Ferroplasma and relatives, recently discovered cell wall-lacking archaea making a living in extremely acid, heavy metal-rich environments

For several decades, the bacterium Acidithiobacillus (previously Thiobacillus) has been considered to be the principal acidophilic sulfur- and iron-oxidizing microbe inhabiting acidic environments rich in ores of iron and other heavy metals, responsible for the metal solubilization and leaching from such ores, and has become the paradigm of such microbes. However, during the last few years, new studies of a number of acidic environments, particularly mining waste waters, acidic pools, etc., in diverse geographical locations have revealed the presence of new cell wall-lacking archaea related to the recently described, acidophilic, ferrous-iron oxidizing Ferroplasma acidiphilum. These mesophilic and moderately thermophilic microbes, representing the family Ferroplasmaceae, were numerically significant members of the microbial consortia of the habitats studied, are able to mobilize metals from sulfide ores, e.g. pyrite, arsenopyrite and copper-containing sulfides, and are more acid-resistant than iron and sulfur oxidizing bacteria exhibiting similar eco-physiological properties. Ferroplasma cell membranes contain novel caldarchaetidylglycerol tetraether lipids, which have extremely low proton permeabilities, as a result of the bulky isoprenoid core, and which are probably a major contributor to the extreme acid tolerance of these cell wall-less microbes. Surprisingly, several intracellular enzymes, including an ATP-dependent DNA ligase have pH optima close to that of the external environment rather than of the cytoplasm. Ferroplasma spp. are probably the major players in the biogeochemical cycling of sulfur and sulfide metals in highly acidic environments, and may have considerable potential for biotechnological applications such as biomining and biocatalysis under extreme conditions.     

Effect of anions on selective solubilization of zinc and copper in bacterial leaching of sulfide ores

Bacterial leaching of sulfide ores using Thiobacillus ferrooxidans, Thiobacillus thiooxidans, or a combination of the two was studied at various concentrations of specific anions. Selective zinc and copper solubilization was obtained by inhibiting iron oxidation without affecting sulfur/sulfide oxidation. Phosphate reduced iron solubilization from a pyrite (FeS(2))-sphalerite (ZnS) mixture without significantly affecting zinc solubilization. Copper leaching from a chalcopyrite (CuFeS(2))-sphalerite mixture was stimulated by phosphate, whereas chloride accelerated zinc extraction. In a complex sulfide ore containing pyrite, chalcopyrite, and sphalerite, both phosphate and chloride reduced iron solubilization and increased copper extraction, whereas only chloride stimulated zinc extraction. Maximum leaching obtained was 100% zinc and 50% copper. Time-course studies of copper and zinc solubilization suggest the possibility of selective metal recovery following treatment with specific anions.     

Biomineralization of metal-containing ores and concentrates

Biomining is the use of microorganisms to extract metals from sulfide and/or iron-containing ores and mineral concentrates. The iron and sulfide is microbially oxidized to produce ferric iron and sulfuric acid, and these chemicals convert the insoluble sulfides of metals such as copper, nickel and zinc to soluble metal sulfates that can be readily recovered from solution. Although gold is inert to microbial action, microbes can be used to recover gold from certain types of minerals because as they oxidize the ore, they open its structure, thereby allowing gold-solubilizing chemicals such as cyanide to penetrate the mineral. Here, we review a strongly growing microbially-based metal extraction industry, which uses either rapid stirred-tank or slower irrigation technology to recover metals from an increasing range of minerals using a diversity of microbes that grow at a variety of temperatures.     

The role of bacteria in copper mining operations

The results of our survey of several mining operations have shown in every instance typical T. ferrooxidans bacteria to be present in the leaching solutions. The numbers of bacteria found are in general what, one would expect considering the chemical reactions occurring in these processes. A major difficulty is encountered when one considers the complex and unknown series of events which occur, as the bacteria-bearing, acidic, ferric-ion solution penetrates the massive ore dumps. Laboratory studies show microbial activity to be the major cause of rapid solubilization of copper sulfide ores. Field studies suggest that microbial activity cannot explain all observations in a typical leaching operation.     

Microbial leaching of zinc concentrate in fresh‐water microcosms: Comparison between aerobic and oxygen‐limited conditions

Microbially mediated leaching and solubilization of zinc ore concentrate by native aquatic microbial communities incubated under aerobic and oxygen‐limited conditions were examined in static microcosms consisting of stream sediment and water. Sterile controls provided information on abiotic sulfide oxidation and leaching of zinc, lead, cadmium, and copper. The flux of these heavy metals from the sediments to the water column was greatest under biotic oxygen‐limited conditions. When calculated as the percentage of total metal available in zinc concentrate‐amended microcosms, the order of metal solubilization under oxygen‐limited conditions was lead, copper, zinc, and cadmium. Under biotic aerobic conditions, the order of solubilization was zinc, lead, cadmium, and copper. This study indicates that aquatic heterotrophs are capable of leaching and solubilizing metallic sulfides under conditions of neutral to slightly acidic pH and are effective in releasing heavy metals to the water column under oxygen‐limited conditions.     

Progress in bioleaching: fundamentals and mechanisms of bacterial metal sulfide oxidation—part A

Bioleaching of metal sulfides is performed by a diverse group of microorganisms. The dissolution chemistry of metal sulfides follows two pathways, which are determined by the mineralogy and the acid solubility of the metal sulfides: the thiosulfate and the polysulfide pathways. Bacterial cells can effect this metal sulfide dissolution via iron(II) ion and sulfur compound oxidation. Thereby, iron(III) ions and protons, the metal sulfide-attacking agents, are available. Cells can be active either in planktonic state or in forming biofilms on the mineral surface; however, the latter is much more efficient in terms of bioleaching kinetics. In the case of Acidithiobacillus ferrooxidans, bacterial exopolymers contain iron(III) ions, each complexed by two uronic acid residues. The resulting positive charge allows an electrostatic attachment to the negatively charged pyrite. Thus, the first function of complexed iron(III) ions is the mediation of cell attachment, while their second function is oxidative dissolution of the metal sulfide, similar to the role of free iron(III) ions in non-contact leaching. In both cases, the electrons extracted from the metal sulfide reduce molecular oxygen via a redox chain forming a supercomplex spanning the periplasmic space and connecting both outer and inner membranes. In this review, we summarize some recent discoveries relevant to leaching bacteria which contribute to a better understanding of these fascinating microorganisms. These include surface science, biochemistry of iron and sulfur metabolism, anaerobic metabolism, and biofilm formation. The study of microbial interactions among multispecies leaching consortia, including cell-to-cell communication mechanisms, must be considered in order to reveal more insights into the biology of bioleaching microorganisms and their potential biotechnological use.     

Siderophores mediate reduced and increased uptake of cadmium by Streptomyces tendae F4 and sunflower (Helianthus annuus), respectively

Aims: As a toxic metal, cadmium (Cd) affects microbial and plant metabolic processes, thereby potentially reducing the efficiency of microbe or plant‐mediated remediation of Cd‐polluted soil. The role of siderophores produced by Streptomyces tendae F4 in the uptake of Cd by bacteria and plant was investigated to gain insight into the influence of siderophores on Cd availability to micro‐organisms and plants. Methods and Results: The bacterium was cultured under siderophore‐inducing conditions in the presence of Cd. The kinetics of siderophore production and identification of the siderophores and their metal‐bound forms were performed using electrospray ionization mass spectrometry. Inductively coupled plasma spectroscopy was used to measure iron (Fe) and Cd contents in the bacterium and in sunflower plant grown in Cd‐amended soil. Siderophores significantly reduced the Cd uptake by the bacterium, while supplying it with iron. Bacterial culture filtrates containing three hydroxamate siderophores secreted by S. tendae F4 significantly promoted plant growth and enhanced uptake of Cd and Fe by the plant, relative to the control. Furthermore, application of siderophores caused slightly more Cd, but similar Fe uptake, compared with EDTA. Bioinoculation with Streptomyces caused a dramatic increase in plant Fe content, but resulted only in slight increase in plant Cd content. Conclusion: It is concluded that siderophores can help reduce toxic metal uptake in bacteria, while simultaneously facilitating the uptake of such metals by plants. Also, EDTA is not superior to hydroxamate siderophores in terms of metal solubilization for plant uptake. Significance and Impact of the Study: The study showed that microbial processes could indirectly influence the availability and amount of toxic metals taken up from the rhizosphere of plants. Furthermore, although EDTA is used for chelator‐enhanced phytoremediation, microbial siderophores would be ideal for this purpose.     

铁还原菌在水资源再生与能源转化领域的研究进展

铁还原菌是一种典型的异化金属还原菌,广泛分布于海洋沉积物、陆地深地层等自然环境,该类细菌可以将铁氧化物中的Fe(Ⅲ)还原为Fe(Ⅱ),在铁、碳的生物地球化学铁循环中发挥重要作用。铁还原菌的末端电子不局限于Fe(Ⅲ),还可以是其他高价金属、有机污染物,可用于土壤、地下水的污染修复和毒性削减。在微生物电化学系统中,铁还原菌氧化有机物产生的电子直接传递给电极,可以产生电能。基于这种独特的胞外电子传递方式,衍生出了微生物燃料电池、微生物电解池、微生物脱盐电池、微生物燃料电池耦合芬顿反应以及光催化微生物燃料电池,常用于微生物发电、生物传感器、生物制氢、定向发酵、海水淡化、生物脱盐和污染物分解矿化。本文从异化铁还原菌的代谢机制、微生态作用、环境修复、水资源再生与能源转化四个方面,综述了铁还原菌的作用原理及国内外研究现状,分析论述了目前亟需解决的关键问题和未来的研究方向,以期为铁还原菌的基础理论研究和应用技术研发提供参考。     

Bioleaching review part B:

This review describes the historical development and current state of metals leaching and sulfide mineral biooxidation by the minerals industries. During the past 20 years commercial processes employing microorganisms for mineral recovery have progressed from rather uncontrolled copper dump leaching to mineral oxidation and leaching in designed bioheaps for oxidation of refractory gold ores and for copper recovery. Also during this period of time, stirred tank bioleaching has been commercialized for cobalt recovery and for biooxidation of refractory gold ores. Chalcopyrite bioleaching in stirred tanks is on the verge of commercialization. Commercial applications of biohydrometallurgy have advanced due to favorable process economics and, in some cases, reduced environmental problems compared to conventional metal recovery processes such as smelting. Process development has included recognition of the importance of aeration of bioheaps, and improvements in stirred tank reactor design and operation. Concurrently, knowledge of the key microorganisms involved in these processes has advanced, aided by advances in molecular biology to characterize microbial populations.     

Bacterial Oxidation of Refractory Sulfide Ores for Gold Recovery

Abstract

The microbiological leaching of refractory sulfide ores (pyrite, arsenopyrite) for recovery of gold is reviewed in this article. The underlying physiological, biochemical, and genetic fundamentals of the bacteria involved (Thiobacillus and Sulfolobus spp.) are complex and have yet to be elucidated in depth. The chemistry of acid and biological leaching of pyrite and arsenopyrite minerals is also complex, and many of the individual reactions are not known in detail. Bacterial leaching is discussed in relation to chemical speciation at acid pH values. Attempts to develop models for a better understanding of bioleaching processes are summarized. The importance of pH, redox potential, temperature, sulfur balance, and toxic metals is evaluated for optimizing conditions for bacterial activity. Gold is finely disseminated in refractory sulfide ores, thereby decreasing Au recoveries upon conventional cyanidation for gold dissolution. In the bioleaching process, bacteria remove the sulfide minerals by oxidative dissolution and thus expose Au to extraction with cyanide solution. Stirred tank reactors appear most suited for this biological leaching process. The overall oxidation of the sulfides is an important variable for gold recovery. Pilot- and commercial-scale bioleaching processes for gold-containing pyrite and arsenopyrite ores are reviewed. This application of mineral biotechnology competes favorably with pressure leaching and roasting processes, both of which are problematic and energy-intensive alternatives for pretreatment of auriferous pyrite/arsenopyrite ores.     

Bioleaching of manganese (IV) oxide and application to its recovery from ores

Summary Bioleaching of manganese (IV) oxide with Thiobacillus thiooxidans has been studied in media with and without sulfur, ferrous sulfide and ferrous sulfate. The knowledge of the role played by the bacteria and the reducing substances suggest that the leaching of manganese (IV) ores through the use of thiobacteria is only justified when suitable amounts of sulfur or metal sulfides are present.     

冶金微生物的铁硫代谢多样性及其与矿物的相互作用

生物冶金是利用微生物铁硫元素代谢活性加速硫化矿物氧化溶解,并对其中有价金属加以提取回收的技术。冶金系统中微生物的代谢多样性及其耦合功能网络,尤其是以铁硫代谢途径为主的功能网络,在硫化矿物加速氧化溶解过程中承担了重要作用,是生物冶金技术理论研究的核心领域。本文归纳了冶金系统中多样化的微生物物种及其铁硫代谢途径,并从微生物代谢耦合角度探讨了微生物代谢多样性与矿物的相互作用。     

Organoheterotrophe Laugung resistenter Materialien

Leaching processes can be classified in chemolithotrophic and organoheterotrophic mechanisms. In the case of chemolithotrophic leaching sulphide minerals, elemental sulphur, ferrous iron and a number of different reduced metals will be oxidized in a solution containing sulphuric acid of bacterial and/or chemical origin. Organoheterotrophic leaching however is connected with the accumulation of microbial metabolites such as organic acids, proteins, peptides and polysaccharides, which are capable to disintegrate ores, minerals or industrial wastes through dissolution, formation of complexes or chelates. Whereas at the present time chemolithotrophic leaching processes are in operation in industrial scale for the winning of copper, uranium and some other special metals, organoheterotrophic processes, their problems and technical applications are still under study. Therefore problems of organoheterotrophic leaching of chemical high resistant materials such as phosphorus furnace slag and zircon from the Baltic shield have been investigated with regard to possible technical applications for the winning of rare earth elements (REE) and other precious metals. Using a strain of Acetobacter methanolicus which is able to accumulate large amounts of gluconic acid on the basis of glucose or glucose containing by-products, leaching effects up to 90% of REE could be realized in the case of phosphorus furnace slag and up to 45% in the case of zircon.     

Bioremediation of toxic heavy metals using acidothermophilic autotrophes

Investigations were carried out to isolate microbial strains from soil, mud and water samples from metallurgically polluted environment for bioremediation of toxic heavy metals. As a result of primary and secondary screening various 72 acidothermophilic autotrophic microbes were isolated and adapted for metal tolerance and biosorption potentiality. The multi-metal tolerance was developed with higher gradient of concentrations of Ag, As, Bi, Cd, Cr, Co, Cu, Hg, Li, Mo, Pb, Sn and Zn. The isolates were checked for their biosolubilization ability with copper containing metal sulfide ores. In case of chalcopyrite 85.82% and in covellite as high as 97.5% copper solubilization occurred in presence of 10(-3) M multi-heavy metals on fifth day at 55 degrees C and pH 2.5. Chemical analyses were carried out by inductively coupled plasma spectroscopy (ICP) for metal absorption. The selected highly potential isolate (ATh-14) showed maximum adsorption of Ag 73%, followed by Pb 35%, Zn 34%, As 19%, Ni 15% and Cr 9% in chalcopyrite.     

Biogeochemical phenomena induced by bacteria within sulfidic mine tailings

Summary Mill tailings resulting from mining and metallurgical processes are usually disposed of into open-air impoundments, where they become subjected to chemical or microbial leaching. At the surface of the tailings, where oxic conditions prevail, acidophilic bacteria, such as thiobacilli, can oxidize sulfidic minerals (e.g. pyrite and pyrrhotite) and generate acidic metal-rich leachates as by-products of their metabolism. This, combined with chemical oxidation, leads to acid mine drainage (AMD). Biomineralization, whereby a proportion of the metal leachate is precipitated, can also occur in the oxidized tailings, often as a result of a close metal-bacteria interaction. Iron-rich precipitates are usually found on bacterial cell walls, and are thought to serve as nucleation sites for further mineralization within the tailings impoundments. As depth increases in mine tailings, oxygen depletion and the presence of water-saturated pores usually lead to anoxic conditions. Under such redox and chemical conditions, populations of sulfate-reducing bacteria (SRBs) can colonize the tailings. As a result of their metabolic activity, sulfate is reduced to hydrogen sulfide, which in turn can react with dissolved metals to form metal sulfide precipitates. Microbial sulfate reduction also generates alkalinity, although chemical dissolution of carbonate and oxide minerals probably also play an important role in the generation of alkaline conditions in mine tailings.     

Genetics of metal resistance in acidophilic prokaryotes of acidic mine environments

Acidophilic bacteria inhabiting acidic mine regions cause natural leaching of sulphidic ores. They are now exploited in industrial operations for leaching of metals and beneficiation of low-grade and recalcitrant ores. Recent trends emphasize application of thermoacidophiles and genetic engineering of ore-leaching bacteria for greater success in this area. This requires an in-depth understanding on the molecular genetics of these bacteria and construction of cloning vectors for them. Metal resistance is considered as the most suitable phenotypic trait for cloning vectors of bio-mining chemolithoautotrophic (viz. Acidithiobacillus ferrooxidans) and heterotrophic (Acidiphilium and Acidocella species) bacteria of mine environments. These bacteria take part in ore-leaching either directly or indirectly, exhibit low to high level of resistance/tolerance to various metals under different conditions. Majority of these bacteria contain one or more plasmids--the genetic elements that usually carry metal resistant genes. But none of the At. ferrooxidans plasmids has been definitely proved to harbour metal-resistant genes which have mostly been found in the chromosome of this bacterium. Plasmids of acidophilic heterotrophs of the genera Acidiphilium and Acidocella, on the other hand, carry metal resistant genes. While genes bestowing arsenic resistance in Acidiphilium multivorum are similar to those analyzed from other sources, the metal (Cd and Zn)-resistance conferring cloned plasmid DNA fragments from Acidiphilium symbioticum KM2 and Acidocella GS19h strains were found to have no sequence similarity with the reported Cd- and Zn-resistant genes. Such observations indicate some novel aspects of metal resistance in acidophilic bacteria.     

Bioleaching review part A:

Bioleaching of metal sulfides is caused by astonishingly diverse groups of bacteria. Today, at least 11 putative prokaryotic divisions can be related to this phenomenon. In contrast, the dissolution (bio)chemistry of metal sulfides follows only two pathways, which are determined by the acid-solubility of the sulfides: the thiosulfate and the polysulfide pathway. The bacterial cell can effect this sulfide dissolution by contact and non-contact mechanisms. The non-contact mechanism assumes that the bacteria oxidize only dissolved iron(II) ions to iron(III) ions. The latter can then attack metal sulfides and be reduced to iron(II) ions. The contact mechanism requires attachment of bacteria to the sulfide surface. The primary mechanism for attachment to pyrite is electrostatic in nature. In the case of Acidithiobacillus ferrooxidans, bacterial exopolymers contain iron(III) ions, each complexed by two uronic acid residues. The resulting positive charge allows attachment to the negatively charged pyrite. Thus, the first function of complexed iron(III) ions in the contact mechanism is mediation of cell attachment, while their second function is oxidative dissolution of the metal sulfide, similar to the role of free iron(III) ions in the non-contact mechanism. In both cases, the electrons extracted from the metal sulfide reduce molecular oxygen via a complex redox chain located below the outer membrane, the periplasmic space, and the cytoplasmic membrane of leaching bacteria. The dominance of either At. ferrooxidans or Leptospirillum ferrooxidans in mesophilic leaching habitats is highly likely to result from differences in their biochemical iron(II) oxidation pathways, especially the involvement of rusticyanin.     

Recent advances in microbial mining

Microbial mining of copper sulphide ores, has been practiced on an industrial scale since the late 1950s. Since then, advances in microbial mining and the role of microorganisms involved in solubilization of metals have assumed commerical importance. The fact that bioleaching processes save energy, have a minimum pollution potential and are able to yield value-added by-products make these processes invaluable. The metal extraction processes using microorganisms, which are currently in active use, concern copper and uranium bioleaching. Biobeneficiation is also applied at an industrial scale for recovery of gold from arsenopyrites. The developments in these processes during the last 15 years, with particular reference to developing nations, are reviewed. Information gathered on molecular genetics of these microorganisms should lead to a better understanding and control of microbial leaching processes. Areas still needing research to sustain economic expansion of microbial mining techniques are indicated.The author is with the Agharkar Research Institute, Agarkar Road, Pune 411 004, India     

Biological leaching of sulfide minerals with the use of shake flask,aerated column,air-lift reactor,and percolation techniques

Four different experimental approaches were used to evaluate the microbiological leaching of ore material containing metal sulfides (Fe, Zn, Ni, Cu, Co) and aluminum silicates. A shake flask technique required the shortest contact time for the complete solubilization of the most readily leachable metals (Ni and Zn). Air-lift reactors and aerated column reactors required longer contact times and complete solubilization of either zinc or nickel was not achieved. The air-lift reactor approach was somewhat more effective than the aerated slurry technique. A percolation system was the least effective and yielded the lowest recoveries. Shake flasks (easily autoclavable) offered the advantage of comparison of the microbiological and solely chemical leaching. Aseptic conditions could not be maintained with the air-lift and aerated column reactors because of contamination via aerosol formation. In a relative scale the leaching patterns were similar in that the precipitation of Fe(III) occurred regardless of the technique; zinc and nickel sulfides were solubilized more quantitatively than those of copper and cobalt; aluminum concentrations, although high, indicated low leaching yields relative to aluminum silicates in the ore material; and the pH reached similar final values in the presence of bacteria.     

Fluorescence study of the interaction of Suwannee River fulvic acid with metal ions and Al3+-metal ion competition

In this study emission and synchronous-scan fluorescence spectroscopy have been used to investigate the interaction of the class A (oxygen seeking 'hard acid') metal Al(3+), with Suwannee River fulvic acid (SRFA), as well as competition between Al(3+) and several other metal ions (Ca(2+), Mg(2+), Cu(2+), Pd(2+), La(3+), Tb(3+) and Fe(3+)) for binding sites on SRFA. Of the four metal ions possessing very similar (and relatively low) ionic indices (Ca(2+), Mg(2+), Cu(2+) and Pd(2+)) only the latter two paramagnetic ions significantly quenched SRFA fluorescence emission intensity. Of the four metal ions possessing very similar (and relatively low) covalent indices (Ca(2+), Mg(2+), La(3+) and Tb(3+)) only the last paramagnetic ion (Tb(3+)) significantly quenched SRFA fluorescence. None of these metals was able to significantly compete with SRFA-bound Al(3+).Fe(3+), which differs substantially from all of the other metals examined in this study in that it possesses a relatively high ionic index (but not as high as Al(3+)) and a relatively low covalent index (but not as low as Al(3+)), was able not only to quench SRFA fluorescence but also to compete (at least to some extent) with SRFA-bound Al(3+). Synchronous-scan fluorescence SRFA spectra taken in the absence and presence of Fe(3+) and/or Al(3+) support the view that these two metal ions can compete for sites on SRFA. The results of these fluorescence experiments further confirm the Al(3+), and metal ions that have electronic properties somewhat similar to Al(3+) (such as Fe(3+)) are somewhat unique in their ability to interact strongly with binding sites on fulvic acids.