Papers by Keyword: Biomining

Abstract: Phage display for discovery of specific binding peptides is nowadays widely used in the pharmaceutical industry and in many biotechnological applications. Using state-of-the-art cloning techniques we developed an easy-to-use cloning and expression system, allowing the fast production of identified peptides while avoiding proteolysis.

Abstract: Since the discovery of bioleaching microorganisms and their role in metal extraction in the 1940s, a number of technical approaches have been developed to enhance microbially catalysed solubilisation of metals from ores, concentrates and waste materials. Biomining has enabled the transformation of uneconomic resources to reserves, and thus help to alleviate the challenges related to continually declining ore grades. The rapid advancement of microbial characterisation methods has vastly increased our understanding of microbial communities in biomining processes. The objective of this paper is to review the recent advances in biomining processes and microbial characterisation.

Abstract: Bioleaching and biooxidation of sulfidic ores and concentrates generate very high acidities and a great of heat, which rise the temperature in the reactors or heaps, and accumulate the sulfur on the surface of the ores. Extremely thermoacidophilic archaea, mainly from the genus of Acidianus, Sulfolobus, Metallosphaera and sulfurisphaera, have great potential to contribute to biomining processes for their inherent tolerance for low pH, high temperature, and high-soluble metal concentrations. Species of the genus Metallosphaera typically grow by aerobic respiration on CO₂ with S⁰, tetrathionate (S₄O₆²⁺), and Fe²⁺ as electron donors, particularly suitble for metal extraction under high temperature by their iron- and sulfur-oxidation ability. Several species from Metallosphaera and Acidianus genera were investigated for their ability and conditions to dissolve various ores under a range of conditions. All of them showed good performance in copper extraction from chalcopyrite, with strain M.cuprina Ar-4 displaying higher activity than others. Surface analysis of chalcopyrite leached with the strain showed the leaching products accumulated on the ores. Our study will cover new understandings on the application of these thermoacidophilic archaea in biomining.

Abstract: Acidophiles have been widely used in heap and dump bioleaching of secondary copper sulfide ores and biooxidation of refractory gold ores. 22 genera of acidophiles have been found in biomining environments. This paper gives a preliminary introduction to the application of mineral biomining in China. Challenges and technical trends for heap bioleaching of primary copper sulfide ores, purification of bioleaching solution of polymetallic sulfide ores and biooxidation of carbonaceous refractory gold ores are also recommended.

Abstract: Extremophiles such as the acidophilic Sulfolobus metallicus (Archaea) and Acidithiobacillus ferrooxidans (Bacteria) can resist Cu (CuSO4) concentrations of 200 mM and 800 mM respectively. These microorganisms are important in biomining processes to extract copper and other metals. A. ferrooxidans grown at low Cu concentrations (5 mM) expressed genes coding for ATPases most likely involved in pumping the metal from the cytoplasm to the periplasm of the bacterium. At 100 mM Cu the previous systems were repressed and there was a great induction in the expression of efflux systems known to use the proton motive force energy to export the metal outside the cell. These Cu-resistance determinants from A. ferrooxidans were found to be functional since when expressed in Escherichia coli they conferred higher Cu tolerance to it. Novel Cu-resistance determinants for A. ferrooxidans were found and characterized. S. metallicus possessed at least 2 CopM metallochaperones and 2 CopA ATPases whose expressions were induced by Cu (5 to 50 mM). Furthermore, we previously reported that both microorganisms accumulate high levels of inorganic polyphosphate (PolyP) and that intracellular Cu concentration stimulates polyP hydrolysis. The resulting Pi would then be transported out of the cell as a metal-Pi complex to detoxify the cells. In addition, our results suggest that at high Cu concentrations polyP could also provide energy for the metal efflux. All the data suggest that both biomining microorganisms use different systems to respond to Cu depending on the extracellular concentrations of the metal and suggest that the presence of different additional systems to respond to Cu may explain the extremely high metal resistance of these extremophiles.

Abstract: Acidithiobacillus ferrooxidans, A. thiooxidans, and A. caldus are acidophilic Gram-negative -proteobacteria involved in the bioleaching of metal sulfides. Bacterial attachment to mineral surface and biofilm development play a pivotal role in this process. Therefore, the understanding of biofilm formation has relevance to the design of biological strategies to improve the efficiency of bioleaching processes. For this reason, our laboratory is focused on the characterization of the molecular mechanisms involved in biofilm formation in biomining bacteria. In many bacteria, the intracellular level of c-di-GMP molecules regulates the transition from the motile planktonic state to sessile community-based behaviors, such as biofilm development. Thus, we recently started the study of c-di-GMP pathway in biomining bacteria. C-di-GMP molecules are synthesized by diguanylate cyclases (DGCs) and degraded by phosphodiesterases (PDEs). So far, two kinds of effectors have been identified, including three protein families (pilZ, PleD and FleQ) and a conserved RNA domain (GEMM) which acts as a riboswitch. We previously reported the existence of different molecular players involved in c-di-GMP pathway in A. ferrooxidans ATCC 23270. Here, we expanded our work to other Acidithioibacillus species: A. thiooxidans ATCC 19377 and A. caldus ATCC 51756. In both, we identified several putative-ORFs encoding DGC, PDE and effector proteins. By using total RNA extracted from A. ferrooxidans and A. caldus cells in RT-PCR and qPCR experiments, we demonstrated that these genes are expressed. In addition, we characterized the presence of c-di-GMP in A. ferrooxidans ATCC 23270 and A. caldus ATCC 51756 cell extracts. Taken together, these results strongly suggest that A. ferrooxidans, A. caldus and A. thiooxidans possess functional c-di-GMP pathways. As it occurs in other Gram-negative bacteria, this pathway should be involved in the regulation of the planktonic/biofilm switch. In the future, we have to integrate this new biological dimension to improve the biological understanding of bioleaching.

Abstract: Acidithiobacillus ferrooxidans is a chemolithoautotrophic acidophile capable of catalyzing the oxidation of ferrous iron and sulfur reducing compounds. During the dissolution of ores A. ferrooxidans adheres to the solid substrate by hydrophobic and hydrophilic interactions and also extracellular polymeric substances may be involved. Nevertheless, many of these agents that participate in the process are unknown. Therefore, the identification and characterization of the extracellular proteome of A. ferrooxidans was addressed during this work. First, by searching the genome of A. ferrooxidans ATCC 23270 the putative genes coding for proteins present in most of the different secretion systems (type I, II, IV, V systems and the Usher pathway) were found. Second, when the secretion signal prediction server SubCel 1.0 was used, 26.7 % of the total open reading frames showed possible signals that would allow these proteins to leave the cytoplasm towards the internal and outer membranes, the periplasm, or the extracellular milieu. By using 2D-NEPHGE and MS sequencing we identified the eleven most abundant proteins present in the extracellular fraction. Proteins related to the transport and binding of solutes and the folding of proteins, among others were identified. Along with the non-classical pathway, the protein secretion systems identified could constitute the secretion machinery for the proteins found in the extracellular sub-proteome of A. ferrooxidans that as a whole constitute its secretome. Further studies of the extracellular proteins from this biomining microorganism will be important to find out their possible role if any, during bacteria-mineral interactions.

Abstract: The metabolic potential of 16 bioleaching microorganisms (Eubacteria and Archaea) has been investigated, allowing the prediction of potential inter- and intra-species physiological interactions (ecophysiology) during spatial and temporal changes that are known to occur within industrial bioleaching heaps. Genome analysis has allowed preliminary models to be built for genes and pathways involved in key processes such as nitrogen and carbon cycling, sulfur and iron uptake and homeostasis, extra-cellular polysaccharide biosynthesis, heavy metal resistance and energy metabolism. This paper will focus on the diverse ways that microorganisms obtain carbon from their environment with a particular emphasis on elucidating how these processes might be expected to vary over space and time during the lifetime of a bioleaching operation. It is anticipated that this knowledge will improve our understanding of fundamental biological processes in extremely acidic environments and it is hoped that it will capture usable knowledge that can be applied to bioleaching. Comparative genomics between two strains of Acidthiobacillus ferrooxidans highlights the importance of lateral gene transfer in increasing genetic and metabolic potential and suggests that classical molecular DNA techniques, such as rDNA typing, significantly underestimate the microbial diversity of bioleaching heaps.

Abstract: Small regulatory RNAs (srRNAs) control gene expression in Bacteria, usually at the posttranscriptional level, by acting as antisense RNAs that bind targeted mRNAs or by interacting with regulatory proteins. srRNAs are involved in the regulation of a large variety of processes such as plasmid replication, transposition and global genetic circuits that respond to environmental changes. Since their discovery a few years ago, it has become apparent that they are prolific and widespread. In this study, we describe bioinformatic approaches to srRNA discovery in the biomining microorganisms Acidithiobacillus ferrooxidans, A. caldus and A. thiooxidans. Intergenic regions of the annotated genomes were extracted and computationally searched for srRNAs. Candidate srRNAs that were associated with predicted sigma 70 promoters and/or rho-independent terminators were chosen for further study. The resulting potential srRNAs include known examples from other microorganisms and some novel candidates and reveal interesting underlying biology of the Acidithiobacillus genus.

Abstract: Biofilm development plays a pivotal role in the bioleaching process. The attachment of the acidophilic chemolithotrophic Acidithiobacillus ferrooxidans to mineral surfaces is mediated by extracellular polymeric substances (EPS) involved in biofilm development. Previous work suggests that EPS composition of A. ferrooxidans is adapted to the energy source and, accordingly, the bacterium must be able to sense the surface to which attachment occurs with the consequent triggering of the expression of different EPS-genes. Quorum sensing (QS) is recognized as one of the main regulators of biofilm formation. A. ferrooxidans possesses a functional QS type AI-1 system and the analysis of culture supernatants revealed us that this bacterium is able to synthesize nine different homoserine lactones (AHLs) whose acyl-chain lengths oscillate between 8 and 16 carbons and include an alcohol or a ketone function at the C3 position. The transcription levels of the afeI gene encoding for the AHL synthase are higher in cells grown in sulfur and thiosulfate media than in iron-grown cells, suggesting that biofilm formation in A. ferrooxidans would be regulated by the QS type AI-1 system. In the present study, the effect of several synthetic AHLs and analogues on the attachment of A. ferrooxidans to pyrite was analyzed. Preliminary results suggest that some of these molecules are changing the bacterial attachment to pyrite.