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,

Differential utilization of cyclic, orthorhombic α- and chain-like polymeric μ-sulfur by Acidithiobacillus ferrooxidans

The differential utilization of cyclic, orthorhombic α-sulfur （α-S） and chain-like polymeric g-sulfur （μ-S） by Acidithiobacillus ferrooxidans was investigated. The growth and sulfur oxidation results indicated that utilization of μ-S by A. ferrooxidans was clearly different from α-S. Even if the planktonic cells were produced, the fall of pH and the rise of sulfate concentration were the same after 300 h on each substrate, the speeds of the planktonic cells increase, pH decrease and sulfate concentration increase in the earlier cultivation stage were faster on polymeric sulfur compared with the orthorhombic form. The adsorption capacity of the cells was higher on μ-S than on α-S, The results of SEM, DRIFTS and XRD analyses indicated that the surfaces of α-S and μ -S were modified differently by cells. Differential expression of 11 selected sulfur adsorption-activation and metabolism relevant genes was detected by RT-qPCR. The results showed that the expression of the hydrophobic substrate transport proteins and the sulfur metabolism related proteins was up-regulated, and the adsorption and activation related proteins were down-regulated when the cells were grown on μS, suggesting that μ-S could be more easily bio-adapted and activated than α-S.

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.

Effect of pH values on the extracellular polysaccharide secreted by Acidithiobacillus ferrooxidans during chalcopyrite bioleaching

The pH value plays an important role in the bioleaching of sulphide minerals. The effect of pH values on the extracellular poly-saccharide secreted by Acidithiobacillus ferrooxidans was investigated in different phases of bacterial growth during chalcopyrite bioleach-ing. It is found that extracellular polysaccharide secretion from the cells attached to chalcopyrite is more efficiently than that of the free cells in the bioleaching solution. Three factors, pH values, the concentration of soluble metal ions, and the bacterial growth and metabolism, affect extracellular polysaccharide secretion in the free cells, and are related to the bacterial growth phase. Extracellular polysaccharide secretion from the attached cells is mainly dependent on the pH value of the bacterial culture.

Acidophilic bacterial community reflecting pollution level of sulphide mine impacted by acid mine drainage

To reveal the impact of mining on bacterial ecology around mining area,bacterial community and geochemical characteristics about Dabaoshan Mine(Guangdong Province,China)were studied.By amplified ribosomal DNA restriction analysis and phylogenetic analysis,it is found that mining pollution greatly impacts the bacterial ecology and makes the habitat type of polluted environments close to acid mine drainage(AMD)ecology.The polluted environment is acidified so greatly that neutrophil and alkaliphilic microbes are massively dead and decomposed.It provided organic matters that can make Acidiphilium sp.rapidly grow and become the most bacterial species in this niche.Furthermore,Acidithiobacillus ferrooxidans and Leptospirillum sp.are also present in this niche.The amount of Leptospirillum sp.is far more than that of Acidithiobacillus ferrooxidans,which indicates that the concentration of toxic ions is very high.The conclusions of biogeochemical analysis and microbiological monitor are identical. Moreover,because the growth of Acidithiobacillus ferrooxidans and Leptospirillum sp.depends on ferrous iron or inorganic redox sulfur compounds which can be supplied by continual AMD,their presence indicates that AMD still flows into the site.And the area is closer to the outfalls of AMD,their biomasses would be more.So the distinction of their biomasses among different areas can help us to find the effluent route of AMD.

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.

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.

Bioleaching of zinc from gold ores using Acidithiobacillus ferrooxidans

The present work investigated microbial leaching of zinc from ore using Acidithiobacillus ferrooxidans （ATCC 14859）. The ore samples, consisted of 13wt% zinc, were obtained from a gold mine in north-eastern Thailand. A shake flask study was performed on the ore samples using a rotary shaker under the following fixed conditions （250 r·min^-1, 30℃ for 16 d）. The influence of various conditions, namely medium type （with and without iron）, particle ore size （〈20, 20-40, 40-60, 60-100, and 〉100 mesh）, ore density （20 kg·m^-3, 50 kg·m^-3, and 100 kg·m^-3）, and pH of the medium （2, 2.5, 2.8, and 3）, were investigated. The microbial leaching was assessed by determining the concentration of zinc in the medium and compared with the initial sample concentration. The results show that Acidithiobacillus ferrooxidans can successfully leach zinc by as much as 6 times compared with the control experiment （without Acidithiobacillus ferrooxidans ferrooxidans）. The maximum efficiency （92.3%） for microbial leaching is obtained in iron-containing medium, 20-40 mesh ore sizes, 20 kg·m^-3 ore density at pH 2.8, and the zinc content is found in the medium at about 120 mg·m^-1.

Bioleaching of soluble phosphorus from rock phosphate containing pyrite with DES-induced Acidithiobacillus ferrooxidans

The effects of Acidithiobacillus ferrooxidans(At. f) mutated with diethyl sulfate(DES) as a mutagen on the bioleaching of soluble phosphorus(P) from rock phosphate(RP) were investigated. The results show that the oxidative activity of At. f is greatly improved by 1.0%(volume fraction) of DES. Correspondingly,the highest leaching rate of soluble P is also obtained to be 14.9% by the At. f mutated,which is 85.8% higher than that of the adapted At. f without mutation. In addition,the SEM images are significantly performed that the corrosion of RP residue surfaces leached by 1.0% DES-induced At.f is much worse than that of leached by the adapted At. f. All the above indicate that the leaching efficiency of soluble P from RP with pyrite can be greatly improved by using DES-induced At. f to a certain extent.

Electrochemical mechanism of rusticyanin (Rus.) isolated from A.ferrooxidans measured by Rus.-ZnS-QDs/L-Cys/Au electrode

Electrochimcal behaviors of rusticyanin （Rus.） isolated from Acidithiobacillus ferrooxidans were investigated through Rus.-ZnS-QDs/L-Cys/Au electrode. The cyclic voltammetric results indicate that rusticyanin immobilized on the surface of Rus.-ZnS-QDs/L-Cys/Au electrode can undergo a direct quasi-reversible electrochemical reaction. The immobilized rusticyanin is not denatured and still retains its activity in the temperature range of 19-43 ℃. The reduction ability of the protein increases and its oxidation ability becomes weak with the increase of pH from 6.0 to 7.8. Fe^2＋ ions in the solution can promote the electron transfer kinetics of the immobilized rusticyanin and make its peak potentials （φp） markedly move negatively.