Phenotypic and genetic characterization of a novel strain of Acidithiobacillus ferrooxidans （AF2）

A comparative study on the phenotypic and genetic characteristics among Acidithiobacillus ferrooxidans (AF2), a typic strain ATCC23270 and a previously isolated strain AF3 was performed. AF2 can use ferrous ion (Fe²⁺) or elemental sulfur (S⁰) as sole energy source, but oxidizes S⁰ more effectively than Fe²⁺, which is different from ATCC23270 and AF3. The G+C content of AF2 is 51.8% (molar fraction), however, ATCC23270 and AF3 strains have G+C content of 63.7% and 64.8% (molar fraction), respectively. The DNA-DNA hybridization results show that AF2 has 41.53% and 52.38% genome similarity to ATCC 23270 and AF3, respectively, but AF3 has a high genome similarity of 89.86% to ATCC 23270 strain. Rusticyanin (rus) and subunit III of aa₃-type cytochrome oxidase (coxC) genes are not detected in AF2, but Fe²⁺ oxidase (iro) gene can be detected. To understand the genomic organization of iro gene, a cosmid library of AF2 genome was constructed and iro gene-containing clone was screened. The sequencing result shows that although the nucleotide sequence of iro gene in AF2 is completely identical to that of ATCC 23270 strain, its genomic organization is different from that of ATCC 23270. In AF2, iro is located at downstream of purA gene, while it is located at downstream of petC-2 gene in ATCC 23270 strain. These results indicate that AF2 is a novel strain of A. ferrooxidans, and that phenotypic differences among the strains of A. ferrooxidans are closely correlated with their genetic polymorphisms.     

Isolation and characterization of YNTC-1, a novel Alicyclobacillus sendaiensis strain

A heterotrophic acidothermophilic bacterial strain, YNTC-1, was isolated from an acidic hot spring in Tengchong, Yunan, China. YNTC-1 grows at pH value of 1.5-8.0 and temperature of 40-70 ℃, with optimal pH and temperature at 3.0 and 55 ℃, respectively. The cells of the strain are in shape of short rod, with 1.0-1.2 μm in length and 0.7-0.8 μm in diameter, and with distinct spores at both poles of each cell. The predominant fatty acids in cellular membrane of the strain are C18：1 ω7c. 16s rRNA gene analysis reveals that this strain is closely related to Alicyclobacillus sendaiensis, with over 99% sequence similarity. Based on phenotypic and genotypic analyses, YNTC-1 is identified as a member ofA. sendaiensis. Considering some important morphological and biochemical differences between strain YNTC-1 and A. sendaiensis ATCC 27009T, YNTC-1 may be proposed to be a novel subspecies of A. sendaiensis. However, this viewpoint has to be confirmed by further studies. Co-bioleaching of pyrite and chalcopyrite with strain YN22, Sulfobacillus thermosulfidooxidans, shows that strain YNTC-1 has no evident influence on bioleaching rates of these two sulphide minerals.     

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. Foundation item: Project(50621063) supported by the Science Fund for Creative Research Groups of China; Project(2004CB619201) supported by the Major State Basic Research Development Program of China     

Micaceous iron oxide prepared from pyrite cinders by hydrothermal method

Micaceous iron oxide (MIO) with a hexagonal flaky shape was prepared by hydrothermal method. The ferric hydroxide used as precursor was obtained by an acidic leaching solution of pyrite cinders reacting with ammonia solution. The optimal experimental conditions for preparing micaceous iron oxide were investigated by orthogonal experiments. Micaceous iron oxide can be successfully prepared when optimal parameters of total iron concentration of 2.0 mol/L, pH value of 8, n(Fe²⁺)/n(Fe³⁺) of 0.1, mass of seed crystal of 1 g, reaction temperature of 260 °C and reaction time of 30 min are applied. X-ray diffractometry (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM) and selected area electron diffractometry (SAEM) were adopted to characterize the hydrothermal products prepared under optimal conditions. The results indicate that highly crystallized α-Fe₂O₃ hexagonal flakes, about 1.0–1.5 μm in diameter and 0.1 μm in thickness, are prepared. Furthermore, the quality of micaceous iron oxide prepared can meet the required characteristics of micaceous iron oxide pigments for paints (ISO10601—2007).     

Thermodynamic study on reaction path of Hg（ II ） with S（ II ） in solution

The mercury sulfidation experiments were conducted in the pH range from 1 to 13. The results show that Hg(Ⅱ) reacted with equimolar S(Ⅱ) has the lowest remained Hg(Ⅱ) concentration (9.7 μg/L) at pH 1.0 and the highest remained concentration (940.8 ug/L) at pH 13.0. Meanwhile, the changes of pH values were monitored exactly, which reveal that solution pH values change when mixing the same pH value solutions of HgCl_2 and Na_2S. In order to explain the phenomena and determine the reaction paths of Hg(Ⅱ) reacting with S(Ⅱ) in the solution, the concerned thermodynamics was studied. Species of S(Ⅱ)-H_2O system and Hg(Ⅱ)-H_2O system at different pH values were calculated, and then the species distribution diagrams of S(Ⅱ)-H_2O system, Hg(Ⅱ)-H_2O system and Hg(Ⅱ)-Cl~-0H~~-H_2O system were drawn. Combining the experimental data and thermodynamic calculation, the mechanism of Hg(Ⅱ) reacting with S(Ⅱ) was deduced. The results indicate that different species of S(Ⅱ) and Hg(Ⅱ) have the diverse reaction paths to form HgS precipitate at different pH values and the standard Gibbs free energies change(△_rG_m~Θ) of those equations are also calculated, which can provide a guidance for mercury-containing wastewater treatment with Na_2S.     

Citrate-stabilized CdSe/CdS quantum dots as fluorescence probe for protein determination

A rapid, ultrasensitive and convenient fluorescence measurement technology based on the enhancement of the fluorescence intensity resulting from the interaction of functionalized CdSe/CdS quantum dots (QDs) with bovine serum albumin (BSA) was proposed. The citrate-stabilized CdSe/CdS (QDs) were synthesized by using Se powder and Na₂S as precursors instead of any pyrophoric organometallic precursors. The modified CdSe/CdS QDs are brighter and more stable against photobleaching in comparison with organic fluorophores. At pH 7.0, the fluorescence signal of CdSe/CdS is enhanced by increasing the concentration of BSA in the range of 0.1–10 μg/mL, and the low detection limit is 0.06 μg/mL. A linear relationship between the enhanced fluorescence peak intensity (ΔF) and BSA concentration (c) is established using equation ΔF=50.7c+16.4 (R=0.996 36). Results of determination for BSA in three synthetic samples are identical with the true values, and the recovery (98.9%–102.4%) and relative standard deviation (RSD, 1.8%–2.5%) are satisfactory.     

Isolation of Leptospirillum ferriphilum by single-layered solid medium

According to physiological and biochemical characteristics of Leptospirillum ferriphilum,a strain of object bacteria was isolated successfully.Bacteria were enriched by selective liquid medium and plated on designed single-layered agar solid medium.Colony was cultured and bacteria were collected.The morphologies of the object bacteria were observed using crystal violet staining,scanning electron microscope(SEM)and transmission electron microscope(TEM).The result of 16S rDNA identification shows that this bacterium belongs to Leptospirillum ferriphilum and it is named as Leptospirillum ferriphilum strain D1.These results indicate that this new single-layered agar solid medium is efficient and simple for isolation of Leptospirillum ferriphilum.Additionally,physiological-biochemical characteristics show that the optimum initial pH value and its growth temperature are 1.68 and 40 ℃,respectively.The culture of it is used to leach a complex concentrate chalcopyrite,the leaching efficiencies of copper and iron are 1.93 % and 13.74 %,respectively,and it is more effective than the A.ferrooxidans culture in the leaching of the complex concentrate chalcopyrite.     

Influence of copper solvent extractant on microbial community structure of acidophilic microorganisms

The influence of different concentrations of copper solvent extractant ZJ 988 on the growth and activity of acidophilic microorganisms was studied and the microbial community structures were compared by 16S rRNA gene clone library analysis. The total bacteria numbers are reduced when 0.5% (volume fraction) extractant is added. The proportions of Acidithiobacillus ferrooxidans and Acidiphilium organovorum are increased, whereas the proportion of Leptospirillum ferriphilum is reduced. When the concentration of extractant is elevated to 1%, growth of all bacteria is inhibited. Clone library results reveal that the dominant bacteria in the culture solution with/without the extractant are At. ferrooxidans, A. organovorum and L. ferriphilum. The sensitivity order of the three bacteria to the extractant from the most to the least is found to be L. ferriphilum>At. ferrooxidans>A. organovorum.     

Screening, identification and desilication of a silicate bacterium

The strain Lv（1- z） isolated from the Henan bauxite was characterized by morphological observation, biochemical and physiological identification, and 16S rDNA sequence analysis. The influences of temperature, initial pH value, the volume of medium, shaking speed and illite concentration on the desilicating ability of the strain Lv（1- z） were investigated. The results show that the bacterium is a Gram-negative rod-shaped bacterium with oval endspores and thick capsule, but without flagellum. The biochemical and physiological tests indicate that the strain Lv（1- z） is similar to Bacillus rnucilaginosus. In GenBank the 16S rDNA sequence similarity of the strain Lv（1- z） and the B. rnucilaginosus YNUCC0001（AY571332） is more than 99%. Based on the above results, the strain Lv（1- z） is identified as B. rnucilaginosus. The optimum conditions for the strain L（1- z） to remove silicon from illite are as follows., temperature is 30℃ ;initial pH value is 7.5; medium volume in 200 mL bottle is 60 mL; shaking speed of rotary shaker is 220 r/m ; illite concentration is 1%.     

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 °C.The reduction ability of the protein increases and its oxidation a...     

Production and characterization of alkaline extracellular lipase from newly isolated strain Aspergillus awamori HB-03

A strain HB-03 to produce alkaline extracellular lipase was isolated from oil-rich soil samples and identified as Aspergillus awamori.The growth conditions and nutritional factors for lipase production by strain HB-03 were optimized,and the maximum lipase production of (45.9±2.3) U/mL was obtained at 30 °C and pH 7.0 after 36 h using olive oil (1%) and sucrose (0.5%) as carbon sources and combination of peptone (2%),yeast extract (0.5%) and ammonium sulfate (0.1%) as nitrogen sources.The lipase was purified...     

Homology modeling and evolutionary trace analysis of superoxide dismutase from extremophile Acidithiobacillus ferrooxidans

The gene sod in Acidithiobacillus ferrooxidans may play a crucial role in its tolerance to the extremely acidic, toxic and oxidative environment of bioleaching. For insight into the anti-toxic mechanism of the bacteria, a three-dimensional (3D) molecular structure of the protein encoded by this gene was built by homology modeling techniques, refined by molecular dynamics simulations, assessed by PROFILE-3D and PROSTAT programs and its key residues were further detected by evolutionary trace analysis. Through these procedures, some trace residues were identified and spatially clustered. Among them, the residues of Asn38, Gly103 and Glu161 are randomly scattered throughout the mapped structure; interestingly, the other residues are all distinctly clustered in a subgroup near Fe atom. From these results, this gene can be confirmed at 3D level to encode the Fe-depending superoxide dismutase and subsequently play an anti-toxic role. Furthermore, the detected key residues around Fe binding site can be conjectured to be directly responsible for Fe binding and catalytic function. Foundation item: Project(2004CB619201) supported by the National Basic Research Program of China; Project (50321402) supported by the National Natural Science Foundation of China     

A Series Piezoelectric Quartz Crystal Response Kinetics for T.ferrooxidans Growth in the Presence of Cu(Ⅱ)

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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.     

Impact of strain relaxation of AlGaN barrier layer on the performance of high Al-content AlGaN/GaN HEMT

Owing to large direct bandgap energy, high saturation drift velocity, large conduction band discontinuities, high thermal stability and strong piezoelectric and spontaneous po- larization[1-4], AlGaN/GaN HEMTs have advantages over electronic devices based on Si, GaAs and their alloys in high-frequency, -temperature and -power applications. Many researches show[5,6] that the characteristics of AlGaN/GaN HEMTs depend on two-dimensional gas (2DEG) in heterostructures which has intimate r…     

高磷赤铁矿两步法生物除磷实验研究

采用嗜酸氧化硫硫杆菌(At.t)直接浸出和两步法浸出,对鄂西高磷鲕状赤铁矿(铁品位43.50%,磷含量0.85%)进行生物除磷的实验研究。结果表明:矿浆浓度为2%时,At.t菌直接浸出除磷率为62.35%,且硫含量高达28.57%;采用两步法摇床培养At.t菌,24d菌液pH值接近0.8,磷含量可降至0.15%,硫含量为1.09%;采用自行设计制作的生物反应器培养At.t菌8d,菌液pH值接近0.98,分离菌液浸出原矿12h磷含量为0.18%。对磁选精矿进行的两步法浸出表明,当矿浆浓度为3%以下时菌液的除磷效果明显。     

Modeling and process optimization for microbial desulfurization of coal by using a two-level full factorial design

The microbial sulfur removal was investigated on high sulfur content (1.9%) coal concentrate from Tabas coal preparation plant. A mixed culture of ferrooxidans microorganisms was isolated from the tailing dam of the plant. Full factorial method was used to design laboratory test and to evaluate the effects of pH, particle size, iron sulfate concentration, pulp density, and bioleaching time on sulfur reduction. Statistical analyses of experimental data were considered and showed increases of pH and particle size had negative effects on sulfur reduction, whereas increases of pulp density and bioleaching time raised microbial desulfurization rate. According to results of designing, and regarding statistical factors, the optimum values for maximum sulfur reduction were obtained; pH (1.5), particle size (?180 μm), iron sulfate concentration (2.7 mmol/L), pulp density (10%) and bioleaching time (14 d), which leaded to 51.5% reduction from the total sulfur of sample.     

Regulation mechanism of EM parameters in natural ferrite and its application in microwave absorbing materials

1 Introduction Microwave absorbing materials (MAM) refer to a kind of materials that can be used to absorb the emitted electromagnetic (EM) energy and to minimize the wave reflected in the direction of an energy radar receiver. When the EM wave incidents into MAM, it is readily absorbed, attenuated, and changed into heat or other energy. MAM are functional materials that possess special performance for absorbing EM wave. Along with the advancement in radar and microwave technologies, MA…     

Identification and fermentation optimization of protopectinase-overproducing strain Aspergillus niger CD-01 for pectin production

In order to solve the citrus peel resource waste problem and minimize the drawbacks of chemical extraction of pectin, a protopectinase-overproducing strain CD-01 for pectin production was isolated from a pit soil dumped with perished orange in Changde City, Hunan Province of China. The strain CD-01 had the same morphology and 28S rRNA gene sequence (FJ184995) as that of Aspergillus niger (ATCC 64028). It was thus identified and named as Aspergillus niger CD-01. The fermentation condition was optimized based on L₉(3⁴) orthogonal experimental design and the variances analyses. The results show that the optimal condition for producing pectin is as follows: time 36 h, temperature 35 °C, pH 5, and urea as the nitrogen source. Under this condition, the pectin yield can reach up to 24.5%. This shows a great potential of Aspergillus niger CD-01 in pectin extraction from citrus. Foundation item: Projects(50621063, 50674101) supported by the National Natural Science Foundation of China     

Effect of tin on the reaction synthesis of ternary carbide Ti3AlC2

The effect of tin on synthesis of Ti₃AlC₂ by spark plasma sintering (SPS) from TiC/Ti/Al powders was investigated. X-ray diffraction (XRD) and scanning electron microscopy (SEM) were used for phase identification and microstructure evaluation. The experimental results show that addition of tin can considerably accelerate the synthesis reaction of Ti₃AlC₂ and fully dense, essentially single-phase polycrystalline Ti₃AlC₂ could be successfully obtained by sintering 2TiC/1Ti/1Al/0.2Sn powders at 1200–1250 °C under a pressure of 30 MPa. SEM images show that Ti₃AlC₂ samples in about 2–5 μm thick and 10–25 μm long platelets can be obtained. The fracture toughness and flexural strength of Ti₃AlC₂ were 6.5±0.2 MPa·m^1/2 and 560±10 MPa, respectively. Funded by the National Natural Science Foundation of China (No.20771088, No.50572080) and Doctoral Foundation of Wuhan University of Technology (No.471-38650142)