典型嗜酸硫氧化菌作用下元素硫的形态及其转化的研究
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摘要
本论文选用代表性的嗜酸硫氧化菌包括嗜酸氧化亚铁硫杆菌(Acidithiobacillus ferrooxidans)和喜温嗜酸硫杆菌(Acidithiobacillus caldus)氧化利用代表性还原型硫化物单质硫和硫代硫酸钠,及Acidithiobacillusferrooxidans、嗜酸氧化硫硫杆菌(Acidithiobacillus thiooxidans)、Acidithiobacilluscaldus、嗜热硫氧化硫化杆菌(Sulfobacillus thermosulfidooxidans)和万座嗜酸菌(Acidianus manzaensis)作用典型金属硫化矿黄铁矿和黄铜矿。旨在通过研究嗜酸硫氧化菌作用下单质硫的形态转化;嗜酸硫氧化细菌硫氧化相关胞外蛋白的分离、鉴定、功能分析及其功能基因探寻;细胞胞内外累积硫球硫的形态;细菌在不同含硫能源底物中表面特性;金属硫化矿浸出过程中矿物表面硫的形态及转化;为阐明硫的生物氧化过程和金属硫化矿的生物浸出机理提供基础数据。具体内容主要包括以下六部分:
(1)硫的K边X射线吸收近边结构(XANES)光谱
本文通过硫的K边XANES方法获取了一系列含硫有机和无机化合物的吸收光谱,首次在国内建立了含硫模式化合物的谱图库。结果表明硫的K边XANES光谱是一种相当灵敏的指纹光谱方法,不同含硫化合物中硫的化学形态与其吸收光谱之间存在相当精确的对应关系。利用模式化合物的吸收光谱采用最小二乘线性组合法对混合样品中硫的吸收光谱进行定量拟合分析。结果表明,如果选取合适的模式化合物,完全可以对复杂样品中硫的赋存形态展开原位分析,得到样品中硫的精确结构,并且可以对样品进行定量分析。
(2)Acidithiobacillus ferrooxidans作用下单质硫的化学形态转变研究
综合利用了SEM、TEM、FT-IR、EDS和硫的K边XANES对A.ferrooxidans作用下单质硫的形态转化展开研究。结果表明,当细菌在单质硫中生长时,细菌首先吸附到单质硫颗粒表面,并且细菌分泌的一些两性分子对硫颗粒表面起到修饰作用。这是第一次在国内外利用实验结果证明单质硫在嗜酸硫氧化菌活化作用下部分环状硫变成链状硫。进一步对单质硫中生长的细胞硫的K边XANES光谱解析表明,富含巯基的蛋白明显参与了单质硫的活化氧化过程。
(3)嗜酸氧化亚铁硫杆菌硫活化相关胞外蛋白质的分离鉴定和功能验证
利用2-D电泳比较了亚铁和单质硫能源基质中A.ferrooxidans差异蛋白质电泳谱,借助MALDI-TOF/MS分析了18个在单质硫中表达上调的胞外蛋白。通过与A.ferrooxidans ATCC 23270基因库中蛋白和基因信息比对发现其中12个是属于未知蛋白质,以及6个分子量较小的多肽序列中含有相对丰富的半胱氨酸残基(Cys)。对10个可能与硫氧化相关的基因进行RT-PCR验证发现它们在单质硫中表达均明显上调,首次证明了A.ferrooxidans中这些基因与单质硫的活化氧化的关联性。对其中一个吸附相关的菌毛蛋白(AFE 2621:pilin,putative)基因AFE 2621进行原核表达。利用硫的K边XANES光谱对胞外蛋白样品及纯化表达的菌毛蛋白样品进行分析,结果表明样品中富含巯基的氨基酸主要是半胱氨酸,从蛋白质、基因和硫的化学形态层面证明了富含巯基蛋白在单质硫的活化氧化过程中扮演重要角色。
(4)Acidithiobacillus ferrooxidans和Acidithiobacillus caldus胞内外硫球硫的形态分析
综合利用SEM、TEM、EDS和硫的K边XANES光谱比较分析了单质硫和硫代硫酸钠中A.ferrooxidans和A.caldus细胞累积胞内外硫球的差异及累积硫球中硫的形态差异。结果发现A.ferrooxidans在单质硫和硫代硫酸钠中生长时能在细胞胞内分别累积链状和环状硫,然而A.caldus在两种能源底物中生长时均不能累积胞内硫球。相比之下,A.ferrooxidans在硫代硫酸钠中生长时累积胞外硫球主要是链状硫,而A.caldus在硫代硫酸中生长时累积胞外硫球主要是环状硫。
(5)极度嗜热菌Acidianus manzaensis在不同的能源底物中生长时细胞胞外特性研究
比较研究了极度嗜热古菌Acidianus manzaensis YN-25在硫酸亚铁,单质硫,黄铁矿和黄铜矿中生长时细胞表面电位、亲疏水性以及表面官能团等细胞胞外特性。结果表明硫酸亚铁中生长时,细胞的亲疏水性和表面电位与固体底物中(单质硫,黄铁矿和黄铜矿)生长的细菌存在很大差别。FT-IR光谱结果表明固体底物中生长的细胞比亚铁中生长的细胞含有更多的蛋白质成份。
(6)典型嗜酸硫氧化菌浸出黄铁矿和黄铜矿过程矿物表面硫的形态分析
针对代表性的浸矿功能菌:中温菌A.ferrooxidans、A.thiooxidans,中度嗜热菌S.thermosulfidooxidans,极度嗜热菌Acidianus manzaensis,综合利用SEM、XRD和硫的K边XANES研究这些细菌浸出黄铁矿和黄铜矿过程中,矿物表面形貌、矿物组成成分和矿物表面硫的形态变化。结果发现这些细菌对金属硫化矿浸出具有选择性,且不同的单一细菌和不同的细菌组合在浸出过程中引起矿物表面硫的形态存在很大差异。Acidianus manzaensis浸出黄铁矿和黄铜矿效果都相当明显;S.thermosulfidooxidans对黄铁矿浸出能力较强;A.ferrooxidans浸出黄铜矿能力较黄铁矿强,并且A.thiooxidans对A.ferrooxidans浸出黄铜矿和黄铁矿促进作用不明显。通过对浸出前后矿物中组成成分和硫的化学形态分析表明,Acidianus manzaensis浸出黄铁矿时矿物表面会产生真绿矾的结晶体,黄铜矿表面会产生黄钾铁矾的沉积层,还可能产生铜蓝中间产物。S.thermosulfidooxidans浸出黄铁矿和黄铜矿时矿物表面均会产生黄钾铁矾的沉积层。单独A.ferrooxidans浸出黄铁矿时,矿物表面会产生单质硫层,A.thiooxidans加入可以有效防止硫层的产生,但是从浸出行为来看硫层对浸出过程不会产生明显的阻碍作用。A.ferrooxidans单独或与A.thiooxidans混合浸出黄铜矿时矿物表面均会产生黄钾铁矾沉积层。在这些菌浸出金属硫化矿过程中产生的沉积层均会阻碍铜离子的浸出。综合考虑各种变量之间的关系发现,体系中增加的酸度抑制细胞生长氧化活性,矿物表面形成黄钾铁矾沉积层又进一步阻碍细菌与矿物作用,阻碍金属硫化矿的浸出。
In present work, we chose the typical acidophilic sulfur oxidizing microbes including Acidithiobacillus ferrooxidans, Acidithiobacillus thiooxidans, Acidithiobacillus caldus, Sulfobacillus thermosulfidooxidans, Acidianus manzaensis to leach pyrite and chalcopyrite, and Acidithiobacillus ferrooxidans, Acidithiobacillus caldus to oxidize thiosulfate and elemental sulfur powder. The aim of this paper is to obtain some basic data to illustrate the biological oxidation process of inorganic sulfur compounds and the mechanism of bioleaching by investigation: the elemental sulfur speciation transformation mediated by A. ferrooxidans; the screen , identification , the verification of gens and proteins of sulfur activation related extracellular proteins; the sulfur chemical speciation of intracellular and extracellular sulfur globules; growth and surface properties of cells grown on different sulfur-containing energy substrates; the sulfur chemical speciation and transformation on the surface of metal sulfides. The primary subjects of which include six parts as follows:
(1) The sulfur K-edge X-ray absorption near edge structure (XANES) spectra
In this part, we investigated the sulfur K-edge XANES spectra of a series of sulfur containing organic and inorganic model compounds with synchrotron radiation X-ray absorption fine structure. And based on these spectra of model compounds, we firsty built a sulfur K-edge XANES spectra gallery in our country. The results showed that sulfur K-edge XANES spectroscopy was a sensitive analytical method. The sulfur chemical speciation of different sulfur containing compounds had exact correlations with its absorption spectra. The defined mixture sample was fitted with model compounds by least square linear combination fitting method. It is possible to analyze and obtain the precise structural information of sulfur in nature sample in situ and the main components of mixture, if the suitable model compounds were involved.
(2) Investigation of elemental sulfur speciation transformation mediated by Acidithiobacillus ferrooxidans
The speciation transformation of elemental sulfur mediated by the leaching bacterium, A. ferrooxidans was firstly investigated with the employment of an integrated approach including: SEM, TEM, FT-IR, EDS and XANES. Our results showed that when grown on elemental sulfur powder, A. ferrooxidans ATCC23270 cells were first attached to sulfur particles, and modified the surface sulfur with some amphiphilic compounds. In addition, part of the elemental sulfur powder might be converted to linear polysulfides. Up to our knowledge, it is the first time to prove the elemental sulfur must be changed to linear sulfur baseing on the experimental results. Furthermore, sulfur globules were accumulated inside the cells. XANES spectra of these cells suggested that these globules consisted of elemental sulfur bound to thiol groups of protein.
(3) The screen, identification and functional investigation of extracellular proteins and genes of A. ferrooxidans
A set of proteins that changed their levels of synthesis during growth of A. ferrooxidans on elemental sulfur/ferrous iron was characterized by using two dimensional polyacrylamide gel electrophoresis. We identified a total of 18 extracellular proteins with apparent higher abundance grown in element sulfur than in ferrous sulfate. Matrix-assisted laser desorption/ ionization time-of-flight/ time -of-flight mass spectroscopy (MALDI-TOF/MS) analysis of these proteins allowed their identification and the localization of the corresponding genes in the available genomic sequence of A. ferrooxidans. Additionally, six hypothetical proteins (or peptides) of which contain abundant of the cysteine residues. Expression analysis of the corresponding genes by real-time PCR showed the constitutive expression of all those genes was highly upregulated by growth on elmental sulfur compounds (and downregulated by growth on ferrous iron). A gene of AFE_2621 from A. ferrooxidans was cloned and expressed in E. coli. The extracellular proteins of A. ferrooxidans grown on elemental sulfur and the purified AFE_2621 (pinlin) were characterized with sulfur K-edge XANES spectroscopy. The results showed that the thiol-containing amino acid mainly cysteine play an important role in elemental sulfur activation and oxidation.
(4) Sulfur species investigation in intra-/extracellular sulfur globules of Acidithiobacillus ferrooxidans and Acidithiobacillus caldus
For the first time, the sulfur chemical speciation in intracellular and extracellular sulfur globules of A. ferrooxidans and A. caldus were investigated with the multiple employment of SEM, TEM, EDS and sulfur K-edge XANES. When respectively grown with elemental sulfur (S ) and thiosulfate, A. ferrooxidans could accumulate both linear sulfur and ring sulfur internally, whereas A. caldus could not accumulate intracellular sulfur globules. When grown with thiosulfate, in contrast, the sulfur species in extracellular sulfur globules produced was polymeric sulfur(with unknown) and homocycles(just like artificial made sediment) A. ferrooxidans and A. caldus, respectively.
(5) Growth and surface properties of new thermoacidophilic Archaea strain Acidianus manzaensis grown on different substrates
The growth characteristics of Acidianus manzaensis YN-25 have been firstly investigated. The cells were cultured with ferrous sulfate, elemental sulfur, pyrite and chalcopyrite mineral respectively, and the hydrophobicity, electrokinetic behaviour and the surface groups of cells grown under above substrate have been investigated. The results showed that the hydrophobicity and the Zeta-potential of cells grown in ferrous sulfate presented distinctly characteristics compared to the cells cultured in solid substrate. The FT-IR spectra indicated that the sulfur, pyrite and chalcopyrite grown cells contained a higher amount of protein component than the ferrous ion-grown cells on the cells surface.
(6) Typical acidophilic sulfur oxidizing microbes leaching pyrite and chalcopyrite
The typical mesophiles A. ferrooxidans、A. thiooxidans, moderate thermophiles S. thermosulfidooxidans and extreme thermophiles Acidianus manzaensis were chosen to leach pyrite and chalcopyrite. For the first time, the characteristics of mineral surface, main components of mineral and the sulfur speciation on the mineral surface were investigated with SEM,XRD and sulfur K-edge XANES spectra. The results showed that there were distinct differences among the sulfur speciation and the main components of leaching residual leached with these pure or mixture of microbes. The extreme thermophiles Acidianus manzaensis had excellent leaching ability with pyrite and chalcopyrite. S. thermosulfidooxidans had better leaching capability on pyrite than chalcopyrite. The mesophiles A. ferrooxidans had better leaching efficiency on chalcopyrite than pyrite. The strains of A. thiooxidans had no oblivious promotion on A. ferrooxidans leaching pyrite and chalcopyrite. During all the leaching experiments, the cells attached to the mineral surfaces firstly. The results of the sulfur speciation and the main components of leaching residual during the Acidianus manzaensis leaching experiments, the crystal of coquimbite and covellite and jarosite were accumulated on the surface of pyrite and chalcopyrite, respectively and the sediments layer of later could negatively affect the dissolution of copper ions. The passivation layer on the surface of metal sulfides were caused by the lowering acidity, in that case, the growth of cells were strongly inhibited. During the leaching of pyrite and chalcopyrite with S. thermosulfidooxidans, the precipitation of jarosite was formed on the surface of metal sulfides. The amount of jarosite produced from the former was much more than the latter, which also form negative barrier between cells and mineral surface. The elemental sulfur sediments were accumulated on the surface of pyrite leached with A. ferrooxidans, and the addition of A. thiooxidans could avoid the accumulation of elemental sulfur. The jarosite sediments on the chalcopyrite were formed with pure A. ferrooxidans and mixture of A. ferrooxidans and A. thiooxidans, which could lead to a gradual passivation of chalcopyrite whereby Cu~(2+) dissolution leveled off. Nevertheless, judging from the leaching curves of pyrite, the precipitation of elemental sulfur could not effected significantly the leaching of iron ions.
(1)硫的K边X射线吸收近边结构(XANES)光谱
本文通过硫的K边XANES方法获取了一系列含硫有机和无机化合物的吸收光谱,首次在国内建立了含硫模式化合物的谱图库。结果表明硫的K边XANES光谱是一种相当灵敏的指纹光谱方法,不同含硫化合物中硫的化学形态与其吸收光谱之间存在相当精确的对应关系。利用模式化合物的吸收光谱采用最小二乘线性组合法对混合样品中硫的吸收光谱进行定量拟合分析。结果表明,如果选取合适的模式化合物,完全可以对复杂样品中硫的赋存形态展开原位分析,得到样品中硫的精确结构,并且可以对样品进行定量分析。
(2)Acidithiobacillus ferrooxidans作用下单质硫的化学形态转变研究
综合利用了SEM、TEM、FT-IR、EDS和硫的K边XANES对A.ferrooxidans作用下单质硫的形态转化展开研究。结果表明,当细菌在单质硫中生长时,细菌首先吸附到单质硫颗粒表面,并且细菌分泌的一些两性分子对硫颗粒表面起到修饰作用。这是第一次在国内外利用实验结果证明单质硫在嗜酸硫氧化菌活化作用下部分环状硫变成链状硫。进一步对单质硫中生长的细胞硫的K边XANES光谱解析表明,富含巯基的蛋白明显参与了单质硫的活化氧化过程。
(3)嗜酸氧化亚铁硫杆菌硫活化相关胞外蛋白质的分离鉴定和功能验证
利用2-D电泳比较了亚铁和单质硫能源基质中A.ferrooxidans差异蛋白质电泳谱,借助MALDI-TOF/MS分析了18个在单质硫中表达上调的胞外蛋白。通过与A.ferrooxidans ATCC 23270基因库中蛋白和基因信息比对发现其中12个是属于未知蛋白质,以及6个分子量较小的多肽序列中含有相对丰富的半胱氨酸残基(Cys)。对10个可能与硫氧化相关的基因进行RT-PCR验证发现它们在单质硫中表达均明显上调,首次证明了A.ferrooxidans中这些基因与单质硫的活化氧化的关联性。对其中一个吸附相关的菌毛蛋白(AFE 2621:pilin,putative)基因AFE 2621进行原核表达。利用硫的K边XANES光谱对胞外蛋白样品及纯化表达的菌毛蛋白样品进行分析,结果表明样品中富含巯基的氨基酸主要是半胱氨酸,从蛋白质、基因和硫的化学形态层面证明了富含巯基蛋白在单质硫的活化氧化过程中扮演重要角色。
(4)Acidithiobacillus ferrooxidans和Acidithiobacillus caldus胞内外硫球硫的形态分析
综合利用SEM、TEM、EDS和硫的K边XANES光谱比较分析了单质硫和硫代硫酸钠中A.ferrooxidans和A.caldus细胞累积胞内外硫球的差异及累积硫球中硫的形态差异。结果发现A.ferrooxidans在单质硫和硫代硫酸钠中生长时能在细胞胞内分别累积链状和环状硫,然而A.caldus在两种能源底物中生长时均不能累积胞内硫球。相比之下,A.ferrooxidans在硫代硫酸钠中生长时累积胞外硫球主要是链状硫,而A.caldus在硫代硫酸中生长时累积胞外硫球主要是环状硫。
(5)极度嗜热菌Acidianus manzaensis在不同的能源底物中生长时细胞胞外特性研究
比较研究了极度嗜热古菌Acidianus manzaensis YN-25在硫酸亚铁,单质硫,黄铁矿和黄铜矿中生长时细胞表面电位、亲疏水性以及表面官能团等细胞胞外特性。结果表明硫酸亚铁中生长时,细胞的亲疏水性和表面电位与固体底物中(单质硫,黄铁矿和黄铜矿)生长的细菌存在很大差别。FT-IR光谱结果表明固体底物中生长的细胞比亚铁中生长的细胞含有更多的蛋白质成份。
(6)典型嗜酸硫氧化菌浸出黄铁矿和黄铜矿过程矿物表面硫的形态分析
针对代表性的浸矿功能菌:中温菌A.ferrooxidans、A.thiooxidans,中度嗜热菌S.thermosulfidooxidans,极度嗜热菌Acidianus manzaensis,综合利用SEM、XRD和硫的K边XANES研究这些细菌浸出黄铁矿和黄铜矿过程中,矿物表面形貌、矿物组成成分和矿物表面硫的形态变化。结果发现这些细菌对金属硫化矿浸出具有选择性,且不同的单一细菌和不同的细菌组合在浸出过程中引起矿物表面硫的形态存在很大差异。Acidianus manzaensis浸出黄铁矿和黄铜矿效果都相当明显;S.thermosulfidooxidans对黄铁矿浸出能力较强;A.ferrooxidans浸出黄铜矿能力较黄铁矿强,并且A.thiooxidans对A.ferrooxidans浸出黄铜矿和黄铁矿促进作用不明显。通过对浸出前后矿物中组成成分和硫的化学形态分析表明,Acidianus manzaensis浸出黄铁矿时矿物表面会产生真绿矾的结晶体,黄铜矿表面会产生黄钾铁矾的沉积层,还可能产生铜蓝中间产物。S.thermosulfidooxidans浸出黄铁矿和黄铜矿时矿物表面均会产生黄钾铁矾的沉积层。单独A.ferrooxidans浸出黄铁矿时,矿物表面会产生单质硫层,A.thiooxidans加入可以有效防止硫层的产生,但是从浸出行为来看硫层对浸出过程不会产生明显的阻碍作用。A.ferrooxidans单独或与A.thiooxidans混合浸出黄铜矿时矿物表面均会产生黄钾铁矾沉积层。在这些菌浸出金属硫化矿过程中产生的沉积层均会阻碍铜离子的浸出。综合考虑各种变量之间的关系发现,体系中增加的酸度抑制细胞生长氧化活性,矿物表面形成黄钾铁矾沉积层又进一步阻碍细菌与矿物作用,阻碍金属硫化矿的浸出。
In present work, we chose the typical acidophilic sulfur oxidizing microbes including Acidithiobacillus ferrooxidans, Acidithiobacillus thiooxidans, Acidithiobacillus caldus, Sulfobacillus thermosulfidooxidans, Acidianus manzaensis to leach pyrite and chalcopyrite, and Acidithiobacillus ferrooxidans, Acidithiobacillus caldus to oxidize thiosulfate and elemental sulfur powder. The aim of this paper is to obtain some basic data to illustrate the biological oxidation process of inorganic sulfur compounds and the mechanism of bioleaching by investigation: the elemental sulfur speciation transformation mediated by A. ferrooxidans; the screen , identification , the verification of gens and proteins of sulfur activation related extracellular proteins; the sulfur chemical speciation of intracellular and extracellular sulfur globules; growth and surface properties of cells grown on different sulfur-containing energy substrates; the sulfur chemical speciation and transformation on the surface of metal sulfides. The primary subjects of which include six parts as follows:
(1) The sulfur K-edge X-ray absorption near edge structure (XANES) spectra
In this part, we investigated the sulfur K-edge XANES spectra of a series of sulfur containing organic and inorganic model compounds with synchrotron radiation X-ray absorption fine structure. And based on these spectra of model compounds, we firsty built a sulfur K-edge XANES spectra gallery in our country. The results showed that sulfur K-edge XANES spectroscopy was a sensitive analytical method. The sulfur chemical speciation of different sulfur containing compounds had exact correlations with its absorption spectra. The defined mixture sample was fitted with model compounds by least square linear combination fitting method. It is possible to analyze and obtain the precise structural information of sulfur in nature sample in situ and the main components of mixture, if the suitable model compounds were involved.
(2) Investigation of elemental sulfur speciation transformation mediated by Acidithiobacillus ferrooxidans
The speciation transformation of elemental sulfur mediated by the leaching bacterium, A. ferrooxidans was firstly investigated with the employment of an integrated approach including: SEM, TEM, FT-IR, EDS and XANES. Our results showed that when grown on elemental sulfur powder, A. ferrooxidans ATCC23270 cells were first attached to sulfur particles, and modified the surface sulfur with some amphiphilic compounds. In addition, part of the elemental sulfur powder might be converted to linear polysulfides. Up to our knowledge, it is the first time to prove the elemental sulfur must be changed to linear sulfur baseing on the experimental results. Furthermore, sulfur globules were accumulated inside the cells. XANES spectra of these cells suggested that these globules consisted of elemental sulfur bound to thiol groups of protein.
(3) The screen, identification and functional investigation of extracellular proteins and genes of A. ferrooxidans
A set of proteins that changed their levels of synthesis during growth of A. ferrooxidans on elemental sulfur/ferrous iron was characterized by using two dimensional polyacrylamide gel electrophoresis. We identified a total of 18 extracellular proteins with apparent higher abundance grown in element sulfur than in ferrous sulfate. Matrix-assisted laser desorption/ ionization time-of-flight/ time -of-flight mass spectroscopy (MALDI-TOF/MS) analysis of these proteins allowed their identification and the localization of the corresponding genes in the available genomic sequence of A. ferrooxidans. Additionally, six hypothetical proteins (or peptides) of which contain abundant of the cysteine residues. Expression analysis of the corresponding genes by real-time PCR showed the constitutive expression of all those genes was highly upregulated by growth on elmental sulfur compounds (and downregulated by growth on ferrous iron). A gene of AFE_2621 from A. ferrooxidans was cloned and expressed in E. coli. The extracellular proteins of A. ferrooxidans grown on elemental sulfur and the purified AFE_2621 (pinlin) were characterized with sulfur K-edge XANES spectroscopy. The results showed that the thiol-containing amino acid mainly cysteine play an important role in elemental sulfur activation and oxidation.
(4) Sulfur species investigation in intra-/extracellular sulfur globules of Acidithiobacillus ferrooxidans and Acidithiobacillus caldus
For the first time, the sulfur chemical speciation in intracellular and extracellular sulfur globules of A. ferrooxidans and A. caldus were investigated with the multiple employment of SEM, TEM, EDS and sulfur K-edge XANES. When respectively grown with elemental sulfur (S ) and thiosulfate, A. ferrooxidans could accumulate both linear sulfur and ring sulfur internally, whereas A. caldus could not accumulate intracellular sulfur globules. When grown with thiosulfate, in contrast, the sulfur species in extracellular sulfur globules produced was polymeric sulfur(with unknown) and homocycles(just like artificial made sediment) A. ferrooxidans and A. caldus, respectively.
(5) Growth and surface properties of new thermoacidophilic Archaea strain Acidianus manzaensis grown on different substrates
The growth characteristics of Acidianus manzaensis YN-25 have been firstly investigated. The cells were cultured with ferrous sulfate, elemental sulfur, pyrite and chalcopyrite mineral respectively, and the hydrophobicity, electrokinetic behaviour and the surface groups of cells grown under above substrate have been investigated. The results showed that the hydrophobicity and the Zeta-potential of cells grown in ferrous sulfate presented distinctly characteristics compared to the cells cultured in solid substrate. The FT-IR spectra indicated that the sulfur, pyrite and chalcopyrite grown cells contained a higher amount of protein component than the ferrous ion-grown cells on the cells surface.
(6) Typical acidophilic sulfur oxidizing microbes leaching pyrite and chalcopyrite
The typical mesophiles A. ferrooxidans、A. thiooxidans, moderate thermophiles S. thermosulfidooxidans and extreme thermophiles Acidianus manzaensis were chosen to leach pyrite and chalcopyrite. For the first time, the characteristics of mineral surface, main components of mineral and the sulfur speciation on the mineral surface were investigated with SEM,XRD and sulfur K-edge XANES spectra. The results showed that there were distinct differences among the sulfur speciation and the main components of leaching residual leached with these pure or mixture of microbes. The extreme thermophiles Acidianus manzaensis had excellent leaching ability with pyrite and chalcopyrite. S. thermosulfidooxidans had better leaching capability on pyrite than chalcopyrite. The mesophiles A. ferrooxidans had better leaching efficiency on chalcopyrite than pyrite. The strains of A. thiooxidans had no oblivious promotion on A. ferrooxidans leaching pyrite and chalcopyrite. During all the leaching experiments, the cells attached to the mineral surfaces firstly. The results of the sulfur speciation and the main components of leaching residual during the Acidianus manzaensis leaching experiments, the crystal of coquimbite and covellite and jarosite were accumulated on the surface of pyrite and chalcopyrite, respectively and the sediments layer of later could negatively affect the dissolution of copper ions. The passivation layer on the surface of metal sulfides were caused by the lowering acidity, in that case, the growth of cells were strongly inhibited. During the leaching of pyrite and chalcopyrite with S. thermosulfidooxidans, the precipitation of jarosite was formed on the surface of metal sulfides. The amount of jarosite produced from the former was much more than the latter, which also form negative barrier between cells and mineral surface. The elemental sulfur sediments were accumulated on the surface of pyrite leached with A. ferrooxidans, and the addition of A. thiooxidans could avoid the accumulation of elemental sulfur. The jarosite sediments on the chalcopyrite were formed with pure A. ferrooxidans and mixture of A. ferrooxidans and A. thiooxidans, which could lead to a gradual passivation of chalcopyrite whereby Cu~(2+) dissolution leveled off. Nevertheless, judging from the leaching curves of pyrite, the precipitation of elemental sulfur could not effected significantly the leaching of iron ions.
引文
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