Biological removal of pyritic sulfur from coal by the thermophilic organism Sulfolobus acidocaldarius

More than 90% of initial pyritic sulfur was removed from bituminous coal samples (containing 2.1% pyritic sulfur) using the thermophilic organism Sulfolobus acidocaldarius. Microbial desulfurization rate was improved nearly ten fold by adjusting the N/P and N/Mg ratios in the nutrient medium. Environmental conditions were optimized. The optimal values of temperature and pH were 70 degrees C and 1.5, respectively. The influence of certain process variables (such as coal pulp density, particle size, and initial cell number density) on the rate of pyritic sulfur removal were determined. A pulp density of 20%, particle size of D (p) < 48 mum, and an initial cell number density of 10(12) cells/g pyrite in coal were found to be optimal. The carbon dioxide enriched air did not improve the rate of pyritic sulfur removal compared to pure air at 10% pulp density of coal samples containing 2.1% pyritic sulfur. The kinetics of microbial leaching of pyritic sulfur from coal was investigated. The rate of leaching was found to be first order with respect to pyritic sulfur concentration in the reaction medium.     

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.     

Continuous microbial desulfurization of coal--application of a multistage slurry reactor and analysis of the interactions of microbial and chemical kinetics

Microbial desulfurization of coal by pyrite oxidizing bacterial enrichment cultures has been studied in air-agitated slurry reactors of 4- and 20-L volumes. Batch experiments showed that inoculation with an active bacterial culture is essential to minimize the lag phase, although a considerable number of pyrite oxidizing bacteria was found on the coal prior to desulfurization. For detailed investigations of kinetics, energy requirements, and technical applicability, a bioreactor equipment consisting of a cascade of eight stages was developed and operated continuously. Microbial desulfurization of coal-monitored by measuring the axial profile of dissolved iron concentration, real and maximum oxygen consumption rates, and cell concentration-at pulp densities to 30% was performed over a period of 200 days without any disturbances concerning the aeration system, fluidization, transport of solids and microbial growth. At a pulp density of 20%, a pyrite conversion of 68% was achieved after the third reactor stage at a total residence time of five days in the first three stages. The kinetics of pyrite degradation were found to be well described by a rate equation of first order in pyrite surface area concentration if the pyrite is directly accessible for microbial attack. Rate constants were determined to 0.48 mg pyrite/(cm(2) day) in the first and to 0.24 mg pyrite/(cm(2) day) in the following reactor stages. Kinetic models taking into account adsorption/desorption as well as growth kinetics failed to describe the observed reaction rates. However, a model treating pyrite degradation and microbial growth kinetics formalistically seems to be applicable when backmixing between the reactor stages can be avoided. The advantage of a multistage reactor in comparison to single-stage equipment was shown by calculation. To obtain a pyrite conversion of 68%, a three-stage reactor would require only 58% of the volume of single-stage equipment.Measurement of oxygen consumption rates proved to provide quickly and easily measurable parameters to observe microbial coal desulfurization in technical scale: the real oxygen consumption rate is correlated to the pyrite oxidation rate and the maximum oxygen consumption rate is correlated to the concentration of viable cells. The Y(o/s) coefficient for the amount of oxygen consumed per mass unit of pyrite oxygen was determined to approximately 0.33 in comparison to 1.0 which can be calculated from stoichiornetry. This could yet not be explained. Chemical leaching experiments as well as sulfur analyses of desulfurized coal samples showed that the microorganisms play the main role in degradation of pyrite from coal and that pyrite oxidation by ferric iron can be neglected.     

Removal of Sulfur Compounds from Coal by the Thermophilic Organism Sulfolobus acidocaldarius

The thermophilic, reduced-sulfur, iron-oxidizing bacterium Sulfolobus acidocaldarius was used for the removal of sulfur compounds from coal. The inclusion of complex nutrients such as yeast extract and peptone, and chemical oxidizing agents, 0.01 M FeCl₃ into leaching medium, reduced the rate and the extent of sulfur removal from coal. The rate of sulfur removal by S. acidocaldarius was strongly dependent on the sulfur content of the coal and on the total external surface area of coal particles. Approximately 96% of inorganic sulfur was removed from a 5% slurry of coal which had an initial total sulfur content of 4% and an inorganic (pyritic S and sulfate) sulfur content of 2.1%. This resulted in removal of 50% of initial total sulfur present in coal.     

An airlift-recycle fermenter for microbial desulfurization of coal

Summary An airlift-external recycle fermenter has been constructed and used for the removal of pyritic sulfur from coal samples (4% initial total sulfur) by the thermophilic, sulfur oxidizing organism Sulfolobus acidocaldarius. The airlift fermenter behaved as a well mixed reactor. Approximately, 30% of initial pyritic sulfur has been removed from a 5% coal slurry of ~125 particle size, at a maximum rate of 1.8 mg S/l.h.     

An investigation of the efficacy of biological additives for the suppression of pyritic sulphur during simulated froth flotation of coal

The biological molecule responsible for the suppression of pyritic sulfur in fine coal simulated froth flotation treated with bacteria was identified. Protein was found to be the most effective agent in pyrite suppression of the three cell components (protein, lipid, and carbohydrate) assayed. Coal recovery and ash removal of the flotation process were only slightly reduced by this treatment. Other protein-containing materials were evaluated for their ability to suppress pyrite flotation. Whey was found to be the most cost-effective flotation additive of those assayed. The sulfur content of the whey-treated float was reduced by 84.0% in a synthetically prepared fractionated coal (10.7% sulfur), by a raw whey dosage of 20 muL/g coal. The inorganic sulfur component of a natural high sulfur coal fraction (10.9%) was completely depressed by this whey addition. The effect of particle size and pulp density upon the process were investigated.     

Organic sulfur removal from coal by microorganisms from extreme environments

Abstract: Microbial samples were collected from sulfurous, near neutral pH, thermal waters of Yellowstone Park. Thermophilic mixed cultures were identified that removed 90% of pyritic and sulfate sulfur and 33% of the organic sulfur from North Dakota lignite. The 30–40% organic desulfurization barrier was studied for possible inhibitors to organic sulfur removal from coal.     

Optimization of Microbial Leaching of Base Metals from a South African Sulfidic Nickel Ore Concentrate by Acidithiobacillus ferrooxidans

X-ray diffraction analysis revealed that pentlandite and chalcopyrite were the prominent mineral phases in a South African sulfidic nickel ore concentrate that hosted nickel and copper. Cobalt was found to be closely associated with the nickel-bearing pentlandite phase of the ore sample. Microbial batch leaching experiments designed according to a central composite design model were run for 15 days in a shaking incubator (150 rpm) at a constant temperature (30°C) with variations in experimental parameters like ore pulp density, particle size, bacterial inoculum, pH of the culture medium, and residence time. Quadratic mathematical models were developed to predict the rate of metal extractions. The suitability of the model of the microbial leaching process was confirmed from normal probability curves. An analysis of variance indicated that the residence time, pulp density of the ore, and particle size were the most significant factors. Bacterial inoculum size hardly showed any effect on the total metal extractions. Maximum nickel (82%), cobalt (76%) and copper (25.6%) extractions were achieved under optimum conditions, operated for 15 days at pulp density of 2% and particle size of ?75 µm at pH 1.5.     

Kinetics of the Removal of Iron Pyrite from Coal by Microbial Catalysis

Different strains of Thiobacillus ferrooxidans and Thiobacillus thiooxidans were used to catalyze the oxidative dissolution of iron pyrite, FeS₂, in nine different coal samples. Kinetic variables and parametric factors that were determined to have a pronounced effect on the rate and extent of oxidative dissolution at a fixed Po₂ were: the bacterial strain, the nitrogen/phosphorus molar ratio, the partial pressure of CO₂, the coal source, and the total reactive surface area of FeS₂. The overall rate of leaching, which exhibited a first-order dependence on the total surface area of FeS₂, was analyzed mathematically in terms of the sum of a biochemical rate, ν₁, and a chemical rate, ν₂. Results of this study show that bacterial desulfurization (90 to 98%) of coal samples which are relatively high in pyritic sulfur can be achieved within a time-frame of 8 to 12 days when pulp densities are ≤20% and particle sizes are ≤74 μm. The most effective strains of T. ferrooxidans were those that were isolated from natural systems, and T. ferrooxidans ATCC 19859 was the most effective pure strain. The most effective nutrient media contained relatively low phosphate concentrations, with an optimal N/P molar ratio of 90:1. These results suggest that minimal nutrient additions may be required for a commercial desulfurization process.     

Influence of pulp density and bioreactor design on microbial desulphurization of coal

Summary Pyrite was microbiologically removed by Thiobacillus ferrooxidans in pure and mixed cultures from German bituminous coal at 10% pulp density with maximum pyrite oxidation rate of 350 mg pyritic S/l per day. However, at pulp densities above 20% bacterial growth and consequently pyrite oxidation were completely prevented both in a conventional airlift reactor and in a stirred-tank reactor. Modifying the airlift reactor by adapting a conical bottom part, bacterial growth and pyrite oxidation could be achieved even at 30% pulp density, resulting in a pyrite removal of more than 90% at a pyrite oxidation rate of 230 mg pyritic S/l per day.Dedicated to Prof. Dr. H. Jüntgen on the occasion of his 60th birthday     

Leaching of Zinc Sulfide by Thiobacillus ferrooxidans: Bacterial Oxidation of the Sulfur Product Layer Increases the Rate of Zinc Sulfide Dissolution at High Concentrations of Ferrous Ions

This paper reports the results of leaching experiments conducted with and without Thiobacillus ferrooxidans at the same conditions in solution. The extent of leaching of ZnS with bacteria is significantly higher than that without bacteria at high concentrations of ferrous ions. A porous layer of elemental sulfur is present on the surfaces of the chemically leached particles, while no sulfur is present on the surfaces of the bacterially leached particles. The analysis of the data using the shrinking-core model shows that the chemical leaching of ZnS is limited by the diffusion of ferrous ions through the sulfur product layer at high concentrations of ferrous ions. The analysis of the data shows that diffusion through the product layer does not limit the rate of dissolution when bacteria are present. This suggests that the action of T. ferrooxidans in oxidizing the sulfur formed on the particle surface is to remove the barrier to diffusion by ferrous ions.     

The removal of pyritic sulphur from coal by Leptospirillum-like bacteria

Summary The microbial oxidation of pyritic sulphur was studied in a 4.5-l airlift fermentor at pH 1.5 and 100 g/l pulp density. By microbial leaching with Leptospirillum-like bacteria 85% of the pyritic sulphur was removed within 40 days; 30% of the removed pyrite was oxidized to elemental sulphur, the rest being transformed to soluble sulphate. Accumulation of elemental sulphur could be avoided by using a mixed culture of Leptospirillum-like bacteria and Thiobacillus ferrooxidans. Apart from oxidation of elemental sulphur neither the pure nor the mixed culture showed a significant difference as to removal of pyrite.     

Suppression of pyritic sulphur during flotation tests using the bacterium Thiobacillus ferrooxidans

Environmental concern about sulphur dioxide emissions has led to the examination of the possibility of removing pyritic sulphur from coal prior to combustion during froth flotation, a routine method for coal cleaning at the pit-head. The bacterium Thiobacillus ferrooxidans was effective in leaching 80% and 63% -53 mum pyrite at 2% and 6% pulp density in shake flasks in 240 and 340 h, respectively.The natural floatability of pyrite was significantly reduced in the Hallimond tube following 2.5 min of conditioning in membrane-filtered bacterial liquor prior to flotation. The suppression effect was greatly enhanced in the presence of Thiobacillus ferrooxidans. A bacterial suspension in pH 2.0 distilled water showed 85% suppression, whereas in spent growth liquor this value was 95%. The optimum bacterial density was 3.25 x 10(10) cells/g pyrite in 230-ml distilled water (2% pulp density) in the Hallimond tube. The degree of suppression by the cells was related to particle size but not to pH or temperature. The sulphur content of a synthetic coal/pyrite mixture was reduced from 10.9 to 2.1% by flotation after bacterial preconditioning. It is postulated that pyrite removal in coals which are cleaned by froth flotation could be significantly reduced using a bacterial preconditioning stage with a short residence time of 2.5 min.     

Temperature Effects on Bacterial Leaching of Sulfide Minerals in Shake Flask Experiments

The microbiological leaching of a sulfide ore sample was investigated in shake flask experiments. The ore sample contained pyrite, pyrrhotite, pentlandite, sphalerite, and chalcopyrite as the main sulfide minerals. The tests were performed at eight different temperatures in the range of 4 to 37°C. The primary data were used for rate constant calculations, based on kinetic equations underlying two simplified models of leaching, i.e., a shrinking particle model and a shrinking core model. The rate constants thus derived were further used for the calculation of activation energy values for some of the sulfide minerals present in the ore sample. The chalcopyrite leaching rates were strongly influenced by the interaction of temperature, pH, and redox potential. Sphalerite leaching could be explained with the shrinking particle model. The data on pyrrhotite leaching displayed good fit with the shrinking core model. Pyrite leaching was found to agree with the shrinking particle model. Activation energies calculated from the rate of constants suggested that the rate-limiting steps were different for the sulfide minerals examined; they could be attributed to a chemical or biochemical reaction rather than to diffusion control.     

Leaching of Pyrites of Various Reactivities by Thiobacillus ferrooxidans

Wide variations were found in the rate of chemical and microbiological leaching of iron from pyritic materials from various sources. Thiobacillus ferrooxidans accelerated leaching of iron from all of the pyritic materials tested in shake flask suspensions at loadings of 0.4% (wt/vol) pulp density. The most chemically reactive pyrites exhibited the fastest bioleaching rates. However, at 2.0% pulp density, a delay in onset of bioleaching occurred with two of the pyrites derived from coal sources. T. ferrooxidans was unable to oxidize the most chemically reactive pyrite at 2.0% pulp density. No inhibition of pyrite oxidation by T. ferrooxidans occurred with mineral pyrite at 2.0% pulp density. Experiments with the most chemically reactive pyrite indicated that the leachates from the material were not inhibitory to iron oxidation by T. ferrooxidans.     

Leaching of zinc sulfide by Thiobacillus ferrooxidans: bacterial oxidation of the sulfur product layer increases the rate of zinc sulfide dissolution at high concentrations of ferrous ions

This paper reports the results of leaching experiments conducted with and without Thiobacillus ferrooxidans at the same conditions in solution. The extent of leaching of ZnS with bacteria is significantly higher than that without bacteria at high concentrations of ferrous ions. A porous layer of elemental sulfur is present on the surfaces of the chemically leached particles, while no sulfur is present on the surfaces of the bacterially leached particles. The analysis of the data using the shrinking-core model shows that the chemical leaching of ZnS is limited by the diffusion of ferrous ions through the sulfur product layer at high concentrations of ferrous ions. The analysis of the data shows that diffusion through the product layer does not limit the rate of dissolution when bacteria are present. This suggests that the action of T. ferrooxidans in oxidizing the sulfur formed on the particle surface is to remove the barrier to diffusion by ferrous ions.     

Evaluation of desorption of proteins adsorbed to hydrophilic surfaces by two-dimensional electrophoresis

The evaluation of the plasma protein adsorption patterns of superparamagnetic iron oxide (SPIO) particles is of high interest concerning their in vivo fate and is carried out by two-dimensional electrophoresis (2-DE). The sample preparation is of great importance, especially the removal of the adsorbed proteins (desorption) from the particle surface for subsequent analysis by 2-DE. The removal is carried out by a desorption solution. In this study, negatively and positively charged SPIO model particles were under investigation concerning the desorption of proteins adsorbed on their surfaces. Firstly, the desorption process was determined quantitatively using the Bradford protein assay. Secondly, the removable or nonremovable protein species, from particles surface were under investigation by 2-DE. Looking at the desorption in a quantitative manner with the Bradford assay, the desorption efficacy from negatively charged particles was about 90%. In the case of the positively charged particles, the desorption efficacy seemed to be reduced, approximately 34% of the proteins remained on the surface. Comparing the protein patterns of the particles evaluated by 2-DE in the desorption solution and the proteins remaining on the particles, they confirmed the results from the protein quantification. After desorption, the IgG gamma-chains were found to be the dominant protein fraction remaining on the negatively charged particles. On the positively charged particles, many more protein species were found after desorption. The more basic the protein fragments, the more ineffective was the desorption from the positively charged model particle, and vice versa. Nevertheless, all protein spots were found qualitatively in the desorption solution, especially when the desorption solutions still containing the particles were used for the 2-DE analysis. In conclusion, 2-DE could be confirmed as the "gold standard" for determining the plasma protein adsorption patterns of nanoparticulate systems.     

Microbial removal of pyrite from coal using a percolation bioreactor

Summary To compare the suspension and the percolation process system for the microbial desulphurization of coal the microbial pyrite oxidation in coal during storage in dumps was investigated in laboratory experiments with Thiobacillus ferrooxidans using a percolation bioreactor and resulted in a removal of 75% of pyrite within 70 days. In the initial desulphurization phase 450 mg pyritic-S/kg coal per day were oxidized at maximum rate, while the overall rate was determined to 130 mg pyritic-S/kg coal per day. During the desulphurization the mean particle size of the coal was reduced from 0.55 mm to 0.175 mm. As shown by microscopy and elemental analyses of the coal the pyrite was completely removed from small coal particles, whereas parts of it remained in the core of the greater particles.     

Influence of Leaching Parameters on the Biological Removal of Uranium from Coal by a Filamentous Cyanobacterium

Axenic cultures of the filamentous cyanobacterium LPP OL3 were incubated with samples of uraniumbearing coal from a German mining area. The influence of leaching parameters such as coal concentration (pulp density), initial biomass, particle size, temperature, and composition of the growth medium on the leaching of uranium from the ore by the cyanobacterial strain was studied. When low pulp densities were applied, the yield of biologically extracted uranium was optimal (reaching 96% at 1% [wt/vol] coal) and all released uranium was found in the culture liquid. Above 10% (wt/vol) coal in the medium, the amount of cell-bound uranium increased. Initial biomass concentration (protein content of the cultures) and particle size were not critical parameters of leaching by LPP OL3. However, temperature and composition of the growth medium profoundly influenced the leaching of uranium and growth of the cyanobacterium. The yield of leached uranium (at 10% [wt/vol] coal) could not be raised with a tank leaching apparatus. Also, coal ashes were not suitable substrates for the leaching of uranium by LPP OL3. In conclusion, the reactions of the cyanobacterium to variations in leaching parameters were different from reactions of acidic leaching organisms.     

The adsorption of Thiobacillus ferrooxidans on coal surfaces

The adsorption of Thiobacillus ferrooxidans to coal surfaces has been studied. Adsorption experiments were conducted on coal samples from eight different Eastern coal fields. In all cases the adsorption process was at least 90% complete within the first two minutes following inoculation. The results of these experiments were used to test the validity of two proposed adsorption models. The first model assumes that bacterial adsorption follows second-order irreversible kinetics of the second kind with respect to the concentration of bacteria and substratum surface area in the system. The second model allows for the contribution of reversible adsorption detected in desorption experiments. It was found that the combined reversible-irreversible model more accurately describes the initial stages of adsorption. Rate constants in both models were calculated for each coal sample. The relation of each of these constants to the pyrite concentration in coal is presented and the significance of these relations is discussed.Scanning electron micrographs of inoculated coal samples sho that Thiobacillus ferrooxidans selectively adsorb to exposed pyrite phases dispersed throughout the organic coal matrix. Preferential attachment was also observed along topographical faults in the caol surface. Mercury contact angle measurements on coal indicate that the selective adsorption of Thiobacillus ferrooxidans may be attributable to the lower surface free energy of pyrite relative to the organic coal matrix.