Differential proteomic analysis of Acidithiobacillus ferrooxidans cells maintained in contact with bornite or chalcopyrite: Proteins involved with the early bacterial response

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estudo da oxidação dos sulfetos sintéticos molibdenita (MoS2) e covelita (CuS) por Acidithiobacillus ferrooxidans via respirometria celular

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Differentiation of Acidithiobacillus ferrooxidans and A-thiooxidans strains based on 16S-23S rDNA spacer polymorphism analysis

Restriction fragment length polymorphism (RFLP) and sequence analyses of the PCR-amplified 16S-23S rDNA intergenic spacer (ITS) were used for differentiating Acidithiobacillus thiooxidans strains from other related acidithiobacilli, including A. ferrooxidans and A. caldus. RFLP fingerprints obtained with AluI, DdeI, HaeIII, HinfI and MspI enabled the differentiation of all Acidithiobacillus reference strains into species groups. The A. thiooxidans strains investigated (metal mine isolates) yielded identical RFLP patterns to the A. thiooxidans type strain (ATCC 19377(T)), except for strain DAMS, which had a distinct pattern for all enzymes tested. Fourteen A. ferrooxidans mine strains were assigned to 3 RFLP groups, the majority of which were grouped with A. ferrooxidans ATCC 23270(T). The spacer region of one representative strain from each of the RFLP groups obtained was subjected to sequence analysis, in addition to eleven additional A. thiooxidans strains isolated from sediment and water samples, and A. caldus DSM 8584(T). The tRNA(IIe) and tRNA(Ala) genes, present in all strains analyzed, showed high sequence similarity. Phylogenetic analysis of the ITS sequences differentiated all three Acidithiobacillus species. Inter- and infraspecific genetic variations detected were mainly due to the size and sequence polymorphism of the ITS3 region. Mantel tests showed no significant correlation between ITS sequence similarity and the geographical origin of strains. The results showed that the 16S-23S rDNA spacer region is a useful target for the development of molecular-based methods aimed at the detection, rapid differentiation and identification of acidithiobacilli. (C) 2004 Elsevier SAS. All rights reserved.

Electrochemical noise analysis of chalcopyrite carbon paste electrodes by Acidithiobacillus ferrooxidans

The electrochemical response of chalcopyrite was studied using electrochemical noise analysis (ENA). The assay was carried out under constant aeration using 30 mL in two electrochemical cells containing iron-free mineral salts solution. These cells were initially monitored for 56 hours, After 72 hours, 7.25x 10(10) cells mL(-1) of A, ferrooxidans strain LR were added in both cells and monitored until 128 h. Subsequent to this period, 0.927 mmol L-1 of silver ions and 400 mmol L-1 of chloride ions were added each one separately. Both conditions were monitored until 168 hours. According to results obtained, it was observed that Cl- ions addition induced an accelerated corrosion process. However, there is a tendency of the system to reach the stationary state due to repassivation of the electrodic surface. In the other side, the Ag+ addition contributed for the maintenance of the oxidant atmosphere, in spite of controversial effect caused by considerable variations in the R-n values, resulting in a instability in the chalcopyrite reactivity.

Protein profile of Acidithiobacillus ferrooxidans strains exhibiting different levels of tolerance to metal sulfates

Strains of Acidithiobacillus ferrooxidans exhibited differences in the inhibition of Fe(2+) oxidation in the presence of 250 mm of cadmium, zinc, and manganese sulfates in respirometric assays. Strains LR and I35 were practically not inhibited, whereas strains SSP and V3 showed significant inhibition (30-70%). Analysis by SDS-PAGE of total proteins from cells grown in the absence of metal sulfates showed different profiles between the more tolerant strains (LR and 135) and the more susceptible ones (SSP and V3). Total proteins of strains LR and V3 were also resolved by two-dimensional polyacrylamide gel electrophoresis (2-DE). A set of major proteins (40, 32, 22, and 20 kDa) could be identified only in the more tolerant strain LR. Our results show that protein profiles analysis could differentiate A. ferrooxidans strains that considerably differ in the tolerance to metal sulfates and present low genomic similarity as revealed by Random Amplified Polymorphic DNA (RAPD) data obtained previously in our laboratory.

Oxidative dissolution of chalcopyrite by Acidithiobacillus ferrooxidans analyzed by electrochemical impedance spectroscopy and atomic force microscopy

The microbiological leaching of chalcopyrite (CuFeS2) is of great interest because of its potential application to many CuFeS2-rich ore materials. However, the efficiency of the microbiological process is very limited because this mineral is one of the most refractory to bacterial attack. Knowledge of bacterial role during chalcopyrite oxidation is very important in order to improve the efficiency of bioleaching operation. The oxidative dissolution of a massive chalcopyrite electrode by Acidithiobacillus ferrooxidans was evaluated by electrochemical impedance spectroscopy (EIS) and atomic force microscopy (AFM). A massive chalcopyrite electrode was utilized in a Tait-type electrochemical cell in acid medium for different immersion times in the presence or absence of bacterium. The differences observed in the impedance diagrams were correlated with the adhesion process of bacteria on the mineral surface. (C) 2004 Elsevier B.V. All rights reserved.

Oxidation of chalcopyrite by Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans in shake flasks

Chalcopyrite oxidation was evaluated with two acidophilic thiobacilli that are important in bioleaching processes. Acidithiobacillus thiooxidans in pure culture did not oxidize CuFeS2 but oxidized externally added S in the presence of CuFeS2. Acidithiobacillus ferrooxidans released Cu2+ and soluble Fe from chalcopyrite, and the time course lead to a gradual passivation of chalcopyrite whereby Cu2+ dissolution leveled off. Fe3+ acted as a chemical oxidant in CuFeS2 leaching and was reduced to Fe2+. Parallel bacterial re-oxidation of Fe2+ contributed to a high Fe3+/Fe2+ ratio and an increase in redox potential. Chemical oxidation of chalcopyrite was slow compared with A. ferrooxidans-initiated solubilization. X-ray analysis revealed new solid phases: (i) jarosite, found in solids from A. ferrooxidans cultures and in chemical controls that initially received Fe2+ or Fe3+, and (ii) S-0, found mostly in iron-amended A. ferrooxidans culture and the corresponding chemical controls. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Electrochemical noise analysis of bioleaching of bornite (Cu5FeS4) by Acidithiobacillus ferrooxidans

Electrochemical noise (EN) is a generic term describing the phenomenon of spontaneous fluctuations of potential or current noise of electrochemical systems. Since this technique provides a non-destructive condition for investigating corrosion processes, it can be useful to study the electrochemical oxidation of mineral sulfides by microorganisms, a process known as bacterial leaching of metals. This technique was utilized to investigate the dissolution of a bornite electrode in the absence (first 79 h) and after the addition of Acidithiobacillus ferrooxidans (next 113 h) in salts mineral medium at pH 1.8, without addition of the energy source (Fe2+ ions) for this chemolithotrophic bacterium. Potential and current noise data have been determined simultaneously with two identical working bornite electrodes which were linked by a zero resistance ammeter (ZRA). The mean potential, E-coup, coupling current, I-coup, standard deviations of potential and current noise fluctuations and noise resistance, R-n, have been obtained for coupled bornite electrodes. Noise measurements were recorded twice a day in an unstirred solution at 30 degrees C. Significant changes in these parameters were observed when the A. ferrooxidans suspension was added, related with bacterial activity on reduced species present in the sulfide moisture (Fe2+, S2-). ENA was a suitable tool for monitoring the changes of the corrosion behavior of bornite due to the presence of bacterium. (C) 2006 Elsevier B.V. All rights reserved.

Oxidation of isochemical FeS2 (marcasite-pyrite) by Acidithiobacillus thiooxidans and Acidithiobacillus ferrooxidans

A research-grade mineral sample that contained marcasite and pyrite (FeS2) was subjected to the oxidation by Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans. Oxidation of FeS2 by A. ferrooxidans produced acid, and the redox potential increased with sulfide dissolution and the oxidation of Fe2+. jarosite was detected in solids from spent cultures. Preferential oxidation of either mineral was not consistently observed across all treatments. Neither iron sulfide was oxidized by A. thiooxidans. (C) 2006 Elsevier Ltd. All rights reserved.

Tolerance to copper and zinc of Acidithiobacillus thiooxidans isolated from sewage sludge

The effect of copper and zinc ions on sulphur oxidation by Acidithiobacillus thiooxidans, strain SFR01, isolated from anaerobic sewage sludge was assessed, resulting in tolerance levels up to 20 and 200 mmol l(-1) for copper and zinc, respectively. The tolerance levels obtained were higher than the concentration of copper and zinc usually found in the collected sewage sludge. The tolerance levels obtained indicate no constraints for sludge bioleaching of those metals due to their toxicities to the indigenous A. thiooxidans.

Immobilized cells of Acidithiobacillus ferrooxidans in PVC strands and sultite removal in a pilot-scale bioreactor

The biooxidation of ferrous ion into ferric ion by Acidithiobacillus ferrooxidans can be potentially used for the removal of H2S from industrial gases. In this work, Fe3+ ions were obtained through the oxidation of Fe2+ using the LR strain of At. ferrooxidans immobilized in PVC stands in a pilot-scale bioreactor, while H2S was removed in an absorption tower equipped with Rasching rings. At. ferrooxidans LR strain cells were immobilized by inoculating the bacterium in a Fe2+-mineral medium and percolating it through the support. After complete Fe2+ oxidation, which took around 90 h, the reactor was washed several times with sulfuric acid (pH 1.7) before a new cycle was started. Four additional cycles using fresh Fe2+ mineral medium were then run. During these colonization cycles, the time required for complete iron oxidation decreased, dropping to about 60 h in the last cycle. The batch experiments in the H2S gas removal trials resulted in a gas removal rate of about 98-99% under the operational conditions employed. In the continuous experiments with the bioreactor coupled to the gas absorption column, a gas removal efficiency of almost 100% was reached after 500 min. Precipitate containing mainly sulfur formed during the experimental trial was identified by EDX. (c) 2005 Elsevier B.V. All rights reserved.

Molecular characterization of Acidithiobacillus ferrooxidans and A-thiooxidans strains isolated from mine wastes in Brazil

Nineteen strains of Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans, including 12 strains isolated from coal, copper, gold and uranium mines in Brazil, strains isolated from similar sources in other countries and the type strains of the two species were characterized together with the type strain of A. caldus by using a combination of molecular systematic methods, namely ribotyping, BOX- and ERIC-PCR and DNA-DNA hybridization assays. Data derived from the molecular fingerprinting analyses showed that the tested strains encompassed a high degree of genetic variability. Two of the Brazilian A. ferrooxidans organisms (strains SSP and PCE) isolated from acid coal mine waste and uranium mine effluent, respectively, and A. thiooxidans strain DAMS, isolated from uranium mine effluent, were the most genetically divergent organisms. The DNA-DNA hybridization data did not support the allocation of Acidithiobacillus strain SSP to the A. ferrooxidans genomic species, as it shared only just over 40% DNA relatedness with the type strain of the species. Acidithiobacillus strain SSP was not clearly related to A. ferrooxidans in the 16S rDNA tree.

Jarosite pseudomorph formation from arsenopyrite oxidation using Acidithiobacillus ferrooxidans

In this work, the oxidizing action of a native strain type A. ferrooxidans on a sulphide containing a predominance of arsenopyrite and pyrite has been evaluated. Incubation of the A. ferrooxidans strain in flasks containing 200 mL of T&K medium with the ore (particle size of 106 mu m) at pulp density 8% (w/v) at 35 degrees C on a rotary shaker at 200 rpm resulted in preferential oxidation of the arsenopyrite and the mobilization of 88% of the arsenic in 25 days. Mineralogical characterization of the residue after biooxidation was carried out with FTIR. XRD and SEM/XEDS techniques. An in situ oxidation of the arsenopyrite is suggested on the basis of the frequent appearance of jarosite pseudomorph replacing arsenopyrite, in which the transformations Fe(2+) -> Fe(3+), S(-2) -> S(+6) and As(-1) -> As(+3) -> As(+5) occur for the most part without formation of soluble intermediates, resulting in a type of jarosite that typically contains high concentrations of arsenic (type A-jarosite). However, during pyrite oxidation, dissolution of the constituent Fe and S predominates, which is evidenced by corrosion of pyrite particles with formation of pits, generating a type of jarosite with high quantities of K (type B-jarosite). Lastly, a third type of jarosite (type C-jarosite) also precipitated forming a thin film that covered the grains of pyrite principally. (C) 2010 Elsevier B.V. All rights reserved.

Expression, purification and spectroscopic analysis of an HdrC: An iron-sulfur cluster-containing protein from Acidithiobacillus ferrooxidans

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Characterization of bornite (Cu5FeS4) electrodes in the presence of the bacterium Acidithiobacillus ferrooxidans

Bornite electrodes were characterized in the absence or in the presence of Acidithiobacillus ferrooxidans, which is an important microorganism involved in metal bioleaching processes. The presence of the bacterium modified the mineral/electrolyte interface, increasing the corrosion rate, as revealed by interferometric, AEM, ICP and EIS analyses. As a consequence of bacterial activity the electrode became porous, increasing its surface heterogeneity. This behavior was correlated with the evolution of impedance diagrams obtained during the time course of experiments. The main difference in these diagrams was the presence of an inductive feature (up to 44 h), which was related to bacterial action on the mineral dissolution, better than to its adhesion on the bornite. The total real impedance measured in presence of the bacterium was about 10 times lower than in its absence, due to the acceleration of the mineral dissolution, because an oxidant environment was maintained.