嗜酸氧化亚铁硫杆菌生长动力学方程的应用

基于Monod模型推导出了A.f的生长动力学方程模型,采用Gauss-Newton算法确定了在不同初始条件下细菌生长的动力学参数,即最大比生长速率μm、Monod常数K及R0。通过在不同初始条件下细菌生长特性的研究,得到了相应初始生长条件下以限制性底物亚铁离子浓度为表征的生长动力学方程,理论上揭示了动力学参数变化对细菌生长的影响规律,其中生长动力学方程的数值模拟与实验数据相吻合。

低温等离子体诱变耐高温氧化亚铁硫杆菌的试验研究

为培育出耐高温的氧化亚铁硫杆菌(Thiobacillus ferrooxidans,简称T.f菌),通过低温等离子体诱变试验探讨T.f菌高温诱变的可行性,研究结果表明:低温等离子诱变过后的T.f菌经PCR鉴定确认为氧化亚铁硫杆;T.f菌生长规律符合在有限环境容量下微生物的"S"型生长曲线;低温等离子的诱变可以提高T.f菌对高温的耐受性,但诱变功率60、70 W的诱变效果无明显差异;T.f菌活性均随温度的升高而减小,诱变过后的菌株也无法逆转这一生长规律;35、38、42℃三个诱变温度相比较,38℃诱变效果最好,诱变组Fe^(2+)氧化率比对照组高13%。

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.

Enhancement of bio-oxidation of refractory arsenopyritic gold ore by adding pyrolusite in bioleaching system

Pyrolusite was added in the bioleaching process to enhance the bio-oxidation process. Bioleaching tests at different dosages of pyrolusite ore, pH and inoculation amounts of Acidithiobacillus ferrooxidans were studied. The results showed that the time of the bio-oxidation process was decreased obviously and the arsenic leaching rate reached 94.4% after the bioleaching. The bio-oxidation of arsenopyrite and the effective extraction of manganese from pyrolusite were achieved by the bioleaching process. After bioleaching, the leaching rate of gold from the reaction residues reached 95.8% by cyanide leaching. In the bio-oxidation process, pyrolusite increased the redox potential of the solution to accelerate the bioleaching rate. The experiment showed that there were two reaction modes in the bioleaching process.

Decomposition mechanism of pentlandite during electrochemical bio-oxidation process

Electrochemical measurements were carried out to elucidate decomposition mechanism of pentlandite using modified powder microelectrode with Acidithiobacillus ferrooxidans attached or without on the mineral powder surface.Cyclic voltammetry（CV） results show that at a low potential of about-0.2 V（vs SCE）,the pentlandite was transformed to an intermediated phase like Fe4.5-yNi4.5-xS8-z when Fe and Ni ions were evacuated from mineral lattice;when the potential was changed from-0.2 V to 0.2 V,the unstable violarite（Fe3Ni3S4） and FeNi2S4 were formed which was accompanied by element sulfur formed on the mineral surface;when the potential increased over 0.2 V,the unstable intermediated phase decomposed entirely;at a higher potential of 0.7 V,the evacuated ferrous ion was oxidized to ferric ion.The presence of Acidithiobacillus ferrooxidans made the oxidation peak current increase with initial peak potential negatively moving,and the bacteria also contributed to the sulfur removing from mineral surface,which was demonstrated by the reduction characteristic at potential ranging from-0.75 to-0.5 V.Leaching experiments and electrochemical results show that the solution acidity increasing when pH2 may impede the oxidation process slightly.

Acidthiobacillus ferrooxidans中磁小体的提取(英文)

At.f和趋磁细菌在生理特性和生长环境有一定的相似性,而且镜检发现At.f具有趋磁性,所以本文采用了趋磁细菌中磁小体的提取方法尝试提取At.f中的磁小体,用超声波破碎At.f后,以磁铁吸取其体内的磁性颗粒,经过检测,发现其体内确实存在含铁元素的磁性颗粒。提取粗样品经过电镜分析,证实其体内存在着少量由脂质包裹的磁小体。磁小体悬浮液经过蔗糖密度梯度离心纯化后,对其作透射电镜,可以清晰的看到磁小体。实验结果表明,At.f体内存在少量的磁小体,正是由于磁小体的存在,才使得At.f在外加磁场作用下发生磁生物效应。这是首次发现从酸性矿坑水分离的At.f具有趋磁性,并从中提取到了磁小体,可以利用At.f的趋磁性将其按照不同磁性进行分离,从而获得活性高的、对不同磁性矿物有特异性的高效浸矿菌种。

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.

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.

Catalytic effect of visible light and Cd^2+on chalcopyrite bioleaching

The effects of visible light and Cd^2+ion on chalcopyrite bioleaching in the presence of Acidithiobacillus ferrooxidans(A.ferrooxidans)were studied by scanning electron microscopy(SEM),synchrotron radiation X-ray diffraction(SR-XRD),and X-ray photoelectron spectroscopy(XPS).The results of bioleaching after 28 days showed that the copper dissolution increased by 4.96%with only visible light,the presence of Cd2+alone exerted slight inhibition effect on chalcopyrite dissolution and the concentration of dissolved copper increased by 14.70%with visible light and 50 mg/L Cd^2+.The results of chemical leaching showed that visible light can promote the circulation of iron.SEM results showed that Cd^2+promoted the attachment of A.ferrooxidans on chalcopyrite surface under visible light.SR-XRD and XPS results indicated that visible light and Cd^2+promoted chalcopyrite dissolution,but did not inhibit the formation of passivation.Finally,a model of synergistic catalysis mechanism of visible light and Cd2+on chalcopyrite bioleaching was proposed.

Photogenerated-hole scavenger for enhancing photocatalytic chalcopyrite bioleaching

The effects of photogenerated-hole scavengers(ascorbic acid,oxalic acid,humic acid and citric acid)on chalcopyrite bioleaching in the presence of visible light were studied using Acidithiobacillus ferrooxidans(A.ferrooxidans).Four sets of bioleaching experiments were performed:(1)visible light+0 g/L scavenger,(2)visible light+0.1 g/L of different scavenger(ascorbic acid,oxalic acid,humic acid and citric acid),(3)dark+0.1 g/L of different scavenger(ascorbic acid,oxalic acid,humic acid and citric acid),and(4)dark+0 g/L scavenger(control group).The results showed that ascorbic acid and oxalic acid could act as photogenerated-hole scavengers and significantly enhance chalcopyrite bioleaching under visible light.The dissolved copper in the light group without scavenger was only 18.7%higher than that of the control group.The copper extraction rates of the light groups with oxalic acid and ascorbic acid were respectively 30.1%and 32.5%higher than those of the control group.Scanning electron microscopy(SEM),X-ray diffraction(XRD)and Fourier transform infrared spectroscopy(FT-IR)analyses indicated that ascorbic acid and oxalic acid as photogenerated-hole scavenger could capture photo-generated holes and inhibit jarosite formation on the chalcopyrite surface,thereby enhancing bioleaching of chalcopyrite under visible light.

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.

Bioleaching of chalcopyrite by UV-induced mutagenized Acidiphilium cryptum and Acidithiobacillus ferrooxidans

The original strains Acidithiobacillusferrooxidans GF and Acidiphilium cryptum DXI-1 were isolated from the drainage of some caves riched in chalcopyrite in Dexing Mine in Jiangxi Province of China. The optimum temperature and pH for growth were 30 ℃ and 3.5 for Ac. cryptum DXI-1, and 30 ℃ and 2.0 for At. ferrooxidans GF, respectively. For Ac. cryptum DXI-1, the optimum UV radiating time was 60 s and the positive mutation rate was 22.5%. The growth curves show that Ac. cryptum after mutagenesis reached stationary phase within 60 h, which was 20 h earlier than the original strain. For At. ferrooxidans GF, the optimum mutation time was 60 s and the positive mutation rate was 35%. The most active UV-mutated strain At. ferrooxidans GF oxidized all the ferrous after 48 h. The bioleaching experiments showed that bioleaching with the mixture of UV-mutated strains of At. ferrooxidans GF and A c. cryptum DX1-1 （1：1） could extract 3.01 g/L of copper after 30 d, while the extracted copper was 2.63 g/L with the mixture of the original strains before UV-mutation. At the end of the bioleaching experiments, the proportion of the cell density in the cultures ofAc. cryptum DXI-1 andAt.ferrooxidans GF was approximately 1：5.

Enhancing recovery of uranium column bioleaching by process optimization and kinetic modeling

This research aimed to enhance the column bioleaching recovery of uranium ore by Acidithiobacillus ferrooxidans.Seven factors were examined for their significance on bioleaching using a Plackett-Burman factorial design.Four significant variables([Fe2+]initial,pH,aeration rate and inoculation percent)were selected for the optimization studies.The effect of these variables on uranium bioleaching was studied using a central composite design(CCD).The optimal values of the variables for the maximum uranium bioleaching recovery(90.27±0.98)%were as follows:[Fe2+]initial=2.89g/L,aeration rate420mL/min,pH1.45and inoculation6%(v/v).[Fe2+]initial was found to be the most effective parameter.The maximum uranium recovery from the predicted models was92.01%.This value was in agreement with the actual experimental value.The analysis of bioleaching residue of uranium ore under optimum conditions confirmed the formation of K-jarosite on the surface of minerals.By using optimal conditions,uranium bioleaching recovery is increased at column and jarosite precipitation is minimized.The kinetic model showed that uranium recovery has a direct relation with ferric ion concentration.

Mechanism of bioleaching chalcopyrite by Acidithiobacillus ferrooxidans in agar-simulated extracellular polymeric substances media

The mechanism of leaching chalcopyrite by Acidithiobacillus ferrooxidans （,4. ferrooxidans） in agar-simulated extracellular polymeric substances （EPS） media was investigated. The results indicate that bacterial EPS can release H＋ and concentrate Fe3＋; Fe2＋ is movable between agar-simulated EPS phase and bulk solution phase, but it is difficult for Fe3＋ to move due to its hydroxylation and EPS complex action; A. ferrooxidans first prefer Fe2＋ as energy to metabolize compared with chalcopyrite, and a suitable simulated EPS environment for bacterial living is at about pH 1.8; the iron precipitates and jarosites formed by a lot of biologically oxidized Fe3 cover the simulated EPS easily and form an impermeable deposit acting as a limited barrier of ion transport that attenuates the aggressiveness of the bioleaching attack. The EPS layer blocked by iron precipitates or jarosites is responsible for the chalcopyrite passivation.

Purification and enzymatic properties of arsenic resistance protein ArsH from heterogeneous expression in E.coli BL21

Four arsenic-resistance genes(arsB,arsC,arsH,arsR) have been discovered in Acidithiobacillus ferrooxidans.Their gene sequences have been identified and three different arsenic-resistance mechanisms have been elucidated.However,the function of the arsH gene in At.ferrooxidans remains unclear.In order to evaluate the function of the arsH gene,we cloned it and expressed it in Escherichia coli.The protein was purified and its relative molecular mass was determined by SDS-PAGE(Sodium dodecyl sulfate-polyacrylamide gel electrophoresis).The results indicated that the relative molecular mass of the purified ArsH was approximately 29 kDa.The purified protein ArsH from E.coli BL21 was a flavoprotein that oxidized in vitro NADPH with an optimal pH of 6.4.

Formation of jarosite and its effect on important ions for Acidithiobacillus ferrooxidans bacteria

The precipitation of jarosite adversely affects the bio-leaching of copper sulfides in the Sarcheshmeh heap bio-leaching process. The variables of the initial concentration of ferrous iron in the growth medium, pH, and temperature were examined in the laboratory to determine how they affect the precipitation of jarosite in the presence of Acidithiobacillus ferrooxidans bacteria. It was found that the maximum ferric precipitate occurred at a ferrous sulfate concentration of 50 g/L, a temperature of 32 ℃, and an initial pH value of 2.2. The effects of the precipitation of ferric iron on the quantities of ions that are important for A. ferrooxidans bacteria in aqueous phase, i.e., ferric, sulfate, potassium, phosphate, and magnesium ions, also were assessed. The results showed relatively similar patterns for the ferric and potassium ions, and then reason might have been the co-precipitation of these ions as constituent elements of jarosite mineral. At pH values greater than 1.6, the solubility of phosphate ions decreased dramatically due to the co-precipitation of phosphate ions with the jarosite precipitate and due to the significant growth rate of A. ferrooxidans bacteria in this pH range. Due to the dissolution of a gangue constituent in the ore, the magnesium levels increased in the first few days of the bio-leaching process;thereafter, it decreased slightly.

Diversity of microbial community at acid mine drainages from Dachang metals-rich mine, China

Two acid mine drainage(AMD)samples TS and WK,which were from the Dachang metals-rich mine in Guangxi province,China,were studied using PCR-based cloning approach.A total of 44 operational taxonomic units(OTUs)were obtained from the two AMD samples.However,only three OTUs(GXDC-9,GXDC-19 and GXDC-50)detected in sample TS can also be observed in sample WK.Phylogenetic analysis revealed that the bacteria in the two samples fell into four putative divisions,which were Nitrospira,Alphaproteobacteria,Gamaproteobacteria,and Acidobacteria.Organisms of genuses Acidithiobacillus and Leptospirillum,which were in gamaproteobacteria class and Nitrospira family,were dominant in two samples,respectively.In sample TS,which was characterized by low pH,high sulfate,high iron,and high arsenide,two species(Acidithiobacillus ferrooxidans and Leptospirillum ferrooxidans)constituted 98.22% of the entire microbial community.Compared with sample TS,the microbial community in sample WK was more diversified according to the observation.Interestedly,the Legionella species,which was rarely observed in the low-pH environment,was detected in sample WK.This work helps us to further understand the diversity of microbial community living in extreme acid mine drainages with unique geochemistry and the tolerance capability of acidophiles to heavy metal.

Bio-decomposition of rock phosphate containing pyrites by Acidithiobacillus ferrooxidans

Leaching soluble phosphorus from rock phosphate containing pyrites by Acidithiobacillus ferrooxidans (A.f.) is feasible, and the reaction mechanism is as follows. Pyrites are oxidized by A.f. to produce H_2SO_4 and FeSO_4; the rock phosphate is decomposed by H_2SO_4, forming soluble phosphorus compounds; and Fe2+ from FeSO_4 is oxidized to Fe^3+, providing energy for the growth of A. f.. In this process, as H_2SO_4 is produced in the reaction, an acidic condition in the culture medium is formed, which benefits the growth of A. f. and aids both continuous oxidation of pyrites and leaching of soluble phosphorus from rock phosphate. The fraction of phosphorous leached can reach the largest in the presence of 1.0 g/L Fe^3+, 200 mg/L Mg^2+ and 400 mg/L NH_4^+. The optimal technological parameters on the fraction of phosphorous leached are as follows: the volume fraction of inocula of A. f., the mass ratio of pyrites to rock phosphate and the pH value are in ranges of 5%-25%, 3:1-5:1 and 1.8-2.2, respectively.