A Plasmid Vector with a Selectable Marker for Halophilic Archaebacteria MELISSA L

Total Page:16

File Type:pdf, Size:1020Kb

A Plasmid Vector with a Selectable Marker for Halophilic Archaebacteria MELISSA L JOURNAL OF BACTERIOLOGY, Feb. 1990, p. 756-761 Vol. 172, No. 2 0021-9193/90/020756-06$02.00/0 Copyright © 1990, American Society for Microbiology A Plasmid Vector with a Selectable Marker for Halophilic Archaebacteria MELISSA L. HOLMES* AND MICHAEL L. DYALL-SMITH Department of Microbiology, University of Melbourne, Parkville, Victoria 3052, Australia Received 25 May 1989/Accepted 10 November 1989 A mutant resistant to the gyrase inhibitor novobiocin was selected from a halophilic archaebacterium belonging to the genus Haloferax. Chromosomal DNA from this mutant was able to transform wild-type ceils to novobiocin resistance, and these transformants formed visible colonies in 3 to 4 days on selective plates. The resistance gene was isolated on a 6.7-kilobase DNA KpnI fragment, which was inserted into a cryptic multicopy plasmid (pHK2) derived from the same host strain. The recombinant plasmid transformed wild-type cells at a high efficiency (>106/pg), was stably maintained, and could readily be reisolated from transformants. It could also transform Halobacterium volcanii and appears to be a useful system for genetic analysis in halophilic archaebacteria. Archaebacteria are phylogenetically distinct from eubac- cle was the lack of selectable traits, such as antibiotic teria and eucaryotes and can be broadly divided into three resistance. groups: methanogens, sulfur-dependent thermoacidophiles, The aim of this study was to find a selectable marker for and extreme halophiles (for a review, see reference 22). use in constructing plasmid vectors suitable for isolating and Numerous halobacteria have been isolated from hypersaline studying genes in halobacteria. We report the successful environments around the world, but it is only recently that construction of such a vector by using a gene conferring their taxonomy has been analyzed in a rigorous and compre- resistance to novobiocin and a cryptic plasmid from Halo- hensive fashion. The studies of Tindall et al. (20) and ferax isolate Aa 2.2. Torreblanca et al. (21) identified six genera: Halobacterium, Halococcus, Haloarcula, Haloferax, Natronobacterium, MATERIALS AND METHODS and Natronococcus. Bacterial strains. The following halobacterial strains were Genetic analysis of halobacteria has been complicated in used in this study: Haloferax phenon K isolate Aa 2.2 from strains such as Halobacterium halobium by the high rate of M. Torreblanca, University of Alicante, Alicante, Spain; spontaneous mutation due to insertion sequences and be- Halobacterium (now Haloferax) volcanii NCMB2012 and cause H. halobium strains have complex nutrient require- Halobacterium halobium NCMB777 from the National Col- ments (15). However, there are halobacteria with low muta- lection of Marine Bacteria, Aberdeen, Scotland; and Halo- tion rates that grow in a mineral salts medium containing a bacterium halobium DSM617 and strain RI from R. Schin- single carbon source (16), and these are obvious choices for zel, Wurzburg, Federal Republic of Germany. studying gene structure and function if only suitable gene Escherichia coli XL1-Blue, used in all E. coli transforma- transfer systems, such as transformation or transduction, tion experiments, was obtained from Stratagene, La Jolla, were available. Fortunately, an efficient DNA transforma- California. tion method was recently described by Cline and Doolittle Media and growth conditions. A 25% (wt/vol) solution of (4), using both H. halobium and Halobacterium volcanii (3), artificial salt water (SW) was prepared by using a slightly which opens the way to methods of contemporary genetic modified formulation of that described by Rodriguez-Valera analysis. Plasmid transformation was possible, but with no et al. and contained liter: 197 g of NaCl, 17 g of selectable marker, the detection of transformants was ex- (16) per MgCl2 .6H20, 26 g of MgSO4 - 7H20, 0.1 g of CaCI2, 5 g of tremely laborious (3). To date, the transformation of other KCI, and 5 g of NH4Cl. All Haloferax media were prepared archaebacteria is either extremely inefficient, such as in by using this stock solution. Growth medium (GM) con- methanogens (1), or not currently possible, as is the case tained SW (18%, wt/vol), yeast extract (0.5%, wt/vol; Oxoid with the sulphur-dependent thermoacidophiles; so, of the Ltd., London, England) and 25 mM Tris hydrochloride three main branches of archaebacteria, the halobacteria buffer (pH 7.5). Solid media were prepared by adding show the most promise for genetic research. Bacto-Agar (15 g/liter; Difco Laboratories, Detroit, Mich.). Members of the genus Haloferax are aerobic chemoorgan- Cultures were incubated at 37°C in an orbital shaker (liquid otrophs, only moderately halophilic, requiring 2.0 to 2.5 M media) or in plastic containers (solid media). NaCl for optimum growth, and are capable of growth on Haloferax transformation medium (TM) contained SW single carbon sources. We chose to use an isolate (Aa 2.2) (18%, wt/vol), sucrose (15%, wt/vol), MnCl2 (1.7 ,uM), Tris belonging to phenon K (21). It grew rapidly in liquid and on hydrochloride (25 mM, pH 7.5), yeast extract (0.3%, wt/vol) solid media, had a low frequency of spontaneous mutation, and tryptone (Oxoid; 0.5%, wt/vol). Novobiocin was in- and contained a small multicopy plasmid. These were con- cluded in GM and TM at 0.1 ,ug/ml when required. sidered to be useful properties for constructing a model All solutions and growth media for H. halobium and H. genetic system in the archaebacteria, but a remaining obsta- volcanii were made up as previously described (3, 4). For the H. halobium strains, novobiocin was included in selective media at 0.2 ,ug/ml. * Corresponding author. E. coli XL1-Blue was grown at 37°C in YT medium (9) 756 VOL. 172, 1990 PLASMID VECTOR WITH SELECTABLE MARKER FOR HALOBACTERIA 757 containing tetracycline (15 ,ug/ml) or on YT-TAXI plates, chasing sequencing reactions with terminal deoxynucleoti- which contained YT medium supplemented with tetracycline dyl transferase (Boehringer Mannheim) (5). (15 ,ug/ml), ampicillin (50 ,ug/ml), 5-bromo-4-chloro-3-in- Plasmid map determination. The 10.5-kilobase (kb) cryptic dolyl-p-D-galactopyranoside (50 ,ug/ml), isopropyl-13-D-thio- plasmid ofHaloferax strain Aa 2.2, pHK2, was isolated from galactopyranoside (0.1 mM), and Bacto-Agar (15 g/liter). an agarose gel by electrophoresis onto a dialysis membrane Transformation of bacteria. Haloferax transformations (9), linearized with HindlIl, and cloned into the E. coli were performed by using the polyethylene glycol (PEG) plasmid pUC18 (23) by using T4 DNA ligase (Bresatec, Ltd., method described by Charlebois et al. (3). Freshly inocu- Adelaide, South Australia). A physical map was determined lated cultures were shaken at 37°C until the cells had reached by restricting 0.5 to 1 ,ug of DNA with various restriction a density (A550) of 1 to 1.5 (corresponding to a viable count endonucleases (both single and double digests) and resolving at 1 x 109 to 2 x 109 CFU/ml) and were then centrifuged the fragments on agarose or polyacrylamide gels. Sizes of (4,000 x g, 10 min) and suspended in a 1/10 volume of fragments were then estimated by using HindIlI-cut lambda buffered spheroplasting solution. The spheroplasting solu- DNA or 1-kilobase-pair-ladder DNA standards (Boehringer tion had reduced NaCl (1 M) but was otherwise the same as Mannheim). described previously (3). After PEG treatment, spheroplasts As some of the Sall and AvaI fragments were rather small were diluted in a solution containing SW (18%, wt/vol), and difficult to place, the nuclease digestion method (9) was sucrose (15%, wt/vol), and Tris hydrochloride (20 mM, pH also used. HindIII-cut pUC18-pHK2 (10 ,ug) was incubated 7.5) and allowed to recover for 5 to 12 h at 37°C, and then with Bal3l nuclease (New England BioLabs, Inc., Beverly, 100-1.l samples were spread onto TM plates containing Mass.; 0.75 U) prior to restriction with Sall or AvaI. novobiocin (0.1 ,ugIml) and incubated for 3 to 5 days at 37°C Samples were removed at various time intervals and ana- in plastic containers. lyzed on 10% polyacrylamide gels. The progressive disap- For transforming H. volcanii and H. halobium strains, we pearance of DNA bands then helped to position the restric- followed exactly the procedures described by Cline and tion fragments within pHK2. Doolittle (4) and Charlebois et al. (3). Determination of plasmid copy number. The copy number Isolation of plasmid DNA. For large-scale plasmid isolation of pHK2 (number of plasmids per cell) was estimated by from Haloferax strain Aa 2.2, DNA was isolated from carefully extracting total DNA from mid-log-phase cultures late-log-phase cultures by the following procedure. Cells of Haloferax cells grown in GM, running serial dilutions of were harvested by centrifugation at 2,800 x g for 15 min and the DNA preparation on an agarose gel (0.5%, wt/vol), and were washed once with 1 M NaCl. The cells were then comparing intensities of DNA bands stained with ethidium suspended in a buffer containing 1 M NaCl and 0.1 M EDTA bromide to those of DNA standards of known concentra- and lysed by the addition of 0.007 M sodium deoxycholate. tions. The number of viable cells used in the DNA prepara- This solution was left on ice for 30 min before spinning down tion was determined by plating dilutions onto GM plates, cellular debris and chromosomal DNA at 28,000 x g for 30 incubating for 2 to 3 days at 37°C, and counting colonies. min. The cleared lysate was transferred to a clean tube, and Selection of novobiocin-resistant Haloferax mutants. The DNA was precipitated by the addition of 10% (wt/vol) PEG MIC of novobiocin was first determined and found to be 6000 and incubation on ice for 60 min.
Recommended publications
  • Biotechnological Potential of Peptides and Secondary Metabolites in Archaea
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Crossref Hindawi Publishing Corporation Archaea Volume 2015, Article ID 282035, 7 pages http://dx.doi.org/10.1155/2015/282035 Review Article Untapped Resources: Biotechnological Potential of Peptides and Secondary Metabolites in Archaea James C. Charlesworth1,2 and Brendan P. Burns1,2 1 School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW 2052, Australia 2Australian Centre for Astrobiology, University of New South Wales, Sydney, NSW 2052, Australia Correspondence should be addressed to Brendan P. Burns; [email protected] Received 5 February 2015; Revised 7 July 2015; Accepted 8 July 2015 Academic Editor: Juergen Wiegel Copyright © 2015 J. C. Charlesworth and B. P. Burns. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Archaea are an understudied domain of life often found in “extreme” environments in terms of temperature, salinity, and a range of other factors. Archaeal proteins, such as a wide range of enzymes, have adapted to function under these extreme conditions, providing biotechnology with interesting activities to exploit. In addition to producing structural and enzymatic proteins, archaea also produce a range of small peptide molecules (such as archaeocins) and other novel secondary metabolites such as those putatively involved in cell communication (acyl homoserine lactones), which can be exploited for biotechnological purposes. Due to the wide array of metabolites produced there is a great deal of biotechnological potential from antimicrobials such as diketopiperazines and archaeocins, as well as roles in the cosmetics and food industry.
    [Show full text]
  • Natronococcus Jeotgali Sp. Nov., a Halophilic Archaeon Isolated from Shrimp Jeotgal, a Traditional Fermented Seafood from Korea
    International Journal of Systematic and Evolutionary Microbiology (2007), 57, 2129–2131 DOI 10.1099/ijs.0.65120-0 Natronococcus jeotgali sp. nov., a halophilic archaeon isolated from shrimp jeotgal, a traditional fermented seafood from Korea Seong Woon Roh,1,2 Young-Do Nam,1,2 Ho-Won Chang,2 Youlboong Sung,2 Kyoung-Ho Kim,2 Ho-Jae Lee,2 Hee-Mock Oh2 and Jin-Woo Bae1,2,3 Correspondence 1University of Science & Technology, 52 Eoeun-dong, Daejeon 305-333, Korea Jin-Woo Bae 2Biological Resource Center, KRIBB, Daejeon 305-806, Korea [email protected] 3Environmental Biotechnology National Core Research Center, Gyeongsang National University, Jinju 660-701, Korea A novel halophilic archaeon (strain B1T) belonging to the genus Natronococcus was isolated from shrimp jeotgal, a traditional fermented food from Korea. Colonies of this strain were orange–red and cells were non-motile cocci that stained Gram-variable. Strain B1T grew in 7.5–30.0 % (w/v) NaCl and at 21–50 6C and pH 7.0–9.5, with optimal growth occurring in 23–25 % (w/v) NaCl and at 37–45 6C and pH 7.5. Strain B1T was most closely related to the type strain of Natronococcus occultus, with which it shared 97.91 % 16S rRNA gene sequence similarity. Within the phylogenetic tree, this novel strain shared a branching point with N. occultus and occupied a phylogenetic position that was distinct from the main Natronococcus branch. The degree of DNA–DNA hybridization with the type strain of N. occultus, the most closely related species phylogenetically, was 16.4 %.
    [Show full text]
  • Diversity of Halophilic Archaea in Fermented Foods and Human Intestines and Their Application Han-Seung Lee1,2*
    J. Microbiol. Biotechnol. (2013), 23(12), 1645–1653 http://dx.doi.org/10.4014/jmb.1308.08015 Research Article Minireview jmb Diversity of Halophilic Archaea in Fermented Foods and Human Intestines and Their Application Han-Seung Lee1,2* 1Department of Bio-Food Materials, College of Medical and Life Sciences, Silla University, Busan 617-736, Republic of Korea 2Research Center for Extremophiles, Silla University, Busan 617-736, Republic of Korea Received: August 8, 2013 Revised: September 6, 2013 Archaea are prokaryotic organisms distinct from bacteria in the structural and molecular Accepted: September 9, 2013 biological sense, and these microorganisms are known to thrive mostly at extreme environments. In particular, most studies on halophilic archaea have been focused on environmental and ecological researches. However, new species of halophilic archaea are First published online being isolated and identified from high salt-fermented foods consumed by humans, and it has September 10, 2013 been found that various types of halophilic archaea exist in food products by culture- *Corresponding author independent molecular biological methods. In addition, even if the numbers are not quite Phone: +82-51-999-6308; high, DNAs of various halophilic archaea are being detected in human intestines and much Fax: +82-51-999-5458; interest is given to their possible roles. This review aims to summarize the types and E-mail: [email protected] characteristics of halophilic archaea reported to be present in foods and human intestines and pISSN 1017-7825, eISSN 1738-8872 to discuss their application as well. Copyright© 2013 by The Korean Society for Microbiology Keywords: Halophilic archaea, fermented foods, microbiome, human intestine, Halorubrum and Biotechnology Introduction Depending on the optimal salt concentration needed for the growth of strains, halophilic microorganisms can be Archaea refer to prokaryotes that used to be categorized classified as halotolerant (~0.3 M), halophilic (0.2~2.0 M), as archaeabacteria, a type of bacteria, in the past.
    [Show full text]
  • Characterization of the First Cultured Representative of Verrucomicrobia Subdivision 5 Indicates the Proposal of a Novel Phylum
    The ISME Journal (2016) 10, 2801–2816 OPEN © 2016 International Society for Microbial Ecology All rights reserved 1751-7362/16 www.nature.com/ismej ORIGINAL ARTICLE Characterization of the first cultured representative of Verrucomicrobia subdivision 5 indicates the proposal of a novel phylum Stefan Spring1, Boyke Bunk2, Cathrin Spröer3, Peter Schumann3, Manfred Rohde4, Brian J Tindall1 and Hans-Peter Klenk1,5 1Department Microorganisms, Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany; 2Department Microbial Ecology and Diversity Research, Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany; 3Department Central Services, Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany and 4Central Facility for Microscopy, Helmholtz-Centre of Infection Research, Braunschweig, Germany The recently isolated strain L21-Fru-ABT represents moderately halophilic, obligately anaerobic and saccharolytic bacteria that thrive in the suboxic transition zones of hypersaline microbial mats. Phylogenetic analyses based on 16S rRNA genes, RpoB proteins and gene content indicated that strain L21-Fru-ABT represents a novel species and genus affiliated with a distinct phylum-level lineage originally designated Verrucomicrobia subdivision 5. A survey of environmental 16S rRNA gene sequences revealed that members of this newly recognized phylum are wide-spread and ecologically important in various anoxic environments ranging from hypersaline sediments to wastewater and the intestine of animals. Characteristic phenotypic traits of the novel strain included the formation of extracellular polymeric substances, a Gram-negative cell wall containing peptidoglycan and the absence of odd-numbered cellular fatty acids. Unusual metabolic features deduced from analysis of the genome sequence were the production of sucrose as osmoprotectant, an atypical glycolytic pathway lacking pyruvate kinase and the synthesis of isoprenoids via mevalonate.
    [Show full text]
  • The Role of Stress Proteins in Haloarchaea and Their Adaptive Response to Environmental Shifts
    biomolecules Review The Role of Stress Proteins in Haloarchaea and Their Adaptive Response to Environmental Shifts Laura Matarredona ,Mónica Camacho, Basilio Zafrilla , María-José Bonete and Julia Esclapez * Agrochemistry and Biochemistry Department, Biochemistry and Molecular Biology Area, Faculty of Science, University of Alicante, Ap 99, 03080 Alicante, Spain; [email protected] (L.M.); [email protected] (M.C.); [email protected] (B.Z.); [email protected] (M.-J.B.) * Correspondence: [email protected]; Tel.: +34-965-903-880 Received: 31 July 2020; Accepted: 24 September 2020; Published: 29 September 2020 Abstract: Over the years, in order to survive in their natural environment, microbial communities have acquired adaptations to nonoptimal growth conditions. These shifts are usually related to stress conditions such as low/high solar radiation, extreme temperatures, oxidative stress, pH variations, changes in salinity, or a high concentration of heavy metals. In addition, climate change is resulting in these stress conditions becoming more significant due to the frequency and intensity of extreme weather events. The most relevant damaging effect of these stressors is protein denaturation. To cope with this effect, organisms have developed different mechanisms, wherein the stress genes play an important role in deciding which of them survive. Each organism has different responses that involve the activation of many genes and molecules as well as downregulation of other genes and pathways. Focused on salinity stress, the archaeal domain encompasses the most significant extremophiles living in high-salinity environments. To have the capacity to withstand this high salinity without losing protein structure and function, the microorganisms have distinct adaptations.
    [Show full text]
  • Biofilm and Quorum Sensing in Archaea
    #0# Acta Biologica 26/2019 | www.wnus.edu.pl/ab | DOI: 10.18276/ab.2019.26-04 | strony 35–44 Biofilm and Quorum Sensing in Archaea Małgorzata Pawlikowska-Warych,1 Beata Tokarz-Deptuła,2 Paulina Czupryńska,3 Wiesław Deptuła4 1 Department of Microbiology, Faculty of Biology, University of Szczecin, Felczaka 3c, 71-412 Szczecin, Poland 2 Department of Immunology, Faculty of Biology, University of Szczecin, Felczaka 3c, 71-412 Szczecin, Poland 3 Department of Microbiology, Faculty of Biology, University of Szczecin, Felczaka 3c, 71-412 Szczecin, Poland 4 Nicolaus Copernicus University in Toruń, Faculty of Biological and Veterinary Sciences, Institute of Veterinary Medicine, Gagarina 7, 87-100 Toruń, Poland Corresponding author, e-mail: [email protected] Keywords Archaea, biofilm, environment, quorum sensing Abstract In the article, we presented a brief description of the Archaea, considering their structure, physiology and systematics. Based on the analysis of the literature, we bring closer the mecha- nism of biofilm formation, including extracellular polymeric substances as well as cellular organelles, such as archaella, pili and ‘hami’. The method of forming a biofilm depends on the type of Archaea and the environment in which it naturally lives. We are also introducing the phenomenon of quorum-sensing, as a mechanism of communication of Archaea in the environment. This phenomenon corresponds to similar molecules as bacteria, namely acylated homoserines lactones, QS peptides, autoinducer-2 and -3 and others. In the case of biofilms and the occurrence of the phenomenon of quorum sensing, it can be concluded that these phenomena are very important for the life of Archaea.
    [Show full text]
  • Across the Tree of Life, Radiation Resistance Is Governed By
    Across the tree of life, radiation resistance is PNAS PLUS + governed by antioxidant Mn2 , gauged by paramagnetic resonance Ajay Sharmaa,1, Elena K. Gaidamakovab,c,1, Olga Grichenkob,c, Vera Y. Matrosovab,c, Veronika Hoekea, Polina Klimenkovab,c, Isabel H. Conzeb,d, Robert P. Volpeb,c, Rok Tkavcb,c, Cene Gostincarˇ e, Nina Gunde-Cimermane, Jocelyne DiRuggierof, Igor Shuryakg, Andrew Ozarowskih, Brian M. Hoffmana,i,2, and Michael J. Dalyb,2 aDepartment of Chemistry, Northwestern University, Evanston, IL 60208; bDepartment of Pathology, Uniformed Services University of the Health Sciences, Bethesda, MD 20814; cHenry M. Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD 20817; dDepartment of Biology, University of Bielefeld, Bielefeld, 33615, Germany; eDepartment of Biology, Biotechnical Faculty, University of Ljubljana, Ljubljana, SI-1000, Slovenia; fDepartment of Biology, Johns Hopkins University, Baltimore, MD 21218; gCenter for Radiological Research, Columbia University, New York, NY 10032; hNational High Magnetic Field Laboratory, Florida State University, Tallahassee, FL 32306; and iDepartment of Molecular Biosciences, Northwestern University, Evanston, IL 60208 Contributed by Brian M. Hoffman, September 15, 2017 (sent for review August 1, 2017; reviewed by Valeria Cizewski Culotta and Stefan Stoll) Despite concerted functional genomic efforts to understand the agents of cellular damage. In particular, irradiated cells rapidly form •− complex phenotype of ionizing radiation (IR) resistance, a genome superoxide (O2 ) ions by radiolytic reduction of both atmospheric sequence cannot predict whether a cell is IR-resistant or not. Instead, O2 and O2 released through the intracellular decomposition of IR- we report that absorption-display electron paramagnetic resonance generated H2O2 ascatalyzedbybothenzymaticandnonenzymatic (EPR) spectroscopy of nonirradiated cells is highly diagnostic of IR •− metal ions.
    [Show full text]
  • Potential Biotechnological Applications of Cyanobacterial Exopolysaccharides
    Vol.64: e21200401, 2021 https://doi.org/10.1590/1678-4324-2021200401 ISSN 1678-4324 Online Edition Review - Agriculture, Agribusiness and Biotechnology Potential Biotechnological Applications of Cyanobacterial Exopolysaccharides Laxmi Parwani1* https://orcid.org/0000-0001-7627-599X Medha Bhatt1 https://orcid.org/0000-0002-2411-8172 Jaspreet Singh2 https://orcid.org/0000-0003-2470-3280 1Banasthali Vidyapith, Department of Bioscience and Biotechnology, P.O. Banasthali, Rajasthan, India; 2 Jaipur National University, SIILAS Campus, School of Life Sciences, Jaipur, Rajasthan, India. Editor-in-Chief: Paulo Vitor Farago Associate Editor: Aline Alberti Received: 2020.06.24; Accepted: 2021.02.24. *Correspondence: [email protected]; Tel.: +91-9982216197 HIGHLIGHTS Illustration of various important fields of biotechnology where cyanobacterial exopolysaccharides are potentially useful. Discussion of research gaps and new opportunities to make the biomaterial suitable for industrial uses. Abstract: The cyanobacterial exopolysaccharides (EPSs) are considered as one of the important group of biopolymers having significant ecological, industrial, and biotechnological importance. Cyanobacteria are regarded as a very abundant source of structurally diverse, high molecular weight polysaccharides having variable composition and roles according to the organisms and the environmental conditions in which they are produced. Due to their structural complexity, versatility and valuable biological properties, they are now emerging as high-value compounds.
    [Show full text]
  • Lipids of Sulfolobus Spp. | Encyclopedia
    Lipids of Sulfolobus spp. Subjects: Biophysics | Biotechnology Contributors: Kerstin Rastaedter , David J. Wurm Submitted by: Kerstin Rastaedter Definition Archaea, and thereby, Sulfolobus spp. exhibit a unique lipid composition of ether lipids, which are altered in regard to the ratio of diether to tetraether lipids, number of cyclopentane rings and type of head groups, as a coping mechanism against environmental changes. Sulfolobales mainly consist of C40-40 tetraether lipids (caldarchaeol) and partly of C20-20 diether lipids (archaeol). A variant of caldarchaeol called glycerol dialkylnonitol tetraether (GDNT) has only been found in Sulfolobus and other members of the Creanarchaeota phylum so far. Altering the numbers of incorporated cyclopentane rings or the the diether to tetraether ratio results in more tightly packed membranes or vice versa. 1. The Cell Membrane and Lipids of Sulfolobus spp. The thermoacidophilic genus Sulfolobus belongs to the phylum Crenarchaeota and is a promising player for biotechnology [1], since it harbors a couple of valuable products, such as extremozymes[ 2], trehalose [3], archaeocins [4] and lipids for producing archaeosomes [5]. Genetic tools for this genus have rapidly advanced in recent years[ 6], generating new possibilities in basic science and for biotechnological applications alike. The cultivation conditions are preferably at around 80 °C and pH 3 [7]. Sulfolobus species are able to grow aerobically and can be readily cultivated on a laboratory scale. These organisms became a model organism for Crenarchaeota and for adaption processes to extreme environments [8][9][10][11][12][13]. Sulfolobus species were found in solfataric fields all over the world[ 14]. A major drawback of cultivating this organism was the lack of a defined cultivation medium.
    [Show full text]
  • Determination of Hydrolytic Enzyme Capabilities of Halophilic Archaea Isolated from Hides and Skins and Their Phenotypic and Phylogenetic Identification by S
    33 DETERMinATION OF HYDROLYTic ENZYME CAPABILITIES OF HALOPHILIC ARCHAEA ISOLATED FROM HIDES AND SKins AND THEIR PHENOTYpic AND PHYLOGENETic IDENTIFicATION by S. T. B LG School of Health, Canakkale Onsekiz Mart University, Terzioglu Campus Canakkale, Turkey, 17100. and B. MER ÇL YaPiCi Biology Department, Faculty of Arts and Science, Canakkale ONSEKIZ MART UNIVERSITY, TERZIOGLU CAMPUS, Canakkale, Turkey, 17100. and İsmail Karaboz Basic and Industrial Microbiology Section, Biology Department, Faculty of Science, Ege University, Bornova, İzmi r, Turkey, 35100. ABSTRACT INTRODUCTION This research aims to isolate extremely halophilic archaea The main constituent of the raw hide is protein, mainly from salted hides, to determine the capacities of their collagen (33% w/w), and remainder is moisture and fat. During hydrolytic enzymes, and to identify them by using phenotypic storage of raw hide, collagen’s excessive proteolysis by and molecular methods. Domestic and imported salted hide lysosomal autolysis or proteolytic bacterial enzymes can lead and skin samples obtained from eight different sources were to the disintegration of the structure of collagen fibers.1 used as the research material. 186 extremely halophilic Biodeterioration is among the major causes of impairment of microorganisms were isolated from salted raw hides and aesthetic, functional and other properties of leather and other skins. Some biochemical, antibiotic sensitivity, pH, NaCl, biopolymers or organic materials and the products made from temperature tolerance and quantitative and qualitative them. Due to the fact that prevention of biological degradation hydrolytic enzyme tests were performed on these isolates. In is very important in conservation and processing of leather, our study, taking into account the phenotypic findings of the great effort is being made for decontamination of these research, 34 of 186 isolates were selected.
    [Show full text]
  • Characterization and Identification of an Isolate of Halobacterium from Soda Springs Mojave Desert
    California State University, San Bernardino CSUSB ScholarWorks Theses Digitization Project John M. Pfau Library 1979 Characterization and identification of an isolate of halobacterium from Soda Springs Mojave Desert Abayomi Odubela Follow this and additional works at: https://scholarworks.lib.csusb.edu/etd-project Part of the Biology Commons, and the Desert Ecology Commons Recommended Citation Odubela, Abayomi, "Characterization and identification of an isolate of halobacterium from Soda Springs Mojave Desert" (1979). Theses Digitization Project. 67. https://scholarworks.lib.csusb.edu/etd-project/67 This Thesis is brought to you for free and open access by the John M. Pfau Library at CSUSB ScholarWorks. It has been accepted for inclusion in Theses Digitization Project by an authorized administrator of CSUSB ScholarWorks. For more information, please contact [email protected]. CHARAG'liERIZATION AND IDENT'IFICATION GF AN ISOLATl OF HALOBACTERIUM FROM SODA:SPRINGS. MOJAVE DESERT —, A Thesis Presented to the Faculty of California State Cblleges San Bernardino in Partial FulfilIment of the.Requirements for the .Degree Master of Science in: • . Biology hy . Abayomi Qdubela April 1979 CHARACTERIZATION AND IDENTIFICATION OF AN ISOLATE OF . HALOBACTERIUM FROM SODA SPRINGS MO J AYE 'DESERT A Thesis .. Preisented to the Faculty of California State College, San Bernardino by Abayomi Odubela April 1979 Approved bys Gh^i»arson Biol _ Date Graduate Committee Corama^tee Member Co mi itt Member Mayor Professor ■ . Representative of^he Graduate^ Dean ABSTRACT : : A strai|i of Halobacterium . isolated from a vond at the California State Universities and Colleges Desert . Studies Center |at, Soda Springs, Mojave Desert, California „ was studied in order to define its taxonomic placeuin the genus Halohactelrium.
    [Show full text]
  • Extreme Environments and High-Level Bacterial Tellurite Resistance
    microorganisms Review Extreme Environments and High-Level Bacterial Tellurite Resistance Chris Maltman 1,* and Vladimir Yurkov 2 1 Department of Biology, Slippery Rock University, Slippery Rock, PA 16001, USA 2 Department of Microbiology, University of Manitoba, Winnipeg, MB R3T 2N2, Canada; [email protected] * Correspondence: [email protected]; Tel.: +724-738-4963 Received: 28 October 2019; Accepted: 20 November 2019; Published: 22 November 2019 Abstract: Bacteria have long been known to possess resistance to the highly toxic oxyanion tellurite, most commonly though reduction to elemental tellurium. However, the majority of research has focused on the impact of this compound on microbes, namely E. coli, which have a very low level of resistance. Very little has been done regarding bacteria on the other end of the spectrum, with three to four orders of magnitude greater resistance than E. coli. With more focus on ecologically-friendly methods of pollutant removal, the use of bacteria for tellurite remediation, and possibly recovery, further highlights the importance of better understanding the effect on microbes, and approaches for resistance/reduction. The goal of this review is to compile current research on bacterial tellurite resistance, with a focus on high-level resistance by bacteria inhabiting extreme environments. Keywords: tellurite; tellurite resistance; extreme environments; metalloids; bioremediation; biometallurgy 1. Introduction Microorganisms possess a wide range of extraordinary abilities, from the production of bioactive molecules [1] to resistance to and transformation of highly toxic compounds [2–5]. Of great interest are bacteria which can convert the deleterious oxyanion tellurite to elemental tellurium (Te) through reduction. Currently, research into bacterial interactions with tellurite has been lagging behind investigation of the oxyanions of other metals such as nickel (Ni), molybdenum (Mo), tungsten (W), iron (Fe), and cobalt (Co).
    [Show full text]