Surface sulphur as promoting agent of pyrite leaching by Thiobacillus ferrooxidans

Abstract: Methanol extraction conducted with a HPLC-Iike device and spectroscopic analysis were used to remove and characterize the sulphur layer (Ss) present on freshly ground pyrite surface after dry grinding. Accurate measurements of ferric and sulphate contents in the leachate showed a significant delay in the lag phase and in the first step of oxidation by Thiobacillus ferrooxidans for the so-cleaned pyrite (without sulphur layer) in comparison to the initial pyrite (with sulphur layer). Voltammetric studies (current-potential curves) showed a modification of the anodic behaviour of the initial pyrite, corresponding to a higher chemical oxidability of the uncleaned pyrite. During the bacterial oxidation, the difference in redox potential between a special pyrite electrode and a platinum standard electrode both placed in the bioleaching reactor was shown to be related to the occurrence of a sulphur layer. This difference, which is more important in the case of the initial pyrite (with sulphur layer), corresponded to an increase in oxidation kinetics of the pyrite by Thiobacillus ferrooxidans .     

Anaerobic leaching of covellite by Thiobacillus ferrooxidans

Thiobacillus ferrooxidans was able to grow under anaerobic conditions on copper sulphide with ferric ion as the electron acceptor. The dissolution of covellite under these conditions (68% after 35 days) was higher than values observed aerobically in cultures with similar media composition and almost as high as under aerobic conditions without iron. From these results we propose a mechanism for anaerobic bioleaching of covellite in the presence of ferric iron and speculate that it may occur in leach dumps where the oxygen concentration is, as reported elsewhere, very low. Received: 3 September 1996 / Received revision: 13 January 1997 / Accepted: 24 January 1997     

Identification of two outer membrane proteins involved in the oxidation of sulphur compounds in Thiobacillus ferrooxidans

Abstract: Shift of three Thiobacillus ferrooxidans strains from Fe(II) to S⁰ or thiosulphate liquid medium caused distinctive changes in the outer membrane protein profile. In addition to a new 55-kDa protein which was synthesized only in the presence of sulphur compounds, a higher expression of a 47-kDa protein was observed. This latter protein appeared to be constitutively synthesized, since it was detectable in small amounts even in tile presence of ferrous iron as sole energy source, but its expression was greatly enhanced when elemental sulphur or thiosulphate were present in the growth medium.     

The reductive reactions of Thiobacillus ferrooxidans on sulphur and selenium

Abstract An investigation into the sulphur oxidation of iron grown Thiobacillus ferrooxidans is reported. A new mechanism for sulphur catabolism in this obligate acidophile, involving the initial reduction of the substrate, is proposed. Evidence to support substrate reduction comes from the evolution of H₂S in both aerobic and anaerobic conditions (O₂ and Fe³⁺ acting as terminal electron acceptors, respectively) and a accurate trapping method of detection is described. Levels of sulphur reduction are such that a role in the full catabolism of sulphur can be envisaged. In addition, red selenium reduction was investigated and the bioenergetic implications for both substrates are discussed.     

Enzymic comparisons of the inorganic sulfur metabolism in autotrophic and heterotrophic Thiobacillus ferrooxidans.

Activities of enzymes which mediate the oxidation of thiosulfate to sulfate and the assimilation of sulfate to sulfide were assayed in various cell-free fractions of Thiobacillus ferrooxidans grown autotrophically on either ferrous iron or thiosulfate or heterotrophically on glucose. There was no activity of the thiosulfate-oxidizing enzyme in extracts of bacteria grown with ferrous iron. Comparable activities for ATP-sulfurylase (EC 2.7.7.4), ADP-sulfurylase (EC 2.7.7.5), and adenylate kinase (EC 2.7.4.3) were found in the bacteria grown autotrophically with either Fe2+ or S2O32- or heterotrophically with glucose.     

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°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.     

Morphology of bacterial leaching patterns by Thiobacillus ferrooxidans on synthetic pyrite

Thiobacillus ferrooxidans has been cultivated on synthetic pyrite (FeS₂) single crystals as the only energy source and the pyrite interface investigated with respect to characteristic morphological changes using scanning electron microscopy. Corrosion patterns of bacterial size were identified in different stages of development and correlated with bacterial activity. It appears that bacterial attack of the sulfide interface starts by secretion of organic substances around the contact area between the bacterial cell and the sulfide energy source. They might either be part of a pseudo capsule which shields the contact area or may form a sulfur absorbing and transporting organic film. Degradation of the sulfide occurs in the contact area below the bacterial cell leading to a corrosion pit which the bacterium may abandon after it has reached a depth of bacterial dimension. Electron spectroscopic (XPS) and X-ray fluorescence studies indicate a layer of organic substances covering the sulfide surface under bacterial leaching conditions, which is sufficiently thick for consideration in interfacial chemical mechanisms.     

Role of cell attachment in leaching of chalcopyrite mineral by Thiobacillus ferrooxidans

Summary Experiments on the leaching of copper from chalcopyrite mineral by the bacterium Thiobacillus ferrooxidans show that, in the presence of adequate amounts of sulphide, iron-grown bacteria preferentially oxidise sulphur in the ore (through direct attachment) rather than ferrous sulphate in solution. At 20% pulp density, the leaching initially takes place by a predominantly direct mechanism. The cell density in the liquid phase increases, but the Fe²⁺ is not oxidised. However, in the later stages when less solid substrate is available and the cell density becomes very high, the bacteria start oxidising Fe²⁺ in the liquid phase, thus contributing to the indirect mechanism of leaching. Contrary to expectations, the rate of leaching increased with increasing particle size in spite of the decreasing specific surface area. This has been found to be due to increasing attachment efficiency with increase in particle size. Offprint requests to: R. Kumar     

Dissolution of sulphur particles by Thiobacillus ferrooxidans: substrate for unattached cells

The growth of Thiobacillus ferrooxidans on sulphur is known to proceed through the attachment of cells to the sulphur particles. Experiments, However, show that the cells in the liquid phase, which are not attached to the sulphur particles, also grow. It has been shown through the use of a two-compartment membrane reactor that this increase is partially due to the release of ions, corresponding to partially oxidized of sulphur, into the solution by the attached cells. The main soluble ion has been found to the thiosulphate, but traces of sulphite have also been detected. (c) 1993 John Wiley & Sons, Inc.     

Suppression of pyritic sulphur during flotation tests using the bacterium Thiobacillus ferrooxidans

Environmental concern about sulphur dioxide emissions has led to the examination of the possibility of removing pyritic sulphur from coal prior to combustion during froth flotation, a routine method for coal cleaning at the pit-head. The bacterium Thiobacillus ferrooxidans was effective in leaching 80% and 63% -53 mum pyrite at 2% and 6% pulp density in shake flasks in 240 and 340 h, respectively.The natural floatability of pyrite was significantly reduced in the Hallimond tube following 2.5 min of conditioning in membrane-filtered bacterial liquor prior to flotation. The suppression effect was greatly enhanced in the presence of Thiobacillus ferrooxidans. A bacterial suspension in pH 2.0 distilled water showed 85% suppression, whereas in spent growth liquor this value was 95%. The optimum bacterial density was 3.25 x 10(10) cells/g pyrite in 230-ml distilled water (2% pulp density) in the Hallimond tube. The degree of suppression by the cells was related to particle size but not to pH or temperature. The sulphur content of a synthetic coal/pyrite mixture was reduced from 10.9 to 2.1% by flotation after bacterial preconditioning. It is postulated that pyrite removal in coals which are cleaned by froth flotation could be significantly reduced using a bacterial preconditioning stage with a short residence time of 2.5 min.     

Novel technique for investigation and quantification of bacteriol leaching by Thiobacillus ferrooxidans

A novel, efficient, and simple technique for the in situ study and quantification of the heterogeneous Bacteriol activity and the Bacteriol degradation of metal sulfides by Thiobacillus ferrooxidans is presented. It consists of exposing an ultrathin (300-2500 A) metal sulfide layer, FeS(2) in the experiments, to Thiobacillus f. grown in Touvinen media and visually following the Bacteriol attack and development of Bacteriol corrosion patterns under a light microscope. The uniform pyrite layer, partially transparent for visible light, permits the optical characterization of Bacteriol attack in remarkable detail. Several open or little understood questions concerning Bacteriol leaching, such as those on the kinetics of adhesion, the interfacial Bacteriol reproduction, the density of surface active bacteria, and the rate and morphology of sulfide degradation can also be studied. The degree of Bacteriol activity can be distinguished on the basis of development of variable sizes of spots and halos around Bacteriol cells produced by light passing through differently sized corrosion pits. The information obtained and identification of microorganisms has additionally been accentuated by immunofluorescence techniques (FA). It is concluded that the described method can be developed as a convenient testing and control technique for use in mine laboratories and bioleaching operations.     

The effect of xanthate floatation reagents on bacterial leaching of chalcopyrite by Thiobacillus ferrooxidans

Summary Xanthate floatation compounds, added to cultures of Thiobacillus ferrooxidans leaching chalcopyrite, inhibited growth in the concentration range 1–10 mM and caused a drop in formation of soluble copper and iron. Maximum (80–90%) growth inhibition was at 10 mM for isobutyl-, amyl- and ethyl xanthate while isopropyl xanthate was innocuous but caused a four-fold increase in lag phase. Copper production was depressed by 30% for isopropyl-, 53% for isobutyl-, ethyl- and 77% for amyl xanthate at 10 mM additions.     

Bioleaching of metallic sulphides withThiobacillus ferrooxidans in the absence of iron (II)

Bioleaching of metallic sulphides withThiobacillus ferrooxidans in the absence of iron (II) was studied using pure sulphides and mixtures. The direct mode of bacterial action was analysed with respect to sulphide solubility, exposed solid surfaces and bacterial attachment to the solids. Bioleaching of mixed sulphides showed enhancement of metal extraction in comparison with pure sulphides which suggests metal extractions would be better from polymetallic sulphide ores than from similar matrices with only one sulphide.     

Preliminary proteomic analysis of Thiobacillus ferrooxidans growing on elemental sulphur and Fe2+ separately

Thiobacillus ferrooxidans is one of the most important bacterium used in bioleaching, and can utilize Fe2+ or sulphide as energy source. Growth curves for Thiobacillus ferrooxidans have been tested, which show lag, logarithmic, stationary and aging phases as seen in other bacteria. The logarithmic phases were from 10 to 32 hours for Thiobacillus ferrooxidans cultivated with Fe2+ and from 4 to 12 days for Thiobacillus ferrooxidans cultivated with elemental sulphur. Differences of protein patterns of Thiobacillus ferrooxidans growing on elemental sulphur and Fe2+ separately were investigated after cultivation at 30 degrees C by the analysis of two-dimensional gel electrophoresis (2-DE), matrix-assisted laser desorption/ ionization (MALDI)-Mass spectrometry and ESI-MS/MS. From the 17 identified protein spots, 11 spots were found more abundant when growing on elemental sulphur. By contrast 6 protein spots were found decreased at elemental cultivation condition. Among the proteins identified, cytochrome C have been previously identified as necessary elements of electron-transferring pathway for Thiobacillus ferrooxidans to oxidize Fe2+; ATP synthase alpha chain and beta are expressed increased when Thiobacillus ferrooxidans cultivated with Fe2+ as energy source. ATP synthase Beta chain is the catalytic subunit, and ATP synthase alpha chain is a regulatory subunit. The function of ATPase produces ATP from ADP in the presence of a proton gradient across the membrane.     

Growth models of the continuous bacterial leaching of iron pyrite by Thiobacillus ferrooxidans

The leaching of iron pyrite by Thiobacillus ferrooxidans was studied in a continuous stirred tank reactor at a variety of dilution rates (0.012-0.22 h(-1)), pyrite surface areas (18-194 m(2)/L), and inlet soluble substrate (Fe(2+)) concentrations (0-3000 ppm). The bacterial leaching rate was found to increase with increasing pyrite surface area, dilution rate, and inlet Fe(2+) concentration. The concentration of bacteria in solution was related to the concentration of bacteria attached to the pyrite surface by a Langmuir-type adsorption-desorption relation. Fitting the experimental data to this relation yielded a value for the area occupied per bacterium of 86 mum(2). This result is consistent with the concept of preferential bacterial attachment of certain sites on the solid. A bacterial growth model was developed that included both bacterial growth in solution and growth of bacteria attached to the pyrite surface. The specific growth rate of the attached bacteria was calculated from this model and was found to increase with increasing solid dilution rate and to decrease with increasing pyrite surface area and soluble substance concentration. An explanation of these results based on an active-inactive site mechanisms was also developed.     

Bacterial leaching of a sulfide ore by Thiobacillus ferrooxidans and Thiobacillus thiooxidans: I. Shake flask studies

Bacterial leaching of a sulfide ore containing pyrite, chalcopyrite, and sphalerite was studied in shake flask experiments using Thiobacillus ferrooxidans and Thiobacillus thiooxidans strains isolated from mine sites. The Fe(2+)grown T. ferrooxidans isolates solubilized sphalerite preferentially over chalcopyrite leaching 7-10% Cu, 68-76% Zn, and 10-22% Fe from the ore in 18 days. The sulfur grown T. thiooxidans isolates leached Zn much more slowly and very little Fe, with a Cu-Zn extraction ratio twice the value obtained with T. ferrooxidans. The ore adapted T. ferrooxidans started solubilizing Cu and Zn without a lag period. The ore-adapted T. thiooxidans extracted Cu as well as T. ferrooxidans, but the extraction of Zn or Fe was still much slower in the low-phosphate medium, while in the high-phosphate medium it approached the value obtained with T. ferrooxidans. A high Cu-Zn extraction ratio of 0.34 was obtained with T. thiooxidans in the low phosphate medium. In the mixed-culture experiments with T. ferrooxidans and T. thiooxidans, the culture behaved as T. thiooxidans in the low-phosphate medium with a higher Cu-Zn extraction ratio and as T. ferrooxidans in the high-phosphate medium with a lower Cu-Zn extraction ratio. It is concluded that T. ferrooxidans and T. thiooxidans solubilize sulfide minerals by different mechanisms.