Flocculation behaviour of hematite-kaolinite suspensions in presence of extracellular bacterial proteins and polysaccharides.

Cells of Bacillus subtilis exhibited higher affinity towards hematite than to kaolinite. Bacterial cells were grown and adapted in the presence of hematite and kaolinite. Higher amounts of mineral-specific proteinaceous compounds were secreted in the presence of kaolinite while hematite-grown cells produced higher amounts of exopolysaccharides. Extracellular proteins (EP) exhibited higher adsorption density on kaolinite which was rendered more hydrophobic. Hematite surfaces were rendered more hydrophilic due to increased adsorption of extracellular polysaccharides (ECP). Significant surface chemical changes were produced due to interaction between minerals and extracellular proteins and polysaccharides. Iron oxides such as hematite could be effectively removed from kaolinite clays using selective bioflocculation of hematite after interaction with EP and ECP extracted from mineral-grown cells.

Microbially induced separation of quartz from calcite using Saccharomyces cerevisiae.

Cells of Saccharomyces cerevisiae and their metabolites were successfully utilized to achieve selective separation of quartz and calcite through microbially induced flotation and flocculation. S. cerevisiae was adapted to calcite and quartz minerals. Adsorption studies and electrokinetic investigations were carried out to understand the changes in the surface chemistry of yeast cells and the minerals after mutual interaction. Possible mechanisms in microbially induced flotation and flocculation are outlined.

Microbially induced separation of quartz from hematite using sulfate reducing bacteria.

Cells and metabolic products of Desulfovibrio desulfuricans were successfully used to separate quartz from hematite through environmentally benign microbially induced flotation. Bacterial metabolic products such as extracellular proteins and polysaccharides were isolated from both unadapted and mineral-adapted bacterial metabolite and their basic characteristics were studied in order to get insight into the changes brought about on bioreagents during adaptation. Interaction between bacterial cells and metabolites with minerals like hematite and quartz brought about significant surface-chemical changes on both the minerals. Quartz was rendered more hydrophobic, while hematite became more hydrophilic after biotreatment. The predominance of bacterial polysaccharides on interacted hematite and of proteins on quartz was responsible for the above surface-chemical changes, as attested through adsorption studies. Surface-chemical changes were also observed on bacterial cells after adaptation to the above minerals. Selective separation of quartz from hematite was achieved through interaction with quartz-adapted bacterial cells and metabolite. Mineral-specific proteins secreted by quartz-adapted cells were responsible for conferment of hydrophobicity on quartz resulting in enhanced separation from hematite through flotation.

Modeling and analysis of nanoscale interaction forces between Acidithiobacillus ferrooxidans and AFM tip.

The specificity exhibited by Acidithiobacillus ferrooxidans, a chemolithoautotrophic acidophile, in their attachment to different sulphide minerals has resulted in effective physical separation of these minerals. This can be explained in terms of surface forces of interaction between the cells and substrates using AFM. In the light of this, the present study reports interaction studies with A. ferrooxidans cells and an AFM silicon nitride tip. The aim of the present investigation is to probe the nanoscale interactions between A. ferrooxidans cells and silicon nitride tip of an AFM, model the approach and retraction forces, and elucidate the effects of pH, ionic strength and surface biopolymers on interfacial forces.

Soft-particle model analysis of effect of LPS on electrophoretic softness of Acidithiobacillus ferrooxidans grown in presence of different metal ions.

The effect of surface lipopolysaccharides (LPS) on the electrophoretic softness and fixed charge density in the ion-penetrable layer of Acidithiobacillus ferrooxidans cells grown in presence of copper or arsenic ions have been discussed. The electrophoretic mobility data were analyzed using the soft-particle electrophoresis theory. Cell surface potentials of all the strains based on soft-particle theory were lower than those estimated using the conventional Smoluchowski theory. Exposure to metal ions increased the surface electrophoretic softness with decrease in the fixed charge density. Effect of cell surface lipopolysaccharides on the model parameters are investigated and discussed.

Biomineralisation of manganese on titanium surfaces exposed to seawater.

A 2-year long study was carried out to isolate and characterise various bacterial species present in the biofilm formed on titanium surfaces exposed to seawater and to assess the manganese oxidizing potential of the marine isolates. The amount of manganese present in the biofilm was also measured using atomic absorption spectrometry (AAS). The results showed that titanium was susceptible to biofouling. More than 50% of the culturable marine bacterial isolates were capable of bringing about oxidation of Mn(II). All these manganese oxidizing bacteria were heterotrophic. Autotrophic manganese oxidizing bacteria such as Leptothrix was not isolated in the present study. The AAS results confirmed that the manganese content in the biofilms increased with increasing exposure time. Hence, the study indicates that the titanium surfaces when exposed to seawater were colonised by a large number of heterotrophic bacteria, which have the ability of bringing about biomineralisation of manganese.

Bioremediation of zinc using Desulfotomaculum nigrificans: bioprecipitation and characterization studies.

Desulfotomaculum nigrificans, a typical sulfate reducing bacterium (SRB) was successfully grown in the presence of 12-210 mg/L of zinc. Complete bioremoval of zinc was achieved in 2 days for 12 mg/L while the bioremoval efficiency was about 70% in 40 days in the presence of 210 mg/L initial concentration of zinc, attesting to the inhibition of bacterial cell growth at higher zinc concentrations. The bioremoval mechanism was predominantly governed by bioprecipitation with biosorption contributing to a minor extent. The amount of protein present in the extracellular secretions obtained by growth of SRB in modified Baars' medium devoid of iron was the highest followed by those obtained in the presence of zinc or iron, in that order. Bioremediation studies carried out using a specially designed set-up, facilitating the transfer of biogenically produced hydrogen sulfide gas to a separate precipitation assembly, confirmed that zinc could be successfully precipitated from its corresponding sulfate solution, varying in concentration from 10 to 20,000 mg/L. Detailed characterization of the various zinc sulfide precipitates by EDAX and X-ray diffraction analysis conformed to wurtzite structure. The isoelectric points of high purity zinc sulfide and that of chemically synthesized, biogenically produced and zinc sulfide precipitated using bacterially produced hydrogen sulfide gas (BPH-ZnS) were located at pH 3, 7.8, 2.8 and 8, respectively.

Surface chemical studies on selective separation of pyrite and galena in the presence of bacterial cells and metabolic products of Paenibacillus polymyxa.

Selective separation of pyrite and galena from mixture of the two minerals was achieved through interaction with cells and metabolic products from a culture of Paenibacillus polymyxa. Adsorption of cells and metabolic products onto minerals and electrokinetic studies of minerals after interaction with cells and metabolic products were carried out to examine the resulting surface modification on the mineral surfaces. Flocculation and flotation techniques were successfully applied in the selective separation of minerals after bacterial interaction. The effect of varying conditions for production of extracellular polysaccharides and protein provided an insight into the possible mechanism involved in microbially induced flocculation and flotation of pyrite and galena.

Mechanisms for solubilization of cobalt, copper and nickel from Indian Ocean nodules at near neutral pH by a marine isolate.

Polymetallic ocean nodules offer an alternative source for extracting valuable strategic metals like Cu, Co and Ni. A novel biodissolution process was carried out, employing the cell-free spent growth medium from a marine organism ( Bacillus M1) isolated from nodules; and Cu, Co and Ni solubilization from the nodules was observed to be beyond the theoretical solubility limits at near neutral pH. Different characterization techniques revealed the presence of phenolic substances in the spent growth medium, which might have formed soluble complexes with the transition metals. The low prevailing E(h) redox value in the medium suggested a strong reducing environment, favoring the reductive dissolution of the oxides. A correlation study of dissolution of Cu, Co and Ni with that of Mn and Fe in the nodules was made to investigate the mechanisms of metal solubilization by the marine isolate. Under the influence of a strong reducing environment coupled with complexation by a phenolic substance present in the spent growth medium, Mn and Fe oxides were solubilized from the nodules, resulting in concomitant dissolution of Cu, Co and Ni associated with them in the nodules.

Selective separation of pyrite and chalcopyrite by biomodulation.

Selective separation of pyrite from other associated ferrous sulphides at acidic and neutral pH has been a challenging problem. This paper discusses the utility of Acidithiobacillus ferrooxidans for the selective flotation of chalcopyrite from pyrite. Consequent to interaction with bacterial cells, pyrite remained depressed even in the presence of potassium isopropyl xanthate collector while chalcopyrite exhibited significant flotability. However, when the minerals were conditioned together, the selectivity achieved was poor due to the activation of pyrite surface by the copper ions in solution. The selectivity was improved when the sequence of conditioning with bacterial cells and collector was reversed, since the bacterial cells were able to depress collector interacted pyrite effectively, while having negligible effect on chalcopyrite. The observed behaviour is analysed and discussed in detail. The separation obtained was significant both at acidic and alkaline pH. This selectivity achieved was retained when the minerals were interacted with both bacterial cells and collector simultaneously.

Selective separation of arsenopyrite from pyrite by biomodulation in the presence of Acidithiobacillus ferrooxidans.

Effective methods for selective separation using flotation or flocculation of arsenopyrite from pyrite by biomodulation using Acidithiobacillus ferrooxidans are presented here. Adhesion of the bacterium to the surface of arsenopyrite was very slow compared to that to pyrite, resulting in a difference in surface modification of the minerals subsequent to interaction with cells. The cells were able to effectively depress pyrite flotation in presence of collectors like potassium isopropyl xanthate and potassium amyl xanthate. On the other hand the flotability of arsenopyrite after conditioning with the cells was not significantly affected. The activation of pyrite by copper sulfate was reduced when the minerals were conditioned together, resulting in better selectivity. Selective separation could also be achieved by flocculation of biomodulated samples.

Microbially induced flotation and flocculation of pyrite and sphalerite.

Cells of Paenibacillus polymyxa and their metabolite products were successfully utilized to achieve selective separation of sphalerite from pyrite, through microbially induced flocculation and flotation. Adsorption studies and electrokinetic investigations were carried out to understand the changes in the surface chemistry of bacterial cells and the minerals after mutual interaction. Possible mechanisms in microbially induced flotation and flocculation are outlined.

Solubilization of cobalt from ocean nodules at neutral pH-a novel bioprocess.

A marine organism ( Bacillus M1) isolated from Indian Ocean manganese nodules was characterized. The organism grew well in artificial seawater medium, at near neutral pH, 30 degrees C and 0.25 M NaCl, and showed MnO(2)-reducing activity. Growing cultures of Bacillus M1 as well as cell-free spent liquor from fully-grown cultures were employed to extract metals from the nodules. The spent liquor of cultures of the organism could dissolve around 45% cobalt (Co) at a pH of 8.2 in 2 h. Co recovery by this treatment was comparable to that in acidic leaching with 2.5 M hydrochloric acid solutions, and was independent of pulp density (w/v ratio). The amount of Co dissolved was beyond the thermodynamic solubility limit in aqueous solution at a pH of 8.2. It is inferred that the metabolites present in the spent liquor played a pivotal role in complexing the Fe (III) phase, solubilizing Co in the process. Partial characterization of spent liquor by spot tests, UV visible spectroscopy and FTIR spectroscopy, showed the presence of siderophore-like phenolic compound(s) with an attached carboxyl group that might form soluble organic complexes with Fe (III).

Modeling and analysis of biooxidation of gold bearing pyrite-arsenopyrite concentrates by Thiobacillus ferrooxidans.

The results of modeling the biooxidation of a mixed sulfidic concentrate by Thiobacillus ferrooxidans is reported here. A kinetic model, which accounts for the dissolution of sulfide matrix due to both bacterial attachment onto the mineral surface and indirect leaching, has been proposed. A comprehensive system approach is employed for modeling the complex biooxidation process by (a) the decomposition of the complete system into several subsystems, (b) modeling individual systems, and (c) integrating the subsystems model in a final system model. The model for subsystems was developed by writing mass balance equations for the different species involved. The bacterial balance accounts for its growth, both on solid substrate and in solution, and for the attachment to and detachment from the surface. The kinetic parameters of the model were determined by designing the experiments in such a manner that only one subsystem was operational. This model was tested in both laboratory scale batch and continuous biooxidation processes. The model predictions agreed with the experimental data reasonably well. A further analysis of the model was carried out to predict the conditions for efficient biooxidation. Studies on the effect of residence time and pulp density on steady-state behavior showed that there is a critical residence time and pulp density below which washout conditions occur. Operation at pulp densities lower than 5% and residence times lower than 72 h was found unfavorable for efficient leaching.

Surface chemical studies on sphalerite and galena using extracellular polysaccharides isolated from Bacillus polymyxa.

Adsorption, electrokinetic, microflotation, and flocculation studies have been carried out on sphalerite and galena minerals using extracellular polysaccharides (ECP) isolated from Bacillus polymyxa. The adsorption density of ECP onto galena is found to be higher than that onto sphalerite. The adsorption of ECP onto sphalerite is found to increase from pH 3 to about pH 7, where a maximum is attained, and thereafter continuously decreases. With respect to galena, the adsorption density of ECP steadily increases with increased pH. The addition of ECP correspondingly reduces the negative electrophoretic mobilities of sphalerite and galena in absolute magnitude without shifting their isoelectric points. However, the magnitude of the reduction in the electrophoretic mobility values is found to be greater for galena compared to that for sphalerite. Microflotation tests show that galena is depressed while sphalerite is floated using ECP in the entire pH range investigated. Selective flotation tests on a synthetic mixture of galena and sphalerite corroborate that sphalerite could be floated from galena at pH 9-9.5 using ECP as a depressant for galena. Flocculation tests reveal that in the pH range 9-11, sphalerite is dispersed and galena is flocculated in the presence of ECP. Dissolution tests indicate release of the lattice metal ions from galena and sphalerite, while co-precipitation tests confirm chemical interaction between lead or zinc ions and ECP. Fourier transform infrared spectroscopic studies provide evidence in support of hydrogen bonding and chemical interaction for the adsorption of ECP onto galena/sphalerite surfaces.

Surface Chemical Studies on Sphalerite and Galena Using Bacillus polymyxa.

The interaction of sphalerite and galena with cells of Bacillus polymyxa was investigated through adsorption, electrokinetic, flotation, and flocculation studies. Adsorption experiments indicated that a higher amount of the cells was adsorbed onto galena compared to sphalerite. The adsorption density of the cells onto galena was almost independent of pH while that onto sphalerite was found to continuously decrease with increasing pH. The adsorption isotherms of the bacterial cells on galena and sphalerite exhibited Langmuirian behavior. Electrokinetic measurements showed that the negative electrophoretic mobilities of the cells were reduced in magnitude in proportion to the time of interaction with either sphalerite or galena. Similar trends were observed in the cases of sphalerite and galena after interaction with the cells. However, the magnitude of the reduction in the electrophoretic mobilities was found to be greater for galena than for sphalerite. Flotation tests revealed that galena was almost completely depressed after interaction with the cells both in the absence and in the presence of the collector. In contrast, the addition of collector and activator to sphalerite, which was initially interacted with the cells, restored the floatability at and beyond pH 8.5. Selective flotation tests on a synthetic mixture of galena and sphalerite confirmed that sphalerite could be preferentially floated from galena, which was depressed by the bacterial cells. Selective flocculation tests further demonstrated that galena could be flocculated from sphalerite, which was dispersed in the presence of cells of B. polymyxa at pH 9-9.5. Copyright 2001 Academic Press.

Surface Chemical Studies on Sphalerite and Galena Using Bacillus polymyxa.

Biodissolution tests reveal the release of lead/zinc species from galena/sphalerite, respectively, while biosorption experiments confirm interaction of cells of Bacillus polymyxa (B. polymyxa) with the metal ions of interest. The amount of exo-polysaccharides is found to be the highest in the case of galena-interacted cells, followed by the Bromfield medium-grown cells while the sphalerite-interacted cells have the least, based on ruthenium red adsorption studies. In contrast, the sphalerite-interacted cells assay the highest amount of protein while the galena-interacted cells have the lowest amount, on a comparative basis. The adsorption of xanthate onto galena is found to be diminished in the presence of the cells whereas the xanthate adsorption density for activated sphalerite is unaffected in the pH range 9-11. Additionally, the cell surface hydrophobicity tests confirm that the sphalerite-interacted cells are more hydrophobic relative to the galena-interacted cells. FTIR spectroscopic data lend support to the higher adsorption density of the cells onto galena vis-à-vis sphalerite. The higher exo-polysaccharide and lower protein contents together with the hydrophilic nature of the galena-interacted cells could be the contributing factors to the selective flocculation and depression of galena. In a similar manner, the higher protein and lower exo-polysaccharide contents as well as the greater hydrophobicity of the sphalerite-interacted cells favor its floatability and dispersion. Copyright 2001 Academic Press.

Surface chemical studies of Thiobacillus ferrooxidans with reference to copper tolerance.

A strain of Thiobacillus ferrooxidans was adapted to grow at higher concentrations of copper by single step culturing in the presence of 20 g/L (0.314 mol/L) cupric ions added to 9K medium. Exposure to copper results in change in the surface chemistry of the microorganism. The isoelectric point of the adapted strain (pI = 4.7) was observed to be at a higher pH than that of the wild unadapted strain (pI = 2.0). Compared to the wild strain, the copper adapted strain was found to be more hydrophobic and showed enhanced attachment efficiency to the pyrite mineral. The copper adsorption ability of the adapted strain was also found to be higher than that of the wild strain. Fourier transform infrared spectroscopy of adapted cells suggested that a proteinaceous new cell surface component is synthesized by the adapted strain. Treatment of adapted cells with proteinase-K, resulted in complete loss of tolerance to copper, reduction in copper adsorption and hydrophobicity of the adopted cells. These observations strongly suggest a role played by cell surface modifications of Thiobacillus ferrooxidans in imparting the copper tolerance to the cells and bioleaching of sulphide minerals.

Stability of copper tolerance in Thiobacillus ferrooxidans.

A strain of Thiobacillus ferrooxidans MAL-4-1 was adapted to grow at higher concentrations of copper by repeated subculturing in the presence of increasing levels of added cupric ions in 9K medium. The strains adapted to copper were found to be more efficient in bioleaching of copper from concentrates. When copper tolerant strains were back cultured repeatedly in 9K medium without cupric ions, the initially developed metal tolerance was observed to be lost. This indicates that the copper tolerance developed is stress-dependent and not a permanent trait of the adapted strain.

Surface Chemistry of Thiobacillus ferrooxidans Relevant to Adhesion on Mineral Surfaces.

Thiobacillus ferrooxidans cells grown on sulfur, pyrite, and chalcopyrite exhibit greater hydrophobicity than ferrous ion-grown cells. The isoelectric points of sulfur-, pyrite-, and chalcopyrite-grown cells were observed to be at a pH higher than that for ferrous ion-grown cells. Microbe-mineral interactions result in change in the surface chemistry of the organism as well as that of the minerals with which it has interacted. Sulfur, pyrite, and chalcopyrite after interaction with T. ferrooxidans exhibited a significant shift in their isoelectric points from the initial values exhibited by uninteracted minerals. With antibodies raised against sulfur-grown T. ferrooxidans, pyrite- and chalcopyrite-grown cells showed immunoreactivity, whereas ferrous ion-grown cells failed to do so. Fourier transform infrared spectroscopy of sulfur-grown cells suggested that a proteinaceous new cell surface appendage synthesized in mineral-grown cells brings about adhesion to the solid mineral substrates. Such an appendage was found to be absent in ferrous ion-grown cells as it is not required during growth in liquid substrates.