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PubMed1997

Characterization of Thiobacillus thioparus LV43 and its distribution in a chemoautotrophically based groundwater ecosystem.

Bacterial strain LV43 was previously isolated from a floating microbial mat located in Movile Cave, the access point to a chemoautotrophically based groundwater ecosystem in southern Romania. This gram-negative, rod-shaped organism grows autotrophically through the oxidation of thiosulfate and sulfide, but it does not grow heterotrophically. Strain LV43 grows over a pH range of 5.0 to 9.0, with an optimum near 7.5 at 28 degrees C. The pH of the medium decreased from 7.5 to 6.5 during growth on thiosulfate. Carbon isotope fractionation values for strain LV43 were within the previously reported range of fractionation values for the overall floating microbial mat in Movile Cave and were similar to values reported for chemoautotrophic sulfur-oxidizing strains of Thiobacillus neapolitanus and Thiomicrospira sp. The 16S rRNA gene sequence of strain LV43 was determined, and phylogenetic analysis indicated that strain LV43 was most closely related to Thiobacillus thioparus and the uncultured bacterial strain Strip2, which is represented by a 16S rRNA clone obtained by direct PCR from the Stripa research mine in Sweden. This identification of strain LV43 is supported by its G+C content of 62%, which is within the range reported for strains of T. thioparus. Fluorescently labeled polyclonal antibodies specific for strain LV43 were used to locate and enumerate this strain at different locations in Movile Cave and in nearby surface-water and groundwater sources. Strain LV43 was found only at aerobic, neutral-pH sites within the cave. Strain LV43 was also found outside Movile Cave in surface waters and in groundwater believed to intercept the same sulfurous aquifer as Movile Cave.

Aerobiosis↗

PubMed1984

Expression of a Thiobacillus ferrooxidans origin of replication in Escherichia coli.

A cryptic plasmid from an autotrophically grown arsenic-resistant strain of Thiobacillus ferrooxidans was isolated and cloned into pBR325. The origin of replication of pBR325 was deleted, and the recombinant plasmid was shown to replicate in Escherichia coli, using an origin of replication located on the Thiobacillus plasmid.

Acidithiobacillus thiooxidans↗

PubMed1998

A Tn7-like transposon is present in the glmUS region of the obligately chemoautolithotrophic bacterium Thiobacillus ferrooxidans.

The region downstream of the Thiobacillus ferrooxidans ATCC 33020 atp operon was examined, and the genes encoding N-acetylglucosamine-1-uridyltransferase (glmU) and glucosamine synthetase (glmS) were found. This atpEFHAGDC-glmUS gene order is identical to that of Escherichia coli. The T. ferrooxidans glmS gene was shown to complement E. coli glmS mutants for growth on minimal medium lacking glucosamine. A Tn7-like transposon, Tn5468, was found inserted into the region immediately downstream of the glmS gene in a manner similar to the site-specific insertion of transposon Tn7 within the termination region of the E. coli glmS gene. Tn5468 was sequenced, and Tn7-like terminal repeat sequences as well as several open reading frames which are related to the Tn7 transposition genes tnsA, tnsB, tnsC, and tnsD were found. Tn5468 is the closest relative of Tn7 to have been characterized to date. Southern blot hybridization indicated that a similar or identical transposon was present in three T. ferrooxidans strains isolated from different parts of the world but not in two Thiobacillus thiooxidans strains or a Leptospirillum ferrooxidans strain. Since T. ferrooxidans is an obligately acidophilic autotroph and E. coli is a heterotroph, ancestors of the Tn7-like transposons must have been active in a variety of physiologically different bacteria so that their descendants are now found in bacteria that occupy very different ecological niches.

Acidithiobacillus thiooxidans↗

PubMed1962

Relation of the iron oxidizer, Thiobacillus ferrooxidans, to thiosulfate.

Colmer, Arthur R. (Louisiana State University, Baton Rouge). Relation of the iron oxidizer, Thiobacillus ferrooxidans, to thiosulfate. J. Bacteriol. 83:761-765. 1962.-A restudy has been made of Thiobacillus ferrooxidans secured from the acid drainage of some northern West Virginia coal mines. Cultures isolated from ferrous iron agar and then subcultured for 85 consecutive transfers in thiosulfate broth during a period of 14 months were able to use ferrous iron as their energy source when returned to ferrous iron broth. Thiosulfate utilization was quantitatively followed by iodine titration and pH change; ferrous iron oxidation was quantitatively determined by use of 1,10-phenanthroline.Characteristic colonial forms were produced by T. ferrooxidans growing on thiosulfate agar irrespective of whether the inoculum was from iron broth or thiosulfate broth.

Acidithiobacillus↗

PubMed1966

Absorption and utilization of organic matter by the strict autotroph, Thiobacillus thiooxidans, with special reference to aspartic acid.

Butler, Richard G. (Rutgers, The State University, New Brunswick, N.J.), and Wayne W. Umbreit. Absorption and utilization of organic matter by the strict autotroph, Thiobacillus thiooxidans, with special reference to aspartic acid. J. Bacteriol. 91:661-666. 1966.-The strictly autotrophic bacterium, Thiobacillus thiooxidans, can be shown to assimilate a variety of organic materials. Aspartic acid can be assimilated into protein and can be converted into CO(2), but even in the presence of sulfur it cannot serve as the sole source of carbon for growth. The reason appears to be that aspartic acid is converted into inhibitory materials.

Amino Acids↗

PubMed1966

Generation of reducing power in chemosynthesis. 3. Energy-linked reduction of pyridine nucleotides in Thiobacillus novellus.

Aleem, M. I. H. (Research Institute for Advanced Studies, Baltimore, Md.). Generation of reducing power in chemosynthesis. III. Energy-linked reduction of pyridine nucleotides in Thiobacillus novellus. J. Bacteriol. 91:729-736. 1966.-Cell-free extracts from Thiobacillus novellus. catalyzed an adenosine triphosphate (ATP)-dependent reduction of pyridine nucleotides anaerobically, or aerobically when the respiratory chain was inhibited by azide. The exogenous electron donor employed for the reduction of nicotinamide adenine dinucleotide or nicotinamide adenine dinucleotide phosphate (NADP) was thiosulfate, formate, or mammalian ferrocytochrome c. In the latter case, the oxidation of ferrocytochrome c was observed with the concomitant reduction of the pyridine nucleotide. Values calculated for the molar ratios of ATP utilized-NADP reduced and of cytochrome c oxidized-NADP reduced were 1:1 and 2:1, respectively. The energy-dependent reduction of the pyridine nucleotides was inhibited by Atabrine or amytal and by low concentrations of the uncouplers of oxidative phosphorylation such as 2,4-dinitrophenol and carbonyl cyanide m-chlorophenyl hydrazone. Evidence is presented showing that the reduced pyridine nucleotides are essential for providing the reducing power for the energy-dependent reduction of carbon dioxide in T. novellus.

Adenosine Triphosphate↗

PubMed1966

Effects of organic matter on the growth of Thiobacillus intermedius.

London, Jack (University of California, Los Angeles), and Sydney C. Rittenberg. Effects of organic matter on the growth of Thiobacillus intermedius. J. Bacteriol. 91:1062-1069. 1966.-Yeast extract, glucose, glutamate, and other organic materials stimulate the rate and extent of growth of Thiobacillus intermedius in thiosulfate broth. Growth did not occur in glucose or glutamate mineral salts medium in the absence of thiosulfate, although a stable variant was obtained which grows on yeast extract alone. Cells harvested from media supplemented with organic matter have a reduced rate of thiosulfate oxidation (20 to 30% of autotrophic), oxidize the organic supplement, and have an additive rate of oxidation in the presence of both the organic substrate and thiosulfate. Carboxydismutase synthesis is repressed, and the incorporation of bicarbonate carbon into cell material is almost completely eliminated by the presence of organic matter in the growth medium. It is concluded that the availability of organic matter eliminates the autotrophic assimilatory mechanisms of T. intermedius but not its autotrophic energy-generating system. The data are discussed in relation to the existence of "obligate" chemoautotrophic bacteria.

Bicarbonates↗

PubMed1966

Membrane-associated sulfur oxidation by the autotroph Thiobacillus thiooxidans.

Adair, Frank W. (Rutgers, The State University, New Brunswick, N. J.). Membrane-associated sulfur oxidation by the autotroph Thiobacillus thiooxidans. J. Bacteriol. 92:899-904. 1966.-Washed cell wall-membrane fragments derived from sulfur-grown cells of the strictly autotrophic bacterium, Thiobacillus thiooxidans, oxidized elemental sulfur to sulfate without the addition of cofactors. The oxidation was optimal at pH 7.0 and was increased by the presence of wetting agents. Oxygen uptake was inhibited by cyanide, azide, and thiol-binding agents. Sulfite was also oxidized, and both the sulfur- and sulfite-oxidizing systems were heat-labile. Neither thiosulfate nor tetrathionate was oxidized by soluble or membrane preparations. The fragments fixed C(14)O(2) in the presence of ribose-5-phosphate, Mg(++), and adenosine triphosphate. Sulfur oxidation did not provide energy for C(14)O(2) fixation in this system.

Adenosine Triphosphate↗

PubMed1978

Characteristics of Thiobacillus thioparus and its thiocyanate assimilation.

Thiocyanate-assimilatig bacterium, TK 21, was isolated from activated sludge used for the treatment of thiocyanate contained in coke-oven liquor. This organism oxidized thiosulfate and elemental sulfur, causing a decrease of pH of the medium. These facts indicated that it belongs to the genus Thiobacillus. Potassium thiocyanate (0.5 g/l) was completely assimilated during 60 h. Thiosulfate inhibited the assimilation of thiocyanate but elemental sulfur did not. This bacterium did not evolve cyanide as its oxidation product after the decomposition of thiocyanate. The isoalted bacterium was identified as Thiobacillus thioparus. Examination of the composition of cellular fatty acid of three strains of T. thioparus showed that they prossessed 3-hydroxy fatty acid of C10 and C12; saturated straight chains of C10, C12, C15, C16, C17, and C18; monounsaturated straight chains of C16 and C18; and cyclopropane acid of C17.

Fatty Acids↗

PubMed1981

Physiological and ultrastructural characterization of a new acidophilic Thiobacillus species (T. kabobis).

A new autotrophic acidophilic Thiobacillus sp. was isolated from acidic soil adjacent to a natural gas processing plant's sulfur stockpile. This isolate metabolized S2O3(2-) to S4O6(2-) during growth and could not reoxidize this product; instead, the remaining S2O3(2-) substrate was oxidized to SO4(2-) in the stationary phase which represented a significant metabolic change as compared with other acidophilic thiobacilli. In contrast the isolate oxidized S0 to H2SO4 during the log-phase growth with no S4O6(2-) being formed. Ultrastructural studies revealed that the isolate's cytoplasm contained unidentified membrane-bound granules, volutin type bodies, and carboxysomes. Cleavage planes within the cell wall revealed a pattern of highly ordered subunits in the outer leaflet of the outer membrane and these ordered subunits were also discerned through the eutectic at the cell surface. This regular structure was not observed at the surface of cells of other acidophilic thiobacilli. The isolate bears a single pilus and one unusually long polar flagellum. The physiological and ultrastructural data are discussed in relation to other known thiobacilli and show this isolate to be a new Thiobacillus sp. named T. kabobis.

Cell Membrane↗

PubMed1984

The role of Thiobacillus albertis glycocalyx in the adhesion of cells to elemental sulfur.

Thiobacillus albertis, a newly characterized acidophilic Thiobacillus sp., was found not to be dependent on physiological conditions such as pH, cellular energy, or peripheral cell envelope sulfhydryl groups for attachment to elemental sulfur (S0). Heat-killed cells or those pretreated with sulfhydryl reagents (iodoacetate or iodoacetamide) were able to adhere to S0 in comparable numbers as assayed by epifluorescence microscopy. In addition, iodoacetate and iodoacetamide were found to be bactericidal, the former more potent than the latter. Sodium lauryl sulfate was found to cause nearly complete detachment of T. albertis cells from glass slides implicating its glycocalyx for this cell-glass attachment. In addition scanning electron microscopy visually revealed T. albertis cellular adhesion to S0 was due to the organism's threadlike glycocalyx material interacting with the sulfur surface. It was concluded that T. albertis glycocalyx plays an important role in the attachment to solid surfaces (glass or S0). In addition T. albertis was shown to colonize S0 surfaces by microcolonies.

Adhesiveness↗

PubMed1992

Cooperation between two Thiobacillus strains for heavy-metal removal from municipal sludge.

A mixed culture of two fast-growing bacterial strains for heavy-metal solubilization of municipal sewage sludge has been developed. Strain VA-7 decreases the initial sludge pH (7-8.5) to a value between 4.0 and 4.5. Then, strain VA-4 begins growing and further reduces the pH to values below 2.0. The rapid decrease of sludge pH by a mixed culture through sulfur oxidation into sulfuric acid solubilizes the toxic metals (Cd 83-96%, Cr 16-54%, Cu 85-87%, Mn 91-94%, Ni 78-79%, Pb 28-46%, Zn 82-96%) to levels recommended for intensive use of residual sludge in agriculture. A study of the physiological and metabolic characteristics of these strains revealed that isolate VA-7 is a strain of Thiobacillus thioparus (ATCC 55127), while isolate VA-4 corresponds to a Thiobacillus thiooxidans (ATCC 55128). These bacterial strains possess distinctive physiological characteristics that allow them to easily grow and solubilize heavy metals in municipal sludge.

Hydrogen-Ion Concentration↗

PubMed1998

Overproduction and substrate specificity of 3-isopropylmalate dehydrogenase from Thiobacillus ferrooxidans.

We constructed an overexpression system in Escherichia coli of the leuB gene coding for 3-isopropylmalate dehydrogenase in Thiobacillus ferrooxidans. E. coli harboring the plasmid we constructed, pKK leuB1, produced 17-fold the enzyme protein of the expression system previously used for purification. The substrate specificity of the enzyme was analyzed with synthetic (2R, 3S)-3-alkylmalates. The 3-isopropylmalate dehydrogenase of Thiobacillus ferrooxidans had broad specificity toward the alkylmalates.

3-Isopropylmalate Dehydrogenase↗

PubMed2004

[Studies of polymorphisms of Thiobacillus Ferrooxidans using RAPD].

Random amplified polymorphic DNA(RAPD) was used in analyzing the polymorphisms of Thiobacillus ferrooxidans from seven different places. Of the 20 primers, four could generate reproducible RAPD profiles, and each one produced 1 approximately 9 bands. The similarity coefficients obtained from profiles generated by four primers among Thiobacillus Ferrooxidans were about 44% approximately 83%.

Cluster Analysis↗

PubMed1991

The structural homology of amicyanin from Thiobacillus versutus to plant plastocyanins.

The complete amino acid sequence of the blue copper protein amicyanin of Thiobacillus versutus, induced when the bacterium is grown on methylamine, has been determined as follows: QDKITVTSEKPVAAADVPADAVVVGIEKMKYLTPEVTIKAGETVYWVNGEVMPHNVA FKKGIVGEDAFRGEMMTKDQAYAITFNEAGSYDYFCTPHPFMRGKVIVE. The four copper ligand residues in this 106-residue-containing polypeptide chain are His54, Cys93, His96, and Met99. The Thiobacillus amicyanin is 52% similar to the amicyanin of Pseudomonas AM1, the only other copper protein known with the same spacing between the second histidine ligand and the methionine ligand. T. versutus amicyanin contains no cysteine bridge and is more closely related to the plant copper protein plastocyanin than to the bacterial copper protein azurin. Alignment of the two known amicyanin sequences with the consensus sequence of the plastocyanins and comparison with the known three-dimensional structure of poplar leaves plastocyanin reveals that the bacterial proteins have the same overall structure with two beta-sheets packed face to face. The major structural differences between the amicyanins and the plastocyanins appear to be located in two of the five loops that connect the six identified beta-strands of the amicyanins. The first of these two loops, connecting strands F and G, contains a ligand histidine and must have a different conformation from the same loop in the plastocyanins because it is shorter by two amino acids. Further differences occur in the loop connecting the strands D and E. This loop contains only 17 residues in amicyanin whereas the corresponding loop of plastocyanin contains 25 residues. Despite these differences the amicyanins appear much closer related to the plastocyanins than to the azurins. The present findings demonstrate that the occurrence of blue copper proteins with clearly plastocyanin-like features is not restricted to photosynthetic redox chains.

Amino Acid Sequence↗

PubMedDate not supplied

[Effect of pH and temperature on the kinetics of Fe2+ oxidation by Thiobacillus ferrooxidans].

A correlation has been established between the action of pH and temperature on the activity of Fe2+ oxidation by Thiobacillus ferrooxidans. The rate of Fe2+ oxidation by the bacterium is inhibited at 12 degrees C mainly due to an increase in the pH. Autooxidation of the medium upon hydrolysis of Fe3+ is inhibited at low temperatures. Apparently, the enzymes of Thiobacillus ferrooxidans involved in Fe2+ oxidation are only slightly susceptible to a decrease of temperature to 10--12 degrees C.

Ferrous Compounds↗

PubMedDate not supplied

[Fractionation of stable isotopes of sulfur during its oxidation by Thiobacillus ferrooxidans].

The fractionation of sulfur isotopes was studied in the process of sulfur oxidation by Thiobacillus ferrooxidans. The isotope effect was observed at all stages of the bacterial growth during sulfur oxidation; as a result, residual sulfur became heavier whereas the fraction of colloidal sulfur and the fraction of sulfates became lighter. The isotope differences between the residual sulfur and the sulfates were greatest at the end of the lag phase (delta 34S = 0.38%) and the beginning of the stationary phase of the cultural growth (delta 34S = 0.34%). As was shown in model experiments, the phospholipids lecithin and cephalin can dissolve sulfur and form products enriched in 32S(delta 34S = -0.51 -1.60%). The mechanism of oxidation of elementary sulfur by Thiobacillus ferrooxidans is discussed.

Culture Media↗

PubMedDate not supplied

[Composition of a nutrient medium for the continuous cultivation of Thiobacillus ferrooxidans].

Thiobacillus ferrooxidans was found to require the following biogenous elements for constructing its biomass: phosphorus, 14 +/- 2; magnesium, 2 +/- 0.5; potassium, 5 +/- 1; nitrogen, 100 +/- 10 (milligrams per gram of dry biomass). It was shown that the concentrations of biogenous elements in the growth medium, which did not limit the biosynthesis, did not exceed 5 divided by 10 mg per litre for all of the studied elements. The medium 9K that is commonly used for growing Thiobacillus ferrooxidans has been found to differ significantly from the calculated one with the balanced ratio of biogenous elements.

Calcium↗