Sulfobacillus Acidophilus Research Articles

Potential of single and designed mixed cultures to enhance the bioleaching of chalcopyrite by oxidation-reduction potential control

Biomining is the extraction of target metals from ores or wastes such as the dissolution of chalcopyrite for copper recovery. A key outstanding topic of study is to improve the rate and total copper released from chalcopyrite that can become ‘passivated’ by surface layers, which hinders oxidative attack on the metal sulfide bond. One strategy to increase chalcopyrite bioleaching is to control of the oxidation-reduction potential in the desired range by using ‘weak’ iron oxidizers. In this study, 15 acidophilic species were evaluated for their ability to catalyze chalcopyrite dissolution that resulted in the addition of Acidithiobacillus ferrianus, Sulfobacillus thermotolerans, and Metallosphaera sedula to the known ‘weak’ iron oxidizing species. Based upon these data, four microbial consortia were designed including mesophiles (25° and 37 °C), moderate thermophiles (49 °C), and thermophiles (70 °C) that increased copper recoveries by up to 32% compared to abiotic controls. The best performing consortium was the moderate thermophiles Sulfobacillus thermotolerans, Sulfobacillus acidophilus, and Ferroplasma acidiphilum that maintained the oxidation-reduction potential in the desired range. However, the consortia also showed evidence of synergistic interactions between ‘weak’ iron oxidizers that increased the efficiency of ferrous iron oxidation that resulted in oxidation-reduction potentials above the desired range and lower copper release. Therefore, while designing microbial consortia is a promising strategy to improve the performance of chalcopyrite bioleaching, care must be taken to ensure synergistic effects do not result in high oxidation-reduction potentials.

Mechanistic insights into iron-sulfur clusters and flavin oxidation of a novel xanthine oxidoreductase from Sulfobacillus acidophilus TPY.

Xanthine oxidoreductase (XOR) catalyzes the oxidation of purines (hypoxanthine and xanthine) to uric acid. XOR is widely used in various therapeutic and biotechnological applications. In this study, we characterized the biophysical and mechanistic properties of a novel bacterial XOR from Sulfobacillus acidophilus TPY (SaXOR). Our results showed that SaXOR is a heterotrimer consisting of three subunits, namely XoA, XoB, andXoC, which denote the molybdenum cofactor (Moco), 2Fe-2S, and FAD-binding domains, respectively. XoC was found to be stable when co-expressed with XoB, forming an XoBC complex. Furthermore, we prepared a fusion of XoB and XoC via a flexible linker (fusXoBC) and evaluated its function in comparison to that of XoBC. Spectroscopic analysis revealed that XoB harbors two 2Fe-2S clusters, whereas XoC bears a single-bound FAD cofactor. Electron transfer from reduced forms of XoC, XoBC, and fusXoBC to molecular oxygen (O2 ) during oxidative half-reaction yielded no flavin semiquinones, implying ultrafast single-electron transfer from 2Fe-2Sred to FAD. In the presence of XoA, XoBC and fusXoBC exhibited comparable XoA affinity and exploited a shared overall mechanism. Nonetheless, the linkage may accelerate the two-step, single-electron transfer cascade from 2Fe-2Sred to FAD while augmenting protein stability. Collectively, our findings provide novel insights into SaXOR properties and oxidation mechanisms divergent from prior mammalian and bacterial XOR paradigms.

An uncommon [K+(Mg2+)2] metal ion triad imparts stability and selectivity to the Guanidine-I riboswitch.

The widespread ykkC-I riboswitch class exemplifies divergent riboswitch evolution. To analyze how natural selection has diversified its versatile RNA fold, we determined the X-ray crystal structure of the Burkholderia sp. TJI49 ykkC-I subtype-1 (Guanidine-I) riboswitch aptamer domain. Differing from the previously reported structures of orthologs from Dickeya dadantii and Sulfobacillus acidophilus, our Burkholderia structure reveals a chelated K+ ion adjacent to two Mg2+ ions in the guanidine-binding pocket. Thermal melting analysis shows that K+ chelation, which induces localized conformational changes in the binding pocket, improves guanidinium-RNA interactions. Analysis of ribosome structures suggests that the [K+(Mg2+)2] ion triad is uncommon. It is, however, reminiscent of metal ion clusters found in the active sites of ribozymes and DNA polymerases. Previous structural characterization of ykkC-I subtype-2 RNAs, which bind the effector ligands ppGpp and PRPP, indicate that in those paralogs, an adenine responsible for K+ chelation in the Burkholderia Guanidine-I riboswitch is replaced by a pyrimidine. This mutation results in a water molecule and Mg2+ ion binding in place of the K+ ion. Thus, our structural analysis demonstrates how ion and solvent chelation tune divergent ligand specificity and affinity among ykkC-I riboswitches.

Study of Ribulose 1, 5-Bisphosphate Carboxylase from Sulfobacillus acidophilus Strain NY-1 Isolated from Lignite Mines

Study of Ribulose 1, 5-Bisphosphate Carboxylase from Sulfobacillus acidophilus Strain NY-1 Isolated from Lignite Mines

Thermostable arginase from Sulfobacillus acidophilus with neutral pH optimum applied for high-efficiency L-ornithine production.

This study aims to use neutral pH optimum arginase as the catalyst for high-efficiency L-ornithine production. Sulfobacillus acidophilus arginase was firstly cloned and overexpressed in Escherichia coli. The purified enzyme was obtained, and the molecular mass determination showed that this arginase was a hexamer. S. acidophilus arginase possessed similarities with the other arginases such as the conserved sequences, purification behavior, and the necessity for Mn2+ as a cofactor. The maximum enzyme activity was obtained at pH7.5 and 70°C. Thermostability and pH stability analysis showed that the arginase was stable at 30-60°C and pH7.0-8.5, respectively. The kinetic parameters suggested that S. acidophilus arginase could efficiently hydrolyze L-arginine. Bioconversion with this neutral pH optimum arginase had the advantages of avoiding producing by-product, high molar yield, and high-level production of L-ornithine. When the bioconversion was performed with a fed-batch strategy and a coupled-enzyme system involving S. acidophilus arginase and Jack bean urease, the final production of 2.87mol/L was obtained with only 1.72mmol/L L-arginine residue, and the molar yield was 99.9%. The highest production record suggests that S. acidophilus arginase has a great prospect in industrial L-ornithine production.

Assessment of microbial diversity and enumeration of metal tolerant autochthonous bacteria from tailings of magnesite and bauxite mines

The magnesite and bauxite mines are a major source for some minerals and metals. The tailings of these magnesite and bauxite mines contains massive amount of Mn. (3221.2 ± 6.51 & 7102.8.4 ± 10.24), Cd (40.95 ± 2.12 & 892.86 ± 8.48), Zn (951.08 ± 4.31 &724.12 ± 3.18), and Pb (425.8 ± 5.21 & 812.13 ± 9.19 mg Kg−1) with alkaline and acid pH respectively. In the microbial diversity analysis, the results 8 (Thiobacillus thiooxidans, Leptospirillum ferrooxidans, Acetobacter methanolicus, T. intermedius, Bacillus cereus, Sulfobacillus acidophilus, Methylobacterium sp, and Thiobacillus ferooxidans) and 12 (Serratia marcescens, Metalogenium symbioticum 1, Metallogenium symbioticum 2, Bacillus alcalophilus, Aminobacter sp, Naumaniella neustonica, Staphylococcus aureus 1, Methylbacillus sp, Pandoraea sputorum, Acenetobacter sp, S. aureus 2, Pseudomonas aeruginosa) bacteria species were isolated from bauxite and magnesite mine tailings. The fungus such as Rhizopus arrhizus, Mucor sp, Aspergillus niger and Penicillium sp were obtained in both acid and alkaline environment. Among this microbial diversity, P. sputorum and B. cereus (identified through 16S rDNA sequencing) were shown better resistance to these four metals for up to 250 mg L−1. The bacterial diversity indexes, concludes that there was least diverse among the three sites. The evenness/equitability index also conform lower level of variation among these sites.

Effects of sulfur dosage and inoculum size on pilot-scale thermophilic bioleaching of heavy metals from sewage sludge

Land application of sewage sludge has received significant attention in recent years but the presence of elevated heavy metals in the sludge limits its land application. The purposes of this study were to investigate the effects of sulfur dosage and inoculum size on the thermophilic bioleaching of heavy metals from sewage sludge in a pilot-scale bioreactor. The microbial communities in this thermophilic bioleaching process were also identified using real-time polymerase chain reaction (real-time PCR). The results showed that the oxidation of sulfur and metal solubilization decreased with the increasing sulfur dosage. When the sulfur dosage was greater than 2% (w/v), the sulfur oxidation and metal solubilization rates decreased, indicating that the thermophilic bioleaching was hindered by high levels of substrate. However, it was found that the efficiency of metal solubilization and solid degradation was increased with the increase of inoculum size in the range from 5% to 20%. At the end of bioleaching, the efficiency of Mn, Zn, Ni, Cu and Cr from the sewage sludge reached 73–100%, 51–60%, 38–52%, 17–43% and 1–38%, respectively, while SS and VSS were degraded by 33–48% and 47–67%, respectively. Based on the analysis of real-time PCR, Sulfobacillus acidophilus was observed to be the predominant species (13–67% of total bacteria), whereas the populations of Sulfobacillus thermosulfidooxidans and Acidithiobacillus caldus were accounted relatively low (<1%).

Cleaner utilization of electroplating sludge by bioleaching with a moderately thermophilic consortium: A pilot study

The semi-pilot scale bioleaching of electroplating sludge by the moderately thermophilic acidophilic consortium was carried out for the first time. During the microbial cultivation, Leptospirillum ferriphilum CS13, Acidithiobacillus caldus S2, and Sulfobacillus acidophilus CS5 could grow rapidly in a 300 L aeration packed reactor, in which the total suspended cell concentration could fluctuate around 3 × 108 cells/mL and the community structure remained relatively stable. During the bioleaching process, the microbial stock solution could effectively leach heavy metals from electroplating sludge in a stirred reactor within a few hours. Meanwhile, the effects of pH, temperature, the quantity of active culture, and liquid-solid ratio on the bioleaching behavior were also investigated. The optimal conditions for electroplating sludge bioleaching were pH 1.5, temperature 45 °C, bacterial liquid ratio 40%, liquid-solid ratio 4:1 L kg−1, and leaching time 5 h. The total removal rate of various heavy metals in electroplating sludge was over 99%. The bioleaching residue was successfully passed the TCLP test, and the total contents of heavy metals in the residue were also well below the regulatory criteria. In addition, the XRD analysis of the bioleaching residue was also confirmed that the moderately thermophilic consortium bioleaching provided a cleaner process than chemical leaching on the removal of the residual fraction metals, which was feasible and attractive for industrial treatment of electroplating sludge.

Electrochemiluminescence for the identification of electrochemically active bacteria

Electrochemically active bacteria (EAB) use extracellular electron transfer (EET) to exchange electron with extracellular acceptors. Previous studies regarding the measurement of EAB were based on either extracellular reduction or oxidation. In this work, we developed a simple electrochemiluminescence (ECL) assay for the identification and detection of EAB. The results of this proposed method revealed that EET of EAB influenced the content of dissolved oxygen and the formation of Ru(bpy)32+• thus leading to qualitative changes of the ECL signal. EAB with the ability of extracellular reduction (such as Shewanella oneidensis MR-1) gave enhanced signal on ECL emission while those displaying the ability of extracellular oxidation (i.e., Sulfobacillus acidophilus) showed the opposite effect on ECL emission, but non-EAB (i.e., Escherichia coli) did not. These changes in ECL intensity were also proportional to the cell density that could be quantitatively detected in the concentration range of (1.1 ± 1) × 105–212 ± 2 CFU/mL (i.e. Shewanella oneidensis MR-1). Moreover, the measurement of the ability of EAB using this approach was in agreement with measurements using the dissimilatory Fe(III) reduction method. Compared to previous reports, this method displayed a continual and steady ECL signal that allowed accurate measurements of EAB. Most important, only a low cell density was needed in this Ru(bpy)32+ - based ECL method, which is beneficial for cell detection.

Characterization of enzymatic properties of two novel enzymes, 3,4-dihydroxyphenylacetate dioxygenase and 4-hydroxyphenylacetate 3-hydroxylase, from Sulfobacillus acidophilus TPY

BackgroundAs an environmental pollutant, 4-hydroxyphenylacetate (4-HPA) was a product of softwood lignin decomposition and was found in industrial effluents from olive oil production. Sulfobacillus acidophilus TPY was a moderately thermoacidophilic bacterium capable of degrading aromatic compounds including 4-HPA. The enzymes involved in the degradation of 4-HPA and the role of this strain in the bioremediation of marine pollutants need to be illustrated.Results3,4-dihydroxyphenylacetate dioxygenase (DHPAO) encoded by mhpB2 and two components of 4-hydroxydroxyphenylacetate (4-HPA) 3-hydroxylase encoded by hpaB and hpaC from S. acidophilus TPY, a moderately thermoacidophilic bacterium, involved in the degradation of 4-HPA possessed quite low amino acid sequence identity (22–53%) with other ever reported corresponding enzymes, which suggest their novelty. These two enzymes were expressed in E. coli and purified to homogeneity. DHPAO activity in E. coli was revealed by spraying with catechol or 3,4-dihydroxyphenylacetate (3,4-DHPA) on the colonies to make them turn brilliant yellow color. DHPAO possessed total activity of 7.81 U and 185.95 U/mg specific activity at the first minute when 3,4-DHPA was served as substrate. DHPAO was a thermophilic enzyme with optimum temperature of 50 °C and optimum substrate of 3,4-DHPA. The small component (HpaC) was a flavoprotein, and both HpaB and HpaC of 4-HPA 3-hydroxylase were NADH-dependent and essential in the conversion of 4-HPA to 3,4-DHPA. 4-HPA 3-hydroxylase possessed 3.59 U total activity and 27.37 U/mg specific activity at the first minute when enzymatic coupled assay with DHPAO was applied in the enzymatic determination.ConclusionsThe ability of this extreme environmental marine strain to degrade catechol and substituted catechols suggest its applications in the bioremediation of catechol and substituted catechols polluted marine environments.

Optimization of Mixed Cultivation of the Moderate Thermophilic Bioleaching Microorganisms for High Cell Density Using Statistical Methodology

The low functional microbial population density in the industrial bioleaching process has been a limiting factor for the high leaching efficiency, making the microbial cultivation and continuous inoculation an alternative for sustaining the microbial activity. In the present experiment, the defined mixed cultivation of Leptospirillum ferriphilum YSK, Sulfobacillus acidophilus TPY, Acidithiobacillus caldus S2, and Ferroplasma thermophilum L1 was evaluated and optimized by Statistical Methodology. Going through the Plackett–Burman experimental design, pH value, temperature, and c(MgSO4·7H2O) were considered as the most significant factors in the defined range. Then, the relationships were analyzed using the steepest ascent design, the central composite design, and finally the response surface methodology. It was suggested that the optimum parameters were pH 1.38, MgSO4·7H2O 0.552 g/L, temperature 44 °C, FeSO4·7H2O 40 g/L, sulfur 8 g/L, yeast 0.02% w/v, (NH4)2SO4 3g/L, K2HPO4 0.5g/L, KCl 0.1g/L, Ca(NO3)2 0.01 g/L, in which allowed total cell density of the microbial community to reach 7.63 × 108 cells/mL in the cultivation period. The lab experiments were routinely undertaken with the expectation that the L. ferriphilum YSK, S. acidophilus TPY, A. caldus S2, F. thermophilum L1 could rapid grown from initial cell density of 0.25 × 107 cells/mL to 2.82 × 108 cells/mL, 1.68 × 108 cells/mL, 2.76 × 108 cells/mL, 2.51 × 107 cells/mL, respectively in 58 h. It demonstrates a possibility to co-culture these microbes in a single reactor, providing an efficient way to regenerate of inoculation for biomining process.

Microbiological Redox Potential Control to Improve the Efficiency of Chalcopyrite Bioleaching

ABSTRACTThe effect of controlling the redox potential (Eh) on chalcopyrite bioleaching kinetics was studied as a new aspect of redox control during chalcopyrite bioleaching, and its mechanism was investigated by employing the “normalized” solution redox potential (Enormal) and the reaction kinetics model. Different Eh ranges were established by use of different acidophiles (Sulfobacillus acidophilus YTF1; Sulfobacillus sibiricus N1; Acidimicrobium ferrooxidans ICP; Acidiplasma sp. Fv-AP). Cu dissolution was very susceptible to real-time change in Eh during the reaction. It was found that efficiency of bioleaching of chalcopyrite can be effectively evaluated on the basis of Enormal, since it is normalized for real-time fluctuations of concentrations of major metal solutes during bioleaching. For steady Cu solubilization during bioleaching at a maximum rate, it was important to maintain a redox potential range of 0 ≤ Enormal ≤ 1 (−0.35 mV optimal) at the mineral surface by employing a “weak” ion-oxidizer. This led to a copper recovery of > 75%. At higher Enormal levels (Enormal > 1 by “strong” microbial Fe2+ oxidation), Cu solubilization was slowed by diffusion through the product film at the mineral surface (< 50% Cu recovery) caused by low reactivity of the chalcopyrite and by secondary passivation of the chalcopyrite surface, mainly by jarosite.

Expression, purification and function of cysteine desulfurase from Sulfobacillus acidophilus TPY isolated from deep-sea hydrothermal vent.

The cysteine desulfurase (SufS) gene of Sulfobacillus acidophilus TPY, a Gram-positive bacterium isolated from deep-sea hydrothermal vent, was cloned and over-expressed in E. coli BL21. The recombinant SufS protein was purified by one-step affinity chromatography. The TPY SufS contained a well conserved motif RXGHHCA as found in that of other microorganisms, suggesting that it belonged to group II of cysteine desulfurase family. The recombinant TPY SufS could catalyze the conversion of l-cysteine to l-alanine and produce persulfide, and the enzyme activity was 95μ/μL of sulfur ion per minute. The growth of E. coli BL21 was promoted by over-expressing TPY SufS in vivo or by directly adding recombinantTPY SufS in the medium (4.3-4.5×108cells/mL vs. 3.2-3.5×108cells/mL). Furthermore, the highest cell density of E. coli BL21 when the TPY SufS was over-expressed was about 3.5 times that of the control groups in the presence of sodium thiosulfate. These results indicate that the SUF system as the only assembly system of iron-sulfur clusters not only has significant roles in survival of S. acidophilus TPY, but also might be important for combating with high content of sulfide.

Structural Basis for Ligand Binding to the Guanidine-I Riboswitch

The guanidine-I riboswitch is a conserved RNA element with approximately 2,000 known examples across four phyla of bacteria. It exists upstream of nitrogen metabolism and multidrug resistance transporter genes and alters expression through the specific recognition of a free guanidinium cation. Here we report the structure of a guanidine riboswitch aptamer from Sulfobacillus acidophilus at 2.7Å resolution. Helices P1, P1a, P1b, and P2 form a coaxial stack that acts as a scaffold for ligand binding. A previously unidentified P3 helix docks into P1a to form the guanidinium binding pocket, which is completely enclosed. Every functional group of the ligand is recognized through hydrogen bonding to guanine bases and phosphate oxygens. Guanidinium binding is further stabilized through cation-π interactions with guanine bases. This allows the riboswitch to recognize guanidinium while excluding other bacterial metabolites with a guanidino group, including the amino acid arginine.

Sulfur Metabolism Pathways in Sulfobacillus acidophilus TPY, A Gram-Positive Moderate Thermoacidophile from a Hydrothermal Vent.

Sulfobacillus acidophilus TPY, isolated from a hydrothermal vent in the Pacific Ocean, is a moderately thermoacidophilic Gram-positive bacterium that can oxidize ferrous iron or sulfur compounds to obtain energy. In this study, comparative transcriptomic analyses of S. acidophilus TPY were performed under different redox conditions. Based on these results, pathways involved in sulfur metabolism were proposed. Additional evidence was obtained by analyzing mRNA abundance of selected genes involved in the sulfur metabolism of sulfur oxygenase reductase (SOR)-overexpressed S. acidophilus TPY recombinant under different redox conditions. Comparative transcriptomic analyses of S. acidophilus TPY cultured in the presence of ferrous sulfate (FeSO4) or elemental sulfur (S0) were employed to detect differentially transcribed genes and operons involved in sulfur metabolism. The mRNA abundances of genes involved in sulfur metabolism decreased in cultures containing elemental sulfur, as opposed to cultures in which FeSO4 was present where an increase in the expression of sulfur metabolism genes, particularly sulfite reductase (SiR) involved in the dissimilatory sulfate reduction, was observed. SOR, whose mRNA abundance increased in S0 culture, may play an important role in the initial sulfur oxidation. In order to confirm the pathways, SOR overexpression in S. acidophilus TPY and subsequent mRNA abundance analysis of sulfur metabolism-related genes were carried out. Conjugation-based transformation of pTrc99A derived plasmid from heterotrophic E. coli to facultative autotrophic S. acidophilus TPY was developed in this study. Transconjugation between E. coli and S. acidophilus was performed on modified solid 2:2 medium at pH 4.8 and 37°C for 72 h. The SOR-overexpressed recombinant S. acidophilus TPY-SOR had a -accumulation increase, higher oxidation/ reduction potentials (ORPs) and lower pH compared with the wild type strain in the late growth stage of S0 culture condition. The transcript level of sor gene in the recombinant strain increased in both S0 and FeSO4 culture conditions, which influenced the transcription of other genes in the proposed sulfur metabolism pathways. Overall, these results expand our understanding of sulfur metabolism within the Sulfobacillus genus and provide a successful gene-manipulation method.

A novel NADPH-dependent reductase of Sulfobacillus acidophilus TPY phenol hydroxylase: expression, characterization, and functional analysis.

The reductase component (MhpP) of the Sulfobacillus acidophilus TPY multicomponent phenol hydroxylase exhibits only 40% similarity to Pseudomonas sp. strain CF600 phenol hydroxylase reductase. Amino acid sequence alignment analysis revealed that four cysteine residues (Cys-X 4 -Cys-X 2 -Cys-X 29-35 -Cys) are conserved in the N terminus of MhpP for [2Fe-2S] cluster binding, and two other motifs (RXYS and GXXS/T) are conserved in the C terminus for binding the isoalloxazine and phosphate groups of flavin adenine dinucleotide (FAD). Two motifs (S/T-R and yXCGp) responsible for binding to reduce nicotinamide adenine dinucleotide phosphate (NADPH) are also conserved in MhpP, although some residues differ. To confirm the function of this reductase, MhpP was heterologously expressed in Escherichia coli BL21(DE3) and purified. UV-visible spectroscopy and electron paramagnetic resonance spectroscopy revealed that MhpP contains a [2Fe-2S] cluster. MhpP mutants in which the four cysteine residues were substituted via site-directed mutagenesis lost the ability to bind the [2Fe-2S] cluster, resulting in a decrease in enzyme-specific oxidation of NADPH. Thin-layer chromatography revealed that MhpP contains FAD. Substrate specificity analyses confirmed that MhpP uses NADPH rather than NADH as an electron donor. MhpP oxidizes NADPH using cytochrome c, potassium ferricyanide, or nitro blue tetrazolium as an electron acceptor, with a specific activity of 1.7±0.36, 0.78±0.13, and 0.16±0.06U/mg, respectively. Thus, S. acidophilus TPY MhpP is a novel NADPH-dependent reductase component of phenol hydroxylase that utilizes FAD and a [2Fe-2S] cluster as cofactors.

Phenol degradation by Sulfobacillus acidophilus TPY via the meta-pathway

Due to its toxicity and volatility, phenol must be cleared from the environment. Sulfobacillus acidophilus TPY, which was isolated from a hydrothermal vent in the Pacific Ocean as a moderately thermoacidophilic Gram-positive bacterium, was capable of aerobically degrading phenol. This bacterium could tolerate up to 1300mg/L phenol and degrade 100mg/L phenol in 40h completely at 45°C and pH 1.8 with a maximal degradation rate of 2.32mg/L/h at 38h. Genome-wide search revealed that one gene (TPY_3176) and 14 genes clustered together in two regions with locus tags of TPY_0628-0634 and TPY_0640-0646 was proposed to be involved in phenol degradation via the meta-pathway with both the 4-oxalocrotonate branch and the hydrolytic branch. Real-time PCR analysis of S. acidophilus TPY under phenol cultivation condition confirmed the transcription of proposed genes involved in the phenol degradation meta-pathway. Degradation of 3-methylphenol and 2-methylphenol confirmed that the hydrolytic branch was utilised by S. acidophilus TPY. Phylogenetic analysis revealed that S. acidophilus TPY was closely related to sulphate-reducing bacteria and some Gram-positive phenol-degrading bacteria. This was the first report demonstrating the ability of S. acidophilus to degrade phenol and characterising the putative genes involved in phenol metabolism in S. acidophilus TPY.

Relationship between microbial community dynamics and process performance during thermophilic sludge bioleaching.

Heavy metals can be removed from the sludge using bioleaching technologies at thermophilic condition, thereby providing an option for biotreatment of wasted sludge generated from wastewater treatment. The purposes of this study were to establish a molecular biology technique, real-time PCR, for the detection and enumeration of the sulfur-oxidizing bacteria during the thermophilic sludge bioleaching. The 16S rRNA gene for real-time PCR quantification targeted the bioleaching bacteria: Sulfobacillus thermosulfidooxidans, Sulfobacillus acidophilus, and Acidithiobacillus caldus. The specificity and stringency for thermophilic sulfur-oxidizing bacteria were tested before the experiments of monitoring the bacterial community, bacterial number during the thermophilic sludge bioleaching and the future application on testing various environmental samples. The results showed that S. acidophilus was identified as the dominant sulfur-oxidizing bacteria, while A. caldus and S. thermosulfidooxidans occurred in relatively low numbers. The total number of the sulfur-oxidizing bacteria increased during the thermophilic bioleaching process. Meanwhile, the decrease of pH, production of sulfate, degradation of SS/VSS, and solubilization of heavy metal were found to correlate well with the population of thermophilic sulfur-oxidizing bacteria during the bioleaching process. The real-time PCR used in this study is a suitable method to monitor numbers of thermophilic sulfur-oxidizing bacteria during the bioleaching process.

Sulfobacillus thermosulfidooxidans strain Cutipay enhances chalcopyrite bioleaching under moderate thermophilic conditions in the presence of chloride ion.

Currently more than 90% of the world’s copper is obtained through sulfide mineral processing. Among the copper sulfides, chalcopyrite is the most abundant and therefore economically relevant. However, primary copper sulfide bioleaching is restricted due to high ionic strength raffinate solutions and particularly chloride coming from the dissolution of ores. In this work we describe the chalcopyrite bioleaching capacity of Sulfobacillus thermosulfidooxidans strain Cutipay (DSM 27601) previously described at the genomic level (Travisany et al. (2012) Draft genome sequence of the Sulfobacillus thermosulfidooxidans Cutipay strain, an indigenous bacterium isolated from a naturally extreme mining environment in Northern Chile. J Bacteriol 194:6327–6328). Bioleaching assays with the mixotrophic strain Cutipay showed a strong increase in copper recovery from chalcopyrite concentrate at 50°C in the presence of chloride ion, a relevant inhibitory element present in copper bioleaching processes. Compared to the abiotic control and a test with Sulfobacillus acidophilus DSM 10332, strain Cutipay showed an increase of 42 and 69% in copper recovery, respectively, demonstrating its high potential for chalcopyrite bioleaching. Moreover, a genomic comparison highlights the presence of the 2-Haloacid dehalogenase predicted-protein related to a potential new mechanism of chloride resistance in acidophiles. This novel and industrially applicable strain is under patent application CL 2013–03335.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-014-0084-1) contains supplementary material, which is available to authorized users.

Newly identified thermostable esterase from Sulfobacillus acidophilus: properties and performance in phthalate ester degradation.

EstS1, a newly identified thermostable esterase from Sulfobacillus acidophilus DSM10332, was heterologously expressed in Escherichia coli and shown to enzymatically degrade phthalate esters (PAEs) to their corresponding monoalkyl PAEs. The optimal pH and temperature of the esterase were found to be 8.0 and 70°C, respectively. The half-life of EstS1 at 60°C was 15 h, indicating that the enzyme had good thermostability. The specificity constant (kcat/Km) of the enzyme for p-nitrophenyl butyrate was as high as 6,770 mM(-1) s(-1). The potential value of EstS1 was demonstrated by its ability to effectively hydrolyze 35 to 82% of PAEs (10 mM) within 2 min at 37°C, with all substrates being completely degraded within 24 h. At 60°C, the time required for complete hydrolysis of most PAEs was reduced by half. To our knowledge, this enzyme is a new esterase identified from thermophiles that is able to degrade various PAEs at high temperatures.