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PubMed2000

Purification and characterization of 3-isopropylmalate dehydrogenase from Thiobacillus thiooxidans.

3-Isopropylmalate dehydrogenase was purified to homogeneity from the acidophilic autotroph Thiobacillus thiooxidans. The native enzyme was a dimer of molecular weight 40,000. The apparent K(m) values for 3-isopropylmalate and NAD+ were estimated to be 0.13 mM and 8.7 mM, respectively. The optimum pH for activity was 9.0 and the optimum temperature was 65 degrees C. The properties of the enzyme were similar to those of the Thiobacillus ferrooxidans enzyme, expect for substrate specificity. T. thiooxidans 3-isopropylmalate dehydrogenase could not utilize malate as a substrate.

Journal Article↗

PubMed1998

Surface chemical studies of Thiobacillus ferrooxidans with reference to copper tolerance.

A strain of Thiobacillus ferrooxidans was adapted to grow at higher concentrations of copper by single step culturing in the presence of 20 g/L (0.314 mol/L) cupric ions added to 9K medium. Exposure to copper results in change in the surface chemistry of the microorganism. The isoelectric point of the adapted strain (pI = 4.7) was observed to be at a higher pH than that of the wild unadapted strain (pI = 2.0). Compared to the wild strain, the copper adapted strain was found to be more hydrophobic and showed enhanced attachment efficiency to the pyrite mineral. The copper adsorption ability of the adapted strain was also found to be higher than that of the wild strain. Fourier transform infrared spectroscopy of adapted cells suggested that a proteinaceous new cell surface component is synthesized by the adapted strain. Treatment of adapted cells with proteinase-K, resulted in complete loss of tolerance to copper, reduction in copper adsorption and hydrophobicity of the adopted cells. These observations strongly suggest a role played by cell surface modifications of Thiobacillus ferrooxidans in imparting the copper tolerance to the cells and bioleaching of sulphide minerals.

Acidithiobacillus thiooxidans↗

PubMed1998

Distribution of thiobacillus ferrooxidans and leptospirillum ferrooxidans: implications for generation of acid mine drainage

Although Thiobacillus ferrooxidans and Leptospirillum ferrooxidans are widely considered to be the microorganisms that control the rate of generation of acid mine drainage, little is known about their natural distribution and abundance. Fluorescence in situ hybridization studies showed that at Iron Mountain, California, T. ferrooxidans occurs in peripheral slime-based communities (at pH over 1.3 and temperature under 30 degreesC) but not in important subsurface acid-forming environments (pH 0.3 to 0.7, temperature 30 degrees to 50 degreesC). Leptospirillum ferrooxidans is abundant in slimes and as a planktonic organism in environments with lower pH. Thiobacillus ferrooxidans affects the precipitation of ferric iron solids but plays a limited role in acid generation, and neither species controls direct catalysis at low pH at this site.

Journal Article↗

PubMed1990

Ferrous Iron and Sulfur Oxidation and Ferric Iron Reduction Activities of Thiobacillus ferrooxidans Are Affected by Growth on Ferrous Iron, Sulfur, or a Sulfide Ore.

Eight strains of Thiobacillus ferrooxidans (laboratory strains Tf-1 [= ATCC 13661] and Tf-2 [= ATCC 19859] and mine isolates SM-1, SM-2, SM-3, SM-4, SM-5, and SM-8) and three strains of Thiobacillus thiooxidans (laboratory strain Tt [= ATCC 8085] and mine isolates SM-6 and SM-7) were grown on ferrous iron (Fe), elemental sulfur (S), or sulfide ore (Fe, Cu, and Zn). The cells were studied for their aerobic Fe - and S-oxidizing activities (O(2) consumption) and anaerobic S-oxidizing activity with ferric iron (Fe) (Fe formation). Fe-grown T. ferrooxidans cells oxidized S aerobically at a rate of 2 to 4% of the Fe oxidation rate. The rate of anaerobic S oxidation with Fe was equal to the aerobic oxidation rate in SM-1, SM-3, SM-4, and SM-5, but was only one-half or less that in Tf-1, Tf-2, SM-2, and SM-8. Transition from growth on Fe to that on S produced cells with relatively undiminished Fe oxidation activities and increased S oxidation (both aerobic and anaerobic) activities in Tf-2, SM-4, and SM-5, whereas it produced cells with dramatically reduced Fe oxidation and anaerobic S oxidation activities in Tf-1, SM-1, SM-2, SM-3, and SM-8. Growth on ore 1 of metal-leaching Fe-grown strains and on ore 2 of all Fe-grown strains resulted in very high yields of cells with high Fe and S oxidation (both aerobic and anaerobic) activities with similar ratios of various activities. Sulfur-grown Tf-2, SM-1, SM-4, SM-6, SM-7, and SM-8 cultures leached metals from ore 3, and Tf-2 and SM-4 cells recovered showed activity ratios similar to those of other ore-grown cells. It is concluded that all the T. ferrooxidans strains studied have the ability to produce cells with Fe and S oxidation and Fe reduction activities, but their levels are influenced by growth substrates and strain differences.

Journal Article↗

PubMed1993

Examination of Lipopolysaccharide (O-Antigen) Populations of Thiobacillus ferrooxidans from Two Mine Tailings.

Net acid-generating capacities of 39.74 kg of H(2)SO(4) per ton (ca. 0.05 kg/kg) (pH 2.68) for the Lemoine copper mine tailings (closed ca. 8 years ago; located 40 km west of Chibougamau, Quebec, Canada) and 16.07 kg of H(2)SO(4) per ton (ca. 0.02 kg/kg) (pH 3.01) for the Copper Rand tailings (in current use and 50 km distant [east] from those of Lemoine) demonstrate that these sulfide tailings can support populations of acidophilic thiobacilli. Oxidized regions in both tailings environments were readily visible, were extremely acidic (Lemoine, pH 2.36; Copper Rand, pH 3.07), and provided natural isolates for our study. A 10% (wt/vol) oxalic acid treatment, which solubilizes both ferric sulfate and ferric hydroxide precipitates (B. Ramsay, J. Ramsay, M. deTremblay, and C. Chavarie, Geomicrobiol. J. 6:171-177, 1988), enabled the recovery of intact bacterial cells from the tailings material and from liquid synthetic medium for lipopolysaccharide analysis. No viable cells could be cultured after this oxalic acid treatment. Sodium dodecyl sulfate-polyacrylamide gel electro-phoretic profiles of lipopolysaccharides extracted from the Lemoine tailings were complex, indicating a heterogeneous population of Thiobacillus ferrooxidans. Six T. ferrooxidans subspecies as identified by lipopolysaccharide analysis (i.e., lipopolysaccharide chemotypes) were eventually isolated from a total of 112 cultures from the Lemoine tailings. Using the same isolate and lipopolysaccharide typing techniques, we identified only a single lipopolysaccharide chemotype from 20 cultures of T. ferrooxidans isolated from the Copper Rand tailings. This homogeneity of lipopolysaccharide chemotype was much different from what was found for the older Lemoine tailings and may reflect a progressive lipopolysaccharide heterogeneity of Thiobacillus isolates as tailings leach and age.

Journal Article↗

PubMed1994

Solubilization of Minerals by Bacteria: Electrophoretic Mobility of Thiobacillus ferrooxidans in the Presence of Iron, Pyrite, and Sulfur.

Thiobacillus ferroxidans is an obligate acidophile that respires aerobically on pyrite, elemental sulfur, or soluble ferrous ions. The electrophoretic mobility of the bacterium was determined by laser Doppler velocimetry under physiological conditions. When grown on pyrite or ferrous ions, washed cells were negatively charged at pH 2.0. The density of the negative charge depended on whether the conjugate base was sulfate, perchlorate, chloride, or nitrate. The addition of ferric ions shifted the net charge on the surface asymptotically to a positive value. When grown on elemental sulfur, washed cells were close to their isoelectric point at pH 2.0. Both pyrite and colloidal sulfur were negatively charged under the same conditions. The electrical double layer around the bacterial cells under physiological conditions exerted minimal electrostatic repulsion in possible interactions between the cell and either of its charged insoluble substrates. When Thiobacillus ferrooxidans was mixed with either pyrite or colloidal sulfur at pH 2.0, the mobility spectra of the free components disappeared with time to be replaced with a new colloidal particle whose electrophoretic properties were intermediate between those of the starting components. This new particle had the charge and size properties anticipated for a complex between the bacterium and its insoluble substrates. The utility of such measurements for the study of the interactions of chemolithotrophic bacteria with their insoluble substrates is discussed.

Journal Article↗

PubMed1998

Development of a laboratory-scale leaching plant for metal extraction from fly ash by thiobacillus strains.

Semicontinuous biohydrometallurgical processing of fly ash from municipal waste incineration was performed in a laboratory-scale leaching plant (LSLP) by using a mixed culture of Thiobacillus thiooxidans and Thiobacillus ferrooxidans. The LSLP consisted of three serially connected reaction vessels, reservoirs for a fly ash suspension and a bacterial stock culture, and a vacuum filter unit. The LSLP was operated with an ash concentration of 50 g liter, and the mean residence time was 6 days (2 days in each reaction vessel). The leaching efficiencies (expressed as percentages of the amounts applied) obtained for the economically most interesting metal, Zn, were up to 81%, and the leaching efficiencies for Al were up to 52%. Highly toxic Cd was completely solubilized (100%), and the leaching efficiencies for Cu, Ni, and Cr were 89, 64, and 12%, respectively. The role of T. ferrooxidans in metal mobilization was examined in a series of shake flask experiments. The release of copper present in the fly ash as chalcocite (Cu(2)S) or cuprite (Cu(2)O) was dependent on the metabolic activity of T. ferrooxidans, whereas other metals, such as Al, Cd, Cr, Ni, and Zn, were solubilized by biotically formed sulfuric acid. Chemical leaching with 5 N H(2)SO(4) resulted in significantly increased solubilization only for Zn. The LSLP developed in this study is a promising first step toward a pilot plant with a high capacity to detoxify fly ash for reuse for construction purposes and economical recovery of valuable metals.

Journal Article↗

PubMed1963

SULFATE REQUIREMENT FOR IRON OXIDATION BY THIOBACILLUS FERROOXIDANS.

Lazaroff, Norman (British Columbia Research Council, Vancouver, B.C., Canada). Sulfate requirement for iron oxidation by Thiobacillus ferrooxidans. J. Bacteriol. 85:78-83. 1963.-The growth of Thiobacillus ferrooxidans is initially inhibited in media containing ferrous chloride in place of ferrous sulfate. This inhibition of growth is due to the requirement of a high relative proportion of sulfate ions to chloride (or other anions) for iron oxidation. Adaptation takes place, producing strains which are able to oxidize iron in media containing an initially unfavorable anionic composition. Adaptation is possibly due to the selection of spontaneous mutants capable of oxidizing iron in high chloride, low sulfate media. Such cells are found at a frequency of 10(-5) of the population of unadapted cultures.

Journal Article↗

PubMed1965

Thiosulfate Oxidation and Electron Transport in Thiobacillus novellus.

Aleem, M. I. H. (Research Institute for Advanced Studies, Baltimore, Md.). Thiosulfate oxidation and electron transport in Thiobacillus novellus. J. Bacteriol. 90:95-101. 1965.-A cell-free soluble enzyme system capable of oxidizing thiosulfate was obtained from Thiobacillus novellus adapted to grow autotrophically. The enzyme systems of autotrophically grown cells brought about the transfer of electrons from thiosulfate to molecular oxygen via cytochromes of the c and a types; the reactions were catalyzed jointly by thiosulfate oxidase and thiosulfate cytochrome c reductase. The levels of both of these enzymes were markedly reduced in the heterotrophically grown organism. Cell-free extracts from the autotrophically grown T. novellus catalyzed formate oxidation and enzymatically reduced cytochrome c with formate. Both formate oxidation and cytochrome c reduction activities were abolished under heterotrophic conditions. The thiosulfate-activating enzyme S(2)O(3) (-2)-cytochrome c reductase, as well as thiosulfate oxidase, was localized chiefly in the soluble cell-free fractions, and the former enzyme was purified more than 200-fold by ammonium sulfate fractionation and calcium phosphate gel adsorption procedures. Optimal activity of the purified enzyme occurred at pH 8.0 in the presence of 1.67 x 10(-1)m S(2)O(3) (-2) and 2.5 x 10(-4)m cytochrome c. The thiosulfate oxidase operated optimally at pH 7.5 and thiosulfate concentrations of 1.33 x 10(-3) to 3.33 x 10(-2)m in the presence of added cytochrome c at a concentration of 5 x 10(-4)m. Both enzymes were markedly sensitive to cyanide and to a lesser extent to some metal-binding agents. Although a 10(-3)m concentration of p-hydroxymercuribenzoate had no effect on S(2)O(3) (-2)-cytochrome c reductase, it caused a 50% inhibition of S(2)O(3) (-2) oxidase, which was completely reversed in the presence of 10(-3)m reduced glutathione. Carbon monoxide also inhibited S(2)O(3) (-2) oxidase; the inhibition was completely reversed by light.

Journal Article↗

PubMed1966

Fine Structure of Thiobacillus thiooxidans.

Mahoney, Robert P. (Skidmore College, Saratoga Springs, N.Y.), and Mercedes R. Edwards. Fine structure of Thiobacillus thiooxidans. J. Bacteriol. 92: 487-495. 1966.-Thin section analysis of the chemosynthetic autotroph Thiobacillus thiooxidans revealed structures comparable to gram-negative heterotrophic bacteria. Although this species is unique in that it oxidizes elemental sulfur for energy, uses carbon dioxide as its sole source of carbon, and can withstand a pH of less than 1, thin sections revealed a profile of the cell envelope (cell wall and plasmalemma) similar to other gram-negative species which have more common physiological traits. The cell wall is composed of five layers with an overall width of approximately 200 A, and the plasmalemma appears as a conventional "unit membrane" with a width of about 85 A. Volutin granules and less-dense bodies of similar shape and size were frequently observed in close association with the nucleoplasm. The nature and function of these bodies are unknown at this time.

Journal Article↗

PubMed1964

LEACHING OF CHALCOPYRITE WITH THIOBACILLUS FERROOXIDANS: EFFECT OF SURFACTANTS AND SHAKING.

The rate of leaching of chalcopyrite by Thiobacillus ferrooxidans has been greatly accelerated by using shaken flasks in place of stationary bottles or percolators. A further increase in rate and extent of leaching was obtained by the use of Tween 20, 40, 60, and 80, Triton X-100, Quaker TT 5386, and Hyamine 2389. Tween 20 was the most effective surfactant. No individual component of the Tween molecule was responsible for the improved leaching. The Tween-to-chalcopyrite ratio is more important than the Tween-to-medium ratio. The effect of the surfactants is probably due to increased contact between the mineral surface and the organism, and shaking provides the necessary oxygen. Rates and yields obtained by use of surfactants and shaking as aids to microbiological leaching approach those obtained with acidified erric sulfate leaching.

Acidithiobacillus↗

PubMed2004

Development of an automated water toxicity biosensor using Thiobacillus ferrooxidans for monitoring cyanides in natural water for a water filtering plant.

An on-line biosensor consisting of immobilized Thiobacillus ferrooxidans and an oxygen electrode was developed for automated monitoring of acute toxicity in water samples. T. ferrooxidans is an obligatory acidophilic, autotrophic bacterium and derives its energy by the oxidation of ferrous ion, elemental sulfur, and reduced sulfur compounds including metal sulfides. The assay is based on the monitoring of a current increase by addition of toxicoids, which is caused by the inhibition of bacterial respiration and decrease in oxygen consumption. Optimum cell number on the membrane was 5.0 x 10(8) cells. The steady-state current was obtained when concentration of FeSO4 was above 3.6 mM at pH 3. The sensor response of T. ferrooxidans immobilized membrane for 5.0 microM KCN was within an error of 10% for 30 membranes. A linear relationship was obtained at KCN concentration in the range of 0.5-3.0 microM in a flow-type monitoring system. Minimum detectable concentrations of KCN, Na2S, and NaN3 were 0.5, 1.2, and 0.07 microM, respectively. The monitoring system contained two biosensors and these sensors were cleaned with sulfuric acid (pH 1.5) twice a day. This treatment could remove fouling on microbial immobilized membrane by natural water and ferrous precipitation in the flow cell. This flow-type monitoring sensor was operated continuously for 5 months. Also, T. ferrooxidans immobilized membrane can be stored for one month at 4 degrees C when preserved with wet absorbent cotton under argon gas.

Biosensing Techniques↗

PubMed1976

Microbiological leaching of a chalcopyrite concentrate by Thiobacillus ferrooxidans.

The microbiological leaching of a chalcopyrite concentrate has been investigated using a pure strain of Thiobacillus ferrooxidans. The optimum leaching conditions regarding pH, temperature, and pulp density were found to be 2.3, 35 degrees C, and 22% respectively. The energy of activation was calculated to be 16.7 kcal/mol. During these experiments the maximum rate of copper dissolution was about 215 mg/liters/hr and the final copper concentration was as high as 55 g/liter. This latter value is in the range of copper concentrations which may be used for direct electrorecovery of copper. Jarosite formation was observed during the leaching of the chalcopyrite concentrate. When the leach residue was reground to expose new substrate surface, subsequent leaching resulted in copper extractions up to about 80%. On the basis of this experimental work, a flow sheet has been proposed for commercial scale biohydrometallurgical treatment of high-grade chalcopyrite materials.

Biodegradation, Environmental↗

PubMed1977

Ferrous iron oxidation and uranium extraction by Thiobacillus ferrooxidans.

The microbiological oxidation of ferrous iron in batch and continuous systems has been investigated in relation to uranium extraction from a low-grade ore by Thiobacillus ferrooxidans. The influence of the parameters, agitation, and aeration on oxygen saturation concentration, rate of oxygen mass transfer, and rate of ferrous iron oxidation was demonstrated. The kinetic values, Vmax and K were determined using an adapted Monod equation for different dilution rates and initial concentrations of ferrous iron. The power requirements for initial leaching conditions were also calculated. Uranium extraction as high as 68% has been realized during nine days of treatment. Regrinding the leach residue and its subsequent leaching yielded 87% uranium solubilization.

Ferrous Compounds↗

PubMed1978

The interpretation of the mass spectrum of an ornithine-containing lipid from Thiobacillus thiooxidans.

The electron impact mass spectrum of a previously identified ornithine-containing lipid from Thiobacillus thiooxidans has been interpreted using exact mass measurements, low and high energy ionization, and defocused metastable studies. The spectrum, which did not contain a molecular ion for the intact lipid, was consistent with cyclization of the ornithine zwitterionic moiety with elimination of water to give 3[3'-(11,12-methylene-2-hydroxyoctadecanoxy)hexadecanylamine]-2-piperidone. Production of this sufficiently volatile species for mass spectral analysis was accomplished by gentle pyrolysis in the mass spectrometer source. The spectrum can be understood to arise by three primary decompositions which serve to separate the two fatty acid constituents. The remainder of the spectrum is consistent with the expected fragmentations of these constituents.

Acidithiobacillus thiooxidans↗

PubMed1976

Degradation of sulphur nucleotides in cell-free extracts of Thiobacillus ferrooxidans.

The cell-free extracts of Thiobacillus ferrooxidans contain hydrolytic enzymes which cleave sulphate from either adenosine 5'-sulphatophosphate or adenosine 3'-phosphate 5'-sulphatophosphate. These sulphatase activities may be retarded by including either phosphates or adenine nucleotides in the incubation mixtures. Adenosine 3'-phosphate 5'-sulphatophosphate and its immediate breakdown product adenosine 3'-phosphate 5'-phosphate were degraded at slower rates than adenosine 5'-sulphatophosphate.

Adenine Nucleotides↗

PubMed1977

Effects of different compounds of metals and of their mixtures on the growth and survival of Thiobacillus ferrooxidans.

The iron oxidation by Thiobacillus ferrooxidans was studied in the presence of different concentrations of metals as sulphates, chlorides, nitrates, and in some other compounds. The bacteriological oxidation of ferro-ions develops in all solutions in nearly the same way, with the only difference that, as the ion concentration increases, the beginning of measurable oxidation is postponed, i.e., with ion concentration increasing, the lag-phase expands. When salts are mixed in different proportions, the toxic concentration of such a mixture corresponds, in a defined way, to the total of salts, i.e., of their ions.

Chlorides↗

PubMed1981

Geomicrobiological leaching of tin minerals by Thiobacillus ferro-oxidans and organic agents.

Laboratory investigations confirm that it is possible to leach tin from synthetic minerals like stannite, kesterite, stannoidite, herzenbergite, ottemannite and berndtite, and from natural tin minerals which include stannite, cassiterite and varlamoffite, in the presence of Thiobacillus ferrooxidans, and with organic agents of biological origin, especially oxalic acid and oxalic-citric acids mixture. Over a leaching period of 35 days with 0.5% pulp density, initial pH of 2.5, using minus 0.16 mm size fraction at 32 degree C, as much as 54.45, 72.66, 97.13, and 31.30% Sn were extracted from synthetic stannite, kesterite, stannoidite and natural stannite, respectively. Varlamoffite, found in the dried, leached residues of the tin sulphides, provides evidence that bacterial action can be responsible for the genesis of supergene varlamoffite.

Citrates↗