scispace - formally typeset

Journal ArticleDOI

Isolation and characterization of a new CO-utilizing strain, Thermoanaerobacter thermohydrosulfuricus subsp. carboxydovorans, isolated from a geothermal spring in Turkey

23 Aug 2009-Extremophiles (Springer Japan)-Vol. 13, Iss: 6, pp 885-894

TL;DR: Strain TLO has the capability to ferment a wide variety of mono-, di-, and polysaccharides and proteinaceous substrates, producing mainly lactate, next to acetate, ethanol, alanine, H2, and CO2 and was able to grow in an atmosphere of up to 25% of CO as sole electron donor.

AbstractA novel anaerobic, thermophilic, Gram-positive, spore-forming, and sugar-fermenting bacterium (strain TLO) was isolated from a geothermal spring in Ayas, Turkey. The cells were straight to curved rods, 0.4–0.6 μm in diameter and 3.5–10 μm in length. Spores were terminal and round. The temperature range for growth was 40–80°C, with an optimum at 70°C. The pH optimum was between 6.3 and 6.8. Strain TLO has the capability to ferment a wide variety of mono-, di-, and polysaccharides and proteinaceous substrates, producing mainly lactate, next to acetate, ethanol, alanine, H2, and CO2. Remarkably, the bacterium was able to grow in an atmosphere of up to 25% of CO as sole electron donor. CO oxidation was coupled to H2 and CO2 formation. The G + C content of the genomic DNA was 35.1 mol%. Based on 16S rRNA gene sequence analysis and the DNA–DNA hybridization data, this bacterium is most closely related to Thermoanaerobacter thermohydrosulfuricus and Thermoanaerobacter siderophilus (99% similarity for both). However, strain TLO differs from Thermoanaerobacter thermohydrosulfuricus in important aspects, such as CO-utilization and lipid composition. These differences led us to propose that strain TLO represents a subspecies of Thermoanaerobacter thermohydrosulfuricus, and we therefore name it Thermoanaerobacter thermohydrosulfuricus subsp. carboxydovorans.

Topics: Thermoanaerobacter (63%)

...read more

Content maybe subject to copyright    Report

Citations
More filters

Journal ArticleDOI
TL;DR: Upon both acid and base hydrolyses of total cell material, the uncommon membrane-spanning lipid 13,16-dimethyl octacosanedioic acid (iso-diabolic acid) was released in substantial amounts (22 to 43% of the total fatty acids) from all of the acidobacteria studied.
Abstract: The distribution of membrane lipids of 17 different strains representing 13 species of subdivisions 1 and 3 of the phylum Acidobacteria, a highly diverse phylum of the Bacteria, were examined by hydrolysis and gas chromatography-mass spectrometry (MS) and by high-performance liquid chromatography-MS of intact polar lipids. Upon both acid and base hydrolyses of total cell material, the uncommon membrane-spanning lipid 13,16-dimethyl octacosanedioic acid (iso-diabolic acid) was released in substantial amounts (22 to 43% of the total fatty acids) from all of the acidobacteria studied. This lipid has previously been encountered only in thermophilic Thermoanaerobacter species but bears a structural resemblance to the alkyl chains of bacterial glycerol dialkyl glycerol tetraethers (GDGTs) that occur ubiquitously in peat and soil and are suspected to be produced by acidobacteria. As reported previously, most species also contained iso-C15 and C16:1ω7C as major fatty acids but the presence of iso-diabolic acid was unnoticed in previous studies, most probably because the complex lipid that contained this moiety was not extractable from the cells; it could only be released by hydrolysis. Direct analysis of intact polar lipids in the Bligh-Dyer extract of three acidobacterial strains, indeed, did not reveal any membrane-spanning lipids containing iso-diabolic acid. In 3 of the 17 strains, ether-bound iso-diabolic acid was detected after hydrolysis of the cells, including one branched GDGT containing iso-diabolic acid-derived alkyl chains. Since the GDGT distribution in soils is much more complex, branched GDGTs in soil likely also originate from other (acido)bacteria capable of biosynthesizing these components.

317 citations


Cites background from "Isolation and characterization of a..."

  • ...thermohydrosulfuricus (3), iso-diabolic acid was also detected only after hydrolysis of total cell material....

    [...]

  • ...iso-Diabolic acid has previously been encountered only in thermophilic Thermoanaerobacter species (3, 24, 32), where it fulfills a role as a membranespanning lipid....

    [...]


Journal ArticleDOI
TL;DR: It is demonstrated that the presence of membrane spanning ether-lipids and methyl branches shows a striking relationship with the growth boundaries of archaea and bacteria.
Abstract: The cytoplasmic membrane of a prokaryotic cell consists of a lipid bilayer or a monolayer that shields the cellular content from the environment. In addition, the membrane contains proteins that are responsible for transport of proteins and metabolites as well as for signalling and energy transduction. Maintenance of the functionality of the membrane during changing environmental conditions relies on the cell’s potential to rapidly adjust the lipid composition of its membrane. Despite the fundamental chemical differences between bacterial ester lipids and archaeal ether lipids, both types are functional under a wide range of environmental conditions. We here provide an overview of archaeal and bacterial strategies of changing the lipid compositions of their membranes. Some molecular adjustments are unique for archaea or bacteria, whereas others are shared between the two domains. Strikingly, shared adjustments were predominantly observed near the growth boundaries of bacteria. Here, we demonstrate that the presence of membrane spanning ether-lipids and methyl branches shows a striking relationship with the growth boundaries of archaea and bacteria.

154 citations


Journal ArticleDOI
TL;DR: Three main types of fermentative CO metabolism can be distinguished: hydrogenogenesis, methanogenesis, and acetogenesis, generating hydrogen, methane and acetate, respectively, with emphasis on the potential enzymatic routes and bio-energetics involved.
Abstract: Carbon monoxide can act as a substrate for different modes of fermentative anaerobic metabolism. The trait of utilizing CO is spread among a diverse group of microorganisms, including members of bacteria as well as archaea. Over the last decade this metabolism has gained interest due to the potential of converting CO-rich gas, such as synthesis gas, into bio-based products. Three main types of fermentative CO metabolism can be distinguished: hydrogenogenesis, methanogenesis, and acetogenesis, generating hydrogen, methane and acetate, respectively. Here, we review the current knowledge on these three variants of microbial CO metabolism with an emphasis on the potential enzymatic routes and bio-energetics involved.

104 citations


Cites background from "Isolation and characterization of a..."

  • ...No growth Kluyver and Schnellen, 1947 Methanosaeta concillii Aceticlastic Enzyme assay N.D. N.D....

    [...]

  • ...Methanothermobacter thermoautotrophicus Hydrogenotrophic Cultivation/enzyme assay 50 kPa Methane, hydrogen ∼200 Daniels et al., 1977; Wasserfallen et al., 2000 Methanosarcina thermophila Aceticlastic Cultivation >2 kPa Hydrogen, Methane N.D. Zinder and Anguish, 1992 Methanothrix sp. Strain CALS-1 Aceticlastic Cultivation <2 kPa Methane No growth Zinder and Anguish, 1992 Archaeoglobus fulgidusB Sulfate reducer Cultivation >136 kPa Acetate, formate ∼10 Henstra et al., 2007a Not determined parameters are marked N.D. AForward arrows (>) indicate inhibitory levels have not been reached; numbers displayed are the maximal level tested....

    [...]

  • ...Moorella thermoautotrophica EcH/ Cytochromes (H+) 7 (UM), 9 (DM) 33 (DM) 3.34 (UM) 2.53 (DM) 0.82 (DM) Savage and Drake, 1986; Savage et al., 1987; Collins et al., 1994 Moorella thermoacetica EcH/ Cytochromes (H+) 9 (DM) 16 (DM) 1.28 (UM) 0.46 (UM) Daniel et al., 1990; Collins et al., 1994; Pierce et al., 2008 Acetobacterium Woodii RnF (Na+ ) 13D (UM), 5.5A (UM + formate) 6.2A (UM) N.D. 1.05C (DM) Balch et al., 1977; Tschech and Pfennig, 1984; Genthner and Bryant, 1987; Hess et al., 2013; Bertsch and Müller, 2015 Blautia producta RnF (Na+ ) 1.5 (UM) 5 (UM) 2.13C (UM) 0.65C (UM) Lorowitz and Bryant, 1984; Geerligs et al., 1989 Clostridium aceticum RnF (N.D.) ∼10B (UM) 20 (UM) N.D. N.D. Braun et al., 1981; Sim et al., 2007...

    [...]

  • ...…2007 Thermosinus carboxydivorans Hot spring, Norris Basin 60 1.15 Sokolova et al., 2004a Thermoanaerobacter thermohydrosulfuricus ssp. carboxydovorans Geothermal spring, Turkey 70 N.D. Balk et al., 2009 Desulfotomaculum carboxydivorans Paper mill wastewater sludge 55 N.D. Parshina et al., 2005b...

    [...]

  • ...Methanobrevibacter arboriphilicus Hydrogenotrophic Enzyme assay N.D. N.D. N.D. Hammel et al., 1984 Methanosarcina acetivorans C2A Aceticlastic Cultivation/enzyme assay >150 kPa Methane, acetate, formate ∼20 Rother and Metcalf, 2004; Oelgeschläger and Rother, 2009 Methanosarcina barkeri Aceticlastic Cultivation/enzyme assay >100 kPa Hydrogen, Methane ∼65 O’Brien et al., 1984; Bott et al., 1986 Methanobacterium formicicum Hydrogenotrophic Cultivation N.D. N.D....

    [...]


Journal ArticleDOI
TL;DR: Examination of lipid composition of seven phylogenetically divergent strains of subdivision 4 of the Acidobacteria, a bacterial group that is commonly encountered in soil, found the presence of ether bonds in the membrane lipids does not seem to be an adaptation to temperature, because the five mesophilic isolates contained a larger amount of ether lipids than the thermophile “Ca. Chloracidobacterium thermophilum.”
Abstract: Recently, iso-diabolic acid (13,16-dimethyl octacosanedioic acid) has been identified as a major membrane-spanning lipid of subdivisions 1 and 3 of the Acidobacteria, a highly diverse phylum within the Bacteria. This finding pointed to the Acidobacteria as a potential source for the bacterial glycerol dialkyl glycerol tetraethers that occur ubiquitously in peat, soil, lakes, and hot springs. Here, we examined the lipid composition of seven phylogenetically divergent strains of subdivision 4 of the Acidobacteria, a bacterial group that is commonly encountered in soil. Acid hydrolysis of total cell material released iso-diabolic acid derivatives in substantial quantities (11 to 48% of all fatty acids). In contrast to subdivisions 1 and 3 of the Acidobacteria, 6 out of the 7 species of subdivision 4 (excepting “Candidatus Chloracidobacterium thermophilum”) contained iso-diabolic acid ether bound to a glycerol in larger fractional abundance than iso-diabolic acid itself. This is in agreement with the analysis of intact polar lipids (IPLs) by high-performance liquid chromatography-mass spectrometry (HPLC-MS), which showed the dominance of mixed ether-ester glycerides. iso-Diabolic acid-containing IPLs were not identified, because these IPLs are not released with a Bligh-Dyer extraction, as observed before when studying lipid compositions of subdivisions 1 and 3 of the Acidobacteria. The presence of ether bonds in the membrane lipids does not seem to be an adaptation to temperature, because the five mesophilic isolates contained a larger amount of ether lipids than the thermophile “Ca. Chloracidobacterium thermophilum.” Furthermore, experiments with Pyrinomonas methylaliphatogenes did not reveal a major influence of growth temperature over the 50 to 69°C range.

84 citations


Cites background from "Isolation and characterization of a..."

  • ...(61), Sarcina ventriculi (62), members of the 304 Thermotogales (2,57,63-65), Thermoanaerobacter species (58,59,62) and Acidobacteria of 305 SD 1, 3 (14), and 4 (this work), respectively....

    [...]

  • ...This lipid was identified 254 previously as an abundant lipid in Acidobacteria of SD 1 and 3 (14) and in thermophilic 255 Thermoanaerobacter species (58-60), in which they fulfill a role as membrane-spanning 256 lipids....

    [...]


Journal ArticleDOI
TL;DR: Recent advances in the isolation of novel phylogenetic groups utilizing formate or CO, the remarkable genetic engineering that enhances H2 productivity, and the practical implementation of H2 production from C1 substrates are focused on.
Abstract: Among four basic mechanisms for biological hydrogen (H2) production, dark fermentation has been considered to show the highest hydrogen evolution rate (HER). H2 production from one-carbon (C1) compounds such as formate and carbon monoxide (CO) is promising because formate is an efficient H2 carrier, and the utilization of CO-containing syngas or industrial waste gas may render the industrial biohydrogen production process cost-effective. A variety of microbes with the formate hydrogen lyase (FHL) system have been identified from phylogenetically diverse groups of archaea and bacteria, and numerous efforts have been undertaken to improve the HER for formate through strain optimization and bioprocess development. CO-dependent H2 production has been investigated to enhance the H2 productivity of various carboxydotrophs via an increase in CO gas-liquid mass transfer rates and the construction of genetically modified strains. Hydrogenogenic CO-conversion has been applied to syngas and by-product gas of the steel-mill process, and this low-cost feedstock has shown to be promising in the production of biomass and H2. Here, we focus on recent advances in the isolation of novel phylogenetic groups utilizing formate or CO, the remarkable genetic engineering that enhances H2 productivity, and the practical implementation of H2 production from C1 substrates.

49 citations


References
More filters

Journal ArticleDOI
TL;DR: This assay is very reproducible and rapid with the dye binding process virtually complete in approximately 2 min with good color stability for 1 hr with little or no interference from cations such as sodium or potassium nor from carbohydrates such as sucrose.
Abstract: A protein determination method which involves the binding of Coomassie Brilliant Blue G-250 to protein is described. The binding of the dye to protein causes a shift in the absorption maximum of the dye from 465 to 595 nm, and it is the increase in absorption at 595 nm which is monitored. This assay is very reproducible and rapid with the dye binding process virtually complete in approximately 2 min with good color stability for 1 hr. There is little or no interference from cations such as sodium or potassium nor from carbohydrates such as sucrose. A small amount of color is developed in the presence of strongly alkaline buffering agents, but the assay may be run accurately by the use of proper buffer controls. The only components found to give excessive interfering color in the assay are relatively large amounts of detergents such as sodium dodecyl sulfate, Triton X-100, and commercial glassware detergents. Interference by small amounts of detergent may be eliminated by the use of proper controls.

214,383 citations


"Isolation and characterization of a..." refers methods in this paper

  • ...The protein content of the cell extracts was determined according to the method of Bradford (1976) with bovine serum albumin as a standard....

    [...]


01 Jan 1991

9,717 citations


"Isolation and characterization of a..." refers methods in this paper

  • ...PCR was performed with the bacterial primers 7f and 1510r (Lane 1991) by using the Taq DNA polymerase kit (Life Technologies) to amplify the bacterial 16S rRNA gene....

    [...]


Journal ArticleDOI
TL;DR: The ARB program package comprises a variety of directly interacting software tools for sequence database maintenance and analysis which are controlled by a common graphical user interface.
Abstract: The ARB (from Latin arbor, tree) project was initiated almost 10 years ago. The ARB program package comprises a variety of directly interacting software tools for sequence database maintenance and analysis which are controlled by a common graphical user interface. Although it was initially designed for ribosomal RNA data, it can be used for any nucleic and amino acid sequence data as well. A central database contains processed (aligned) primary structure data. Any additional descriptive data can be stored in database fields assigned to the individual sequences or linked via local or worldwide networks. A phylogenetic tree visualized in the main window can be used for data access and visualization. The package comprises additional tools for data import and export, sequence alignment, primary and secondary structure editing, profile and filter calculation, phylogenetic analyses, specific hybridization probe design and evaluation and other components for data analysis. Currently, the package is used by numerous working groups worldwide.

6,569 citations


Book
01 Jan 1991
TL;DR: Isolation and purification of nucleic acids DNA reassociation experiments DNA-rRNA hybridization and methods DNA sequencing in bacterial systematics direct sequence analysis of small RNAs 16S/23S rRNA sequencing the polymerase chain reaction development and application of nucleics acid probes DNA fingerprinting from macromolecules to trees.
Abstract: Isolation and purification of nucleic acids DNA reassociation experiments DNA-rRNA hybridization and methods DNA sequencing in bacterial systematics direct sequence analysis of small RNAs 16S/23S rRNA sequencing the polymerase chain reaction development and application of nucleic acid probes DNA fingerprinting from macromolecules to trees.

5,041 citations


"Isolation and characterization of a..." refers methods in this paper

  • ...PCR was performed with the bacterial primers 7f and 1510r (Lane 1991) by using the Taq DNA polymerase kit (Life Technologies) to amplify the bacterial 16S rRNA gene....

    [...]

  • ...The use of the electron acceptors was examined by following the optical density (600 nm) of the culture, detection of sulfide production (for sulfate, thiosulfate, sulfite and elemental sulfur), change of visible color (for AQDS) and measurements of the reduction of Fe(III) or formation of a…...

    [...]


Journal ArticleDOI
TL;DR: High-performance liquid chromatography is a promising alternative for determining the G+C content of bacterial deoxyribonucleic acid (DNA) and may also be more accurate than indirect methods, such as the buoyant density and thermal denaturation methods.
Abstract: High-performance liquid chromatography is a promising alternative for determining the G+C content of bacterial deoxyribonucleic acid (DNA). The method which we evaluated involves enzymatic degradation of the DNA to nucleosides by P1 nuclease and bovine intestinal mucosa alkaline phosphatase, separation of the nucleosides by high-performance liquid chromatography, and calculation of the G+C content from the apparent ratios of deoxyguanosine and thymidine. Because the nucleosides are released from the DNA at different rates, incomplete degradation produces large errors in the apparent G+C content. For partially purified DNA, salts are a major source of interference in degradation. However, when the salts are carefully removed, the preparation and degradation of DNA contribute little error to the determination of G+C content. This method also requires careful selection of the chromatographic conditions to ensure separation of the major nucleosides from the nucleosides of modified bases and precise control of the flow rates. Both of these conditions are achievable with standard equipment and C18 reversed-phase columns. Then the method is precise, and the relative standard deviations of replicate measurements are close to 0.1%. It is also rapid, and a single measurement requires about 15 min. It requires small amounts of sample, and the G+C content can be determined from DNA isolated from a single bacterial colony. It is not affected by contamination with ribonucleic acid. Because this method yields a direct measurement, it may also be more accurate than indirect methods, such as the buoyant density and thermal denaturation methods. In addition, for highly purified DNA, the extent of modification can be determined.

4,667 citations


"Isolation and characterization of a..." refers methods in this paper

  • ...C content of strain TLO was determined using the HPLC method described by Mesbah et al. (1989); unmethylated lambda DNA (Sigma) was used as the standard....

    [...]


Related Papers (5)