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The chemical attack of ore by ferric iron and/or sulfuric acid releases valuable metals. The products of these reactions are recycled by iron and sulfur oxidizing microorganisms. These acidophilic chemolithotrophic prokaryotes, among which Acidithiobacillus ferrooxidans, grow at the expense of the energy released from the oxidation of ferrous iron and/or inorganic sulfur compounds (ISCs). In At. ferrooxidans, it has been shown that the expression of the genes encoding the proteins involved in these respiratory pathways is dependent on the electron donor and that the genes involved in iron oxidation are expressed before those responsible for ISCs oxidation when both iron and sulfur are present. Since the redox potential increases during iron oxidation but remains stable during sulfur oxidation, we have put forward the hypothesis that the global redox responding two components system RegB/RegA is involved in this regulation. To understand the mechanism of this system and its role in the regulation of the aerobic respiratory pathways in At. ferrooxidans, the binding of different forms of RegA (DNA binding domain, wild-type, unphosphorylated and phosphorylated-like forms of RegA) on the regulatory region of different genes/operons involved in ferrous iron and ISC oxidation has been analyzed. We have shown that the four RegA forms are able to bind specifically the upstream region of these genes. Interestingly, the phosphorylation of RegA did not change its affinity for its cognate DNA. The transcriptional start site of these genes/operons has been determined. In most cases, the RegA binding site(s) was (were) located upstream from the −35 (or −24) box suggesting that RegA does not interfere with the RNA polymerase binding. Based on the results presented in this report, the role of the RegB/RegA system in the regulation of the ferrous iron and ISC oxidation pathways in At. ferrooxidans is discussed.

The chemical attack of ore by ferric iron and/or sulfuric acid releases valuable metals. The products of these reactions are recycled by iron and sulfur oxidizing microorganisms. These acidophilic chemolithotrophic prokaryotes, among which Acidithiobacillus ferrooxidans, grow at the expense of the energy released from the oxidation of ferrous iron and/or inorganic sulfur compounds (ISCs). In At. ferrooxidans, it has been shown that the expression of the genes encoding the proteins involved in these respiratory pathways is dependent on the electron donor and that the genes involved in iron oxidation are expressed before those responsible for ISCs oxidation when both iron and sulfur are present. Since the redox potential increases during iron oxidation but remains stable during sulfur oxidation, we have put forward the hypothesis that the global redox responding two components system RegB/RegA is involved in this regulation. To understand the mechanism of this system and its role in the regulation of the aerobic respiratory pathways in At. ferrooxidans, the binding of different forms of RegA (DNA binding domain, wild-type, unphosphorylated and phosphorylated-like forms of RegA) on the regulatory region of different genes/operons involved in ferrous iron and ISC oxidation has been analyzed. We have shown that the four RegA forms are able to bind specifically the upstream region of these genes. Interestingly, the phosphorylation of RegA did not change its affinity for its cognate DNA. The transcriptional start site of these genes/operons has been determined. In most cases, the RegA binding site(s) was (were) located upstream from the -35 (or -24) box suggesting that RegA does not interfere with the RNA polymerase binding. Based on the results presented in this report, the role of the RegB/RegA systemin the regulation of the ferrous iron and ISC oxidation pathways in At. ferrooxidans is discussed. [ABSTRACT FROM AUTHOR]

Recent understanding of microbial retention within heap bioleaching systems has highlighted the importance of quantifying microbial growth and activity in both the bulk flowing solution and in the ore-associated phases. Typically, industrial heap bioleaching operations report variations in process conditions such as inoculum preparation and concentration and elevated copper concentrations in the recycled irrigation solution. In this paper, a mini-column reactor system containing pre-constructed and agglomerated, low-grade ore samples representing grab samples from a larger heap, were used to investigate the effect of a selection of physico-chemical and operating conditions on microbial growth, colonisation and substrate utilisation kinetics, considering both the planktonic and sessile populations of Acidithiobacillus ferrooxidans . The factors studied included inoculum size, inoculum cultivation conditions, availability of ferrous iron in the bulk flowing solution and copper concentration in the bulk flowing solution. The microbial population in the interstitial phase, i.e. associated with, but not bound to, the ore, remained the most abundant within the heap under all physico-chemical conditions considered. A comparison of the tests with different inoculum sizes found that a smaller inoculum size resulted in an increased delay in microbial growth and ferrous iron oxidation, but similar apparent maximum specific growth rates and iron oxidation rates. In contrast to the microbial culture grown on pyrite, a delay in microbial activity was observed for the culture grown on ferrous iron. However, greater microbial cell densities were reached, in the interstitial and attached phases compared with the pyrite-grown culture. The introduction of 6 g L − 1 cupric ions into the feed solution containing 0.2 g L − 1 ferric iron resulted in decreased microbial growth rate in the interstitial phase but not in the attached phase. Where the pyrite culture was pre-exposed to cupric ion, the microbial growth rate in the interstitial and attached phases was significantly enhanced. Nevertheless, the presence of cupric ion in the irrigation solution resulted in a decrease in microbial ferrous iron oxidation rate, irrespective of pre-culture conditioning. This study emphasises the important role played by the stagnant interstitial phase during the colonisation of a low-grade heap, particularly under adverse conditions for microbial growth and activity. It also highlights the role of inoculum culture conditions on the potential trade-off between increased heap colonisation and increased lag periods in microbial activity during heap start-up. [ABSTRACT FROM AUTHOR]

Acidithiobacillus ferrooxidans plays an important role in bioleaching in reproducing the mineral oxidant of ferric iron ( Fe3+) by oxidization of ferrous iron ( Fe2+). The high-molecular-weight c-type cytochrome Cyc2 that is located in the external membrane is postulated as the first electron carrier in the Fe2+ oxidation respiratory pathway of A. ferrooxidans. To increase ferrous iron oxidation activity, a recombinant plasmid p TCYC2 containing cyc2 gene under the control of Ptac promoter was constructed and transferred into A. ferrooxidans ATCC19859. The transcriptional level of cyc2 gene was increased by 2.63-fold and Cyc2 protein expression was observed in the recombinant strain compared with the control. The ferrous iron oxidation activity and the arsenic stressed cell growth of the recombinant strain were also elevated. [ABSTRACT FROM AUTHOR]

Abstract: A novel Acidithiobacillus ferrooxidans strain QXS-1 was isolated from the acid mine drainage of Qixiashan Pb–Zn–Ag mine area in China. The isolate QXS-1 is a motile, rod-shaped Gram-negative bacterium with an optimum growth at 30°C and pH 2.0. The phylogenetic analysis has demonstrated that QXS-1 is a new strain of A. ferrooxidans. The strain QXS-1 exhibited an unusual trait on the utilization of ferrous iron and sulfur as energy sources. The sulfur-domesticated QXS-1 cells only utilized sulfur as energy source for growth when both ferrous iron and sulfur are available, whereas ferrous-domesticated QXS-1 cells prefer to simultaneously utilize both ferrous iron and sulfur as energy source for growth in the presence of both substrates. The QXS-1 cells accumulated intracellular hydrophilic sulfur during growth. Based on the ultrastructure study, we deduced that the accumulated sulfur in QXS-1 exists in both octasulfur ring form and polysulfide form. The strain QXS-1 exhibited remarkable marmatite-bioleaching capacity. It is able to extract 6585mg/L zinc from marmatite within 18days in shake flask. [Copyright &y& Elsevier]

In contrast to iron-oxidizing Acidithiobacillus ferrooxidans, A. ferrooxidans from a stationary phase elemental sulfur-oxidizing culture exhibited a lag phase in pyrite oxidation, which is similar to its behaviour during ferrous iron oxidation. The ability of elemental sulfur-oxidizing A. ferrooxidans to immediately oxidize ferrous iron or pyrite without a lag phase was only observed in bacteria obtained from growing cultures with elemental sulfur. However, these cultures that shifted to ferrous iron oxidation showed a low rate of ferrous iron oxidation while no growth was observed. Two-dimensional gel electrophoresis was used for a quantitative proteomic analysis of the adaptation process when bacteria were switched from elemental sulfur to ferrous iron. A comparison of total cell lysates revealed 39 proteins whose increase or decrease in abundance was related to this phenotypic switching. However, only a few proteins were closely related to iron and sulfur metabolism. Reverse-transcription quantitative PCR was used to further characterize the bacterial adaptation process. The expression profiles of selected genes primarily involved in the ferrous iron oxidation indicated that phenotypic switching is a complex process that includes the activation of genes encoding a membrane protein, maturation proteins, electron transport proteins and their regulators. [ABSTRACT FROM AUTHOR]

In this study, a proteomic analysis of Acidithiobacillus ferrooxidans by two-dimensional electrophoresis identified 24 proteins that were differentially expressed when the cells were grown on ferrous iron (Fe) or elemental sulfur (S°). Sixteen of these proteins were upregulated by growth on S° or downregulated by growth on Fe, including four proteins involved in disulfide bond reduction such as pyridine nucleotide-disulfide oxidoreductase, heterodisulfide reductase subunit B, thioredoxin-disulfide reductase, and cysteine desulfurase IscS, and three proteins involved in saccharide metabolism. A total of eight proteins were upregulated by growth on Fe or downregulated by S°. Northern blots further confirmed the differences in transcription for these differentially expressed proteins. We functionally characterized cysteine desulfurase IscS, and found that its overexpression in E. coli promoted the growth of the cells in LB containing 2.5 % sodium thiosulfate. Our results provide new insights into the molecular basis for S° and Fe oxidation by this extreme acidophile. [ABSTRACT FROM AUTHOR]

Abstract: A few researchers have reported on work concerning bioleaching of heavy-metal-contaminated soil using Acidithiobacillus ferrooxidans, since this acidophile is sensitive to dissolved low molecular weight (LMW) organic acids. Iron oxidation by A. ferrooxidans R2 as well as growth on ferrous iron was inhibited by a variety of dissolved LMW organic acids. Growth experiments with ferrous iron as an oxidant showed that the inhibition capability sequence was formic acid>acetic acid>propionic acid>oxalic acid>malic acid>citric acid. The concentrations that R2 might tolerate were formic acid 0.1mmolL−1 (2mmolkg−1 soil), acetic and propionic acids 0.4mmolL−1 (8mmolkg−1 soil), oxalic acid 2.0mmolL−1 (40mmolkg−1 soil), malic acid 20mmolL−1 (400mmolkg−1 soil), citric acid 40mmolL−1 (800mmolkg−1 soil), respectively. Although R2 was sensitive to organic acids, the concentrations of LMW organic acids in the contaminated soils were rather lower than the tolerable levels. Hence, it is feasible that R2 might be used for bioleaching of soils contaminated with metals or metals coupled with organic compounds because of the higher concentrations of LMW organic acids to which R2 is tolerant. [Copyright &y& Elsevier]

The oxidation of ferrous iron by Acidithiobacillus ferrooxidans in the pH range 2.5–7.0 was characterized. In order to measure the rate of bacterial oxidation of ferrous iron with A. ferrooxidans in this high pH range, a novel experimental methodology was developed. The results showed that the inhibition of ferrous iron oxidation activity by A. ferrooxidans observed at pH values above 3.0 is partially linked to the formation of ferric iron precipitates, which apparently hinder transport processes on the cell surface. By carefully controlling the amount of iron precipitates formed during the oxidation of ferrous iron in this pH range, enhanced bacterial activity was obtained. [Copyright &y& Elsevier]

This study used bacterial cellulose (BC) as novel support material for Acidithiobacillus ferrooxidans immobilization, with the objective of improving the performance of bioleaching and increasing its sustainability. The BC was synthesized in the laboratory, which allowed selecting the desired size of this highly porous and mechanically resistant material. After bacterial immobilization, the biologically active material (BAM) was used to assess the effect that several operating parameters had on the process (shaking mode and speed, the nutrient medium volume-to-exposed surface area ratio –NMV:ESA-, and Fe2+ concentration). The influence of dissolved copper and BAM storage was also determined. The results showed that the NMV:ESA ratio conditioned Fe2+ bio-oxidation time: after 33 h, 60% and 90% of the initial Fe2+ in the samples was converted, with a 1:0.1 and 1:0.6 ratio, respectively. In addition, Fe3+ productivity increased 16% when raising the initial Fe2+ content from 6 to 9 g L−1. A considerable increase in productivity (from 190 to 223 mg L−1 h−1) was also recorded when the shaking speed was increased from 130 to 170 rpm. BAM successfully adapted to the presence of dissolved copper and oxidized Fe2+ in the presence of up to 30 g Cu2+ L−1. Finally, the integrity of the material was not affected by the demanding operating conditions, and bacterial activity was successfully recovered after storage at 4 °C and 22 °C. In sum, biocellulose has proven to be a suitable candidate for A. ferrooxidans immobilization, which could contribute to improving the efficiency of bioleaching. [Display omitted] • Bacterial cellulose is a new suitable support for A. ferrooxidans immobilization. • The optimum operating conditions for reducing bio-oxidation time were established. • Fe3+ productivity of 223 mg L−1 h−1 was achieved at selected operating conditions. • The immobilized bacteria were resistant to copper concentrations up to 30 g L−1. • The use of bacterial cellulose enhanced the sustainability of bioleaching process. [ABSTRACT FROM AUTHOR]

Acidithiobacillus ferrooxidans YNTRS-40 (A. ferrooxidans) is a chemolithoautotrophic aerobic bacterium isolated from Tengchong hot springs, Yunnan Province, China, with a broad growth pH range of 1.0–4.5. This study reports the genome sequence of this strain and the information of genes related to the adaptation of diverse stresses and the oxidation of ferrous iron and sulfur. Results showed that YNTRS-40 possesses chromosomal DNA (3,209,933-bp) and plasmid DNA (47,104-bp). The complete genome of 3,257,037-bp consists of 3,349 CDS genes comprising 6 rRNAs, 52 tRNAs, and 6 ncRNAs. There are many encoded genes associated with diverse stresses adaptation and ferrous iron and sulfur oxidation such as rus operon, res operon, petI, petII, sqr, doxDA, cydAB, and cyoABCD. This work will provide essential information for further application of A. ferrooxidans YNTRS-40 in industry. [ABSTRACT FROM AUTHOR]