Development and evaluation of whole-genome oligonucleotide array for Acidithiobacillus ferrooxidans ATCC 23270

To effectively monitor the characteristic of Acidithiobacillus ferrooxidans ATCC 23270 at the whole-genomic level,a whole-genome 50-mer-based oligonucleotide microarray was developed based on the 3 217 ORFs of A.ferrooxidans ATCC 23270 genome.Based on artificial oligonucleotide probes,the results showed that the optimal hybridization temperature was 45℃. Specificity tests with the purified PCR amplifications of 5 genes(Sulfide-quinone reductase,Cytochrome C,Iron oxidase,Mercuric resistance protein,Nitrog...     

Repetitive sequence based polymerase chain reaction to differentiate close bacteria strains in acidic sites

To study the diversity of bacteria strains newly isolated from several acid mine drainage（AMD） sites in China, repetitive sequence based polymerase chain reaction （rep-PCR）, a well established technology for diversity analysis of closely related bacteria strains, was conducted on 30 strains of bacteria Leptospirillum ferriphilium, 8 strains of bacteria Acidithiobacillus ferrooxidans, as well as the Acidithiobacillusferrooxidans type strain ATCC （American Type Culture Collection） 23270. The results showed that, using ERIC and BOX primer sets, rep-PCR produced highly discriminatory banding patterns. Phylogenetic analysis based on ERIC-PCR banding types was made and the results indicated that rep-PCR could be used as a rapid and highly discriminatory screening technique in studying bacterial diversity, especially in differentiating bacteria within one species in AMD.     

Mutagenic breeding of silver-resistant Acidithiobacillusferrooxidans and exploration of resistant mechanism

The silver-resistant Acidithiobacillus ferrooxidans were isolated from 22 acid mine drainage （AMD） samples collected from Dexing Copper Mine and Chengmen Mountain Mine, Jiangxi Province, China. Isolate DX16 is obtained from the sample taken from Dexing Copper Mine and still carries out ferrous ion oxidation when incubated in 9K medium containing silver nitrate （240 mg/L）. While isolate HI, a less resistant strain taken from Yin Mountain Mine, has a tolerate level of only 60 mg/L. Based on 16S ribosomal DNA sequencing, both bacterial 16SrDNA sequences are 100% similar to Acidithiobacillus ferrooxidans ATCC 23270. Through ultraviolet irradiation induced mutations, isolate mDX16 that is obtained from DX16 carries out ferrous ion oxidation when incubated in 9K medium containing higher concentration of silver nitrate （250 mg/L）. When silver-resistant gene （SilC） analysis is carried out on the two isolates, it is seen that this gene was absent in both.     

表面活性剂Tween-80对Acidithiobacillus ferrooxidans硫氧化和硫代谢相关蛋白质基因表达的影响(英文)

研究了表面活性剂 Tween-80 对 Acidithiobacillus ferrooxidans ATCC 23270 生长、硫氧化和硫代谢相关典型基因表达的影响。结果表明,当培养基中含有 10 2g/L Tween-80 时,A. ferrooxidans 的生长以及其对不溶性底物(S0和 CuFeS2)的代谢得到了促进。在该条件下,经过 24 d 的生物浸出,黄铜矿的铜离子浸出率比对照组(不含 Tween-80)高 16%。FT-IR 光谱分析表明,这可能是由于 Tween-80 的存在而导致胞外多聚物成分发生变化而引起的。用 RT-qPCR 来分析 17 个硫代谢相关基因在 Tween-80 存在时的表达差异。胞外蛋白质基因表达下调表明了Tween-80 对细菌硫吸附作用的影响。硫代谢相关酶基因表达水平的变化为硫代谢的研究提供了参考。     

Liquid-nitrogen cryopreservation of three kinds of autotrophicbioleaching bacteria

Three kinds of autotrophic bioleaching bacteria strains, including mesophilic and acidophilic ferrous ion-oxidizing bacteria Acidithiobacillus ferrooxidans （A. ferrooxidans）, mesophilic and acidophilic sulfur-oxidizing bacteria Acidithiobacillus thiooxidans （A. thiooxidans）, and moderately thermophilic sulfur-oxidizing bacteria Acidianus brierleyi, were cryopreserved in liquid nitrogen and their ferrous ion- or sulfur-oxidizing activities were investigated and compared with the original ones. The results revealed that ferrous ion/sulfur oxidation activities of the strains were almost equal before and after cryopreservation. Glycerin was used as cryoprotective agent. In conclusion, liquid-nitrogen cryopreservation is a simple and effective method for autotrophic bioleaching microorganisms.     

Purification and enzymatic characteristics of cysteine desulfurase, IscS, in Acidithiobacillus ferrooxidans ATCC 23270

A cysteine desulfurase protein,IscS,was encoded by the operon iscSUA in Acidithiobacillus ferrooxidans.The gene of IscS from Acidithiobacillus ferrooxidans ATCC 23270 was cloned and expressed in Escherichia coli.The protein was purified by one-step affinity chromatography to homogeneity.The final protein yield after affinity chromatography was 12.9%.The enzyme was characterized for thermal stability,pH and kinetic parameters.The molecular mass of recombinant IscS was 46 ku by SDS-PAGE.The optimum pH was 8.0-8.5.The enzyme had a temperature optimum at 30 ℃ and was relatively stable at 40 ℃,with 67% loss of activity.1,5-I-AEDANS significantly inhibited IscS activity.Kinetic parameters Km and Vmax were found to be 0.11 mmol/L and 2.57 μmol/(L-min).     

镍黄铁矿电化学生物氧化过程的分解机理(英文)

应用表面粘附和没有粘附Acidithiobacillus ferrooxidans的镍黄铁矿粉末微电极进行电化学测试,以说明镍黄铁矿氧化分解的机理。循环伏安CV结果显示,在-0.2V的低电位区域,在镍、铁离子析出时镍黄铁矿转变为中间相Fe4.5-yNi4.5-xS8-z;当电位在-0.2V到0.2V区间时,有不稳定的紫硫镍矿Fe3Ni3S4和FeNi2S4形成并在表面伴有元素硫的产生;当电位增加到0.2V以上时,不稳定相将全部分解;在高电位0.7V时,析出的亚铁离子被氧化为高铁离子。嗜酸氧化亚铁硫杆菌Acidithiobacillus ferrooxidans的存在使峰电位增高,反应起始电位负移,并对表面形成的元素硫有氧化去除作用。这一过程可通过-0.75到-0.5V电位区间发生的的还原反应证实。生物浸出和电化学实验结果均表明当pH2时溶液酸度的增加对氧化过程有轻度的阻碍作用。     

Bioleaching of anilite by Acidithiobacillus ferrooxidans

In order to characterize the efficiency of copper bioleaching from anilite using pure cultures of Acidithiobacillus ferrooxidans in the absence and presence of ferrous sulphate,the experiments were carried out in shake flasks with or without 4 g/L ferrous sulphate(FeSO_4·7H_2O)at pH 2.0,150 r/min and 35℃.The tests show that Acidithiobacillus ferrooxidans is unable to attack anilite in iron-free 9K medium.Anilite is rapidly oxidized by bacterial leaching when ferrous sulphate is added.Chemical oxidation o...     

Isolation and characterization of acidophilic bacterium from Gaofeng Mine in China

An acidophilic, chemolithotrophic and ferrous oxidizing bacterium strain GF was isolated from the acid mine drainage （AMD） of Gaofeng Mine, Guangxi Province, China using 9K enrichment medium, and then purified on solid ferrous-agarose medium. The physiological experiments show that it can use ferrous or sulfur as sole energy and a low level （0.1%, w/v） of peptone can accelerate the growth of the isolated strain. The optimum pH and temperature for growth are 2.0 and 30 ℃, respectively. The isolated strain shares 99.64% identities of 16S rRNA gene with the type strain Acidthiobacillusferrooxidans ATCC 23270 and 100% identities of iro gene （CDS） with A.ferrooxidans strain Fe-1. These results show that the strain can be considered as A cidthiobacillus ferrooxidans. Because of the high activity of oxidizing ferrous and sulfide mineral, strain GF was used in bioleaching of marmatite. The Zn concentration is 0.273 g/L under the steriled control and 7.30 g/L with adapted GF strain incubated after 29 d in leaching marmatite. The isolated strain GF can be used to leach marmatite in industry application.     

Microbial aspects of acid mine drainage and its bioremediation

The role of chemolithotrophs such as Acidithiobacillus ferrooxidans, Acidithiobacillus thiooxidans and Leptospirillum ferrooxidans which were isolated from some abandoned mines and processed waste tailings in the generation of acid mine drainage and toxic metal dissolution was discussed. Mechanisms of acid formation and dissolution of copper, zinc, iron and arsenic from copper, lead-zinc and arsenopyrite-bearing sulfide ores and railings were established in the presence of Acidithiobacillus group of bacteria. Sulphate Reducing Bacteria（SRB） isolated from the above mine sites could be used to precipitate dissolved metals such as copper, zinc, iron and arsenic. Arsenic bioremediation was demonstrated through the use of native microorganisms such Thiomonas spp. which could oxidize arsenite to arsenate. Bioremoval of arsenic through the use of jarosite precipitates generated by Acidithiobacillus ferrooxidans and Leptospirillum ferrooxidans was also found to be very effective. Biotechnological processes hold great promise in the remediation of acid mine drainage and efficient removal of toxic metal ions such as copper, zinc and arsenic.     

生物浸铀中影响嗜酸氧化亚铁硫杆菌活性的氟毒物活性形态(英文)

为了确定浸矿菌耐氟的机制,在氟化物存在的条件下,驯化铀矿浸出菌嗜酸氧化亚铁硫杆菌ATCC23270,研究溶液中含不同氟浓度、不同pH值时铀矿浸出菌的活性变化,以及有无蛋白酶K处理时铀矿浸出菌细胞内氟浓度的变化情况。采用铂电极和Ag/AgCl参比电极测量氧化还原电位,以作为细菌不同活性的参照指标,采用氟离子选择性电极测定细胞内的氟浓度。结果表明,真正影响铀矿浸出菌活性的是HF,溶液pH值增加以及溶液中与氟有较强络合能力的离子浓度的变化,也会引起耐氟菌假象的出现。浸矿菌的耐氟能力可能与细胞壁和细胞膜上的一些蛋白密切相关。     

浸矿微生物的研究进展

本文综述了主要浸矿微生物的性质、分类地位、培养条件以及在生物冶金方面的应用及研究进展,涉及到的微生物有嗜酸氧化亚铁硫杆菌、嗜酸氧化硫硫杆菌、氧化亚铁钩端螺旋菌、硫化叶菌属及硫化芽孢杆菌属.     

黄钾铁矾的生成机理及其对嗜酸氧化亚铁硫杆菌有重要作用的离子的影响(英文)

黄钾铁矾的生成对Sarcheshmeh生物堆浸硫化铜矿有不利影响。实验研究了在嗜酸氧化亚铁硫杆菌存在的情况下,生长介质中Fe(II)的初始浓度、pH及温度影响黄钾铁矾沉淀形成的机理。产生最多Fe(III)沉淀的条件为:硫酸亚铁浓度50 g/L、初始pH 2.2、温度32°C。Fe(III)沉淀的生成影响了对嗜酸氧化亚铁硫杆菌有重要作用的离子的浓度,比如:Fe3+、SO 2?4、K+、PO 3?4、Mg2+。对于Fe3+和K+,他们有相似的模式,这些离子共沉淀而形成黄钾铁矾的组分。在pH高于1.6时,由于PO 3?4与黄钾铁矾共沉淀以及嗜酸氧化亚铁硫杆菌较快的生长速度而导致合PO 3?4的化合物的溶解度急剧降低。在生物堆浸的初期,由于脉石的溶解,Mg2+浓度增大,随后缓慢降低。     

Extraction and purification of magnetic nanoparticles from strain of Leptospirillum ferriphilum

The magnetic nanoparticles were extracted from Leptospirillum ferriphilum, strain YSK, isolated from acid mine drainages by treatment with sodium dodecyl sulfate(SDS) and centrifugation through a sucrose density gradient. Transmission electron microscopy(TEM) indicates that the nanoparticles are approximately spherical with a mean diameter of 44 nm, and magnetite crystals in this size range are single magnetic domains. Energy-dispersive X-ray analysis shows that the nanoparticles primarily contain two kinds of elements, iron and oxygen. Thus it can be concluded that the magnetic particles are magnetosomes. Generally, it is thought that cellular magnetotaxis is a direct consequence of the cell possessing magnetosomes. The discovery of magnetosomes in strain YSK can provide the theoretical basis for screening efficient bioleaching bacteria which are specific to different magnetic minerals under an outer magnetic field.     

Bioleaching of chalcopyrite by pure and mixed culture

The bioleaching of chalcopyrite in shake flasks was investigated by using pure Acidithiobacillusferrooxidans and mixed culture isolated from the acid mine drainage in Yushui and Dabaoshan Copper Mine in China, marked as YS and DB, respectively. The mixed culture consisted mainly of Acidithiobacillus fOrrooxidans, Acidithiobacillus thiooxidans, and Leptospirillum spp. （Leptospirillum ferriphilum and Leptospirillum ferrooxians）. The results show that the mixed culture is more efficient than the pure Acidithiobacillus ferrooxidans because of the presence of the sulfur-oxidizing cultures that positively increase the dissolution rate and the recovery of copper from chalcopyrite. The pH value decreases with the decrease of chalcopyrite leaching rate, because of the formation ofjarosite as a passivation layer on the mineral surface during bioleaching. In the bioleaching using the mixed culture, low pH is got from the sulfur oxidizing inhibiting, the formation ofjarosite. The copper extraction reaches 46.27% in mixed culture and 30.37% in pure Acidithiobacillusferrooxidans after leaching for 75 d.     

Extraction and purification of magnetic nanoparticles from strain of Leptospirillum ferriphilum

1Introduction From1975,there is much interest in finding magne-totactic bacteria[1],because bacterial magnetosome particles,unlike those produced chemically,have a consistent shape,a unique crystallography,a narrow size distribution within the single magn…     

EPS对Acidithiobacillus.ferrooxidans在黄铜矿与黄铁矿表面吸附的影响

运用超声波结合离心方法提取Acidithiobacillus.ferrooxidans的胞外多聚物（EPS）,用2-酮基-3-脱氧辛酸（KDO）作为表征EPS含量的指标,采用分光光度法对该提取方法进行评估。通过一系列对比性实验研究EPS对Acidithiobacillus ferrooxidans在黄铜矿与黄铁矿表面吸附的影响。将未处理的Acidithiobacillus ferrooxidans与经处理脱去EPS层的Acidithiobacillus ferrooxidans分别与EPS悬液、Fe^2＋和Fe^3＋重新混合,在2h的反应过程中,实时检测混合液中游离的细菌含量。结果表明：细菌表面EPS的存在是其吸附于黄铁矿和黄铜矿表面的一个重要因素。当缺失EPS层时,Acidithiobacillus ferrooxidans吸附于矿物表面的能力有所下降,但当重新加入EPS混合液时这种能力能大部分恢复,这种恢复程度在黄铁矿中较黄铜矿中更加明显。当加入EPS和Fe3＋时其细菌吸附于黄铜矿表面的程度有所升高,而加入Fe2＋时吸附程度明显降低,这个结果表明静电的相互作用也许是细菌最初吸附于矿物表面的一个主要原因,并且这也许是细菌生产EPS的一种驱动力以使细菌吸附于硫化铜矿物后重新获得其吸附能力。