacus (photosynthesis/bacteriochlorophyll/reaction center/redox potentiometry/evolution) BARRY D. BRUCE*, R. CLINTON FULLER*t, AND ROBERT E. BLANKENSHIPtt *Department of Biochemistry, University of Massachusetts; and MDepartment ofChemistry, Amherst College, Amherst, Massachusetts 01002 Communicated by William Arnold, August 18, 1982 ABSTRACT Photochemical activity was examined in mem- chemistry of Chloroflexus, and in this paper we present evi- brane fragments and a purified membrane preparation from dence that the primary photoact is nearly identical to that found Chloroflexu8. Flash-induced absorption difference spectroscopy in the purple bacteria and unlike that found in other green strongly suggests a primary donor (P8m) that is more similar to the bacteria. P870 bacteriochlorophyll a dimer found in the purple photosyn- thetic bacteria than it is to PNO found in the anaerobic green bac- MATERIALS AND METHODS teria. Redox measurements on PW5 and an early acceptor also in- dicate a photochemical system characteristic of the purple bac- Preparative Methods. Organism and growth conditions were teria. The membrane preparation contains a tightly bound type the same as described by Sprague et aL (14). The isolation of c cytochrome, c5u, that is closely coupled to the reaction center whole membranes and purified cytoplasmic membranes used as indicated by its ability to rereduce photooxidized P8. Chioro- the procedures described by Feick et aL (13). flexus thus appears to be distinct photochemically from other fam- Single-Flash-Induced Absorption Change Measurements. ilies ofphotosynthetic bacteria and may occupy an important role Measurements were made on a homemade single-beam laser in photosynthetic evolution.
    [Show full text]
  • The Cyanobacterial Genome Core and the Origin of Photosynthesis
    The cyanobacterial genome core and the origin of photosynthesis Armen Y. Mulkidjanian*†‡, Eugene V. Koonin§, Kira S. Makarova§, Sergey L. Mekhedov§, Alexander Sorokin§, Yuri I. Wolf§, Alexis Dufresne¶, Fre´ de´ ric Partensky¶, Henry Burdʈ, Denis Kaznadzeyʈ, Robert Haselkorn†**, and Michael Y. Galperin†§ *School of Physics, University of Osnabru¨ck, D-49069 Osnabru¨ck, Germany; ‡A. N. Belozersky Institute of Physico–Chemical Biology, Moscow State University, Moscow 119899, Russia; §National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, Bethesda, MD 20894; ¶Station Biologique, Unite´Mixte de Recherche 7144, Centre National de la Recherche Scientifique et Universite´Paris 6, BP74, F-29682 Roscoff Cedex, France; ʈIntegrated Genomics, Inc., Chicago, IL 60612; and **Department of Molecular Genetics and Cell Biology, University of Chicago, 920 East 58th Street, Chicago, IL 60637 Contributed by Robert Haselkorn, July 14, 2006 Comparative analysis of 15 complete cyanobacterial genome se- to trace the conservation of these genes among other taxa. We quences, including ‘‘near minimal’’ genomes of five strains of analyzed the phylogenetic affinities of genes in this set and Prochlorococcus spp., revealed 1,054 protein families [core cya- identified previously unrecognized candidate photosynthetic nobacterial clusters of orthologous groups of proteins (core Cy- genes. We further used this gene set to address the identity of the OGs)] encoded in at least 14 of them. The majority of the core first phototrophs, a subject of intense discussion in recent years CyOGs are involved in central cellular functions that are shared (8, 9, 12–33). We show that cyanobacteria and plants share with other bacteria; 50 core CyOGs are specific for cyanobacteria, numerous photosynthesis-related genes that are missing in ge- whereas 84 are exclusively shared by cyanobacteria and plants nomes of other phototrophs.
    [Show full text]
  • Mutagenesis and Cloning of Photosynesis Genes of the Filamentous Bacterium, Chloroflexus Aurantiacus
    Illinois Wesleyan University Digital Commons @ IWU Honors Projects Biology 4-28-2000 Mutagenesis and Cloning of Photosynesis Genes of the Filamentous Bacterium, Chloroflexus aurantiacus Kristi L. Berger '00 Illinois Wesleyan University Follow this and additional works at: https://digitalcommons.iwu.edu/bio_honproj Part of the Biology Commons Recommended Citation Berger '00, Kristi L., "Mutagenesis and Cloning of Photosynesis Genes of the Filamentous Bacterium, Chloroflexus aurantiacus" (2000). Honors Projects. 7. https://digitalcommons.iwu.edu/bio_honproj/7 This Article is protected by copyright and/or related rights. It has been brought to you by Digital Commons @ IWU with permission from the rights-holder(s). You are free to use this material in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s) directly, unless additional rights are indicated by a Creative Commons license in the record and/ or on the work itself. This material has been accepted for inclusion by faculty at Illinois Wesleyan University. For more information, please contact [email protected]. ©Copyright is owned by the author of this document. Mutagenesis and Cloning of Photosynthesis Genes of the Filamentous Bacterium, Chloroflexus aurantiacus Kristi L. Berger Illinois Wesleyan University Research Honors Designated: April 14, 2000 Thesis Submitted: April 28, 2000 K. Berger 2 ABSTRACT Chloroflexus aurantiacus is a green non-sulfur bacterium that preferentially grows photosynthetically under lighted, anaerobic conditions. The biosynthetic pathway for one of the photosynthetic pigments, bacteriochlorophyll c, is not well understood but may share common steps with the better understood bacteriochlorophyll a pathway.
    [Show full text]
  • Chloroflexus Aurantiacus
    Tang et al. BMC Genomics 2011, 12:334 http://www.biomedcentral.com/1471-2164/12/334 RESEARCHARTICLE Open Access Complete genome sequence of the filamentous anoxygenic phototrophic bacterium Chloroflexus aurantiacus Kuo-Hsiang Tang1, Kerrie Barry2, Olga Chertkov3, Eileen Dalin2, Cliff S Han3, Loren J Hauser4, Barbara M Honchak1, Lauren E Karbach1,7, Miriam L Land4, Alla Lapidus5, Frank W Larimer4, Natalia Mikhailova5, Samuel Pitluck2, Beverly K Pierson6 and Robert E Blankenship1* Abstract Background: Chloroflexus aurantiacus is a thermophilic filamentous anoxygenic phototrophic (FAP) bacterium, and can grow phototrophically under anaerobic conditions or chemotrophically under aerobic and dark conditions. According to 16S rRNA analysis, Chloroflexi species are the earliest branching bacteria capable of photosynthesis, and Cfl. aurantiacus has been long regarded as a key organism to resolve the obscurity of the origin and early evolution of photosynthesis. Cfl. aurantiacus contains a chimeric photosystem that comprises some characters of green sulfur bacteria and purple photosynthetic bacteria, and also has some unique electron transport proteins compared to other photosynthetic bacteria. Methods: The complete genomic sequence of Cfl. aurantiacus has been determined, analyzed and compared to the genomes of other photosynthetic bacteria. Results: Abundant genomic evidence suggests that there have been numerous gene adaptations/replacements in Cfl. aurantiacus to facilitate life under both anaerobic and aerobic conditions, including duplicate genes and gene clusters for the alternative complex III (ACIII), auracyanin and NADH:quinone oxidoreductase; and several aerobic/ anaerobic enzyme pairs in central carbon metabolism and tetrapyrroles and nucleic acids biosynthesis. Overall, genomic information is consistent with a high tolerance for oxygen that has been reported in the growth of Cfl.
    [Show full text]
  • Phototrophic Methane Oxidation in a Member of the Chloroflexi Phylum
    bioRxiv preprint doi: https://doi.org/10.1101/531582; this version posted January 26, 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. Phototrophic Methane Oxidation in a Member of the Chloroflexi Phylum Lewis M. Ward1,2, Patrick M. Shih3,4, James Hemp5, Takeshi Kakegawa6, Woodward W. Fischer7, Shawn E. McGlynn2,8,9 1. Department of Earth & Planetary Sciences, Harvard University, Cambridge, MA USA. 2. Earth-Life Science Institute, Tokyo Institute of Technology, Ookayama, Meguro-ku, Tokyo, Japan. 3. Department of Plant Biology, University of California, Davis, Davis, CA USA. 4. Department of Energy, Joint BioEnergy Institute, Emeryville, CA USA. 5. School of Medicine, University of Utah, Salt Lake City, UT USA. 6. Department of Geosciences, Tohoku University, Sendai City, Japan 7. Division of Geological & Planetary Sciences, California Institute of Technology, Pasadena, CA USA. 8. Biofunctional Catalyst Research Team, RIKEN Center for Sustainable Resource Science, Wako-shi Japan 9. Blue Marble Space Institute of Science, Seattle, WA, USA Abstract: Biological methane cycling plays an important role in Earth’s climate and the global carbon cycle, with biological methane oxidation (methanotrophy) modulating methane release from numerous environments including soils, sediments, and water columns. Methanotrophy is typically coupled to aerobic respiration or anaerobically via the reduction of sulfate, nitrate, or metal oxides, and while the possibility of coupling methane oxidation to phototrophy (photomethanotrophy) has been proposed, no organism has ever been described that is capable of this metabolism.
    [Show full text]
  • Enzymatic Activity of the Alternative Complex III As a Menaquinol:Auracyanin Oxidoreductase in the Electron Transfer Chain of Chloroflexus Aurantiacus
    FEBS Letters 583 (2009) 3275–3279 journal homepage: www.FEBSLetters.org Enzymatic activity of the alternative complex III as a menaquinol:auracyanin oxidoreductase in the electron transfer chain of Chloroflexus aurantiacus Xinliu Gao, Yueyong Xin, Robert E. Blankenship * Department of Biology, Washington University in St. Louis, United States Department of Chemistry, Washington University in St. Louis, United States article info abstract Article history: The surprising lack of the cytochrome bc1 complex in the filamentous anoxygenic phototrophic bac- Received 24 July 2009 terium Chloroflexus aurantiacus suggests that a functional replacement exists to link the cyclic elec- Revised 5 September 2009 tron transfer chain. Earlier work identified the alternative complex III (ACIII) as a substitute of Accepted 8 September 2009 cytochrome bc complex. Herein, the enzymatic activity of ACIII is studied. The results strongly sup- Available online 13 September 2009 1 port the view that the ACIII functions as menaquinol:auracyanin oxidoreductase in the C. aurantia- Edited by Richard Cogdell cus electron transfer chain. Among all the substrates tested, auracyanin is the most efficient electron acceptor of ACIII, suggesting that ACIII directly transfers the electron to auracyanin instead of cyto- chrome c-554. The lack of sensitivity to common inhibitors of the cytochrome bc complex indicates Keywords: 1 Alternative complex III a different catalytic mechanism for the ACIII complex. Photosynthesis Ó 2009 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved. Electron transfer Auracyanin Chloroflexus aurantiacus 1. Introduction this group is the Chloroflexus aurantiacus. This thermophilic photo- synthetic bacterium possesses an unusual electron transfer path- The five phyla of bacteria that possess chlorophyll-based photo- way due to the lack of both the cytochrome bc1 complex and a synthesis are purple bacteria, cyanobacteria, heliobacteria, green soluble c-type cytochrome [5].
    [Show full text]
  • Temporal Metatranscriptomic Patterning in Phototrophic Chloroflexi Inhabiting a Microbial Mat in a Geothermal Spring
    The ISME Journal (2013) 7, 1775–1789 & 2013 International Society for Microbial Ecology All rights reserved 1751-7362/13 www.nature.com/ismej ORIGINAL ARTICLE Temporal metatranscriptomic patterning in phototrophic Chloroflexi inhabiting a microbial mat in a geothermal spring Christian G Klatt1,6, Zhenfeng Liu2, Marcus Ludwig2, Michael Ku¨ hl3,4, Sheila I Jensen3, Donald A Bryant2,5 and David M Ward1 1Department of Land Resources and Environmental Sciences, Montana State University, Bozeman, MT, USA; 2Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, USA; 3Marine Biology Section, Department of Biology, University of Copenhagen, Helsingør, Denmark; 4Plant Functional Biology and Climate Change Cluster, Department of Environmental Science, University of Technology, Sydney, New South Wales, Australia and 5Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT, USA Filamentous anoxygenic phototrophs (FAPs) are abundant members of microbial mat communities inhabiting neutral and alkaline geothermal springs. Natural populations of FAPs related to Chloroflexus spp. and Roseiflexus spp. have been well characterized in Mushroom Spring, where they occur with unicellular cyanobacteria related to Synechococcus spp. strains A and B0. Metatranscriptomic sequencing was applied to the microbial community to determine how FAPs regulate their gene expression in response to fluctuating environmental conditions and resource availability over a diel period. Transcripts for genes involved in the biosynthesis of bacteriochlor- ophylls (BChls) and photosynthetic reaction centers were much more abundant at night. Both Roseiflexus spp. and Chloroflexus spp. expressed key genes involved in the 3-hydroxypropionate (3-OHP) carbon dioxide fixation bi-cycle during the day, when these FAPs have been thought to perform primarily photoheterotrophic and/or aerobic chemoorganotrophic metabolism.
    [Show full text]