Bacterium Acidithiobacillus Ferrooxidans Research Articles

Synergistic promotion of antimony transformation in the interaction of Acidithiobacillus ferrooxidans and pyrite by driving the formation of reactive oxygen species and secondary minerals

As one of the important microorganisms in the mining area, the role of iron-sulfur oxidizing microorganisms in antimony (element symbolized as Sb) migration and transformation in mining environments has been largely neglected for a long time. Therefore, the processes of the typical iron-sulfur oxidizing bacterium Acidithiobacillus ferrooxidans (A. ferrooxidans) and pyrite interaction coupled with the migration and transformation of Sb were investigated in this paper. The bio-oxidation process of pyrite by A. ferrooxidans not only accelerates the oxidation rate of Sb(III) to Sb(V) (62.93% of 10 mg L−1 within 4 h), but also promotes the adsorption and precipitation of Sb (32.89 % of 10 mg L−1 within 96 h), and changes in the dosage of minerals, Sb concentration, and pH value affect the conversion of Sb. The characterization results show that the interaction between A. ferrooxidans and pyrite produces a variety of reactive species, such as H2O2 and •OH, resulting in the oxidation of Sb(III). In addition, A. ferrooxidans mediates the formation of stereotyped iron-sulfur secondary minerals that can act as a major driver of Sb (especially Sb(V)) adsorption or co-precipitation. This study contributes to the further understanding of the diversified biogeochemical processes of iron-sulfur oxidizing bacteria-iron-sulfur minerals-toxic metals in mining environments and provides ideas for the development of in-situ treatment technologies for Sb.

Evaluation of Mesophilic Species Performance in Extraction of Cobalt and Manganese from Hot Filter Cake of Zinc Processing Plant

ABSTRACT Cobalt, recognized as one of the scarcest industrial metals, is typically discarded during the zinc production process in the high-temperature refining section, along with manganese. This waste product is colloquially referred to as ‘hot cake.’ The present study endeavored to segregate cobalt and manganese from the hot cake and transition them into the liquid phase via bioleaching, a method that is both economically and environmentally superior to pyrometallurgical techniques. In the hot cake, cobalt exists as Co (III) and manganese as Mn (IV). Given their oxidized states, both cobalt and manganese are insoluble in acid and necessitate reduction for dissolution. The study employed Acidithiobacillus thiooxidans and Acidithiobacillus ferrooxidans bacteria to create an environment conducive to the reduction of cobalt and manganese. Under optimal conditions, this biotechnological approach facilitated the recovery of 62.8% of cobalt and 91.6% of manganese. Subsequently, a model was developed to predict the optimal method for processing cobalt from hot cake, based on the recovery rates of cobalt and manganese. This research contributes to the broader goal of resource recovery and waste minimization in the metal industry.

Siderite and vivianite as energy sources for the extreme acidophilic bacterium Acidithiobacillus ferrooxidans in the context of mars habitability

Past and present habitability of Mars have been intensely studied in the context of the search for signals of life. Despite the harsh conditions observed today on the planet, some ancient Mars environments could have harbored specific characteristics able to mitigate several challenges for the development of microbial life. In such environments, Fe2+ minerals like siderite (already identified on Mars), and vivianite (proposed, but not confirmed) could sustain a chemolithoautotrophic community. In this study, we investigate the ability of the acidophilic iron-oxidizing chemolithoautotrophic bacterium Acidithiobacillus ferrooxidans to use these minerals as its sole energy source. A. ferrooxidans was grown in media containing siderite or vivianite under different conditions and compared to abiotic controls. Our experiments demonstrated that this microorganism was able to grow, obtaining its energy from the oxidation of Fe2+ that came from the solubilization of these minerals under low pH. Additionally, in sealed flasks without CO2, A. ferrooxidans was able to fix carbon directly from the carbonate ion released from siderite for biomass production, indicating that it could be able to colonize subsurface environments with little or no contact with an atmosphere. These previously unexplored abilities broaden our knowledge on the variety of minerals able to sustain life. In the context of astrobiology, this expands the list of geomicrobiological processes that should be taken into account when considering the habitability of environments beyond Earth, and opens for investigation the possible biological traces left on these substrates as biosignatures.

Development of Technology for the Bioleaching of Uranium in a Solution of Bacterial Immobilization

This study presents findings regarding the kinetics of ferrous iron oxidation in solution mediated by Acidithiobacillus ferrooxidans bacteria within a continuous-flow bioreactor employing diverse types of immobilizers. The objective is to augment the rate of ferrous iron oxidation in solutions utilizing an immobilizer for Acidithiobacillus ferrooxidans strains. Immobilization represents a promising avenue for enhancing the efficiency of Fe2⁺ oxidation via acidophilic ferrooxidizing bacteria, leading to a several-fold increase in oxidation rate. A comparative analysis was conducted to evaluate the efficacy of different types of immobilizer in facilitating iron oxidation within a continuous-flow bioreactor, including the application of wood chips coated with Fe(OH)3. The results indicate that wood chips coated with iron hydroxide serve as effective type of immobilizer, facilitating the robust attachment of Acidithiobacillus ferrooxidans via electrostatic interactions between negatively charged bacteria and positively charged surfaces. Experimental investigations were conducted using novel immobilization matrices in pilot-scale tests simulating the underground borehole leaching (UBL) of uranium. The bioactivation of leaching solutions enhances the efficiency and environmental compatibility of UBL compared to conventional chemical oxidation methods. The relationships between redox potential and ferric iron content in bioactivated solutions during the UBL of uranium were delineated. The significance of this study lies in its elucidating the pivotal role of Fe2⁺ oxidation in uranium extraction processes, particularly in the context of UBL. By employing bioactivation mediated by Acidithiobacillus ferrooxidans, the study demonstrates not only enhanced uranium extraction efficiency, but also markedly improved environmental sustainability compared to traditional chemical oxidation methods. The findings reveal crucial correlations between redox potential and ferric iron concentration in bioactivated solutions.

Insight into the stabilization/solidification of Cd, Pb and Zn in MSWI FA by three stages: Vulcanization, precipitation and bio-oxidation

Bio-oxidation has been perceived as a promising treatment for the solidification/stabilization (S/S) of heavy metals (HMs) in mining soils. Nonetheless, no research exists on the integration of the iron-sulfur oxidizing bacterium Acidithiobacillus ferrooxidans as a microbial agent into the treatment of municipal solid waste incineration-fly ash (MSWI-FA). The effect of vulcanization (Stage I), precipitation (Stage II) and bio-oxidation (Stage III) on the stabilization of HMs in MSWI-FA was unexplored. This study investigated the stabilization effects of Stage I and Stage II by utilizing Na2S and FeSO4 to adjust the Fe/S ratio. The findings contributed to identifying suitable methods to enhance the stabilization effect of the microbial agent on MSWI-FA (Stage III). The results indicated that the vulcanization (Stage I) increased the compressive strength of MSWI-FA to 1.87 MPa when the concentration of Na2S reached 0.03 mM. The leaching toxicity of Cd, Pb, and Zn at 28 days after Stage II was 0.12 mg/L, 0.99 mg/L, and 356 mg/L, respectively, which improved by 96.3%, 94.8%, and 26.9% compared to the untreated samples. The compressive strength of the samples peaked at 1.97 MPa when the Fe/S ratio was 0.25 (precipitation (Stage II)). Despite the inhibitory effect of MSWI-FA on microbial activity, the leaching toxicity of Cd, Pb, and Zn in the presence of Acidithiobacillus ferrooxidans decreased to 0.07 mg/L, 0.17 mg/L and 97.3 mg/L at 28 days after Stage III, with no significant further increase in compressive strength. This study suggested that bio-oxidation utilizing iron sources may change the formation of iron oxides, thereby impacting the mechanical and chemical properties of MSWI-FA-based materials. The treatment sequence of vulcanization, precipitation and bio-oxidation proved effective in immobilizing HMs, particularly Pb, Cd, and Zn in MSWI FA. This research highlighted the potential of employing this technique to treat MSWI FA, yet it is essential to identify suitable reaction systems with appropriate additives to further enhance the long-term effectiveness and mechanical properties of MSWI-FA-based materials.

Research on the decomposition mechanisms of lithium silicate ores with different crystal structures by autotrophic and heterotrophic bacteria

Ores serve as energy and nutrient sources for microorganisms. Through complex biochemical processes, microorganisms disrupt the surface structure of ores and release metal elements. However, there is limited research on the mechanisms by which bacteria with different nutritional modes act during the leaching process of different crystal structure ores. This study evaluated the leaching efficiency of two types of bacteria with different nutritional modes, heterotrophic bacterium Bacillus mucilaginosus (BM) and autotrophic bacterium Acidithiobacillus ferrooxidans (AF), on different crystal structure lithium silicate ores (chain spodumene, layered lepidolite and ring elbaite). The aim was to understand the behavioral differences and decomposition mechanisms of bacteria with different nutritional modes in the process of breaking down distorted crystal lattices of ores. The results revealed that heterotrophic bacterium BM primarily relied on passive processes such as bacterial adsorption, organic acid corrosion, and the complexation of small organic acids and large molecular polymers with metal ions. Autotrophic bacterium AF, in addition to exhibiting stronger passive processes such as organic acid corrosion and complexation, also utilized an active transfer process on the cell surface to oxidize Fe2+ in the ores for energy maintenance and intensified the destruction of ore lattices. As a result, strain AF exhibited a greater leaching effect on the ores compared to strain BM. Regarding the three crystal structure ores, their different stacking modes and proportions of elements led to significant differences in structural stability, with the leaching effect being highest for layered structure, followed by chain structure, and then ring structure. These findings indicate that bacteria with different nutritional modes exhibit distinct physiological behaviors related to their nutritional and energy requirements, ultimately resulting in different sequences and mechanisms of metal ion release from ores after lattice damage.

Acidithiobacillus ferrooxidans Leaching of Silica-Sulfide Gold Ores from May-Hibey Deposits, Tigray, Ethiopia

Oxidative leaching is an inexpensive alternative to using chemical cyanide extraction methods for gold from low-grade gold sulfide. Oxidation of finely ground gold-bearing ore by Acidithiobacillus ferrooxidans was evaluated in terms of cell density, pH, and leaching efficiency of Fe and Au in shake flask experiments. The compositional and elemental analyses of the beneficiated ore were analyzed using XRD and EDXRF spectroscopy. The ore’s primary constituents are gold (4.356 mg/L), silicon, iron, and sulfur (62.456, 15.441, and 7.912 wt%, respectively). XRD spectra, the main phases of the concentrated ore, showed that the major components of the ore were quartz, syn, silicon sulfide, pyrite, and polymetallic elements such as silderenrite, gismondine, siderenikite, hematite, and syn. The experimental results, with Acidithiobacillus ferrooxidans bacteria and blank, were evaluated. The pH of the blank remained nearly constant, and the pH of the bioleached was occasionally lowered. The A. ferrooxidans strain always grew better throughout the bioleaching process. For the A. ferrooxidans strain, the cell density of cells reached a maximum of 90.00 × 106 cells/mL after the 11th week and decreased to 87.00 × 106 cells/mL after the 12th week. The decrease in cell density may be due to the presence of polymetallic elements such as Al, Cr, Ti, and Ni, leading to reduced metal tolerance of the A. ferrooxidans strain. In the A. ferrooxidans leaching process, the maximum total iron and gold extraction reached 92.16% (14.23 mg/L) and 99.97% (4.355 ppm), respectively, after the 11th week, and leaching tends to decrease up to 14 weeks, which may be due to the formation of secondary minerals. More research will be performed to optimize the procedure and leaching kinetic, examine the impact of metal content, and take into account the potential for bioleaching process pollution in addition to the amount of gold recovered.

Biologicalisation of surface structuring: Comparative environmental assessment of microbial biomachining vs. chemical etching

In 2015, the United Nations adopted the 2030 Agenda for Sustainable Development in response to growing global concern about climate change, resource scarcity and environmental impacts on ecosystems. This agenda also places high demands on industrial manufacturing systems to promote sustainability. Biologicalisation of manufacturing processes can be a promising approach to contribute to the sustainability goals in industrial production systems. In recent years, biomachining has emerged as a field of research in biologicalisation with potential advantages compared to conventional surface treatment methods of metals, such as chemical etching. Surface structuring of metals plays a crucial role in various industrial applications, such as pre-coating preparation, hierarchical structuring, superhydrophobic surfaces and geometries. To assess the environmental impacts of biomachining, this paper focuses on environmental impacts of utilizing the bacterium Acidithiobacillus ferrooxidans compared to a conventional chemical etching process with cupric chloride solution for surface structuring of a metal specimen. The study examines the environmental impact of both processes, considering all relevant material and energy flows from cradle to gate for multiple environmental impact categories. In doing so, the environmental impacts of each process can be comprehensively evaluated and compared, and hotspots for future improvement of the biomachining process can be identified. On current lab-scale, the results indicate higher impacts for the biomachining process in most categories compared to the chemical etching process, mainly due to the energy demand for pre-cultivation and preparation of the biomachining solution.

Enhancing sludge dewaterability in sequential bioleaching: Degradation of dissolved organic matter (DOM) by filamentous fungus Mucor sp. ZG-3 and the influence of energy source.

This study aimed to enhance sludge dewatering through sequential bioleaching, employing the filamentous fungus Mucor sp. ZG-3 and the iron-oxidizing bacterium Acidithiobacillus ferrooxidans LX5. The mechanism by which Mucor sp. ZG-3 alleviates sludge dissolved organic matter (DOM) inhibition of A. ferrooxidans LX5 was investigated, and the optimal addition of energy source for enhanced sludge dewaterability during sequential bioleaching was determined. Sludge dissolved organic carbon (DOC) decreased to 272 mg/L with a 65.2% reduction by Mucor sp. ZG-3 in 3 days, and the degraded fraction of sludge DOM was mainly low-molecular-weight DOM (L-DOM) which inhibited the oxidization of Fe2+ by A. ferrooxidans LX5. By degrading significant inhibitory low-molecular-weight organic acids, Mucor sp. ZG-3 alleviated DOM inhibition of A. ferrooxidans LX5. In the sequential bioleaching process, the optimal concentration of FeSO4·7H2O for A. ferrooxidans LX5 was 4 g/L, resulting in the minimum specific resistance to filtration (SRF) of 2.60×1011 m/kg, 40.0% lower than that in the conventional bioleaching process with 10 g/L energy source. Moreover, the sequential bioleaching process increased the sludge zeta potential (from -31.8 to -9.47 mV) and median particle size (d50) of the sludge particle (from 17.90 to 27.44 μm), contributing to enhanced sludge dewaterability. Inoculation of Mucor sp. ZG-3 during the bioleaching process reduced the demand for energy sources by A. ferrooxidans LX5 while improving sludge dewaterability performance.

Beyond the coupled distortion model: structural analysis of the single domain cupredoxin AcoP, a green mononuclear copper centre with original features.

Cupredoxins are widely occurring copper-binding proteins with a typical Greek-key beta barrel fold. They are generally described as electron carriers that rely on a T1 copper centre coordinated by four ligands provided by the folded polypeptide. The discovery of novel cupredoxins demonstrates the high diversity of this family, with variations in terms of copper-binding ligands, copper centre geometry, redox potential, as well as biological function. AcoP is a periplasmic cupredoxin belonging to the iron respiratory chain of the acidophilic bacterium Acidithiobacillus ferrooxidans. AcoP presents original features, including high resistance to acidic pH and a constrained green-type copper centre of high redox potential. To understand the unique properties of AcoP, we undertook structural and biophysical characterization of wild-type AcoP and of two Cu-ligand mutants (H166A and M171A). The crystallographic structures, including native reduced AcoP at 1.65 Å resolution, unveil a typical cupredoxin fold. The presence of extended loops, never observed in previously characterized cupredoxins, might account for the interaction of AcoP with physiological partners. The Cu-ligand distances, determined by both X-ray diffraction and EXAFS, show that the AcoP metal centre seems to present both T1 and T1.5 features, in turn suggesting that AcoP might not fit well to the coupled distortion model. The crystal structures of two AcoP mutants confirm that the active centre of AcoP is highly constrained. Comparative analysis with other cupredoxins of known structures, suggests that in AcoP the second coordination sphere might be an important determinant of active centre rigidity due to the presence of an extensive hydrogen bond network. Finally, we show that other cupredoxins do not perfectly follow the coupled distortion model as well, raising the suspicion that further alternative models to describe copper centre geometries need to be developed, while the importance of rack-induced contributions should not be underestimated.

Different fates of Sb(III) and Sb(V) during the formation of jarosite mediated by Acidithiobacillus ferrooxidans

Secondary iron-sulfate minerals such as jarosite, which are easily formed in acid mine drainage, play an important role in controlling metal mobility. In this work, the typical iron-oxidizing bacterium Acidithiobacillus ferrooxidans ATCC 23270 was selected to synthesize jarosite in the presence of antimony ions, during which the solution behavior, synthetic product composition, and bacterial metabolism were studied. The results show that in the presence of Sb(V), Fe2+ was rapidly oxidized to Fe3+ by A. ferrooxidans and Sb(V) had no obvious effect on the biooxidation of Fe2+ under the current experimental conditions. The presence of Sb(III) inhibited bacterial growth and Fe2+ oxidation. For the group with Sb(III), products with amorphous phases were formed 72 hr later, which were mainly ferrous sulfate and pentavalent antimony oxide, and the amorphous precursor was finally transformed into a more stable crystal phase. For the group with Sb(V), the morphology and structure of jarosite were changed in comparison with those without Sb. The biomineralization process was accompanied by the removal of 94% Sb(V) to form jarosite containing the Fe-Sb-O complex. Comparative transcriptome analysis shows differential effects of Sb(III) and Sb(V) on bacterial metabolism. The expression levels of functional genes related to cell components were much more downregulated for the group with Sb(III) but much more regulated for that with Sb(V). Notably, cytochrome c and nitrogen fixation-relevant genes for the A.f_Fe2+_Sb(III) group were enhanced significantly, indicating their role in Sb(III) resistance. This study is of great value for the development of antimony pollution control and remediation technology.

Isolation and Characterization of A Novel Iron–Sulfur Oxidizing Bacterium Acidithiobacillus Ferrooxidans YQ-N3 and its Applicability in Coal Biodesulfurization

Acidithiobacillus ferrooxidans is a chemotrophic, aerobic, acidophilic, and Gram-negative bacterium that plays a key role in iron and sulfur cycling and has a wide range of applications in the industrial field. A novel A. ferrooxidans strain, hereinafter referred to as strain “YQ-N3”, was isolated from sediments of a river polluted by acid mine drainage (AMD) of an abandoned mine in Shanxi, China. The whole genome sequencing results revealed that A. ferrooxidans YQ-N3 has a 3,217,720 bp genome, which is comprised of one circular chromosome and five circular plasmids (Plasmid A, Plasmid B, Plasmid C, Plasmid D, Plasmid E). Plasmid E, a new plasmid, had not been annotated in the reference database. A. ferrooxidans YQ-N3 had a close evolutionary relationship with A. ferrooxidans ATCC23270 and A. ferridurans JCM18981 and exhibited higher similarity in its genomic structure with A. ferrooxidans ATCC23270. Multiple genes related to environmental resistance and iron and sulfur metabolism were predicted from its genome. A. ferrooxidans YQ-N3 can remarkably increase the oxidation rate of Fe2+ and S0 and enhance the hydrophilicity of S0, which was supported by functional gene analysis and laboratory experiments. The biological desulfurization experiment demonstrated that A. ferrooxidans YQ-N3 can reduce the sulfur content in coal by removing pyrite sulfur and organic sulfur.

Application of Acidithiobacillus ferrooxidans VKM B-3655 for bioleaching silicate ore

Acid metal bioleaching is common and classical for nickel recovery from the sulfide refractory ores: various microorganisms can oxidize sulfides as energetic substrates. Silicate nickel ores are widespread in the world but their bioleaching is more problematic because silicates cannot serve as energetic substrates. Meanwhile iron in the silicate nickel ores presents a significant part and can be used by some acidophilic autotrophic microorganisms for the ore destruction. In model experiments, we studied application of acidophilic autotrophic sulfur-/ iron-oxidizing bacteria Acidithiobacillus ferrooxidans VKM B-3655 for the nickel recovery from the nickel-bearing silicate ore with high content of iron. The strain was selected by its ability of iron oxidation and resistance to arsenic which also presented in the ore. We also evaluated possibility to stimulate the bioleaching with formate as additional energetic substrates or with persulfate for increasing the medium redox. It was shown that low concentrations of sodium formate (0.3%) and persulfate (0.1%) stimulated growth of A. ferrooxidans while higher persulfate concentration (1.0%) stimulated the ore bioleaching.

Characterization and genomic analysis of two novel psychrotolerant Acidithiobacillus ferrooxidans strains from polar and subpolar environments.

The bioleaching process is carried out by aerobic acidophilic iron-oxidizing bacteria that are mainly mesophilic or moderately thermophilic. However, many mining sites are located in areas where the mean temperature is lower than the optimal growth temperature of these microorganisms. In this work, we report the obtaining and characterization of two psychrotolerant bioleaching bacterial strains from low-temperature sites that included an abandoned mine site in Chilean Patagonia (PG05) and an acid rock drainage in Marian Cove, King George Island in Antarctic (MC2.2). The PG05 and MC2.2 strains showed significant iron-oxidation activity and grew optimally at 20°C. Genome sequence analyses showed chromosomes of 2.76 and 2.84 Mbp for PG05 and MC2.2, respectively, and an average nucleotide identity estimation indicated that both strains clustered with the acidophilic iron-oxidizing bacterium Acidithiobacillus ferrooxidans. The Patagonian PG05 strain had a high content of genes coding for tolerance to metals such as lead, zinc, and copper. Concordantly, electron microscopy revealed the intracellular presence of polyphosphate-like granules, likely involved in tolerance to metals and other stress conditions. The Antarctic MC2.2 strain showed a high dosage of genes for mercury resistance and low temperature adaptation. This report of cold-adapted cultures of the At. ferrooxidans species opens novel perspectives to satisfy the current challenges of the metal bioleaching industry.

Attempts to Stimulate Leaching Activity of Acidithiobacillus ferrooxidans Strain TFBk

Autotrophic acidophilic bacteria Acidithiobacillus ferrooxidans is a model species for studying metal bioleaching from low-grade sulfide ores and concentrates. Arsenopyrite gold-bearing concentrates are refractory and often processed using biohydrometallurgical approaches; therefore, it is important to develop methods to improve arsenopyrite bioleaching. In the present work, we have studied the possibility of improving arsenopyrite concentrate bioleaching by the strain of A. ferrooxidans. For this purpose, we have analyzed the genome of the strain A. ferrooxidans TFBk to reveal the genes potentially important in the bioleaching process. Genes determining resistance to arsenic, as well genes involved in the utilization of C1-compounds and resistance to oxidative stress, were revealed. Therefore, the possibility of increasing the rate of arsenopyrite concentrate bioleaching using C1-compounds (methanol and formate) was studied. Formate was able to increase both the biomass yield of the strain A. ferrooxidans TFBk as well as the bioleaching rate. In addition, the effect of redox potential increase by means of the addition of sodium persulfate in the medium on arsenopyrite concentrate bioleaching was studied. It was shown that the addition of 0.1% sodium persulfate stimulated strain growth, while a higher concentration inhibited it. Despite this, the rate of concentrate bioleaching increased in the presence of 0.5–1.0% of persulfate, which may be explained by the interactions of added oxidizer with concentrate components.

Potential and whole-genome sequence-based mechanism of elongated-prismatic magnetite magnetosome formation in Acidithiobacillus ferrooxidans BYM.

A magnetosome-producing bacterium Acidithiobacillus ferrooxidans BYM (At. ferrooxidans BYM) was isolated and magnetically screened. The magnetosome yield from 0.5896 to 13.1291mg/g was achieved under different aeration rates, ferrous sulfate, ammonium sulfate, and gluconic acid concentrations at 30℃. TEM observed 6-9 magnetosomes in size of 20-80nm irregularly dispersed in a cell. STEM-EDXS and HRTEM-FFT implied that the elongated-prismatic magnetite magnetosomes with {110} crystal faces grown along the [111] direction. Whole-genome sequencing and annotation of BYM showed that 3.2Mb chromosome and 47.11kb plasmid coexisted, and 322 genes associated with iron metabolism were discovered. Ten genes shared high similarity with magnetosome genes were predicted, providing sufficient evidence for the magnetosome-producing potential of BYM. Accordingly, we first proposed a hypothetic model of magnetosome formation including vesicle formation, iron uptake and mineralization, and magnetite crystal maturation in At. ferrooxidans. These indicated that At. ferrooxidans BYM would be used as a commercial magnetosome-producing microorganism.

Evaluation of Cyc1 protein stability in Acidithiobacillus ferrooxidans bacterium after E121D mutation by molecular dynamics simulation to improve electron transfer.

Cyc1 (Cytochrome c552) is a protein in the electron transport chain of the Acidithiobacillus ferrooxidans (Af) bacteria which obtain their energy from oxidation Fe2+ to Fe3+. The electrons are directed through Cyc2, RCY (rusticyanin), Cyc1 and Cox aa3 proteins to O2. Cyc1 protein consists of two chains, A and B. In the present study, a novel mutation (E121D) in the A chain of Cyc1 protein was selected due to electron receiving from Histidine 143 of RCY. Then, the changes performed in the E121D mutant were evaluated by MD simulations analyzes. Cyc1 and RCY proteins were docked by a Patchdock server. By E121D mutation, the connection between Zn 1388 of chain B and aspartate 121 of chain A weaken. Asp 121 gets farther from Zn 1388. Therefore, the aspartate gets closer to Cu 1156 of the RCY leading to the higher stability of the RCY/Cyc1 complex. Further, an acidic residue (Glu121) becomes a more acidic residue (Asp121) and improves the electron transfer to Cyc1 protein. The results of RMSF analysis showed further ligand flexibility in mutation. This leads to fluctuation of the active site and increases redox potential at the mutation point and the speed of electron transfer. This study also predicts that in all respiratory chain proteins, electrons probably enter the first active site via glutamate and exit histidine in the second active site of each respiratory chain protein.

Development of composite bactericides for controlling acidification pollution in coal gangue piles and their mechanisms of bacterial inhibition

Acidification pollution from coal gangue is one of the main pollution sources in coal mining areas, and it is produced by the catalytic oxidation of sulfide in coal gangue by oxidizing bacteria. Inhibiting the activity of oxidizing bacteria is an effective method to control the acidic pollution from coal gangue. The bacterium Acidithiobacillus ferrooxidans was are considered to be the main microorganisms causing the oxidation of coal gangue. Three bactericides, sodium dodecyl sulfate (SDS), Kathon and triclosan, were selected as building blocks for composite bactericides. Our findings indicated that (1) the bacteriostatic effects of composite bactericides were better than those of single bactericides and reduced the oxidation rate of Fe2＋ to less than 7%, (2) the best formulation of the composite bactericides was 10 mg L−1 SDS mixed with 30 mg L−1 Kathon at a 1:1 ratio or 10 mg L−1 SDS mixed with 16 mg L−1 triclosan at a 1:5 ratio, and (3) there was a difference in the synergistic mechanisms in the two composite bactericides. By destroying the cell structure and DNA, the composite bactericides caused irreversible damage to A. ferrooxidans, depriving oxidizing bacteria of the ability to recover their activity.

Building efficient biocathodes with Acidithiobacillus ferrooxidans for the high current generation

The development of biocathodes is highly fascinating in microbial electrochemical technologies research. In this study, iron-oxidizing bacterium Acidithiobacillus ferrooxidans -based biocathodes were developed under the constant polarization of the electrochemical reactors at −0.2 V vs. Ag/AgCl with a pH of 2. On the 15 th day of the 21-day batch experiment, A. ferrooxidans -based biocathode produced a maximum current density of −38.61 ± 13.16 A m −2 when the reactors were supplemented with 125 mM Fe 2+ ions as an electron donor and 9 mM citrate as an iron chelator to buffer the iron-rich medium. Oxidation of Fe 2+ to Fe 3+ by A. ferrooxidans and its electrochemical regeneration at the cathode were mainly responsible for the high current generation. Furthermore, in the presence of iron, A. ferrooxidans develop a multi-layer biofilm on the cathode surface, which could potentially perform an indirect electron transfer mechanism. • Highly improved Acidithiobacillus ferrooxidans -based biocathodes were developed. • Biocathodes had shown the highest current density of −38.61 ± 13.16 A m −2 . • Fe 2+ /Fe 3+ redox transformation is mainly responsible for the highest current densities. • A. ferrooxidans develop a multi-layer biofilm in the presence of iron. • A. ferrooxidans could perform both direct and indirect electron transfer mechanisms.

Potential of Acidithiobacillus ferrooxidans to Grow on and Bioleach Metals from Mars and Lunar Regolith Simulants under Simulated Microgravity Conditions.

The biomining microbes which extract metals from ores that have been applied in mining processes worldwide hold potential for harnessing space resources. Their cell growth and ability to extract metals from extraterrestrial minerals under microgravity environments, however, remains largely unknown. The present study used the model biomining bacterium Acidithiobacillus ferrooxidans to extract metals from lunar and Martian regolith simulants cultivated in a rotating clinostat with matched controls grown under the influence of terrestrial gravity. Analyses included assessments of final cell count, size, morphology, and soluble metal concentrations. Under Earth gravity, with the addition of Fe3+ and H2/CO2, A. ferrooxidans grew in the presence of regolith simulants to a final cell density comparable to controls without regoliths. The simulated microgravity appeared to enable cells to grow to a higher cell density in the presence of lunar regolith simulants. Clinostat cultures of A. ferrooxidans solubilised higher amounts of Si, Mn and Mg from lunar and Martian regolith simulants than abiotic controls. Electron microscopy observations revealed that microgravity stimulated the biosynthesis of intracellular nanoparticles (most likely magnetite) in anaerobically grown A. ferrooxidans cells. These results suggested that A. ferrooxidans has the potential for metal bioleaching and the production of useful nanoparticles in space.