Effect of EPS on adhesion of Acidithiobacillus ferrooxidans on chalcopyrite and pyrite mineral surfaces

Extracellular polymeric substances （EPS） were extracted from Acidithiobacillus ferrooxidans through sonication method associated with centrifugation, which was evaluated tentatively with 2-keto-3-deoxyoctonic acid （KDO） as the indicator of EPS by spectrophotometry. Then the effect of EPS of A. ferrooxidans on the adhesion on chalcopyrite and pyrite surfaces was studied through a series of comparative experiments. The untreated cells and EPS-free cells of A. ferrooxidans were mixed with EPS suspension, Fe^2＋ or Fe^3＋, respectively. The planktonic cells were monitored in 2 h during bioleaching. The results indicate that the presence of EPS on the cell is an important factor for the adhesion to chalcopyrite and pyrite. A decrease of attachment of A. ferrooxidans to minerals was produced by the deficiency of EPS, which can recover mostly when the EPS was re-added into the EPS-free cells. The restoring extent is more obvious in pyrite than in chalcopyrite. The extent of cell adhesion to chalcopyrite increased when EPS and Fe^3＋ added, and decreased when Fe^2＋ added, which imply the electrostatic interaction plays a main role in initial adhesion between bacteria and minerals and it is a driving force for bacteria to produce EPS probably as a result of regaining their attachment ability to copper sulphides.

Effect of surfactant Tween-80 on sulfur oxidation and expression of sulfur metabolism relevant genes of Acidithiobacillus ferrooxidans

The effects of surfactant Tween-80 on the growth, sulfur oxidation, and expression of selected typical sulfur metabolism relevant genes of Acidithiobacillus ferrooxidans ATCC 23270 were investigated. The results showed that in the presence of 10-2 g/L Tween-80, the growth of A. ferrooxidans and its metabolism on the insoluble substrate S0 and CuFeS2 were promoted. After 24 d of bioleaching, the copper extraction yield of chalcopyrite at 10-2 g/L Tween-80 increased by 16% compared with the bioleaching experiment without Tween-80. FT-IR spectra analysis revealed that the result was probably caused by the extracellular polymeric substances whose composition could be changed by the surfactant addition. RT-qPCR was used to analyze the differential expressions of 17 selected sulfur metabolism relevant genes in response to the addition of Tween-80. Down-regulation of the extracellular protein genes indicated the influence of Tween-80 on bacteria-sulfur adsorption. Variation of the expression level of the enzymes provided a supplement to sulfur metabolism investigation.

Differential utilization and speciation transformation of orthorhombic α-S_8 and amorphous μ-S by substrate-acclimated mesophilic Acidithiobacillus ferrooxidans

The utilization and speciation transformation of α-S8 and μ-S by the typical mesophilic acidophilic strain Acidithiobacillus ferrooxidans ATCC 23270 were investigated.A.ferrooxidans cells first acclimated to the energy source α-S8 or μ-S,respectively.The results of cell growth and sulfur oxidation behavior showed that the strain grown on α-S8 entered slowly（about 1 d later） into the exponential phase,while grew faster in the exponential phase and attained higher maximal cell density and lower p H value than that on μ-S.After bio-corrosion,both of the two sulfur samples were evidently eroded and modified by A.ferrooxidans cells.After growth of A.ferrooxidans,the surface composition of amorphous μ-S became 63.1% μ-S and 36.9% α-S8,and that of orthorhombic α-S8 became 68.3% α-S8 and 31.7% μ-S,while the surface compositions of α-S8 and μ-S in sterile experiment were not changed,indicating that these two elemental sulfur species can be interconverted by A.ferrooxidans.

Effects of Acidiphilium cryptum on biosolubilization of rock phosphate in the presence of Acidithiobacillus ferrooxidans

The bioleaching of pyrite and biosolubilization of rock phosphate （RP） in 9K basal salts medium were compared by the following strains of an autotrophic acidophilic bacterium, Acidithiobacillus ferrooxidans, a heterotrophic acidophilic bacterium, Acidiphilium cryptum, and mixed culture of At. ferrooxidans and A. cryptum. The results show that A. cryptum is effective in enhancing the bioleaching of pyrite and biosolubilization of RP in the presence of At. ferrooxidans, although it could not oxidize pyrite and solubilize RP by itself. This effect is demonstrated experimentally that A. cryptum enhances a decrease in pH and an increase in redox potential, concentration of total soluble iron and planktonic part bacterial number in the broth during pyrite bioleaching processes by At. ferrooxidans. The mixed culture of At. ferrooxidans and A. cryptum leads to the most extensive soluble phosphate released at 30 °C. Pulp density exceeding 3% is shown to adversely influence the release of soluble phosphate by the consortium of At. ferrooxidans and A. cryptum. It is essential to add pyrite to the 9K basal salts medium for the biosolubilization of RP by the mixed culture of At. ferrooxidans and A. cryptum, and the percentage of soluble phosphate released is the greatest when the mass ratio of RP to pyrite is 1：2 or 1：3.

Bioleaching of low-grade copper sulfide ores by Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans

The grown conditions of Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans were investigated,and then experiments were conducted to research the bioleaching behaviors of crude ore of copper sulfide and hand-picked concentrates of chalcopyrite by Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans.The experimental results show that the bacteria grow best when the temperature is(30±1) °C and the pH value is 2.0.The bacteria concentration is 2.24×107 mL-1 in this condition.It is found that the copper extraction yield is affected by the inoculum size and the pulp density and the extraction yield increases as the inoculum size grows.The bioleaching rates reach their highest point in sulfide copper and chalcopyrite with a pulp density of 5% and 10%,respectively.Column flotation experiments of low-grade copper sulfide ores show that the bioleaching recovery reaches nearly 45% after 75 days.

Preservation efficiency of new cryoprotectant used for Acidithiobacillus ferrooxidans in liquid nitrogen

The efficiency of a new cryoprotectant,GP,for the preservation of Acidithiobacillus ferrooxidans（A.ferrooxidans） strain DC in liquid nitrogen was investigated.The optimal concentration of this new cryoprotectant for the maximal viable cell recovery and the highest ferrous ion oxidation activity was determined.The results show that 30%（volume fraction） GP is optimal for the cryopreservation with 84.4% of cells surviving,completely oxidizing ferrous ions within 120 h,and growing to a final density of 5.8×107 cell/mL after 6 d in the culture.Furthermore,the optimal residual GP concentration for viable cell recovery after culture of thawed cells in 9K medium for 6 d is 0.6%（volume fraction）.At this concentration,strain DC completely oxidizes ferrous ions within 108 h and grows to a final cell density of 6.8×107 mL-1.Thus,GP is a simple,effective cryoprotectant for the preservation of A.ferrooxidans strain DC in liquid nitrogen.

固定化Acidithiobacillus ferrooxidans对Fe^(2+)氧化及其生物反应器研究进展

Acidithiobacillus ferrooxidans对Fe2+的生物氧化是一个非常重要的反应过程,在生物浸矿、H2S等废气的脱硫、含重金属污泥和酸性矿坑废水的处理等领域有着重要的应用。近些年来,大量的研究主要集中A.ferrooxidans及其反应过程等方面,然而,A.ferrooxidans对Fe2+的催化氧化速率缓慢和稳定性欠佳等问题仍然限制了其商业应用。因此,对A.ferrooxidans的固定化及其生物反应器研究是该技术进一步发展的关键。评述了A.ferrooxidans最新应用、存在的问题和解决办法,重点比较了目前文献中报道的各种A.ferrooxidans固定材料、方法,并对目前采用的各种固定化A.ferrooxidans生物反应系统的效率和结构等方面进行了讨论和分析。

Effect of pH values on extracellular protein and polysaccharide secretions of Acidithiobacillus ferrooxidans during chalcopyrite bioleaching

The effect of pH values on the extracellular protein and polysaccharide secretions of Acidithiobacillus ferrooxidans was comparatively investigated in different phases of bacterial growth during chalcopyrite bioleaching. The results indicate that the extracellular protein is always more than the extracellular polysaccharide secreted by attached cells on the chalcopyrite, on the contrary, and is always less than the extracellular polysaccharide secreted by free cells in the solution at bacterial adaptive phase, logarithmic phase and stationary phase whenever pH value is at 1.0, 1.5, 2.0 or 2.5; free cells are mainly through the secretion of extracellular polysaccharide rather than the extracellular protein to fight against disadvantageous solution environment, such as high concentration of metal ions and unsuitable pH solution; both amounts of polysaccharide and protein secreted by attached cells are mainly positively related to the solution acidity rather than the total concentration of soluble metal ions. The experimental results imply that bacteria are mainly through secreting more extracellular polysaccharide to fight against disadvantageous environment and the extracellular protein perhaps plays an important role in oxidation？reduction reactions in the bioleaching system.

Comparison of Fe^(2+) oxidation by Acidithiobacillus ferrooxidans in rotating-drum and stirred-tank reactors

Fe2＋ oxidation by Acidithiobacillus ferrooxidans（At.ferrooxidans） under different solid contents by adding inert Al2O3 powder was examined in rotating-drum and stirred-tank reactors.The results show that the bioactivity of At.ferrooxidans in the stirred-tank is higher than that in the rotating-drum in the absence of Al2O3 powder,but the biooxidation rate of Fe2＋ decreases markedly from 0.23 g/（L·h） to 0.025 g/（L·h） with increasing the content of Al2O3 powder from 0 to 50%（mass fraction） in the stirred-tank probably due to the deactivation of At.ferrooxidans resulting from the collision and friction of solid particles.The increase in Al2O3 content has a little adverse effect on the bioactivity of At.ferrooxidans in the rotating-drum due to different mixing mechanisms of the two reactors.The biooxidation rate of Fe2＋ in the rotating-drum is higher than that in the stirred-tank at the same content of Al2O3 powder,especially at high solid content.The higher bioactivity of At.ferrooxidans can be maintained for allowing high solid content in the rotating-drum reactor,but its application potential still needs to be verified further by the sulfide bioleaching for the property differences of Al2O3 powder and sulfide minerals.

Really active form of fluorine toxicity affecting Acidithiobacillus ferrooxidans activity in bioleaching uranium

In order to determine the mechanism of bacterial tolerance to fluorine,Acidithiobacillus ferrooxidans ATCC 23270 was domesticated and studied under the conditions of different fluorine concentrations and pH values with or without treatment by Proteinase K.The bacterial activities were observed through measuring the changes of solution potentials by platinum electrode with Ag/AgCl reference electrode and the intracellular fluorine was determined by-uorine ion-selective electrode.The results indicated that the tolerance of Acidithiobacillus ferrooxidans ATCC 23270 to fluorine could be obviously improved by domestication,HF was the effective form of fluorine to affect the bacterial activity,and pH increase or concentration change of ions of strong complex ability with fluorine ions in solution could result in false appearance of high fluorine-resistant strain.Some proteins located in cell wall or cell membrane were intimately relative with the bacterial fluorine tolerance.

Isolation and characterization of Acidithiobacillus caldus from several typical environments in China

Six strains of moderately thermophilic sulfur-oxidizing bacteria were isolated from several different typical environments in China. The identities of the isolates were confirmed by analyses of their 16S rRNA genes, and some key physiological traits. The isolates are Gram negative, rod-shaped bacteria, their optimal temperature and pH value for growth are 45-50℃ and 2.5-3.5 respectively. They are autotrophic and used elemental sulfur, sodium thiosulfate and potassium tetrathionate as electron donor, while a little glucose stimulated their growth. 16S rDNA sequences analysis reveals that the strains are phylogenetically clustered to Acidithiobacillus caldus.

Catalytic mechanism of manganese ions and visible light on chalcopyrite bioleaching in the presence of Acidithiobacillus ferrooxidans

The bioleaching of chalcopyrite is low cost and environmentally friendly,but the leaching rate is low.To explore the mechanism of chalcopyrite bioleaching and improve its leaching rate,the effect and mechanism of manganese ions(Mn^(2+))and visible light on chalcopyrite mediated by Acidithiobacillus ferrooxidans(A.ferrooxidans)were discussed.Bioleaching experiments showed that when both Mn^(2+)and visible light were present,the copper extraction was 14.38%higher than that of the control system(without Mn^(2+)and visible light).Moreover,visible light and Mn^(2+)promoted the growth of A.ferrooxidans.Scanning electron microscopy(SEM)and energy dispersive spectrometer(EDS)analysis revealed that Mn^(2+)promoted the formation of extracellular polymeric substance(EPS)on the surface of chalcopyrite,changed the morphology of A.ferrooxidans,enhanced the adsorption of bacteria on chalcopyrite surface with light illumination,and thus promoted the bioleaching of chalcopyrite.UV–vis absorbance spectra indicated that Mn^(2+)promoted the response of chalcopyrite to visible light and enhanced the catalytic effect of visible light on chalcopyrite bioleaching.Based on X-ray photoelectron spectroscopy(XPS),the relevant sulfur speciation of chalcopyrite before and after bioleaching were analyzed and the results revealed that visible light and Mn^(2+)promoted chalcopyrite bioleaching by reducing the formation of passivation layer(S_(n)^(2-)/S0).Investigation into electrochemical results further indicated that Mn^(2+)and visible light improved the electrochemical activity of chalcopyrite,thus increasing the bioleaching rate.

Cu^2+胁迫对Acidithiobacillus ferrooxidans亚铁氧化代谢及胞外电子传递影响

为了查明Cu^2+胁迫影响Acidithiobacillus ferrooxidans(At.ferrooxidans)亚铁氧化代谢机制,本研究设计了由低到高Cu^2+浓度(0~25 g/L)胁迫环境,利用邻菲啰啉分光光度法、实时荧光定量PCR和循环伏安法等分析手段分别探究了不同Cu^2+浓度胁迫下At.ferrooxidans亚铁氧化率、亚铁氧化电子传递链rus operon相关基因表达及胞外电子传递的响应情况。结果表明:Cu^2+胁迫浓度由0 g/L增加至25 g/L时,At.ferrooxidans菌亚铁氧化活性显著降低(25 g/L Cu^2+,对应下24 h亚铁氧化抑制率约为50%),rus operon相关基因表达显著下调(尤其是细胞内膜蛋白编码基因Cyc1和细胞色素c氧化酶编码基因CoxBACD分别下调12.4倍和4~6倍),Fe2+氧化胞外传递电子能力也降低(循环伏安曲线还原峰值29.0μA(对照组,0 g/L Cu^2+)和26.5μA(25 g/L Cu^2+))。综上所述,高浓度Cu^2+胁迫引起At.ferrooxidans菌亚铁氧化代谢Rus operon相关基因表达显著下调,造成胞外电子传递速率降低,最终导致At.ferrooxidans菌亚铁氧化率降低。

Microbiologically influenced corrosion of cable bolts in underground coal mines:The effect of Acidithiobacillus ferrooxidans

Reports on corrosion failure of cable bolts,used in mining and civil industries,have been increasing in the past two decades.The previous studies found that pitting corrosion on the surface of a cable bolt can initiate premature failure of the bolt.In this study,the role of Acidithiobacillus ferrooxidans(A.ferrooxidans)bacterium in the occurrence of pitting corrosion in cable bolts was studied.Stressed coupons,made from the wires of cable bolts,were immersed in testing bottles containing groundwater collected from an underground coal mine and a mixture of A.ferrooxidans and geomaterials.It was observed that A.ferrooxidans caused pitting corrosion on the surface of cable bolts in the near-neutral environment.The presence of geomaterials slightly affected the p H of the environment;however,it did not have any significant influence on the corrosion activity of A.ferrooxidans.This study suggests that the common bacterium A.ferrooxidans found in many underground environments can be a threat to cable bolts'integrity by creating initiation points for other catastrophic failures such as stress corrosion cracking.

Acidithiobacillus ferrooxidans enhanced heavy metals immobilization efficiency in acidic aqueous system through bio-mediated coprecipitation

This study investigated the promotion effect of A.ferrooxidans on complex heavy metals coprecipitation process.A.ferrooxidans significantly enhanced the ferrous oxidation,which also promoted the formation of iron-oxyhydroxysulphate.Cu(II)concentration reduced to0.058mmol/L in A.ferrooxidans inoculated system,and Cd also reduced to the lowest concentration(0.085mmol/L).Pb was mainly immobilized as anglesite and iron-oxyhydroxysulphate promoted the removal of remanent Pb in solution.The precipitates are characterized by XRD,SEM,and FTIR analysis.The main component of the iron-oxyhydroxysulphate was well crystallized jarosite.A.ferrooxidans contributed to the formation of schwertmannite in later monovalent cation lack stage.Higher ferrous iron oxidation rate and Fe(III)supply rate in A.ferrooxidans inoculated system facilitated polyhedron crystal formation and the increase of particle diameter.Complex heavy metals could be incorporated into iron oxyhydroxysulphate crystal,and efficiently removed from acidic wastewater through A.ferrooxidans mediated coprecipitation.

Surface Mineralogy of Oxidized Pyrite by Acidithiobacillus Ferrooxidans

Sulfide oxidation by microbial activities play an important role in the release of heavy metals. An important source of contamination and formation of AMD is the heavy metals convey to soil, rivulet and groundwater. Pyrite is a commonly sulfide minerals in mine wastes, so it is vitally to prove up the microbial oxidation process.

Effect of iron transformation on Acidithiobacillus ferrooxidans bio-leaching of clay vanadium residue

The acid bio-leaching process of vanadium extraction from clay vanadium water-leached residue was studied and the effect of the performance of iron transformation was investigated.Acidithiobacillus ferrooxidans affects the dissolution of vanadium through the catalytic effect on Fe^3+/Fe^2+couple and material exchange.The passivation of iron settling correlates with ferrous ion content in bio-leaching solution.In medium containing A.ferrooxidans and Fe(Ⅲ),the increment in Fe(Ⅱ)concentration leads to the formation of jarosite,generating a decline in vanadium extraction efficiency.Analysis of cyclic voltammetry shows that Fe(Ⅱ)ion is apt to be oxidized and translated into precipitate by A.ferrooxidans,which strongly adsorbed to the surface of the residue.Fe(Ⅲ)ion promotes the vanadium extraction due to its oxidizing activity.Admixing A.ferrooxidans to Fe(Ⅲ)medium elevates the reduction of low valence state vanadium and facilitates the exchange of substance between minerals and solution.This motivates 3.8%and 21.8%increments in recovery ratio and leaching rate of vanadium compared to the Fe(Ⅲ)exclusive use,respectively.Moreover,Fe(Ⅱ)ion impacts vanadium extraction slightly in sterile medium but negatively influences vanadium leaching in the presence of bacteria.

Characterization of acylated homoserine lactone derivatives and their influence on biofilms of Acidithiobacillus ferrooxidans BY-3 under arsenic stress

Quorum sensing is one kind of cell-to-cell signalling system among microorganisms that works in response to their population density via autoinducers exemplified by AHL and oligopeptides. In this study, fourteen AHL derivatives were synthesised by a chemical synthesis method, and two types of AHL derivatives were measured and screened by crystal violet staining assay, which have more obvious inhibitory effects on A. ferrooxidans biofilms under arsenic environment. Their structures were verified through IR and NMR identification. The morphological changes of A. ferrooxidans under the influence of the AHL derivatives were compared. In addition, the effects of AHL derivatives(0.1 μg/mL and 1 μg/mL) on membrane formation of A. ferrooxidans under high concentration of arsenic resistance(1,600 mg/L) were explored. Solid experimental data firstly showed that a portion of logarithmic microorganisms were ruptured under the effect of high arsenic concentration. Secondly, the volume of the cell shrank and the number of extracellular polymeric substances decreased after the addition of the AHL derivatives at high concentrations. Therefore, we found here that two derivatives used at concentrations of 0.1 μg/mL and 1 μg/m L accompanied with high concentration of arsenic can both effectively restrict biofilms formation by A. ferrooxidans.

Homology modeling and docking studies of IscS from extremophile Acidithiobacillus ferrooxidans

The gene iscS-3 from ,4cidithiobacillus ferrooxidans may play a central role in the delivery of sulfur to a variety of metabolic pathways in this organism. For insight into the sulfur metabolic mechanism of the bacteria, an integral three-dimensional （3D） molecular structure of the protein encoded by this gene was built by homology modeling techniques, refined by molecular dynamics simulations, assessed by PROFILE-3D and PROSTAT programs and further used to search bind sites, carry out flexible docking with cofactor pyridoxal 5＇-phosphate（PLP） and substrate cysteine and hereby detect its key residues. Through these procedures, the detail conformations of PLP-IscS（P-I） and cysteine-PLP-IscS（C-P-I） complexes were obtained. In P-I complex, the residues of Lys208, His106, Thr78, Ser205, His207, Asp182 and Gln185 have large interaction energies and/or hydrogen bonds fixation with PLP. In C-P-I complex, the amino group in cysteine is very near His106, Lys208 and PLP, the interaction energies for cysteine with them are very high. The above results are well consistent with those experimental facts of the homologues from other sources. Interestingly, the four residues of Glul05, Glu79, Ser203 and Hisl80 in P-I docking and the residue of Lys213 in C-P-I docking also have great interaction energies, which are fitly conservation in IscSs from all kinds of sources but have not been identified before. From these results, this gene can be confirmed at 3D level to encode the iron-sulfur cluster assembly protein IscS and subsequently play a sulfur traffic role. Furthermore, the substrate cysteine can be presumed to be effectively recruited into the active site. Finally, the above detected key residues can be conjectured to be directly responsible for the bind and/or catalysis of PLP and cysteine.

Homology modeling and evolutionary trace analysis of superoxide dismutase from extremophile Acidithiobacillus ferrooxidans

The gene sod in Acidithiobacillusferrooxidans may play a crucial role in its tolerance to the extremely acidic, toxic and oxidative environment of bioleaching. For insight into the anti-toxic mechanism of the bacteria, a three-dimensional （3D） molecular structure of the protein encoded by this gene was built by homology modeling techniques, refined by molecular dynamics simulations, assessed by PROFILE-3D and PROSTAT programs and its key residues were further detected by evolutionary trace analysis. Through these procedures, some trace residues were identified and spatially clustered. Among them, the residues of Asn38, Glyl03 and Glul61 are randomly scattered throughout the mapped structure; interestingly, the other residues are all distinctly clustered in a subgroup near Fe atom. From these results, this gene can be confirmed at 3D level to encode the Fe-depending superoxide dismutase and subsequently play an anti-toxic role. Furthermore, the detected key residues around Fe binding site can be conjectured to be directly responsible for Fe binding and catalytic function.