Electrochemical oxidation behavior of pyrite bioleaching by Acidthiobacillus ferrooxidans

The electrochemical oxidation behavior of pyrite in bioleaching system of Acidthiobacillusferrooxidans was investigated by cyclic voltammetry （CV）, polarization curve and electrochemical impedance spectroscopy （EIS）. The results show that in the presence or absence of A. ferrooxidans, the oxidation reaction of pyrite is divided into two steps： the first reaction step involves the oxidation of pyrite to S, and the second reaction step is the oxidation of S to SO4^2-. The oxidation mechanism of pyrite is not changed in the presence of A. ferrooxidans, but the oxidation rate of pyrite is accelerated. With the extension of reaction time of A. ferrooxidan with pyrite, the polarization current density of pyrite increases and the breakdown potential at which the passive film dissolves decreases. The impedance in the presence ofA. ferrooxidans is obviously lower than that in the absence of A. ferrooxidans, further indicating that microorganism accelerates the corrosion process of pyrite.

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.

Acidithiobacillus ferrooxidans ATCC23270与矿物吸附过程的热量变化及不同条件下代谢热的基础研究

用微量热技术研究了不同条件下Acidithiobacillus ferrooxidans ATCC23270（以下简写为A.ferrooxidans ATCC23270）与硫化矿吸附过程的热量变化以及不同细菌浓度、不同的初始pH值及不同的培养条件下细菌的代谢产热情况。研究发现,矿浆浓度0.03g/mL,细菌浓度1.7×10^8个/mL的时候细菌与矿物的吸附放热最大。不同条件培养的细菌胞外多聚物的组成不同,与矿物吸附的反应热也不相同,黄铁矿培养细菌胞外多聚物含量最高,反应热也最高,说明细菌胞外多聚物在吸附过程中起重要作用。用微量热法能够很好地反映出细菌生长代谢过程中每个微观时刻的热量变化。对于A.ferrooxidans ATCC23270,最佳的代谢产热条件为：pH值为2.0和2.3,细菌浓度为3.0×10^8个/mL。不同培养条件下的细菌的生长代谢热不同,2039＋FeSO4培养的代谢热最大、放热最快。

浸矿菌Leptospirillum ferrooxidans氧化Fe^(2+)的电化学动力学

研究Leptospirillum ferrooxidans溶液氧化还原电位φ与Fe2+氧化的内在联系,得到φ随溶液ln[c(Fe3+)/c(Fe2+)]的增加呈线性增长关系;研究Fe2+、细菌浓度、温度变化时Fe2+氧化反应速率的变化规律。在给定条件下,当溶液中Fe2+浓度较低时,随着Fe2+浓度的上升,Fe2+的氧化速率加快,而当Fe2+浓度在5 kg/m3以上时,Fe2+浓度的增加不但不会促使氧化速率加快,反而会抑制Fe2+的氧化;细菌浓度的增加,氧化速率随之上升,当细菌浓度在1.25×108cells/dm3以上时,氧化速率随细菌浓度的增加有较大幅度的增长;当温度在20~35℃范围时,温度的升高会加快细菌氧化Fe2+的速率,当温度再升高,则会抑制Fe2+的氧化。通过一系列电化学与生物化学分析,导出了Leptospirillum ferrooxidan生长动力学方程,并计算出活化能Ea、频率因子K0、表观饱和常数Km和单元附着系数Ki′等动力学参数的值;动力学模型能很好说明实验结果。

Experimental measurements of short-term adsorption of Acidithiobacillus ferrooxidans onto chalcopyrite

The influencing factors in adsorption such as adsorption time, pulp concentration, bacterial concentration, pH as well as ionic strength were investigated to explore the relationship among them and bacterial adsorption. The adsorption was a rapid process for bacterial adhesion to chalcopyrite. The extent of adsorption increased with increasing initial bacterial concentration and pulp concentration. The optimal pH for Acidithiobacillusferrooxidans adsorption onto chalcopyrite surfaces was in the range of pH 1-3. The increase of ionic strength led to decrease in bacterial adsorption, which can be well explained by electric double layer theory. The adsorption behavior appeared to be controlled by both hydrophobic and electrostatic interactions at the interface of bacteria and mineral,

海藻酸钙固定化At.ferrooxidans的研究

采用海藻酸钙固定化包埋氧化亚铁硫杆菌(Acidithiobacillus ferrooxidans,At.ferrooxidans),制备海藻酸钠-氯化钙固定化凝胶珠。以Fe2+的氧化率为主要监测指标,通过对海藻酸钠、氯化钙、硝酸钙及细胞浓度的筛选,最终确定了At.ferrooxidans固定化凝胶珠的最佳制备条件为:2.5%海藻酸钠,2%氯化钙,1.2×107cell.mL-1细胞浓度。整个固定化操作简单,凝胶珠大小均匀,机械强度高,不粘连,稳定性好,具有很好的应用价值。

A.ferrooxidans对高硫煤矸石去除酸性水体中Cr(Ⅵ)的强化作用

本研究考察了Acidithiobacillus ferrooxidans(A.ferrooxidans)联合高硫煤矸石(富含FeS2)对模拟煤矿酸性水体中Cr(Ⅵ)的去除效果.结果表明,处理Cr(Ⅵ)初始浓度为50mg/L的模拟煤矿酸性废水(pH=2.5)时,投配率为6.67~33.33g/L高硫煤矸石可使Cr(Ⅵ)去除达到良好效果.50mg/LCr(Ⅵ)在24h内即可完全被高硫煤矸石中的FeS2还原成Cr(Ⅲ),且在反应终点时(120h),6.67,13.33,33.33g/L高硫煤矸石对还原产物Cr(Ⅲ)的吸附去除率分别为7.1%、20.2%、29.1%.然而,在高硫煤矸石的还原和吸附作用下,大部分的Cr仍以Cr(Ⅲ)形式残留在酸性水体中,且高硫煤矸石的大量投加也给水体带来了Fe^2+、Fe^3+、SO4^2-等二次污染物.在高硫煤矸石-Cr(Ⅵ)体系中引入A.ferrooxidans和9K培养基后,A.ferrooxidans介导的Fe^2+生物氧化及产物Fe^3+水解矿化过程可促进部分Fe^2+、Fe^3+、SO42-等向次生铁矿物(包括施氏矿物和黄钾铁矾)转变,从而使模拟酸性水体中残留的Cr(Ⅲ)通过次生铁矿物的吸附或共沉淀作用被清除.在A.ferrooxidans强化作用下,模拟煤矿酸性废水中Cr(Ⅵ)在96h即可达到99.4%的去除率.

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。

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.

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.

Fe^(2+)浓度对Acidithiobacillus ferrooxidans耐铜性的影响

嗜酸性氧化亚铁硫杆菌(Acidithiobacillus ferrooxidans)是铜矿生物冶金中应用最广泛的微生物之一。但在冶金过程中淋滤出来的Cu2+等重金属逐渐积累,达到一定的浓度后就会抑制A.ferrooxidans的生长,从而降低冶金的效率。本文着重研究了Fe2+初始浓度对A.ferrooxidans耐铜性的影响。实验中ρ(Cu2+)变化范围为0～5.0 g/L。结果表明,当ρ0(Fe2+)为6.7 g/L时,A.ferrooxidans仅在Cu2+为0～0.4 g/L的体系中能显著地氧化Fe2+进行生长;当Cu2+≥0.5 g/L时,A.ferrooxidans生长完全受到抑制。将ρ0(Fe2+)增加到8.9 g/L,A.ferroox-idans在0.5、1.0、2.0和3.0 g/L Cu2+的培养基中也能明显氧化Fe2+,并最终将其完全氧化,Cu2+对A.ferrooxidans生长抑制作用仅出现在4.0和5.0 g/L Cu2+的体系中。因此提高体系中亚铁离子的浓度能提高菌体对Cu2+的耐受力。研究结果对铜矿的生物冶金具有重要意义。

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.

Bioleaching and electrochemical property of marmatite by Leptospirillum ferrooxidans

The effects of several variables on the bioleaching of marmatite with pure L. ferrooxidans were investigated. The results show that zinc extraction increases with the decrease of pulp density. Adjusting pH tol.6 during the bioleaching process has a positive effect to the dissolution of marmatite. External addition of Fe^3＋ ions accelerates the bioleaching, while the concentration of additional Fe^3＋ over 2.5 g/L weakens the acceleration effect due to the inhibition effect on bacteria growth and the promotion of jarosite production. The electrochemical measurements were used to make further understanding on the dissolution of marmatite with and without additional Fe^3＋ in the presence of L. ferrooxidans. The experimental data illustrate that additional Fe^3＋ ions could increase the corrosion current density, which is favorable to zinc extraction. The EIS spectra show that rate-limiting step does not change when Fe^3＋ is added.

冶金模式微生物Acidithiobacillus ferrooxidans表面质子吸附特性的研究

为了探究生物冶金过程及矿山酸性废水中的关键微生物Acidithiobacillus ferrooxidans的表面质子吸附机理,该研究通过酸碱滴定、ProtoFit模拟、电泳迁移率测定及傅里叶变换衰减全反射红外光谱法(ATR-FTIR)系统地揭示不同能源(S0、Fe2+、FeS2)和离子强度(0.001~0.1 mol/L NaNO3)对A.ferrooxidans表面性质的影响及参与质子化/去质子化反应的关键官能团。结果表明:三位点的Donnan壳静电模型能较好地适用于分析A.ferrooxidans的表面酸碱性质;能源和离子强度均影响A.ferrooxidans的表面酸碱性质;在广泛的pH值范围内(2~10),不同能源培养的A.ferrooxidans在不同离子强度下均呈负电性;羧基、磷酸基和酰胺基是质子化/去质子化过程的关键官能团。该研究表明了A.ferrooxidans在吸附重金属方面的重要应用潜力,并对阐明A.ferrooxidans的表面质子吸附机理具有重要意义。

Acidithiobacillus ferrooxidans氧化分解毒砂的次生产物研究

毒砂是常见的含砷硫化物矿物。在金属硫化物矿山环境中,含毒砂矿石和尾矿的风化会导致严重的重金属污染,在其风化过程中,微生物能够显著促进毒砂的氧化分解。本文实验研究了Acidithiobacillus ferrooxidans氧化分解毒砂矿物的现象,利用X射线衍射和扫描电子显微镜,分析了毒砂微生物氧化作用形成的次生矿物类型,发现毒砂表面存在As含量明显不同的2类次生产物,观察到黄钾铁矾、臭葱石、自然硫和施威特曼石等矿物;借助光电子能谱仪重点分析了微生物作用前后毒砂晶面的表面化学组成,基本查明了Fe、As和S三种元素的价态变化,初步探讨了毒砂表面次生矿物成因和As的化学态转化。

Pyrite Surface after Thiobacillus ferrooxidans Leaching at 30℃

In order to investigate the effect of Thiobacillusferrooxidans on the oxidation of pyrite, two parallel experiments, which employed H2SO4 solutions and acidic solutions inoculated with ThiobaciUus ferrooxidans, were designed and carried out at 30℃. The initial pH of the two solutions was adjusted to 2.5 by dropwise addition of concentrated sulphuric acid. The surfaces of pyrite before exposure to leaching solutions and after exposure to the H2SO4 solutions and acidic solutions inoculated with Thiobacillus ferrooxidans were observed by scanning electron microscopy （SEM）. There were a variety of erosion patterns by Thiobacillusferrooxidans on the bio-leached pyrite surfaces. A conclusion can be drawn that the oxidation of pyrite might have been caused by erosion of the surfaces. Attachment of the bacteria to pyrite surfaces resulted in erosion pits, leading to the oxidation of pyrite. It is possible that the direct mechanism plays the most important role in the oxidation of pyrite. The changes in iron ion concentrations of both the experimental solutions with time suggest that ThiobaciUus ferrooxidans can enhance greatly the oxidation of pyrite.

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.