• Journal of Microbiology and Biotechnology
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• v.11 no.6
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• pp.1046-1054
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• 2001

Bioleaching is the process in which insoluble metal sulfide is oxidized by specialized iron- and/or sulfur-oxidizing lithotrophic bacteria in acidic, metal-rich environments. Most of these processes are carried out by the genus Thiobacillus. Three novel Thiobacillus strains (Thiobacillus thiooxidans AZ11, Thiobacillus thiooxidans MET, and thiobacillus thiooxidans TAS) associated with bioleaching have been isolated from soil and sludge (Korean patent No. 1999-0073060 for T. thiooxidans AZ11, Korean patent No. 1999-0005798 for T. thiooxidans MET, and Korean patent No. 1999-0073059 for T. thiooxidans TAS). A partial sequence of 16S ribosomal RNA gene (16S rDNA) and the entire sequence of 16S/23S intergenic spacer region (ISR) were determined in the three above novel strains and in Thiobacillus ferrooxidans ATCC19859 as a reference strain. When phylogenetic analysis was performed based on G+C contents and sequence alignments, T. ferroxidans ATCC19859 was found to be closely related to previously registered T. ferrooxidans strains in a monophyletic manner, while the three novel T. thiooxidans strains were classified in a paraphyletic manner. Close examination on the base composition of 16S/23S ISR revealed that the 5\` part (nucleotide residues 21-200) was specific for the genus Thiobacillus. On the other end, the 3\` part (nucleotide residues 201-520) showed specificity in T. ferrooxidans strains, but not in T. thiooxidans strains. These results suggest that the proximal and distal halves of 16S/23S could be used as a genetic marker for the identification of the genus Thiobacillus and the species T. ferrooxidans, respectively.

• Journal of Microbiology and Biotechnology
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• v.14 no.1
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• pp.56-61
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• 2004

A plasmid pSDK-l containing the Escherichia coli phosphofructokinase-l gene (pfkA) was constructed, and transferred into extremely acidophilic Acidithiobacillus thiooxidans Tt-7 by conjugation with the aid of plasmid RP4 at a frequency of $10^{-5}$ per recipient. This plasmid was stable in A. thiooxidans. The pfkA gene from E. coli could be expressed in this obligately autotrophic bacterium, but the enzyme activity (21.6 U/g protein) was lower than that in E. coli (K12: 85.9 Dig protein; DF1010 carrying plasmid pSDK-l: 96.6 U/g protein). In the presence of glucose, the Tt-7 transconjugants consumed glucose, leading to a better growth yield.

• Microbiology and Biotechnology Letters
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• v.34 no.4
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• pp.338-341
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• 2006

The growth characteristics of sulfur-oxidizing bacteria, Acidithiobacillus thiooxidans AZ11, MET, and TAS were investigated in mineral salt media supplemented with elemental sulfur of 1$\sim$50 g $L^{-1}$. The sulfur oxidation rates of A. thiooxidans. MET and TAS increased highly with increasing sulfur concentration up to 10 g L$^{-1}$, but the rates increased slowly in sulfur concentration over 10 g L$^{-1}$. A. thiooxidans AZ11 showed the parallel increase of sulfur oxidation rate until sulfur concentration increased up to 40 g L$^{-1}$. The maximum sulfur oxidation rates (V$_{max}$) of AZl1, MET and TAS were 1.88, 1.38 and 0.43 g S L$^{-1}$ d$^{-1}$, respectively. The maximum specific growth rates (${\mu}_{max}$) of AZ11, MET, and TAS were 0.33 d$^{-1}$, 0.30 d$^{-1}$ and 0.45 d$^{-1}$, respectively. Although MET and TAS couldn't grow at sulfate concentration of 40 g L$^{-1}$, AZ11 could grow in the presence of 58 g L$^{-1}$ sulfate, the final oxidation product of elemental sulfur.

• KSBB Journal
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• v.16 no.4
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• pp.389-397
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• 2001

The effect of sludge digestion on the leaching efficiency of heavy metals from sludge by bioleaching with Thiobacillus thiooxidans MET was investigated. The used sludges were a non- and anaerobically digested. The leaching efficiency of heavy metals was strongly dependent on the pH of the sludge solution rather than the sludge condition and stolid concentration. The lower the pH the more heavy metal was leached from 3.0 of pH. The sequent orders of leaching heavy metals were Zn, Cu, and Cr. Although the buffering capacity of non-digested sludge was similar to anaerobically digested sludge, the pH decrease rate of the anaerobically digested sludge solution was faster than that of the non-digested sludge solution due to the higher sulfur oxidation rate of T. thiooxidans MET in the anaerobically digested sludge. The amount of leached heavy metals from the anaerobically digested sludge showed higher than that of non-digested sludge at the same pH value. This result might be caused by the difference of the insoluble metal types, which were contained in the sludge. An increase in sludge solids concentration decreased the leaching efficiency of heavy metals in the range of solids concentration 10 g/L to 70 g/L. The optimum ratio of S° to sludge stolid was 0.1 in both the sludge. The bioleaching process of heavy metals with T. thiooxidans MET showed the disinfecting effect over 90% as well as the reduction effect in sludge weight of 20%.

• Journal of Microbiology and Biotechnology
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• v.34 no.3
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• pp.525-537
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• 2024

Pilins are protein subunits of pili. The pilins of type IV pili (T4P) in pathogenic bacteria are well characterized, but anything is known about the T4P proteins in acidophilic chemolithoautotrophic microorganisms such as the genus Acidithiobacillus. The interest in T4P of A. thiooxidans is because of their possible role in cell recruitment and bacterial aggregation on the surface of minerals during biooxidation of sulfide minerals. In this study we present a successful ad hoc methodology for the heterologous expression and purification of extracellular proteins such as the minor pilin PilV of the T4P of A. thiooxidans, a pilin exposed to extreme conditions of acidity and high oxidation-reduction potentials, and that interact with metal sulfides in an environment rich in dissolved minerals. Once obtained, the model structure of A. thiooxidans PilV revealed the core basic architecture of T4P pilins. Because of the acidophilic condition, we carried out in silico characterization of the protonation status of acidic and basic residues of PilV in order to calculate the ionization state at specific pH values and evaluated their pH stability. Further biophysical characterization was done using UV-visible and fluorescence spectroscopy and the results showed that PilV remains soluble and stable even after exposure to significant changes of pH. PilV has a unique amino acid composition that exhibits acid stability, with significant biotechnology implications such as biooxidation of sulfide minerals. The biophysics profiles of PilV open new paradigms about resilient proteins and stimulate the study of other pilins from extremophiles.

• Korean Journal of Environmental Biology
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• v.17 no.2
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• pp.183-190
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• 1999

The bioleaching of heavy metals from fly ash was performed by Thiobacillus thiooxidans MET isolated from the enrichment culture of an anaerobically digested sludge. The effect of solid concentrations on the efficiency of metal leaching was studied in shaken flasks. In the range of solid concentrations 20 g.L­$^1$to 100 g.L­$^1$T. thiooxidans MET oxidized S$^{0}$ to sulfate without any lag period. The final pH of slurry solution was decreased to below pH 1, and the final oxide-redox potential (ORP) was increased to over 420 mV in the solid concentrations below 100 g.L­$^1$. However, the initial lag period of 4 to 8 days was required to obtain the pH reduction and ORP increase of the slurry solutions in the range of solid concentrations 150 g.L­$^1$to 300 g.L­$^1$. The sulfur oxidation rate of T. thiooxidans MET in 20~100 g.L­$^1$solid concentrations was 0.70~0.75 g-S.L­$^1$ㆍ d­$^1$, but its sulfur oxidation activity was remarkably inhibited with increasing solid concentration over 150 g.L­$^1$. Increasing fly ash solids concentration in the range of solids concentration 20 g.L­$^1$ to 200 g.L­$^1$decreased the removal efficiency of Zn, Cu, Mn, Cr and Pb. The solubilization of heavy metals from fly ash was strongly correlated with the pH value of slurry solution. When the pH of slurry solution was reduced to 3, the solubilization process of Zn, Cu and Mn started, and their solubilization efficiency of Zn, Cu and Mn was progressively increased below pH 2. However, the solubilization process of Cr and Pb started at pH 2.5 and 2.0, respectively.

• KSBB Journal
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• v.15 no.2
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• pp.145-149
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• 2000

Methyl mercaptan oxidase was isolated and purified from Thiobacillus thiooxidans KCTC2505 for the detection of mercaptans. T The produre of purification involved DEAE-Sephacel and Superose 12 column chromatographies with recovery yields of 40 a and 6.3%, and specific activity of 19.7 and 80.1 units/mg-protein, respectively. The molecular weight of purified methyl m mercaptan oxidase was determined to be 68.1 kDa by SDS-PAGE. The extract from DEAE-Sephacel column chromatography h had a high activity in oxidizing methyl mercaptan to produce formaldehyde which can be easily detected by purpald-coloring m method. Optimum temperature for activity was observed at $43^{\circ}C$. This enzyme was activated by $NH_4CI and (NH_4)_2S0_4$, and | inhibited by KCI and NaC!.

• Resources Recycling
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• v.30 no.2
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• pp.14-23
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• 2021

To investigate the potential for leaching of heavy metals by bacteria from ores stacked on actual mining sites, leaching tests of a complex metallic ore (Pb-Zn-As ore) were conducted over 60 days using acidophile bacteria Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans under initial acidic conditions. Initially, a small amount of heavy metals was leached due to the initial acidic conditions. After 20 days, when A. thiooxidans in the reactor was adapted to the ore, the amount of leached heavy metals rapidly increased; the concentrations of leached arsenic, iron, and zinc reached a maximum of 2800, 3700, and 2500 mg/L, respectively. On the other hand, in the presence of A. ferrooxidans or in the control test without bacteria, heavy metals, except zinc, were barely detected in leaching. Through this study, it was confirmed that (i) bacteria could leach heavy metals at mining sites under acidic conditions and (ii) leaching of heavy metals from a high arsenic-containing ore by A. thiooxidans was more significant than that by A. ferrooxidans.

• Bulletin of the Korean Chemical Society
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• v.28 no.10
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• pp.1777-1780
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• 2007

The synthesis of well-defined dendrons based on aliphatic polyether dendritic cores and glassy polystyrene peripheries is described. The synthetic route involves a combination of living anionic polymerization and a stepwise convergent method consisting of iterative Williamson etherification and hydroboration/oxidation reactions. On the basis of molecular weight, as characterized by gel permeation chromatography (GPC), the first generation dendron (Generation-1) shows a random coil conformation like a linear polystyrene, while higher generations (Generation-2 and 3) reveal globular forms in solution.

• Korean Journal of Microbiology
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• v.10 no.4
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• pp.155-166
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• 1972

With respect to a bacterium isolated some of the similarities and differences previously indicated between Thiobacillus thiooxidans and Thiobacillus concretivorus wee confirmed and the bacteria also provided considerable diferences compared to the two species in some repects. Severe precipitation of sulfur occurred in colonies and in liquid media of the organism isolated. The organism isolated utilized nitrate and asparagine as well as ammonium sulfate as a sole nitrogen source and grew well without being nearly inhibited by citrate and malate. This organism also showed the resistance to heats and external physical stimulations. Owing to some characters decribed above and the reports proposed, the organism isolated could be concluded to be Thiobacillus concretivorus and it was suggested that Thiobacillus concretivorus might be an apparently different species from Thiobacillus thiooxidans.