The role of exopolymers in the bioleaching of a non-ferrous metal sulphide

Exocellular polysaccharides were extracted from Thiobacillus ferrooxidans cells grown in the presence of iron. Cells without these compounds could not adhere to covellite. The loss of the layer of exocellular polysaccharides also affected the direct mechanism of bioleaching of covellite in a negative way. This ability to attach to and leach covellite was restored within a few hours when exopolymeric material was produced again. The addition of exocellular compounds to cells stripped of exocellular polymers also restored their ability to the same level as that of untreated cells. Thiobacillus thiooxidans was not able to attach to and leach covellite even when exocellular compounds from Thiobacillus ferrooxidans were added. Received 2 June 1998/ Accepted in revised form 8 January 1999     

Effect of particle-particle shearing on the bioleaching of sulfide minerals

The biological leaching of sulfide minerals, used for the production of gold, copper, zinc, cobalt, and other metals, is very often carried out in slurry bioreactors, where the shearing between sulfide particles is intensive. In order to be able to improve the efficiency of the bioleaching, it is of significant importance to know the effect of particle shearing on the rate of leaching. The recently proposed concept of ore immobilization allowed us to study the effect of particle shearing on the rate of sulfide (pyrite) leaching by Thiobacillus ferrooxidans. Using this concept, we designed two very similar bioreactors, the main difference between which was the presence and absence of particle-particle shearing. It was shown that when the oxygen mass transfer was not the rate-limiting step, the rate of bioleaching in the frictionless bioreactor was 2.5 times higher than that in a bioreactor with particle friction (shearing). The concentration of free suspended cells in the frictionless bioreactor was by orders of magnitude lower than that in the frictional bioreactor, which showed that particle friction strongly reduces the microbial attachment to sulfide surface, which, in turn, reduces the rate of bioleaching. Surprisingly, it was found that formation of a layer of insoluble iron salts on the surface of sulfide particles is much slower under shearless conditions than in the presence of particle-particle shearing. This was explained by the effect of particle friction on liquid-solid mass transfer rate. The results of this study show that reduction of the particle friction during bioleaching of sulfide minerals can bring important advantages not only by increasing significantly the bioleaching rate, but also by increasing the rate of gas-liquid oxygen mass transfer, reducing the formation of iron precipitates and reducing the energy consumption. One of the efficient methods for reduction of particle friction is ore immobilization in a porous matrix.     

Bioleaching of zinc sulfide concentrate by Thiobacillus ferrooxidans

The kinetics of the bioleaching of ZnS concentrate by Thiobacillus ferrooxidans was studied in a well-mixed batch reactor. Experimental studies were made at 30 degrees C and pH 2.2 on adsorption of the bacteria to the mineral, ferric iron leaching, and bacterial leaching. The adsorption rate of the bacteria was fairly rapid in comparison with the bioleaching rate, indicating that the bacterial adsorption is at equilibrium during the leaching process. The adsorption equilibrium data were correlated by the Langmuir isotherm, which is a useful means for predicting the number of bacteria adsorbed on the mineral surface. The rate of chemical leaching varied with the concentration of ferric iron, and the first-order reaction rate constant was determined. Bioleaching in an iron-containing medium was found to take place by both direct bacterial attack on the sulfide mineral and indirect attack via ferric iron. In this case, the ferric iron was formed from the reaction product (ferrous iron) through the biological oxidation reaction. To develop rate expressions for the kinetics of bacterial growth and zinc leaching, the two bacterial actions were considered. The key parameters appearing in the rate equations, the growth yield and specific growth rate of adsorbed bacteria, were evaluated by curve fitting using the experimental data. This kinetic model allowed us to predict the liquid-phase concentrations of the leached zinc and free cells during the batch bioleaching process.     

Modeling continuous bioleach reactors

The results of recent research have shown that the bioleaching of sulfide minerals occurs via a two‐step mechanism. In this mechanism, the sulfide mineral is chemically oxidized by the ferric‐iron in the bioleaching liquor. The ferrous‐iron produced is subsequently oxidized to ferric‐iron by the microorganism. Further research has shown that the rates of both the ferric leaching and ferrous‐iron oxidation are governed by the ferric/ferrous‐iron ratio (i.e., the redox potential). During the steady‐state operation of a bioleach reactor, the rate of iron turnover between the chemical ferric leaching of the mineral and the bacterial oxidation of the ferrous‐iron will define the rate and the redox potential at which the system will operate. The balance between the two rates will in turn depend on the species used, the microbial concentration, the residence time employed, the nature of the sulfide mineral being leached, and its active surface area. The model described proposes that the residence time and microbial species present determine the microbial growth rate, which in turn determines the redox potential in the bioleach liquor. The redox potential of the solution, in turn, determines the degree of leaching of the mineral; that is, conversion in the bioleach reactor. © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 64: 671–677, 1999.     

Thermophilic vs. mesophilic bioleaching process performance

Abstract: Mesophilic and thermophilic mineral oxidising microbial cultures were compared for their capacity to leach both a complex pyritic and an arsenopyritic ore, aiming at copper and gold recovery, respectively. The mesophilic cultures are primarily based on the activity of Thiobacillus -type microorganisms, while the thermophile characteristics place them in the sulphur metabolizers branch df Archaebacteria. Study of key process variables and determination of their optimum values were carried out to provide a basis for significant physiological and technical comparison. Each microbial culture finds a preferential process application, depending on the choice of the reactor and ore.     

Quantification of the intragranular porosity formed in bioleaching of pyrite by Thiobacillus ferroxidans

During the bacterial oxidation of a pure pyrite by Thiobacillus ferrooxidans, a great number of corrosion tunnels appear that are easily revealed by scanning electron microscopy observations. This involves an increase in the surface area without significant granulometric reduction of mineral grains. Thus, the evaluation of intragranular porosity, determined by elution front analysis, allows one to estimate accurately the fraction of oxidized sulphide, because of the development of deep holes (propagating pore mechanism). After 60 days of bioleaching, the intragranular porosity represents about 34% of the initial sulphide volume, which corresponds to 25 km of tunnels (2 mum i.d.) per gram of pyrite. On other hand, the granulometric reduction ( approximately 7%) is responsible for a 23% decrease of the initial sulphide volume. The elution front analysis appears as a nondestructive method for measuring the intragranular porosity of the bioleached pyrite.     

Effect of carbon dioxide concentration on the bioleaching of a pyrite-arsenopyrite ore concentrate

The effect of carbon dioxide concentration on the bacterial leaching of a pyrite-arsenopyrite ore concentrate was studied in continuous-flow reactors. Steady-state operation with two feed slurry densities, 6 wt% and 16 wt% solids, were tested for the effect of carbon dioxide concentration. Bacterial growth rates were estimated via the measurement of carbon dioxide consumption rates. Aqueous-phase carbon dioxide concentrations in excess of 10 mg/L were found to be inhibitory to bacterial growth. (c) 1993 John Wiley & Sons, Inc.     

Tank bioleaching of copper from combined flotation concentrate and smelter dust

Bioleaching studies were conducted to evaluate the extraction of copper from combined flotation concentrate and smelter dust using two different dust-addition strategies, non-fractional (simultaneously with concentrate) and fractional (step-wise dust addition) in stirred tank reactors. The highest copper extraction obtained with the fractional dust addition was up to 94% while extraction using the non-fractional method leveled off at approximately 44%. Thus, an increased copper recovery of about 50% was achieved with the fractional dust-addition method. It seems that the main reason for increased copper recovery using fractional dust addition could be the control and prevention of chalcopyrite passivation (according to SEM/EDS, XRD, and quantitative mineralogy data). This leads to its improved dissolution.     

Dissolution of sulphur particles by Thiobacillus ferrooxidans: substrate for unattached cells

The growth of Thiobacillus ferrooxidans on sulphur is known to proceed through the attachment of cells to the sulphur particles. Experiments, However, show that the cells in the liquid phase, which are not attached to the sulphur particles, also grow. It has been shown through the use of a two-compartment membrane reactor that this increase is partially due to the release of ions, corresponding to partially oxidized of sulphur, into the solution by the attached cells. The main soluble ion has been found to the thiosulphate, but traces of sulphite have also been detected. (c) 1993 John Wiley & Sons, Inc.     

The extremophile Acidithiobacillus ferrooxidans possesses a c-di-GMP signalling pathway that could play a significant role during bioleaching of minerals

Aims: The primary goal of this study was to characterize the existence of a functional c‐di‐GMP pathway in the bioleaching bacterium Acidithiobacillus ferrooxidans. Methods and Results: A bioinformatic search revealed that the genome sequence of At. ferrooxidans ATCC 23270 codes for several proteins involved in the c‐di‐GMP pathway, including diguanylate cyclases (DGC), phosphodiesterases and PilZ effector proteins. Overexpression in Escherichia coli demonstrated that four At. ferrooxidans genes code for proteins containing GGDEF/EAL domains with functional DGC activity. MS/MS analysis allowed the identification of c‐di‐GMP in nucleotide preparations obtained from At. ferrooxidans cells. In addition, c‐di‐GMP levels in cells grown on the surface of solid energetic substrates such as sulfur prills or pyrite were higher than those measured in ferrous iron planktonic cells. Conclusions: At. ferrooxidans possesses a functional c‐di‐GMP pathway that could play a key role in At. ferrooxidans biofilm formation during bioleaching processes. Significance and Impact of the Study: This is the first global study about the c‐di‐GMP pathway in an acidophilic bacterium of great interest for the biomining industry. It opens a new way to explore the regulation of biofilm formation by biomining micro‐organisms during the bioleaching process.     

Kinetics of growth and elemental sulfur oxidation in batch culture of thiobacillus ferrooxidans

The kinetics of oxidation of elemental sulfur by Thiobacillus ferrooxidans in a batch reactor was followed by measuring the concentration of adsorbed cells on the sulfur surface, the concentration of free cells in liquid medium, and the amount of sulfur oxidized. As the elemental sulfur was oxidized to sulfate, the liquid-phase concentration of free cells continued to increase with time, whereas the surface concentration of adsorbed cells per unit weight of sulfur approached a limiting value, i.e., the maximum adsorption capacity. During sulfur oxidation, there was a close correlation between the concentrations of adsorbed and free cells, and these data were well correlated with the Langmuir isotherm. The observed rates of batch growth and sulfur oxidation were consistent with a kinetic model, assuming that the growth rate of batch growth and sulfur oxidation were consistent with a kinetic model. Assuming that the growth rate of adsorbed bacteria is proportional to the product of the concentration of adsorbed cells and the fraction of adsorption sites unoccupied by cells. The kinetic and stoichiometric parameters appearing in the model were evaluated using the experimental data and were compared with parameters determined previously for a few metal sulfides. (c) 1994 John Wiley & Sons, Inc.     

氧化亚铁硫杆菌氧化亚硫酸盐的动力学研究

实验用Ms培养基,利用去除铁离子的氧化亚铁硫杆菌(Thiobacillus ferrooxidans)进行了细菌亚硫酸盐的生长代谢研究。实验结果表明氧化亚铁硫杆菌对亚硫酸根具有一定的氧化能力。用Origin 7.0对实验数据进行拟合处理,表明了氧化亚铁硫杆菌催化氧化亚硫酸盐的动力学方程符合Hill方程。氧化亚铁硫杆菌催化氧化亚硫酸盐是一个底物抑制的细胞反应,其KS值随pH值和底物浓度的改变而变化。pH值对反应有很大的影响,pH值越接近中性KS就越小,反应速率就越大。     

Desulfurization of coal by microbial column flotation

Twenty-three strains capable of oxidizing iron were isolated from coal and ore storage sites as well as coal and ore mines, volcanic areas, and hot spring. Four strains were found to have high iron-oxidizing activity. One strain (T-4) was selected for this experiment since the strain showed the fastest leaching rate of iron and sulfate from pyrite among the four strains. The T-4 strain was assigned for Thiobacillus ferrooxidans from its cultural and morphological characteristics.Bacterial treatment was applied to column flotation. An increase of cell density in the microbial column flotation resulted in the increase of pyrite removal from a coal-pyrite mixture (high sulfur imitated coal) with corresponding decrease of coal recovery. The addition of kerosene into the microbial column flotation increased the recovery of the imitated coal from 55% (without kerosene) to 81% (with 50 muL/L kerosene) with the reduction of pyrite sulfur content from 11% (feed coal) to 3.9% (product coal). The kerosene addition could reduce the pyritic sulfur content by collecting the coal in the recovery. However, the addition could not enhance separation of pyrite from the coal-pyrite mixture, since pyrite rejection was not affected by the increase of the kerosene addition. An excellent separation was obtained by the microbial flotation using a long column which had a length-diameter (L/D) ratio of 12.7. The long column flotation reduced the pyritic sulfur content from 11% (feed coal) to 1.8% (product coal) when 80% of the feed coal was recovered without the kerosene addition. The long column flotation not only attained an excellent separation but also reduced the amount of cells for desulfurization to as little as one-tenth of the reported amount. (c) 1994 John Wiley & Sons, Inc.     

Bacterial leaching of uranium ore from Figueira—PR, Brazil, at laboratory and pilot scale

Abstract: A bacterial leaching program was carried out in order to evaluate the potential of applying this process to leach uranium from the ore of Figueira-PR, Brazil. The experiments were carried out in shake flasks, column percolation (laboratory and scmipilot scale)and in heap leaching. In shake flasks and in column percolation experiments at laboratory scale, bacterial activity on Ihe ore was confirmed: approximately 60% of uranium was leached, against around 30% in sterilized controls. Colunm percolation experiments at semipilot scale and heap leaching (850 tons of ore) showed uranium extractions of approximately 50%. In both experiments, a complementary sulfuric acid attack, after the bacterial leaching phase, was neccssaw to reach this level of uranium extraction.     

异养微生物在金属生物淋滤技术中的应用

生物淋滤技术主要应用于低品位矿石金属选矿、煤气脱硫、废弃物中金属回收和污染介质中金属离子毒性的去除等方面。作为生物淋滤技术中的主体微生物之一,异养微生物可通过其产生的酸性代谢物还原、酸化及络合,提取或者溶解非硫化矿、固体废弃物、污水污泥及土壤中的金属,有助于解决目前的资源短缺问题,还可对污染环境治理提供技术支持,具有重要的理论意义和实践价值。应用于异养微生物淋滤技术中的常见微生物包括细菌(以假单胞菌为主)和真菌(以曲霉菌和青霉菌应用最为广泛)。淋滤过程涉及酸解、络合、还原及碱化等。目前,异养微生物淋滤技术主要应用于生物冶金、固体废弃物处理、污水处理和污染土壤修复等。本文分析了异养微生物金属淋滤过程中的问题,并提出了未来研究的发展方向。     

Analysis of muramic acid in holocene microbial environments by gas chromatography,electron impact,and fast atom bombardment mass spectrometry

Analytical procedures have been modified to determine the abundance of muramic acid in four different Holocene sediment samples. Muramic acid is specific to the peptidoglycan moiety of the cell walls of most eubacterial pro‐karyotic organisms. The following procedure seemed to be the most appropriate for the detection of muramic acid and amino acids, including diaminopimelic acid. Hydrolysis of the samples (in 6 N HCl, 4.5 h, at 100°C) was followed by separation and purification of amino sugars and amino acids using Amberlite XAD‐2 and then Bio‐Rad AG 50W‐X8 resins. The N,O‐heptafluorobutyryl‐n‐butyl ester derivatives were prepared by esterification in acidified (3 N HCl) n‐butanol for 3 h at 100°C, followed by acylation by refluxing with heptafluorobutyric anhydride in acetonitrile (2:1 v/v) for 12 min at 150°C. The derivatives were analyzed by gas chromatography (GC) and gas chromatography‐mass spectrometry. Fast atom bombardment (FAB) ionization was used for the muramic acid derivative to determine its molecular weight and structure, d‐and l‐amino acids were separated by GC and a capillary chiral column. By using this technique a stable N,O‐heptafluo‐robutyryl‐n‐butyl ester derivative of muramic acid was identified at picogram levels in Holocene sedimentary microbial communities. It has been reported previously that microorganisms in sediments rapidly degrade muramic acid from cell walls of dead prokaryotes. Kinetic experiments revealed that muramic acid was relatively stable in intact cell walls but decomposed rapidly in the free form. These investigations noted above showed that the concentration of muramic acid may be used as an indicator of the presence of the intact cell walls of cyanobacteria and most other bacteria in Holocene microbial communities, and of microbial contamination in samples older than the Holocene.     

Relationship between the nitrogenase activity and dark respiration rate of Synechococcus RF-1

Growth of three different anaerobic rumen fungi Neocallimastix frontalis, Piromonas communis and Sphaeromonas communis was assessed in vitro at regular intervals by measurements of protein and chitin content and of chitin synthase activity of the cell free extracts. Similar trends and a comparable amount of protein and chitin were observed in the three species. However, chitin synthase activity was higher in S. communis and contrary to the activity of the other two strains did not decrease after maximum enzyme activity was reached. There were positive correlations between chitin content, protein content and chitin synthase activity during the active growth phase of the fungi indicating that they could be confidently used to determine in vitro growth phase and biomass concentration.     

Anaerobic growth on elemental sulfur using dissimilar iron reduction by autotrophic Thiobacillus ferrooxidans

Abstract Anaerobic growth on elemental sulfur using dissimilar iron reduction by Thiobacillus ferrooxidans has been demonstrated. The ferric ion reducing activity (FIR) of the anaerobic cells was double that of the aerobic cells. Significant differences in inhibition of FIR by respiratory inhibitors were observed between aerobic and anaerobic cells. A higher amount of cytochrome was detected in anaerobic cells compared to aerobic cells. Absorption minima developed with the addition of ferric sulfate in the dithionite reduced cell suspension demonstrated that the ferric ion could accept electrons from the cytochrome system of this bacterium. The possibility of two different electron transport chains in ferric ion reduction is discussed.     

生物浸出系统中Acidithiobacillus ferrooxidans对雄黄表面改性研究

【目的】研究Acidithiobacillus ferrooxidans BY-3对雄黄表面改性作用,为进一步研究雄黄的生物炮制技术提供实验基础与理论依据。【方法】在4组生物浸出体系中(每组包含100 mL无亚铁离子的9K培养基和0.500 g雄黄):第1组无添加;第2组添加4.469 g硫酸亚铁;第3组添加0.100 g硫粉;第4组加入4.469 g硫酸亚铁和0.100 g硫粉。在上述4组中使用A.ferrooxidans BY-3对雄黄进行生物浸出。浸出前后雄黄表面形貌及元素变化,使用扫描电镜(SEM)与能谱仪(EDS)、X-射线衍射(XRD)、拉曼光谱(Raman)、电感耦合等离子体原子发射光谱仪(ICP-AES)进行分析。【结果】4组浸出体系均发现A.ferrooxidans BY-3粘附于雄黄表面以此来产生直接作用。含Fe2+的浸出体系中雄黄表面产生非常明显的变化,含硫的浸出体系中雄黄表面变化不明显;只有Fe2+存在的浸出体系中As/S比率增高,而其余3组浸出体系中As/S比率均明显下降;另外,改性雄黄的表面存在黄钾铁矾、硫、赤铁矿、针铁矿和磁铁矿等,但未检测到砷华(As2O3)与副雄黄(Pararealgar)。【结论】A.ferrooxidans对雄黄改性具有重要作用。Fe2+对雄黄的改性具有促进作用,而硫对雄黄的改性具有抑制作用。雄黄改性前后的物化分析结果证实了生物浸出技术可有效解决传统方法制备雄黄及贮存过程中氧化和光化问题。     

Conversion of Hypoxanthine Derivatives to Succinyl Adenine Derivatives by Bacillus subtilis

A mutant strain HA-2 was derived from an adenine-requiring mutant of Bacillus subtilis IAM 1145 by transduction with bacteriophage SP-10 and subsequent N-methyl-N′-nitro-N-nitrosoguanidine treatment. The strain could convert exogenously supplemented hypoxanthine or inosine to succinyladenine, succinyladenosine and succinyladenosine 5′-monophosphate (succinyl AMP), and accumulate these compounds in the culture fluid. The strain, however, failed to convert exogenously supplemented inosine 5′-monophosphate to these succinyl compounds. The yields of succinyladenine, succinyladenosine and succinyl-AMP in a production medium supplemented with 17 mm of hypoxanthine were about 4.4 mm, 5.2 mm and 2.2 mm, respectively, after 4 days growth.