<I>Thauera Aminoaromatica</I> Strain MZ1T
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Heterologous Expression and Identification of the Genes Involved
Heterologous Expression and Identification of the Genes Involved in Anaerobic Degradation of 1,3-Dihydroxybenzene (Resorcinol) in Azoarcus anaerobiusᰔ Paula I. Darley,1† Jutta A. Hellstern,1†‡ Javier I. Medina-Bellver,2 Silvia Marque´s,2 Bernhard Schink,1 and Bodo Philipp1* Fachbereich Biologie, Universita¨t Konstanz, D-78457 Constance, Germany,1 and Estacio´n Experimental del Zaidı´n, C/. Profesor Albareda 1, E-18008 Granada, Spain2 Received 9 November 2006/Accepted 9 March 2007 Azoarcus anaerobius, a strictly anaerobic, gram-negative bacterium, utilizes resorcinol as a sole carbon and energy source with nitrate as an electron acceptor. Previously, we showed that resorcinol degradation by this bacterium is initiated by two oxidative steps, both catalyzed by membrane-associated enzymes that lead to the formation of hydroxyhydroquinone (HHQ; 1,2,4-benzenetriol) and 2-hydroxy-1,4-benzoquinone (HBQ). This study presents evidence for the further degradation of HBQ in cell extracts to form acetic and malic acids. To identify the A. anaerobius genes required for anaerobic resorcinol catabolism, a cosmid library with genomic DNA was constructed and transformed into the phylogenetically related species Thauera aromatica, which cannot grow with resorcinol. By heterologous complementation, a transconjugant was identified that gained the ability to metabolize resorcinol. Its cosmid, designated R؉, carries a 29.88-kb chromosomal DNA fragment containing 22 putative genes. In cell extracts of T. aromatica transconjugants, resorcinol was degraded to HHQ, HBQ, and acetate, suggesting that cosmid R؉ carried all of the genes necessary for resorcinol degradation. On the basis of the physiological characterization of T. aromatica transconjugants carrying transposon insertions -in different genes of cosmid R؉, eight open reading frames were found to be essential for resorcinol miner alization. -
Resolving Electron Transport Pathways in the Selenate Respiring Bacterium Thauera Selenatis
RESOLVING ELECTRON TRANSPORT PATHWAYS IN THE SELENATE RESPIRING BACTERIUM THAUERA SELENATIS. Submitted by Elisabeth Clare Lowe To the University of Exeter as a thesis for the degree of Doctor of Philosophy in Biological Sciences, July 2008. This thesis is available for Library use on the understanding that it is copyright material and that no quotation from the thesis may be published without proper acknowledgement. I certify that all material in this thesis which is not my own work has been identified and that no material has previously been submitted and approved for the award of a degree by this or any other University. Elisabeth Clare Lowe Acknowledgements I would like to thank Clive Butler for all his help, support, encouragement and enthusiasm, especially during the move to Exeter and my last few months in the lab. Thanks to Ian Singleton for help and enthusiasm during my time in Newcastle. I also owe huge thanks to everyone in Team Butler, past and present, Carys Watts for helping me get started and all the help with the Thauera preps, Auntie Helen for being brilliant, Jim Leaver for rugby, beer, tea-offs and 30 hour growth curves and Lizzy Dridge for being a great friend and scientific team member, in and out of the lab. Thanks also to everyone in M3013 at the start of my PhD and everyone in the Biocat at the end, for lots of help, borrowing of equipment and most importantly, tea. Thanks to AFP for help, tea breaks and motivation, especially during the writing period. Thank you to everyone who has been generous with their time and equipment, namely Prof. -
Anaerobic Degradation of Bicyclic Monoterpenes in Castellaniella Defragrans
H OH metabolites OH Article Anaerobic Degradation of Bicyclic Monoterpenes in Castellaniella defragrans Edinson Puentes-Cala 1, Manuel Liebeke 2 ID , Stephanie Markert 3 and Jens Harder 1,* 1 Department of Microbiology, Max Planck Institute for Marine Microbiology, Celsiusstr. 1, 28359 Bremen, Germany; [email protected] 2 Department of Symbiosis, Max Planck Institute for Marine Microbiology, Celsiusstr. 1, 28359 Bremen, Germany; [email protected] 3 Pharmaceutical Biotechnology, University Greifswald, Felix-Hausdorff-Straße, 17489 Greifswald, Germany; [email protected] * Correspondence: [email protected]; Tel.: +49-421-2028-750 Received: 23 January 2018; Accepted: 2 February 2018; Published: 7 February 2018 Abstract: The microbial degradation pathways of bicyclic monoterpenes contain unknown enzymes for carbon–carbon cleavages. Such enzymes may also be present in the betaproteobacterium Castellaniella defragrans, a model organism to study the anaerobic monoterpene degradation. In this study, a deletion mutant strain missing the first enzyme of the monocyclic monoterpene pathway transformed cometabolically the bicyclics sabinene, 3-carene and α-pinene into several monocyclic monoterpenes and traces of cyclic monoterpene alcohols. Proteomes of cells grown on bicyclic monoterpenes resembled the proteomes of cells grown on monocyclic monoterpenes. Many transposon mutants unable to grow on bicyclic monoterpenes contained inactivated genes of the monocyclic monoterpene pathway. These observations suggest that the monocyclic degradation pathway is used to metabolize bicyclic monoterpenes. The initial step in the degradation is a decyclization (ring-opening) reaction yielding monocyclic monoterpenes, which can be considered as a reverse reaction of the olefin cyclization of polyenes. Keywords: monoterpene; anaerobic metabolism; ring-opening reactions; carbon–carbon lyase; isoprenoid degradation 1. -
Thauera Selenatis Gen
INTERNATIONALJOURNAL OF SYSTEMATICBACTERIOLOGY, Jan. 1993, p. 135-142 Vol. 43, No. 1 0020-7713/93/010135-08$02.00/0 Copyright 0 1993, International Union of Microbiological Societies Thauera selenatis gen. nov., sp. nov., a Member of the Beta Subclass of Proteobacteria with a Novel Type of Anaerobic Respiration J. M. MACY,’” S. RECH,’? G. AULING,2 M. DORSCH,3 E. STACKEBRANDT,3 AND L. I. SLY3 Department of Animal Science, University of California, Davis, Davis, Cali ornia 95616’; Institut fur Mikrobiologie, Universitat Hannover, 3000 Hannover 1, Germany4 and Centre for Bacterial Diversity and Identification, Department of Microbiology, The University of Queensland, Brisbane, Australia 40723 A recently isolated, selenate-respiring microorganism (strain AXT [T = type strain]) was classified by using a polyphasic approach in which both genotypic and phenotypic characteristics were determined. Strain AXT is a motile, gram-negative, rod-shaped organism with a single polar flagellum. On the basis of phenotypic characteristics, this organism can be classified as a Pseudomonas sp. However, a comparison of the 16s rRNA sequence of strain AXT with the sequences of other organisms indicated that strain AXT is most similar to members of the beta subclass (level of similarity, 86.8%) rather than to members of the gamma subclass (level of similarity, 80.2%) of the Pruteobacteriu. The presence of the specific polyamine 2-hydroxyputrescine and the presence of a ubiquinone with eight isoprenoid units in the side chain (ubiquinone Q-8) excluded strain AXT from the authentic genus Pseudumnas and allowed placement in the beta subclass of the Pruteobacteria. Within the beta subclass, strain AXT is related to ZodobacterjZuvatiZe.The phylogenetic distance (level of similarity, less than 90%), as well as a lack of common phenotypic characteristics between these organisms, prevents classification of strain AXT as a member of the genus Zodobacter. -
A Bacterial Process for Selenium Nanosphere Assembly
A bacterial process for selenium nanosphere assembly Charles M. Debieuxa, Elizabeth J. Dridgea,1, Claudia M. Muellera,1, Peter Splatta, Konrad Paszkiewicza, Iona Knighta, Hannah Florancea, John Lovea, Richard W. Titballa, Richard J. Lewisb, David J. Richardsonc, and Clive S. Butlera,2 aBiosciences, College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, United Kingdom; bInstitute for Cell and Molecular Biosciences, University of Newcastle, Newcastle upon Tyne NE2 4HH, United Kingdom; and cSchool of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, United Kingdom Edited by Dianne K. Newman, California Institute of Technology/Howard Hughes Medical Institute, Pasadena, CA, and accepted by the Editorial Board July 1, 2011 (received for review April 13, 2011) Thauera selenatis 2− − þ 2− During selenate respiration by , the reduction of SeO4 þ 2e þ 2H ⇆ SeO3 þ H2O [1] selenate results in the formation of intracellular selenium (Se) deposits that are ultimately secreted as Se nanospheres of approxi- and mately 150 nm in diameter. We report that the Se nanospheres 2− − þ 0 SeO þ 4e þ 6H ⇆ Se þ 3H2O: [2] are associated with a protein of approximately 95 kDa. Subsequent 3 experiments to investigate the expression and secretion profile of Thauera selenatis (a β-proteobacterium) is by far the best-studied this protein have demonstrated that it is up-regulated and secreted selenate respiring bacterium (5–8). The selenate reductase in response to increasing selenite concentrations. The protein was (SerABC) isolated from T. selenatis (6) is a soluble periplasmic purified from Se nanospheres, and peptide fragments from a tryp- enzyme. -
Identification and Characterization of Two Thauera Aromatica Strain T1 Genes Induced
Identification and Characterization of Two Thauera aromatica Strain T1 Genes Induced by p-Cresol A dissertation presented to the faculty of the College of Arts and Sciences of Ohio University In partial fulfillment of the requirements for the degree Doctor of Philosophy Mohor Chatterjee August 2012 © 2012 Mohor Chatterjee. All Rights Reserved. 2 This dissertation titled Identification and Characterization of Two Thauera aromatica Strain T1 Genes Induced by p-Cresol by MOHOR CHATTERJEE has been approved for the Program of Molecular and Cellular Biology and the College of Arts and Sciences by Peter W. Coschigano Associate Professor of Biomedical Sciences Howard Dewald Interim Dean, College of Arts and Sciences 3 ABSTRACT CHATTERJEE, MOHOR, Ph.D., August 2012, Molecular and Cellular Biology Identification and Characterization of Two Thauera aromatica Strain T1 Genes Induced by p-Cresol (109 pp) Director of Dissertation: Peter W. Coschigano p-Cresol is a toxic aromatic compound found in the environment and is a constituent of many disinfectants and preservatives. It may act as a tumor promoter and the US Environmental Protection Agency has listed it as a possible human carcinogen. Thauera aromatica strain T1 is a facultative anaerobic, denitrifying, Gram-negative bacterium that is able to degrade many aromatic compounds including toluene and p- cresol. A proteomics approach was used to identify proteins from T. aromatica strain T1 that have differential expression when cells are induced by p-cresol in comparison to benzoate, a common downstream metabolic intermediate in the degradation of many aromatic compounds. Sequences of peptides from proteins selectively up-regulated by p- cresol in comparison to benzoate were obtained by MS analysis and compared against databases of known proteins from other microorganisms. -
Microbial Community of a Gasworks Aquifer and Identification of Nitrate
Water Research 132 (2018) 146e157 Contents lists available at ScienceDirect Water Research journal homepage: www.elsevier.com/locate/watres Microbial community of a gasworks aquifer and identification of nitrate-reducing Azoarcus and Georgfuchsia as key players in BTEX degradation * Martin Sperfeld a, Charlotte Rauschenbach b, Gabriele Diekert a, Sandra Studenik a, a Institute of Microbiology, Friedrich Schiller University Jena, Department of Applied and Ecological Microbiology, Philosophenweg 12, 07743 Jena, Germany ® b JENA-GEOS -Ingenieurbüro GmbH, Saalbahnhofstraße 25c, 07743 Jena, Germany article info abstract Article history: We analyzed a coal tar polluted aquifer of a former gasworks site in Thuringia (Germany) for the Received 9 August 2017 presence and function of aromatic compound-degrading bacteria (ACDB) by 16S rRNA Illumina Received in revised form sequencing, bamA clone library sequencing and cultivation attempts. The relative abundance of ACDB 18 December 2017 was highest close to the source of contamination. Up to 44% of total 16S rRNA sequences were affiliated Accepted 18 December 2017 to ACDB including genera such as Azoarcus, Georgfuchsia, Rhodoferax, Sulfuritalea (all Betaproteobacteria) Available online 20 December 2017 and Pelotomaculum (Firmicutes). Sequencing of bamA, a functional gene marker for the anaerobic benzoyl-CoA pathway, allowed further insights into electron-accepting processes in the aquifer: bamA Keywords: Environmental pollutions sequences of mainly nitrate-reducing Betaproteobacteria were abundant in all groundwater samples, Microbial communities whereas an additional sulfate-reducing and/or fermenting microbial community (Deltaproteobacteria, Bioremediation Firmicutes) was restricted to a highly contaminated, sulfate-depleted groundwater sampling well. By Box pathway conducting growth experiments with groundwater as inoculum and nitrate as electron acceptor, or- Functional gene marker ganisms related to Azoarcus spp. -
Nitrite Reductase Genes in Halobenzoate Degrading Denitrifying Bacteria
FEMS Microbiology Ecology 43 (2003) 349^357 www.fems-microbiology.org Nitrite reductase genes in halobenzoate degrading denitrifying bacteria Bongkeun Song Ã, Bess B. Ward Department of Geosciences, Princeton University, Princeton, NJ 08544-1003, USA Received 5 July 2002;received in revised form 9 October 2002;accepted 18 October 2002 First published online 14 November 2002 Abstract Diversity of the functional genes encoding dissimilatory nitrite reductase was investigated for the first time in denitrifying halobenzoate degrading bacteria and in two 4-chlorobenzoate degrading denitrifying consortia. Nitrite reductase genes were PCR-amplified with degenerate primers (specific to the two different types of respiratory nitrite reductase, nirS and nirK), cloned and sequenced to determine which type of nitrite reductase was present in each isolate and consortium. Halobenzoate degrading isolates belonging to the genera Ochrobactrum, Ensifer and Mesorhizobium, as well as Pseudomonas mendocina CH91 were found to have nirK genes, which were closely related to the previously published nirK genes of Ochrobactrum anthropi, Achromobacter cycloclastes, Alcaligenes faecalis and Pseudomonas aureofaciens, respectively. The isolates assigned to the genera Acidovorax, Azoarcus and Thauera as well as all other species in the genera Thauera and Azoarcus contained nirS genes, which were closely related to the nirS genes from Pseudomonas stutzeri with some exceptions. In addition, only nirS genes were found in 4-chlorobenzoate degrading denitrifying consortia. Three different major terminal restriction fragments from the nirS genes were detected by terminal restriction fragment length polymorphism analysis of the consortia, and five different nirS genes were cloned from one consortium. Three nirS gene clones were closely related to nirS genes from Thauera chlorobenzoica, Azoarcus tolulyticus and Pseudomonas aeruginosa, respectively. -
Thauera Aminoaromatica MZ1T Identified As A
bioengineering Article Thauera aminoaromatica MZ1T Identified as a Polyhydroxyalkanoate-Producing Bacterium within a Mixed Microbial Consortium 1, 1, 1 2 3 Dana I. Colpa y, Wen Zhou y, Jan Pier Wempe , Jelmer Tamis , Marc C. A. Stuart , Janneke Krooneman 1 and Gert-Jan W. Euverink 1,* 1 Products and Processes for Biotechnology Group, Engineering and Technology Institute Groningen, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands; [email protected] (D.I.C.); [email protected] (W.Z.); [email protected] (J.P.W.); [email protected] (J.K.) 2 Paques Technology B.V., Tjalke de Boerstrjitte 24, 8561 EL Balk, The Netherlands; [email protected] 3 Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 7, 9747 AG Groningen, The Netherlands; [email protected] * Correspondence: [email protected]; Tel.: +31-50-363-9203 These authors contributed equally to this work. y Received: 22 January 2020; Accepted: 19 February 2020; Published: 21 February 2020 Abstract: Polyhydroxyalkanoates (PHAs) form a highly promising class of bioplastics for the transition from fossil fuel-based plastics to bio-renewable and biodegradable plastics. Mixed microbial consortia (MMC) are known to be able to produce PHAs from organic waste streams. Knowledge of key-microbes and their characteristics in PHA-producing consortia is necessary for further process optimization and direction towards synthesis of specific types of PHAs. In this study, a PHA-producing mixed microbial consortium (MMC) from an industrial pilot plant was characterized and further enriched on acetate in a laboratory-scale selector with a working volume of 5 L. -
Isolation, Growth, and Metabolism of an Obligately Anaerobic, Selenate-Respiring Bacterium, Strain SES-3 RONALD S
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, Aug. 1994, p. 3011-3019 Vol. 60, No. 8 0099-2240/94/$04.00+0 Copyright X 1994, American Society for Microbiology Isolation, Growth, and Metabolism of an Obligately Anaerobic, Selenate-Respiring Bacterium, Strain SES-3 RONALD S. OREMLAND,* JODI SWITZER BLUM, CHARLES W. CULBERTSON, PIETER T. VISSCHER, LAURENCE G. MILLER, PHILLIP DOWDLE, AND FRANCES E. STROHMAIER U.S. Geological Survey, Menlo Park, Califomia 94025 Received 6 April 1994/Accepted 24 May 1994 A gram-negative, strictly anaerobic, motile vibrio was isolated from a selenate-respiring enrichment culture. The isolate, designated strain SES-3, grew by coupling the oxidation of lactate to acetate plus CO2 with the concomitant reduction of selenate to selenite or of nitrate to ammonium. No growth was observed on sulfate or selenite, but cell suspensions readily reduced selenite to elemental selenium (Seo). Hence, SES-3 can carry out a complete reduction of selenate to Seo. Washed cell suspensions of selenate-grown cells did not reduce nitrate, and nitrate-grown cells did not reduce selenate, indicating that these reductions are achieved by separate inducible enzyme systems. However, both nitrate-grown and selenate-grown cells have a constitutive ability to reduce selenite or nitrite. The oxidation of ['4C] lactate to '4Co2 coupled to the reduction of selenate or nitrate by cell suspensions was inhibited by CCCP (carbonyl cyanide m-chlorophenylhydrazone), cyanide, and azide. High concentrations of selenite (5 mM) were readily reduced to Seo by selenate-grown cells, but selenite appeared to block the synthesis of pyruvate dehydrogenase. Tracer experiments with [75Se] selenite indicated that cell suspensions could achieve a rapid and quantitative reduction of selenite to Seo. -
Comparative Genomics Provides Insights Into the Taxonomy of Azoarcus and Reveals Separate Origins of Nif Genes in the Proposed Azoarcus and Aromatoleum Genera
G C A T T A C G G C A T genes Article Comparative Genomics Provides Insights into the Taxonomy of Azoarcus and Reveals Separate Origins of Nif Genes in the Proposed Azoarcus and Aromatoleum Genera Roberto Tadeu Raittz 1,*,† , Camilla Reginatto De Pierri 2,† , Marta Maluk 3 , Marcelo Bueno Batista 4, Manuel Carmona 5 , Madan Junghare 6, Helisson Faoro 7, Leonardo M. Cruz 2 , Federico Battistoni 8, Emanuel de Souza 2,Fábio de Oliveira Pedrosa 2, Wen-Ming Chen 9, Philip S. Poole 10, Ray A. Dixon 4,* and Euan K. James 3,* 1 Laboratory of Artificial Intelligence Applied to Bioinformatics, Professional and Technical Education Sector—SEPT, UFPR, Curitiba, PR 81520-260, Brazil 2 Department of Biochemistry and Molecular Biology, UFPR, Curitiba, PR 81531-980, Brazil; [email protected] (C.R.D.P.); [email protected] (L.M.C.); [email protected] (E.d.S.); [email protected] (F.d.O.P.) 3 The James Hutton Institute, Invergowrie, Dundee DD2 5DA, UK; [email protected] 4 John Innes Centre, Department of Molecular Microbiology, Norwich NR4 7UH, UK; [email protected] 5 Centro de Investigaciones Biológicas Margarita Salas-CSIC, Department of Biotechnology of Microbes and Plants, Ramiro de Maeztu 9, 28040 Madrid, Spain; [email protected] 6 Faculty of Chemistry, Biotechnology and Food Science, NMBU—Norwegian University of Life Sciences, 1430 Ås, Norway; [email protected] 7 Laboratory for Science and Technology Applied in Health, Carlos Chagas Institute, Fiocruz, Curitiba, PR 81310-020, Brazil; helisson.faoro@fiocruz.br 8 Department of Microbial Biochemistry and Genomics, IIBCE, Montevideo 11600, Uruguay; [email protected] Citation: Raittz, R.T.; Reginatto De 9 Laboratory of Microbiology, Department of Seafood Science, NKMU, Kaohsiung City 811, Taiwan; Pierri, C.; Maluk, M.; Bueno Batista, [email protected] M.; Carmona, M.; Junghare, M.; Faoro, 10 Department of Plant Sciences, University of Oxford, South Parks Road, Oxford OX1 3RB, UK; H.; Cruz, L.M.; Battistoni, F.; Souza, [email protected] E.d.; et al. -
Cross-Feeding Between Thauera Aminoaromatica and Rhodococcus
bioRxiv preprint doi: https://doi.org/10.1101/2020.01.31.929745; this version posted February 2, 2020. 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-ND 4.0 International license. 1 Cross-feeding between Thauera aminoaromatica and Rhodococcus 2 pyridinivorans drove quinoline biodegradation in a denitrifying 3 bioreactor 4 5 Xinxin Wu, Xiaogang Wu, Ji Li, Qiaoyu Wu, Yiming Ma, Weikang Sui, Liping Zhao, 6 Xiaojun Zhang* 7 8 State Key Laboratory of Microbial Metabolism, Joint International Research 9 Laboratory of Metabolic & Developmental Sciences, and School of Life Sciences & 10 Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240, China 11 12 Running title: Syntrophic denitrifying degradation of quinoline 13 14 * Corresponding author: Dr. Xiaojun Zhang, 15 Tel: +86 21 34204878, E-mail: [email protected] 16 800 Dongchuan Road, Minhang District, Shanghai, 200240, China 17 bioRxiv preprint doi: https://doi.org/10.1101/2020.01.31.929745; this version posted February 2, 2020. 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-ND 4.0 International license. 18 Abstract 19 The complex bacterial community is predominated by several taxa, such as 20 Thauera and Rhodococcus, in a quinoline-degrading denitrifying bioreactor. Yet it 21 remains unclear about how the interactions between the different bacteria mediate the 22 quinoline metabolism in denitrifying condition.