柱式反应器处理黄铜矿过程中等嗜热微生物群落变化

从中国的多个铜矿取样,在45°C条件下富集获得了一种高效的中等嗜热浸矿富集物,探讨了该富集物在柱式反应器中浸出低品位黄铜矿的pH变化以及与Cu2+浸出的关系,并采用限制性片段长度多态性(RFLP)技术分析了微生物的群落结构和种群动态变化规律。结果表明在整个浸出过程中pH变化较为明显,且一直在1.8以上,60 d内回收了13.6%的铜。RFLP结果表明:在初期,嗜铁钩端螺旋菌(Leptospirillum ferriphilum)在浸出前期占有很高比例(81%),随后逐渐降低,至后期只有13%,而耐温氧化硫化杆菌(Sulfobacillus thermotolerans)和喜温硫杆菌(Acidithiobacillus caldus)的比例逐渐升高,在中期分别达到32%和23%;至末期,耐温氧化硫化杆菌达到了79%,成为优势种群。研究加深了对中等嗜热微生物浸矿特性的了解,也为中等嗜热菌处理低品位黄铜矿的工业应用提供了可供借鉴的数据。     

一株嗜酸硫氧化菌的分离

从江西德兴分离得到一株嗜酸硫氧化杆菌DX-2,采用双层固体培养基进行分离纯化,对分离菌株进行了形态、生理生化特性研究及16SrRNA序列分析.该菌株为革兰氏阴性细菌,短杆状,菌体大小（0.4～0.5）μm×（1～2）μm,化能自养,可利用硫磺和硫代硫酸盐为能源生长,不能利用亚铁进行生长.以16SrRNA序列同源性为基础构建了相关种属在内的系统发育树,结果表明,DX-2与喜温硫杆菌（Acidithiobacillus caldus）处于同一进化树分支中,相似性达99%以上.考察了不同重金属离子对DX-2的生长的影响.     

嗜酸性硫杆菌群体感应系统研究进展

嗜酸性硫杆菌(Acidithiobacillus spp.)是一类重要的极端环境微生物与工业微生物。该类细菌通过氧化硫或亚铁获得电子以固定二氧化碳进行自养生长,是驱动矿山环境酸化和重金属溶出的关键菌群,也是生物冶金等微生物浸出技术中的核心菌群。群体感应(quorum sensing, QS)系统是细菌种内及种间信息交流的重要方式,广泛分布于嗜酸性硫杆菌等化能自养微生物中,比如类似于LuxI/R的AfeI/R系统。系统介绍近年来嗜酸性硫杆菌菌体感应系统研究成果,尤其是在AfeI/R种群分布、生物学功能、调节机制及其应用研究中的新发现与新理论。讨论今后嗜酸性硫杆菌群体感应系统研究的主要方向及需要解决的关键科学问题,以促进极端微生物群体感应系统理论研究的开展与产业应用技术的开发。     

极端嗜酸硫杆菌高效筛选、高密度发酵及保藏方法的研究

【目的】针对嗜酸硫杆菌极端特殊的生化特性,分别建立双层平板培养高效筛选方法和补料分批高密度发酵策略,并优选最佳保藏方法,以强化对该类菌种资源的利用和储备效率。【方法】分别采用以异养型微生物Sacchromyces ellipsoideu和Rhodotorula sp.为底层培养物的双层平板培养嗜酸硫杆菌,并结合透射电子显微镜技术(TEM)考察细胞形态差异。结合硫化矿培养基设计及单质硫补料培养策略,延长Acidithiobacillus thiooxidans对数期,提高比生长速率。分析不同保藏方法对嗜酸硫杆菌细胞存活率的影响。【结果】采用异养微生物——Rhodotorula sp.作为底层培养物的双层平板培养法在缩减1/3检出周期的同时将Acidithiobacillus ferrooxidans和Acidithiobacillus thiooxidans的检出率提高了3倍左右。TEM结果表明双层培养中细胞形态更为规则。采用基于Starkey-硫化矿培养基的补料分批发酵策略提高了Acidithiobacillus thiooxidans平均比生长速率,硫对生物量转化率和生产强度分别比分批培养提高31.1%和187.9%。4°C低温保藏方式更适于嗜酸硫杆菌的保藏,有效保藏期1–3月。【结论】Rhodotorula sp.为辅助培养物的双层平板培养法可有效提高嗜酸硫杆菌的筛选效率。设计的Starkey-硫化矿培养基结合补料分批培养策略可实现Acidithiobacillus thiooxidans高密度培养。简单高效的4°C低温保藏方式更适合于嗜酸硫杆菌的中短期保藏。     

嗜酸硫化杆菌(Sulfobacillus sp.)的分离鉴定及其在黄铁矿浸取中的应用北大核心CSCD

【目的】分离、培养获得中高温硫化矿浸取菌,为利用其进行生物冶金研究及应用奠定基础。【方法】利用二价铁或单质硫为底物,对酸性热泉泥水样品进行富集培养,以获得能够氧化二价铁或还原性硫的菌株;根据形态学、生理生化特点及系统发育分析对分离菌株进行分类鉴定;通过分析黄铁矿的铁氧化速率评估菌种在生物冶金中应用的潜力。【结果】从哥斯达黎加酸性热泉泥水样品中分离得到两株好氧嗜酸兼性自养细菌Costa C和Costa E。两株菌革兰氏染色反应为阳性,细胞大小相近,分别为(0.4-0.6)μm×(2.5-4.0)μm和(0.4-0.7)μm×(2.4-4.9)μm,端生芽胞,生长温度范围均为30℃-55℃,最适生长温度分别为50℃和40℃,生长pH范围分别为1.2-5.0和1.4-5.0,最适生长pH均为2.8。可以利用Fe(Ⅱ)、S、K2S4O6等为能源进行自养生长,也可利用酵母浸粉等有机物生长。两株菌的16S rRNA基因与硫化杆菌属(Sulfobacillus)其他菌种的最高相似性大于99%,(G+C)%含量分别为56.1 mol%和56.7 mol%。【结论】形态学、生理生化特点及系统发育分析表明,研究中筛选获得的两株菌均属于Sulfobacillus属,分别定名为Sulfobacillus sp.strain Costa C和Sulfobacillus sp.strain Costa E。浸矿结果显示两株菌在45℃时可以氧化黄铁矿,氧化速率分别为63.0 mg/L·d与56.8 mg/L·d,表明两株菌具有在中高温条件下浸取硫化矿的能力。     

嗜酸硫杆菌在工农业中的应用

嗜酸硫杆菌(属)(Acidithiobacillus spp.)能够氧化亚铁、硫或还原性无机硫化合物(reduced inorganic sulfur compounds,RISCs)获得能量,固定二氧化碳,是一类典型的嗜酸性化能自养微生物。嗜酸硫杆菌广泛分布于酸性矿水、热泉等酸性环境中,是地球生态系统硫和铁元素循环的主要推动者。嗜酸硫杆菌独特的生理代谢特征和极端环境适应性,使其广泛应用于生物浸出领域。本文综述了嗜酸硫杆菌的生理代谢特征和极端环境下的适应机制,阐述了嗜酸硫杆菌在工农业中的应用,讨论了面向国家重大需求,嗜酸硫杆菌在今后的主要研究方向和需要解决的关键科学问题,为嗜酸硫杆菌在生理代谢、环境适应和工农业应用的研究提供重要的线索和启示。     

三株氧化硫硫杆菌的分离与鉴定

从煤堆废水中分离得到3株嗜温嗜酸硫氧化细菌.这3株菌株为革兰氏阴性、菌体大小0.4～0.7 μm×1～2 μm、短杆状运动细菌,其最适生长温度为 30 ℃和最适生长pH 2.0～2.5.它们能够利用元素硫,硫代硫酸钠和连四硫酸钾为能源进行自养生长,不能利用有机物质以及硫酸亚铁、黄铁矿和黄铜矿等无机物质作为能源生长.细菌的形态、生理生化特性研究以及基于16S rRNA序列同源性构建的系统发育树结果表明,这3株细菌初步鉴定为氧化硫硫杆菌.氧化硫硫杆菌能够通过产酸有效促进黄铜矿的浸出速率和浸出率.     

23株不同地理来源嗜酸硫杆菌的系统发育及其遗传差异

【目的】研究不同地理来源嗜酸硫杆菌的系统发育及其遗传差异,以及基因指纹图谱技术聚类与嗜酸硫杆菌地理来源的相关性。【方法】采用16S-23S r RNA间隔区(ITS)序列建立系统发育树,并结合ERIC和BOXAIR两种引物进行rep-PCR,以及rus基因扩增,对不同地理来源嗜酸硫杆菌进行分析。【结果】分离自不同样点的23株嗜酸硫杆菌遗传差异显著,依据ITS序列系统发育树被划分为5个大类群,与rep-PCR指纹图谱的分类结果较为接近,其中Acidithiobacillus ferrooxidans在ITS系统发育和BOXAIR-PCR指纹聚类分析中被划分为2个类群,但在ERIC-PCR中归为1个类群,rus基因分组中,在系统发育和聚类分析中处于同一类群的菌株拥有不同类型的rus基因,说明嗜酸硫杆菌的亚铁氧化途径与系统发育类群无明显相关性;ITS基因拥有区分近缘种或亚种的能力,且BOXAIR-PCR的分辨能力较强,非常适于嗜酸硫杆菌的遗传差异分析。     

一株中度嗜热嗜酸铁氧化细菌特性研究*

从我国煤矿废石堆分离到一株中度嗜热嗜酸铁氧化细菌MLY菌株,最适生长温度50℃－54℃,最适pH1.2-1.4。MLY菌株是兼性化能自养菌,能利用酵母粉异养生长。在自养和混合营养条件下,能氧化Fe^2 、黄铁矿(FeS2)和元素硫(S^0)。自养营养时,氧化元素硫较弱。对比研究MLY菌株和氧化亚铁硫杆菌(Thiobacillus ferrooxidans)A10菌株对Fe^2 和黄铁矿的氧化作用,结果表明,MLY比A10的氧化速度快1倍多。     

污泥中重金属的生物淋滤

生物淋滤法(Bioleaching)是指利用自然界中一些微生物(硫细菌)的直接作用或其代谢产物的间接作用,产生氧化、还原、络合、吸附或溶解反应,将固相中某些不溶性成分(如重金属、硫及其他金属)分离浸提出来的技术.在生物淋滤中,嗜酸性氧化亚铁硫杆菌(Acidithiobacillus ferrooxidans,A.f)和嗜酸性氧化硫硫杆菌(Acidithiobacillus thiooxidans,A.f)被用作有效的淋滤载体[1].这种嗜酸性的化能自养型细菌以大气中的CO2为碳源,以无机物铁或硫为能源来维持生长,不需要提供外来的碳源和电子供体.另外,由于pH值很低,抑制了其他细菌的生长,所以在实际的操作过程中不需要严格的无菌条件.氧化亚铁硫杆菌和氧化硫硫杆菌去除重金属适宜于污水处理厂的开放系统,采用土著嗜酸性氧化亚铁硫杆菌(A.f)和氧化硫硫杆菌(A.f)进行重金属去除.也就是说,处理什么地方的污泥,就在什么地方分离A.f和A.t,这样分离的微生物在生物淋滤过程中能发挥较好的作用.这也是微生物在自然界生长繁殖的特点之一. 本期介绍了王聪、宋存江等[2]从剩余活性污泥中分离得到两株土著硫杆菌,对两株菌进行了分类鉴定,确定二者分别为嗜酸性氧化亚铁硫杆菌杆(Acidithiobacillus ferrooxidans,A.f)和嗜酸性氧化硫硫杆菌(Acidithiobacillus thiooxidans,A.t),将二者的单菌和混合菌分别接种于剩余活性污泥中,进行了为期9 d的生物淋滤,对淋滤过程中的pH变化、氧化还原电位(ORP)以及重金属含量进行了检测.结果表明,生物淋滤9 d的混合菌对于As、Cr、Cu、Ni和Zn的去除效果最好,去除率分别达到了96.09%、93.47%、98.32%、97.88%和98.60%.混合菌生物淋滤对于Cd和Pb的去除率在第6天之后迅速下降,但是A.t单菌淋滤保持较高的去除率,此结果为进一步的应用打下了良好的基础.     

Isolation and characterization of Ferroplasma thermophilum sp. nov., a novel extremely acidophilic, moderately thermophilic archaeon and its role in bioleaching of chalcopyrite

Aims:  To isolate Ferroplasma thermophilum L1^T from a low pH environment and to understand its role in bioleaching of chalcopyrite.
Methods and Results:  Using serial dilution method, a moderately thermophilic and acidophilic ferrous iron-oxidizing archaeon, named L1^T, was isolated from a chalcopyrite-leaching bioreactor. The morphological, biochemical and physiological characteristics of strain L1^T and its role in bioleaching of chalcopyrite were studied. Strain L1^T was a nonmotile coccus that lacked cell wall. Strain L1^T had a temperature optimum of 45°C and the optimum pH for growth was 1·0. Strain L1^T was capable of chemomixotrophic growth on ferrous iron and yeast extract. Results of fatty acid analysis, DNA–DNA hybridization, G+C content, and analysis based on 16S rRNA gene sequence indicated that strain L1^T should be grouped in the genus Ferroplasma , and represented a new species, Ferroplasma thermophilum . Ferroplasma thermophilum in combination with Acidithiobacillus caldus and Leptospirillum ferriphilum could improve the copper dissolution in bioleaching of chalcopyrite.
Conclusions:  A novel extremely acidophilic, moderately thermophilic archaeon isolated from a bioleaching reactor has been identified as F. thermophilum that played an important role in bioleaching of chalcopyrite at low pH.
Significance and Impact of the Study:  This study contributes to understand the characteristics of F. thermophilum L1^T and its role in bioleaching of sulfide ores.     

中等嗜热细菌浸出黄铜矿过程中pH对群落结构的影响

为了优化浸出工艺,研究了pH对浸矿过程主要微生物种群结构的影响。用中度嗜热混合菌槽浸黄铜矿精矿,在不控制pH,控制pH为2．5及控制pH为1．2时,应用PCR-RFLP（限制性酶切片段长度多态性）方法对上述浸出条件下的细菌群落动态变化进行研究。结果表明,浸出体系只有两种微生物,一种为Acidithiobacillus Caldus,一种为Leptospirillum ferriphilum。pH对群落结构有明显影响。不控制pH时,浸出开始阶段At．caldus是优势种群,占群落的96％,随着浸出的进行,L．ferriphilum增多,在浸出后期代替At．caldus成为优势菌种,占69％。控制pH时,L．ferriphilum始终占主导地位,同时发现pH为2．5时At．caldus在群落中的丰度比pH为1．2时高。     

Isolation of a strain of <Emphasis Type="Italic">Acidithiobacillus caldus</Emphasis> and its role in bioleaching of chalcopyrite

A moderately thermophilic and acidophilic sulfur-oxidizing bacterium named S₂, was isolated from coal heap drainage. The bacterium was motile, Gram-negative, rod-shaped, measured 0.4 to 0.6 by 1 to 2 μm, and grew optimally at 42–45°C and an initial pH of 2.5. The strain S₂ grew autotrophically by using elemental sulfur, sodium thiosulfate and potassium tetrathionate as energy sources. The strain did not use organic matter and inorganic minerals including ferrous sulfate, pyrite and chalcopyrite as energy sources. The morphological, biochemical, physiological characterization and analysis based on 16S rRNA gene sequence indicated that the strain S₂ is most closely related to Acidithiobacillus caldus (>99% similarity in gene sequence). The combination of the strain S₂ with Leptospirillum ferriphilum or Acidithiobacillus ferrooxidans in chalcopyrite bioleaching improved the copper-leaching efficiency. Scanning electron microscope (SEM) analysis revealed that the chalcopyrite surface in a mixed culture of Leptospirillum ferriphilum and Acidithiobacillus caldus was heavily etched. The energy dispersive X-ray (EDX) analysis indicated that Acidithiobacillus caldus has the potential role to enhance the recovery of copper from chalcopyrite by oxidizing the sulfur formed during the bioleaching progress.     

Bioleaching of sulfidic tailing samples with a novel, vacuum-positive pressure driven bioreactor

This study presents a design for a novel bioreactor that uses alternating vacuum and positive pressure cycles to transfer acidic leach solution in and out of contact with finely ground sulfidic mine tailings. These tailings constitute an environmental problem that needs experimental data to support the development of management and control strategies. A conventional stirred tank bioreactor was used as a reference system. Both bioreactors were inoculated with mixed cultures of acidophilic iron and sulfur oxidizers. The rate of the bioleaching of tailings was 0.50 +/- 0.14 g Fe/L . day in the stirred tank bioreactor and 0.17 +/- 0.05 g Fe/L . day in the novel bioreactor. Microbial populations were identified in the two-bioreactor systems by analysis of 16S rRNA genes involving amplification, denaturing gradient gel electrophoresis (DGGE), cloning, and sequencing. The inoculum contained sulfur-oxidizing Acidithiobacillus caldus and Acidithiobacillus thiooxidans, iron oxidizers from the genera Leptospirillum and Ferroplasma, and a chemoorganotrophic Alicyclobacillus sp. During bioleaching of the tailings, the microbial populations in both bioreactors were similar to the inoculum culture, except that At. thiooxidans outgrew At. caldus. Sequences consistent with a Sulfobacillus sp. were amplified from both bioreactor samples although this bacterium was initially below the level of detection in the inoculum. After prolonged operation, Ferroplasma acidiphilum and an uncultured bacterium related to the CFB group were also detected in the novel bioreactor, whereas Sulfobacillus sp. was no longer detected. The novel bioreactor has potential uses in other areas of environmental biotechnology that involves periodic contact of liquids with solid substrates.     

Analysis of Community Composition during Moderately Thermophilic Bioleaching of Pyrite,Arsenical Pyrite,and Chalcopyrite.

An analysis of the community composition of three previously undefined mixed cultures of moderately thermophilic bioleaching bacteria grown at 45°C on pyrite, arsenical pyrite, and chalcopyrite has been carried out. The bacterial species present were identified by comparative sequence analysis of the 16S rRNA gene isolated from the bioleaching vessels and analyzed by denaturing gradient gel electrophoresis, cloning, and sequencing. The mixed cultures leached all three minerals, as shown by the increase in iron released from the mineral concentrates. The species identified from the mixed cultures during bioleaching of pyrite, arsenical pyrite, and chalcopyrite were clones closely related to Acidithiobacillus caldus C-SH12, Sulfobacillus thermosulfidooxidans AT-1, Sulfobacillus montserratensis L15, and an uncultured thermal soil bacterium YNP. It was also found that the same mixed culture maintained for over a year on chalcopyrite mineral selected approximately the same consortia of bacteria as the original mixed culture grown on chalcopyrite.     

高矿浆浓度下黄铜矿浸出过程中微生物群落结构变化规律

目的：为阐明微生物群落演替及功能与浸出效率之间关系奠定基础,以及如何提高黄铜矿生物浸出效率和铜回收率提供理 论依据。方法：通过连续传代培养进行驯化,使得复合菌群的矿浆浓度耐受能力达到25 %（w/v）。采用该复合菌群在25 %矿浆浓 度下浸出黄铜矿,同时利用变性梯度凝胶电泳和克隆文库技术分析浸出过程中的微生物多样性。最后,采用实时荧光定量PCR 对 浸出过程中微生物群落结构进行定量解析。结果：28天内黄铜矿浸出率能够达到95.1 %,而驯化前的浸出率只有51.5%。该复合 菌群主要由Acidithiobacillus caldus, Sulfobacillus acidophilus,和Fereoplasma theroplasma thermophilum组成,其中Acidithbacillus caldus是浸出前期和后期的优势种群,而Sulfobacillus acidophilus在浸出中期均有竞争优势, Ferroplasma thermophilum在整个浸出过程中占 据整个群落的比例均较低。结论：本研究获得的复合菌群具有较强的浸出黄铜矿能力, Acidithiobacillus caldus和Sulfobacillus acidophilus在浸出过程中起着重要的作用,pH 值和铜浸出率与群落结构相关性较高。     

Biooxidation of pyrite by defined mixed cultures of moderately thermophilic acidophiles in pH-controlled bioreactors: significance of microbial interactions

The oxidative dissolution of pyrite (FeS2) by pure and mixed cultures of moderately thermophilic acidophiles was studied in shake flask cultures and in pH-controlled bioreactors, incubated at 45 degrees C. Various combinations of seven eubacteria (a Leptospirillum sp. (MT6), Acidimicrobium ferrooxidans, Acidithiobacillus caldus, an Alicyclobacillus sp. (Y004), and three Sulfobacillus spp.) and one archaeon (Ferroplasma sp. MT17) were examined. Pyrite dissolution was determined by measuring changes in soluble iron and generation of acidity, and microbial populations were monitored using a combined culture-dependent (plate counts) and culture-independent (fluorescent in situ hybridization) approach. In pure cultures, the most efficient pyrite-oxidizing acidophile was Leptospirillum MT6, which was unique among the prokaryotes used in being obligately autotrophic. Mixed cultures of Leptospirillum MT6 and the sulfur-oxidizer At. caldus generated more acidity than pure cultures of the iron-oxidizer, though this did not necessarily enhance pyrite dissolution. In contrast, a mixed culture of Leptospirillum MT6 and the obligate heterotroph Alicyclobacillus Y004 oxidized pyrite more rapidly and more completely than a pure culture of Leptospirillum MT6, in synchronized bioreactors. Although the autotroph, At. caldus, and the "heterotrophically inclined" iron-oxidizer, Am. ferrooxidans, were both ineffective at leaching pyrite in pure culture, a mixed culture of the two bacteria was able to accelerate dissolution of the mineral. Concentrations of dissolved organic carbon accumulated to >100 mg/L in some mixed cultures, and the most effective bioleaching systems were found to be consortia containing both autotrophic and heterotrophic moderate thermophiles. A mixed culture comprising the autotrophs Leptospirillum MT6 and At. caldus, and the heterotroph Ferroplasma MT17, was the most efficient of all of those examined. Mutualistic interactions between physiologically distinct moderately thermophilic acidophiles, involving transfer of organic and inorganic carbon and transformations of iron and sulfur, were considered to have critical roles in optimizing pyrite dissolution.     

Investigation of energy gene expressions and community structures of free and attached acidophilic bacteria in chalcopyrite bioleaching

In order to better understand the bioleaching mechanism, expression of genes involved in energy conservation and community structure of free and attached acidophilic bacteria in chalcopyrite bioleaching were investigated. Using quantitative real-time PCR, we studied the expression of genes involved in energy conservation in free and attached Acidithiobacillus ferrooxidans during bioleaching of chalcopyrite. Sulfur oxidation genes of attached A. ferrooxidans were up-regulated while ferrous iron oxidation genes were down-regulated compared with free A. ferrooxidans in the solution. The up-regulation may be induced by elemental sulfur on the mineral surface. This conclusion was supported by the results of HPLC analysis. Sulfur-oxidizing Acidithiobacillus thiooxidans and ferrous-oxidizing Leptospirillum ferrooxidans were the members of the mixed culture in chalcopyrite bioleaching. Study of the community structure of free and attached bacteria showed that A. thiooxidans dominated the attached bacteria while L. ferrooxidans dominated the free bacteria. With respect to available energy sources during bioleaching of chalcopyrite, sulfur-oxidizers tend to be on the mineral surfaces whereas ferrous iron-oxidizers tend to be suspended in the aqueous phase. Taken together, these results indicate that the main role of attached acidophilic bacteria was to oxidize elemental sulfur and dissolution of chalcopyrite involved chiefly an indirect bioleaching mechanism.     

Culturable and molecular phylogenetic diversity of microorganisms in an open-dumped,extremely acidic Pb/Zn mine tailings

A combination of cultivation-based and molecular-based approaches was used to reveal the culturable and molecular diversity of the microbes inhabiting an open-dumped Pb/Zn mine tailings that was undergoing intensive acid generation (pH 1.9). Culturable bacteria found in the extremely acidic mine tailings were Acidithiobacillus ferrooxidans, Leptospirillum ferriphilum, Sulfobacillus thermotolerans and Acidiphilium cryptum, where the number of acidophilic heterotrophs was ten times higher than that of the iron- and sulfur-oxidizing bacteria. Cloning and phylogenetic analysis revealed that, in contrast to the adjacent AMD, the mine tailings possessed a low microbial diversity with archaeal sequence types dominating the 16S rRNA gene library. Of the 141 clones examined, 132 were represented by two sequence types phylogenetically affiliated with the iron-oxidizing archaea Ferroplasma acidiphilum and three belonged to two tentative groups within the Thermoplasma lineage so far represented by only a few environmental sequences. Six clones in the library were represented by the only bacterial sequence type and were closely related to the well-described iron-oxidizer L. ferriphilum. The significant differences in the prokaryotic community structures of the extremely acidic mine tailings and the AMD associated with it highlights the importance of studying the microbial communities that are more directly involved in the iron and sulfur cycles of mine tailings.     

Evidence of cell surface iron speciation of acidophilic iron-oxidizing microorganisms in indirect bioleaching process

While indirect model has been widely accepted in bioleaching, but the evidence of cell surface iron speciation has not been reported. In the present work the iron speciation on the cell surfaces of four typically acidophilic iron-oxidizing microorganism (mesophilic Acidithiobacillus ferrooxidans ATCC 23270, moderately thermophilic Leptospirillum ferriphilum YSK and Sulfobacillus thermosulfidooxidans St, and extremely thermophilic Acidianus manzaensis YN25) grown on different energy substrates (chalcopyrite, pyrite, ferrous sulfate and elemental sulfur (S⁰)) were studied in situ firstly by using synchrotron-based micro- X-ray fluorescence analysis and X-ray absorption near-edge structure spectroscopy. Results showed that the cells grown on iron-containing substrates had apparently higher surface iron content than the cells grown on S⁰. Both ferrous iron and ferric iron were detected on the cell surface of all tested AIOMs, and the Fe(II)/Fe(III) ratios of the same microorganism were affected by different energy substrates. The iron distribution and bonding state of single cell of A. manzaensis were then studied in situ by scanning transmission soft X-ray microscopy based on dual-energy contrast analysis and stack analysis. Results showed that the iron species distributed evenly on the cell surface and bonded with amino, carboxyl and hydroxyl groups.