两株嗜铁菌对土壤有效铁浓度及嗜铁素活性单位的影响

为解决植物缺铁变黄现象,利用嗜铁菌为植物提供铁营养、促进植物生长的研究受到各国学者的广泛关注.该研究利用CAS(铬天青)平板法,从蔬菜根际土壤中分离获得7株嗜铁细菌,铬天青(CAS)法定量检测其中E7和W7菌株产嗜铁素能力较强,分别为71.48%、61.17%,16S rRNA基因序列初步鉴定其分别为苍白杆菌属(Ochrobactrum sp.)、假单胞菌属(Pseudomonas sp.).以E7、W7及其混合菌(E7+W7)为供试菌株,研究对绿豆种子萌发以及土壤中有效铁浓度和嗜铁素活性单位的影响.结果显示,与清水对照相比,菌液处理绿豆种子后能显著增加胚芽、幼根、胚轴长度,以复合菌液(E7+W7)100×稀释液处理效果最佳,胚轴、幼根、胚芽分别显著增加了41.38%、60.65%、55.08%.各菌液处理5天后均能显著增加土壤中亚铁离子浓度,其中以混合菌液处理效果最佳,增加了65.77%,其次是E7和W7,分别增加了57.45%、54.39%;E7+W7和E7处理组在第3天时嗜铁素活性单位分别达到90.71%和85.31%,而W7组在第5天时达到最大值79.07%.

嗜酸铁还原菌JF-5改性镧膨润土吸附砷及其矿物颗粒迁移研究

利用嗜酸铁还原菌(Acidiphilium cryputum JF-5),通过生物还原技术释放膨润土中的Fe^(2+)以强化镧改性,从而得到一种新型生物镧改性膨润土。通过等温吸附实验考察生物镧改性膨润土对As(Ⅴ)的吸附效果,并与国际上商用的锁磷剂(Phoslock)进行对比,通过柱实验探讨生物镧改性膨润土在石英砂柱中的迁移行为。结果表明:(1)JF-5能够强化膨润土还原释放Fe^(2+),从而加强La^(3+)负载,低固液比(1.6 g/L)下制备生物镧改性膨润土具有最高的La^(3+)负载量(370 mg/g)。(2)生物镧改性膨润土对As(Ⅴ)的吸附行为符合Freundlich模型,最大吸附量为7.58 mg/g,与Phoslock的最大吸附量(3.08 mg/g)相比大幅提高;对As(Ⅴ)的吸附动力学过程与Phoslock相似,均能在4 h内快速去除As(Ⅴ)。(3)生物镧改性膨润土在石英砂柱中的迁移行为可以用对流扩散模型描述,在0.38、0.77、1.54 mL/min的流速下,生物镧改性膨润土在石英砂柱中的滞留量分别为3.68、3.77、2.29 mg,可见中等流速条件下溶液在石英柱中能够更充分接触介质从而提高滞留量。

硫酸盐还原菌与铁还原菌联合释放含砷水铁矿中砷及渗透性研究

水铁矿是土壤中常见的铁氧化物矿物,也是主要的载砷矿物。从水铁矿中释放砷,是缓解土壤砷污染的一种途径。本文拟通过硫酸盐还原菌(SRB)和嗜酸铁还原菌(JF-5)生物还原含砷水铁矿(As-Fh),促进其砷释放,以达到土壤修复目的。为此,利用采集自湖南石门矿区的典型含砷水铁矿,进行两种微生物摇瓶实验和生物注入水铁矿柱实验,探究水铁矿释放砷浓度及形态、生物溶解过程中悬浮颗粒物变化以及体系渗透性问题。研究结果表明:①在两个循环周期下,从释放的砷总量可看出,JF-5+SRB操作>JF-5+JF-5操作>SRB+JF-5操作>SRB+SRB操作,对应这些操作每克水铁矿能够分别释放16.4、16.2、7.3和3.3 mg的砷;②水化学分析也表明同等操作下SRB能够提供比JF-5更强的还原性;③生物柱实验表明,相比于SRB来说,JF-5的注入能够产生平均浓度1419 mg/L的铁系悬浮颗粒物,是SRB的7.25倍;同时两种生物操作过程下,SRB柱体系渗透性(平均水压差为0.51 psi)显著优于JF-5(平均水压差为2.22 psi),高悬浮颗粒的分散是引起渗透性变化的主因。上述结果证明连续注入SRB于水铁矿体系,是一种有效的降低土壤中砷浓度的技术。

氧化亚铁硫杆菌与中度嗜热铁氧化菌浸出铁闪锌矿的比较

考察了采用氧化亚铁硫杆菌和中度嗜热铁氧化菌浸出铁闪锌矿浮选精矿的浸出效果.结果表明,两种细菌的驯化菌株都可以显著提高铁闪锌矿的溶解速率,且接种中度嗜热铁氧化菌时浸出效果更好.扫描电镜、X射线粉晶衍射和能谱分析等表明,接种不同菌种浸出铁闪锌矿时所产浸渣具有类似的表面形貌特征和矿物组成.可以证明,采用中度嗜热铁氧化菌浸出铁闪锌矿浮选精矿可以在较高的温度和酸度下进行且具有更好的浸出效果,比氧化亚铁硫杆菌具有更好的工业应用前景.

嗜酸铁/硫氧化菌浸出煤矸石中硫的形态变化

利用嗜酸铁/硫氧化微生物Acidithiobacillus thiooxidans SOB-1和Acidithiobacillus ferrooxidans SOB-2浸出煤矸石,利用XRD和硫的K边X-ray Absorption Near Edge Spectroscopy(XANES)分析煤矸石浸出前后物相组成和硫的形态变化。XRD结果表明:经浸出处理后矸石中黄铁矿消失,其他矿物组成变化不明显。XANES结果表明,原始矸石中硫形态主要是黄铁矿硫,氧化型谷胱甘肽硫,硫代硫酸盐硫,单质硫和少量噻吩硫。与空白对照相比,SOB-1对矸石浸出效果较差,样品中各种形态硫的含量变化不明显;而SOB-2及SOB-1和SOB-2的混合菌对矸石中总硫和黄铁矿均有较好的脱除效果,其脱除率最高可达51.08%和71.5%。矸石样品中的黄铁矿硫经氧化转变成硫代硫酸盐硫和硫酸盐硫,SOB-2和混合菌明显促进了硫的形态变化,并且两株菌及其混合菌可能对噻吩硫也有一定脱除效果。

中温及嗜热菌对不同成因黄铜矿浸出行为的影响

以斑岩型黄铜矿和矽卡岩型黄铜矿为研究对象,考察了嗜酸氧化亚铁微螺菌(L f)和嗜热硫氧化硫化杆菌(S t)对不同成因黄铜矿浸出行为的影响。结果表明,在2种不同细菌浸出体系中矽卡岩型黄铜矿均表现出比斑岩型黄铜矿浸出率高;S t浸出2种不同成因黄铜矿的效率均比L f的好。通过对不同浸矿时间黄铜矿浸出渣的XRD检测并结合黄铜矿浸出过程反应步骤的分析表明,2种细菌浸出不同成因黄铜矿的机制相同,细菌的代谢途径及反应温度是影响同类成因黄铜矿浸出率和代谢产物差异的主要原因。S t作用下浸出后期黄铜矿表面有黄钾铁矾生成,而L f浸出体系黄铜矿表面主要是S的不断积累。同种浸矿菌种浸出不同成因黄铜矿时,矽卡岩型黄铜矿在浸出第15 d有S生成,斑岩型黄铜矿在S t浸出体系S生成的时间更晚,在L f浸出体系S的生成量则更少,推测矿物性质是引起其差异的主要原因。

采用中温菌和常温菌浸出含砷金精矿

考察了不同矿浆浓度和接种量对细菌浸出含砷金精矿的影响。结果表明:中度嗜热嗜酸铁氧化菌MLY和嗜酸氧化亚铁硫杆菌(Acidthiobacillus ferrooxidans,At.f)的浸矿效果不同。MLY和At.f对含砷金精矿的浸出机理也不同;接种MLY的矿浆中,可能主要是溶液中的细菌将Fe2+氧化为Fe3+,再由Fe3+氧化溶解矿物,即以间接浸出作用为主;而接种At.f可能首先是At.f被吸附到矿物表面,通过细菌与矿物之间的某种界面作用引起矿物溶解,矿物溶解产生的Fe2+被游离菌氧化成为Fe3+,从而溶解矿物,即At.f浸出可能包括直接作用和间接作用的共同效应。

微生物浸出磁黄铁矿的试验研究

采用矿驯化以及矿坑的嗜酸嗜铁混合菌(Acidthiobacillus ferrooxidans与Leptospirillum ferriphilum)进行了磁黄铁矿浸出试验,并探讨了混合菌浸出磁黄铁矿的机理。结果表明,与矿驯化菌相比,矿坑混合菌对磁黄铁矿具有更强的吸附能力;细菌吸附后,磁黄铁矿的等电点朝细菌的等电点方向偏移;由于矿坑混合菌对磁黄铁矿的作用更强,使其得到的矿物接触角下降幅度均大于矿驯化混合菌作用下的接触角;磁黄铁矿浸出初期是以酸溶为主,随着浸矿的进行,混合菌发挥作用,整个浸矿过程,体系中会发生铁离子的相对富集,致使矿物动电位值增大,而由于铁离子不能稳定性存在,易在体系中生成亲水性的铁沉淀覆盖于矿物表面,从而引起磁黄铁矿表面亲水性增强,接触角变小。在嗜酸嗜铁混合菌浸出磁黄铁矿过程中,磁黄铁矿主要是被溶液中的铁离子氧化。

海底热液环境中嗜中性微需氧铁氧化菌的多样性、生物矿化作用及其代谢特征

铁元素是深海热液活动产物的主要成分之一,也是热液喷口处化能自养微生物生态系统的重要驱动元素。以Zetaproteobacteria为典型代表的嗜中性微需氧铁氧化菌是海底喷口及其周围环境中生物介导的Fe^(2+)氧化这一生物矿化作用的主要驱动者。这些铁氧化菌通过氧化Fe^(2+)获取维持自身代谢所必需的能量,同时分泌有机质将氧化后的不溶铁(氧化物或氢氧化物)沉淀于细胞外,形成具有螺旋丝带状、中空长杆状、分叉管状以及其他具有特殊形貌特征的显微结构体,进而堆积成广泛分布于海底的富铁氧化物/氢氧化物。越来越多的研究表明,编码细胞色素孔蛋白的cyc2基因是Zetaproteobacteria铁氧化菌进行Fe^(2+)氧化的关键基因,而细胞色素c或其他周质细胞色素则是Fe^(2+)氧化过程中的关键电子传递载体。基于宏基因组分析的系列研究揭示了Zetaproteobacteria普遍具有多种与氮、硫、氢以及砷元素循环密切相关的功能基因与代谢途径,暗示了其在上述元素循环过程中的潜在作用。本文系统地总结了海底热液喷口及其周围环境中发现的嗜中性微需氧铁氧化菌的多样性和生理特征以及它们进行Fe^(2+)氧化生物矿化作用而形成的显微结构记录、介导Fe^(2+)氧化的关键基因与电子传递路径等,有利于全面、系统地认识该类微生物在海底热液喷口关键成矿元素的迁移与富集、维系喷口生态以及微生物成矿等方面所起的作用与贡献。

Decomposition mechanism of pentlandite during electrochemical bio-oxidation process

Electrochemical measurements were carried out to elucidate decomposition mechanism of pentlandite using modified powder microelectrode with Acidithiobacillus ferrooxidans attached or without on the mineral powder surface.Cyclic voltammetry（CV） results show that at a low potential of about-0.2 V（vs SCE）,the pentlandite was transformed to an intermediated phase like Fe4.5-yNi4.5-xS8-z when Fe and Ni ions were evacuated from mineral lattice;when the potential was changed from-0.2 V to 0.2 V,the unstable violarite（Fe3Ni3S4） and FeNi2S4 were formed which was accompanied by element sulfur formed on the mineral surface;when the potential increased over 0.2 V,the unstable intermediated phase decomposed entirely;at a higher potential of 0.7 V,the evacuated ferrous ion was oxidized to ferric ion.The presence of Acidithiobacillus ferrooxidans made the oxidation peak current increase with initial peak potential negatively moving,and the bacteria also contributed to the sulfur removing from mineral surface,which was demonstrated by the reduction characteristic at potential ranging from-0.75 to-0.5 V.Leaching experiments and electrochemical results show that the solution acidity increasing when pH2 may impede the oxidation process slightly.

Surface characterization of chalcopyrite interacting with Leptospirillum ferriphilum

The alteration of surface properties of chalcopyrite after biological conditioning with Leptospirillum ferriphilum was studied by adsorption,zeta-potential,contact angle and bioleaching tests.The strains of L.ferriphilum cultured using different energy sources（either soluble ferrous ion or chalcopyrite） were used.The adhesion of bacteria to the chalcopyrite surface was a fast process.Additionally,the adsorption of substrate-grown bacteria was greater and faster than that of liquid-grown ones.The isoelectric point（IEP） of chalcopyrite moved toward that of pure L.ferriphilum after conditioning with bacteria.The chalcopyrite contact angle curves motioned diversely in the culture with or without energy source.The results of X-ray diffraction patterns（XRD）,scanning electron microscopy（SEM） and energy-dispersive X-ray spectroscopy（EDS） analysis indicate that the surface of chalcopyrite is covered with sulfur and jarosite during the bioleaching process by L.ferriphilum.Furthermore,EDS results imply that iron phase dissolves preferentially from chalcopyrite surface during bioleaching.The copper extraction is low,resulting from the formation of a passivation layer on the surface of chalcopyrite.The major component of the passivation layer that blocked continuous copper extraction is sulfur instead of jarosite.

Preservation efficiency of new cryoprotectant used for Acidithiobacillus ferrooxidans in liquid nitrogen

The efficiency of a new cryoprotectant,GP,for the preservation of Acidithiobacillus ferrooxidans（A.ferrooxidans） strain DC in liquid nitrogen was investigated.The optimal concentration of this new cryoprotectant for the maximal viable cell recovery and the highest ferrous ion oxidation activity was determined.The results show that 30%（volume fraction） GP is optimal for the cryopreservation with 84.4% of cells surviving,completely oxidizing ferrous ions within 120 h,and growing to a final density of 5.8×107 cell/mL after 6 d in the culture.Furthermore,the optimal residual GP concentration for viable cell recovery after culture of thawed cells in 9K medium for 6 d is 0.6%（volume fraction）.At this concentration,strain DC completely oxidizes ferrous ions within 108 h and grows to a final cell density of 6.8×107 mL-1.Thus,GP is a simple,effective cryoprotectant for the preservation of A.ferrooxidans strain DC in liquid nitrogen.

油藏嗜热菌与膨润土的相互作用及其对储层防膨的意义

【目的】本文从胜利油田沾3区块的高温油藏的原油采出液中分离得到一株嗜热菌,通过其与膨润土的相互作用,尝试探讨油藏微生物作为油藏储层中水敏性矿物(如蒙皂石)改性剂的可能性。【意义】研究结果将在降低水敏矿物的膨胀性能,为解决水驱采油中遇到的水敏效应的瓶颈问题提供微生物的新途径。【结果】所得菌株为革兰氏阳性菌,呈杆状,具芽孢,兼性厌氧,鉴定为Geobacillus icigianus SL-1。菌株SL-1在厌氧条件下能够还原蒙皂石的结构铁。扫描电镜(SEM)结果显示,无菌对照体系中,蒙皂石呈不规则薄片状。而经微生物作用后,除薄片状蒙皂石外,另有板状次生矿物的生成。进一步能谱(EDS)分析表明,与薄片状蒙皂石相比,板状矿物含有较高的Al/Si比值,且含有明显的K^+信号。XRD结果显示,经过微生物作用后,固相物质中蒙皂石的百分比下降至47.7%,伊利石百分比上升至29.1%,而无菌对照组中蒙皂石的百分含量则为70.4%,伊利石的百分比则为19.8%。XRD物相分析和EDS结果均证实经过微生物作用后,部分蒙皂石转化成了伊利石。膨胀性能的分析进一步揭示菌株SL-1作用后,矿物膨胀性较初始矿物显著降低,缩膨率达到25.9%。以上结果为油藏储层防膨提供了重要的实验依据。

Effect of pH values on extracellular protein and polysaccharide secretions of Acidithiobacillus ferrooxidans during chalcopyrite bioleaching

The effect of pH values on the extracellular protein and polysaccharide secretions of Acidithiobacillus ferrooxidans was comparatively investigated in different phases of bacterial growth during chalcopyrite bioleaching. The results indicate that the extracellular protein is always more than the extracellular polysaccharide secreted by attached cells on the chalcopyrite, on the contrary, and is always less than the extracellular polysaccharide secreted by free cells in the solution at bacterial adaptive phase, logarithmic phase and stationary phase whenever pH value is at 1.0, 1.5, 2.0 or 2.5; free cells are mainly through the secretion of extracellular polysaccharide rather than the extracellular protein to fight against disadvantageous solution environment, such as high concentration of metal ions and unsuitable pH solution; both amounts of polysaccharide and protein secreted by attached cells are mainly positively related to the solution acidity rather than the total concentration of soluble metal ions. The experimental results imply that bacteria are mainly through secreting more extracellular polysaccharide to fight against disadvantageous environment and the extracellular protein perhaps plays an important role in oxidation？reduction reactions in the bioleaching system.

Really active form of fluorine toxicity affecting Acidithiobacillus ferrooxidans activity in bioleaching uranium

In order to determine the mechanism of bacterial tolerance to fluorine,Acidithiobacillus ferrooxidans ATCC 23270 was domesticated and studied under the conditions of different fluorine concentrations and pH values with or without treatment by Proteinase K.The bacterial activities were observed through measuring the changes of solution potentials by platinum electrode with Ag/AgCl reference electrode and the intracellular fluorine was determined by-uorine ion-selective electrode.The results indicated that the tolerance of Acidithiobacillus ferrooxidans ATCC 23270 to fluorine could be obviously improved by domestication,HF was the effective form of fluorine to affect the bacterial activity,and pH increase or concentration change of ions of strong complex ability with fluorine ions in solution could result in false appearance of high fluorine-resistant strain.Some proteins located in cell wall or cell membrane were intimately relative with the bacterial fluorine tolerance.

Comparative genome analysis on intraspecific evolution and nitrogen fixation of Leptospirillum ferriphilum isolates

To reveal the intraspecific evolution of Leptospirillum ferriphilum isolates which thrived in industrial bioleaching ecosystems and acid mine drainages,genome sequences of L.ferriphilum YSK,L.ferriphilum DX and L.ferriphilum ZJ were determined to compare with complete genome of L.ferriphilum ML-04.The genome comparisons reveal that extensive intraspecific variation occurs in their genomes,and that the loss and insertion of novel gene blocks of probable phage origin may mostly contribute to heterogeneity of gene content among L.ferriphilum genomes.Surprisingly,a nif gene cluster is identified in L.ferriphilum YSK and L.ferriphilum ZJ genomes.Intensive analysis and further experiments indicate that the nif gene cluster in L.ferriphilum YSK inherits from ancestor rather than lateral gene transfer.Overall,results suggest that the population of L.ferriphilum undergoes frequent genetic recombination,resulting in many closely related genome types in recent evolution.The combinatorial processes profoundly shape their physiologies and provide the basis for adaptation to different niches.

Formation of jarosite and its effect on important ions for Acidithiobacillus ferrooxidans bacteria

The precipitation of jarosite adversely affects the bio-leaching of copper sulfides in the Sarcheshmeh heap bio-leaching process. The variables of the initial concentration of ferrous iron in the growth medium, pH, and temperature were examined in the laboratory to determine how they affect the precipitation of jarosite in the presence of Acidithiobacillus ferrooxidans bacteria. It was found that the maximum ferric precipitate occurred at a ferrous sulfate concentration of 50 g/L, a temperature of 32 ℃, and an initial pH value of 2.2. The effects of the precipitation of ferric iron on the quantities of ions that are important for A. ferrooxidans bacteria in aqueous phase, i.e., ferric, sulfate, potassium, phosphate, and magnesium ions, also were assessed. The results showed relatively similar patterns for the ferric and potassium ions, and then reason might have been the co-precipitation of these ions as constituent elements of jarosite mineral. At pH values greater than 1.6, the solubility of phosphate ions decreased dramatically due to the co-precipitation of phosphate ions with the jarosite precipitate and due to the significant growth rate of A. ferrooxidans bacteria in this pH range. Due to the dissolution of a gangue constituent in the ore, the magnesium levels increased in the first few days of the bio-leaching process;thereafter, it decreased slightly.

Photogenerated-hole scavenger for enhancing photocatalytic chalcopyrite bioleaching

The effects of photogenerated-hole scavengers(ascorbic acid,oxalic acid,humic acid and citric acid)on chalcopyrite bioleaching in the presence of visible light were studied using Acidithiobacillus ferrooxidans(A.ferrooxidans).Four sets of bioleaching experiments were performed:(1)visible light+0 g/L scavenger,(2)visible light+0.1 g/L of different scavenger(ascorbic acid,oxalic acid,humic acid and citric acid),(3)dark+0.1 g/L of different scavenger(ascorbic acid,oxalic acid,humic acid and citric acid),and(4)dark+0 g/L scavenger(control group).The results showed that ascorbic acid and oxalic acid could act as photogenerated-hole scavengers and significantly enhance chalcopyrite bioleaching under visible light.The dissolved copper in the light group without scavenger was only 18.7%higher than that of the control group.The copper extraction rates of the light groups with oxalic acid and ascorbic acid were respectively 30.1%and 32.5%higher than those of the control group.Scanning electron microscopy(SEM),X-ray diffraction(XRD)and Fourier transform infrared spectroscopy(FT-IR)analyses indicated that ascorbic acid and oxalic acid as photogenerated-hole scavenger could capture photo-generated holes and inhibit jarosite formation on the chalcopyrite surface,thereby enhancing bioleaching of chalcopyrite under visible light.

Enhancing recovery of uranium column bioleaching by process optimization and kinetic modeling

This research aimed to enhance the column bioleaching recovery of uranium ore by Acidithiobacillus ferrooxidans.Seven factors were examined for their significance on bioleaching using a Plackett-Burman factorial design.Four significant variables([Fe2+]initial,pH,aeration rate and inoculation percent)were selected for the optimization studies.The effect of these variables on uranium bioleaching was studied using a central composite design(CCD).The optimal values of the variables for the maximum uranium bioleaching recovery(90.27±0.98)%were as follows:[Fe2+]initial=2.89g/L,aeration rate420mL/min,pH1.45and inoculation6%(v/v).[Fe2+]initial was found to be the most effective parameter.The maximum uranium recovery from the predicted models was92.01%.This value was in agreement with the actual experimental value.The analysis of bioleaching residue of uranium ore under optimum conditions confirmed the formation of K-jarosite on the surface of minerals.By using optimal conditions,uranium bioleaching recovery is increased at column and jarosite precipitation is minimized.The kinetic model showed that uranium recovery has a direct relation with ferric ion concentration.