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Acidithiobacillus thiooxidans secretome containing a newly described lipoprotein Licanantase enhances chalcopyrite bioleaching rate.

ABSTRACT: The nature of the mineral-bacteria interphase where electron and mass transfer processes occur is a key element of the bioleaching processes of sulfide minerals. This interphase is composed of proteins, metabolites, and other compounds embedded in extracellular polymeric substances mainly consisting of sugars and lipids (Gehrke et al., Appl Environ Microbiol 64(7):2743-2747, 1998). On this respect, despite Acidithiobacilli-a ubiquitous bacterial genera in bioleaching processes (Rawlings, Microb Cell Fact 4(1):13, 2005)-has long been recognized as secreting bacteria (Jones and Starkey, J Bacteriol 82:788-789, 1961; Schaeffer and Umbreit, J Bacteriol 85:492-493, 1963), few studies have been carried out in order to clarify the nature and the role of the secreted protein component: the secretome. This work characterizes for the first time the sulfur (meta)secretome of Acidithiobacillus thiooxidans strain DSM 17318 in pure and mixed cultures with Acidithiobacillus ferrooxidans DSM 16786, identifying the major component of these secreted fractions as a single lipoprotein named here as Licanantase. Bioleaching assays with the addition of Licanantase-enriched concentrated secretome fractions show that this newly found lipoprotein as an active protein additive exerts an increasing effect on chalcopyrite bioleaching rate.

SUBMITTER: Bobadilla Fazzini RA

PROVIDER: S-EPMC3023857 | biostudies-literature | 2011 Feb

REPOSITORIES: biostudies-literature

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Acidithiobacillus thiooxidans secretome containing a newly described lipoprotein Licanantase enhances chalcopyrite bioleaching rate.

Bobadilla Fazzini Roberto A RA Levican Gloria G Parada Pilar P

Applied microbiology and biotechnology 20101230 3

The nature of the mineral-bacteria interphase where electron and mass transfer processes occur is a key element of the bioleaching processes of sulfide minerals. This interphase is composed of proteins, metabolites, and other compounds embedded in extracellular polymeric substances mainly consisting of sugars and lipids (Gehrke et al., Appl Environ Microbiol 64(7):2743-2747, 1998). On this respect, despite Acidithiobacilli-a ubiquitous bacterial genera in bioleaching processes (Rawlings, Microb ...[more]

PMID: 21191788

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Project description:Acidithiobacillus thiooxidans is one of the most studied biomining species, highlighting its ability to oxidize reduced inorganic sulfur compounds, coupled with its elevated capacity to live under an elevated concentration of heavy metals. In this work, using an in silico semi-automatic genome scale approach, two biological networks for A. thiooxidans Licanantay were generated: (i) An affinity transcriptional regulatory network composed of 42 regulatory family genes and 1,501 operons (57% genome coverage) linked through 2,646 putative DNA binding sites (arcs), (ii) A metabolic network reconstruction made of 523 genes and 1,203 reactions (22 pathways related to biomining processes). Through the identification of confident connections between both networks (V-shapes), it was possible to identify a sub-network of transcriptional factor (34 regulators) regulating genes (61 operons) encoding for proteins involved in biomining-related pathways. Network analysis suggested that transcriptional regulation of biomining genes is organized into different modules. The topological parameters showed a high hierarchical organization by levels inside this network (14 layers), highlighting transcription factors CysB, LysR, and IHF as complex modules with high degree and number of controlled pathways. In addition, it was possible to identify transcription factor modules named primary regulators (not controlled by other regulators in the sub-network). Inside this group, CysB was the main module involved in gene regulation of several bioleaching processes. In particular, metabolic processes related to energy metabolism (such as sulfur metabolism) showed a complex integrated regulation, where different primary regulators controlled several genes. In contrast, pathways involved in iron homeostasis and oxidative stress damage are mainly regulated by unique primary regulators, conferring Licanantay an efficient, and specific metal resistance response. This work shows new evidence in terms of transcriptional regulation at a systems level and broadens the study of bioleaching in A. thiooxidans species.

Project description:The introduction of Acidithiobacillus thiooxidans A01 strengthens the positive interactions between physiologically distinct microorganisms and enhances the bioleaching ability of the consortium. However, the effect of introducing an exogenous strain, A. thiooxidans A01 on the structure and function of the adsorbed and planktonic microbial consortia during bioleaching of low-grade copper sulfide remains unclear. In this study, A. thiooxidans A01 was introduced into an indigenous leaching microbial community on the 0th (group B), 24th (group C), and 36th day (group D). Results revealed that the copper leaching efficiency was highest in group D, in which the Cu2+ concentration in the solution reached 251.5 mg/L on day 48, which was 18.5% higher than that of the control (group A, no addition of A. thiooxidans A01). Restriction fragment length polymorphism (RFLP) analysis of the microbial community in group D revealed the presence of Leptospirillum ferriphilum, Acidithiobacillus ferrooxidans, Acidithiobacillus caldus, Sulfobacillus sp., Acidiphilium spp., and Acidithiobacillus albertensis before introduction of A. thiooxidans A01 on the 36th or 48th day; however, A. albertensis was absent on day 48 in group A. Further, the proportion of dominant A. caldus, L. ferriphilum, and A. ferrooxidans became altered. The results of real-time PCR in group D showed that A. thiooxidans A01 was primarily adsorbed on the surface of the ore, with the adsorption reaching the maxima on day 42; while the free A. thiooxidans A01 in solution grew slowly, reaching its maximum concentration on day 45. Compared with that in the control group, the abundance of both free and attached A. caldus and Sulfobacillus sp. decreased following the introduction of A. thiooxidans A01, while that of L ferriphilum, A. ferrooxidans, and Acidiphilium sp. increased. Functional gene arrays data indicated that the abundance of genes involved in sulfide and iron oxidation in L. ferriphilum and A. ferrooxidans, as well as that of the metal (loid) resistance genes of A. ferrooxidans, L. ferriphilum, and Acidiphilium sp. increased, while the abundance of genes involved in sulfur metabolism in A. caldus and Sulfolobus spp. decreased. Taken together, these results provide useful information for application of bioleaching of copper sulfide in industry.

Project description:BACKGROUND:Acidithiobacillus thiooxidans (A. thiooxidans), a chemolithoautotrophic extremophile, is widely used in the industrial recovery of copper (bioleaching or biomining). The organism grows and survives by autotrophically utilizing energy derived from the oxidation of elemental sulfur and reduced inorganic sulfur compounds (RISCs). However, the lack of genetic manipulation systems has restricted our exploration of its physiology. With the development of high-throughput sequencing technology, the whole genome sequence analysis of A. thiooxidans has allowed preliminary models to be built for genes/enzymes involved in key energy pathways like sulfur oxidation. RESULTS:The genome of A. thiooxidans A01 was sequenced and annotated. It contains key sulfur oxidation enzymes involved in the oxidation of elemental sulfur and RISCs, such as sulfur dioxygenase (SDO), sulfide quinone reductase (SQR), thiosulfate:quinone oxidoreductase (TQO), tetrathionate hydrolase (TetH), sulfur oxidizing protein (Sox) system and their associated electron transport components. Also, the sulfur oxygenase reductase (SOR) gene was detected in the draft genome sequence of A. thiooxidans A01, and multiple sequence alignment was performed to explore the function of groups of related protein sequences. In addition, another putative pathway was found in the cytoplasm of A. thiooxidans, which catalyzes sulfite to sulfate as the final product by phosphoadenosine phosphosulfate (PAPS) reductase and adenylylsulfate (APS) kinase. This differs from its closest relative Acidithiobacillus caldus, which is performed by sulfate adenylyltransferase (SAT). Furthermore, real-time quantitative PCR analysis showed that most of sulfur oxidation genes were more strongly expressed in the S0 medium than that in the Na2S2O3 medium at the mid-log phase. CONCLUSION:Sulfur oxidation model of A. thiooxidans A01 has been constructed based on previous studies from other sulfur oxidizing strains and its genome sequence analyses, providing insights into our understanding of its physiology and further analysis of potential functions of key sulfur oxidation genes.

Dataset Information

Acidithiobacillus thiooxidans secretome containing a newly described lipoprotein Licanantase enhances chalcopyrite bioleaching rate.

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Acidithiobacillus thiooxidans secretome containing a newly described lipoprotein Licanantase enhances chalcopyrite bioleaching rate.

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