A Large-Scale Multiple Genome Comparison of Acidophilic Archaea (pH ≤ 5.0) Extends Our Understanding of Oxidative Stress Responses in Polyextreme Environments

Acidophilic archaea thrive in anaerobic and aerobic low pH environments (pH < 5) rich in dissolved heavy metals that exacerbate stress caused by the production of reactive oxygen species (ROS) such as hydrogen peroxide (H2O2), hydroxyl radical (OH) and superoxide (O2−). ROS react with lipids, proteins and nucleic acids causing oxidative stress and damage that can lead to cell death. Herein, genes and mechanisms potentially involved in ROS mitigation are predicted in over 200 genomes of acidophilic archaea with sequenced genomes. These organisms are often be subjected to simultaneous multiple stresses such as high temperature, high salinity, low pH and high heavy metal loads. Some of the topics addressed include: (1) the phylogenomic distribution of these genes and what this can tell us about the evolution of these mechanisms in acidophilic archaea; (2) key differences in genes and mechanisms used by acidophilic versus non-acidophilic archaea and between acidophilic archaea and acidophilic bacteria and (3) how comparative genomic analysis predicts novel genes or pathways involved in oxidative stress responses in archaea and likely horizontal gene transfer (HGT) events.

Extraction of Selected Metals from Waste Printed Circuit Boards by Bioleaching Acidophilic Bacteria

Technological innovations and increased demand for electronic devices resulted in production of more and more waste with highmetal content. Worldwide, 50 million tons of WEEE (Waste from Electrical and Electronic Equipment) are generated each year. Giventhe metal content present in electrical waste (e-waste), it is considered to be an urban mine and, if properly treated, can serve as analternative secondary source of metals. Waste printed circuit boards (WPCBs) that constitute approx. 3-5% of WEEE by weight areof particular importance. They contain, on average, 30-40% of metals by weight, with higher purity than in minerals. With environmental and economic benefits in mind, increasing attention is being paid to the development of processes to recover metals and othervaluable materials from WPCBs. The research presented in the article aimed at assessing the usefulness of the biotechnological methodfor leaching of selected metals from e-waste. The results indicate that it is possible to mobilize metals from WPCBs using microorganisms such as Acidithiobacillus ferroxidans bacteria

Intensification of Nickel Bioleaching with Neutrophilic Bacteria Guyparkeria halophila as an Approach to Limitation of Sulfuric Acid Pollution

Hydrometallurgical production of valuable and non-ferrous metals is traditionally accompanied with acid waste effluents/acid mine drainage leading to acidification of the mining areas. The traditional cause of this pollution is the well-known technology based on the recovery of metals with acid solutions and the application of strong acidophilic leaching bacteria for the oxidation of sulfide ores. In our experiments, we used neutrophilic autotrophic bacteria (NAB) stimulated with formic acid or coupled with acidophilic bacteria. The first approach was based on using formic acid as an energetic substrate by autotrophic bacteria. In the second case, the NAB provided initial biogenic acidification for the following growth of the inoculated acidophilic bacteria. Our experiments resulted in increased nickel recovery from the low-grade sulfide ores, which was provided by the NAB in a medium supplemented with formic acid. Bioleaching resulted in 1116 mg Ni/L (69.75%) in the medium with formate and only 35.4 mg Ni/L without formate in 43 days. As a whole, our bench scale experiments showed that the stimulated NAB can be effective at pH 7–5. Partially replacing sulfuric acid with formic acid could also give benefits via the following natural degradation of acid wastes. As a whole, this approach is more environmentally friendly than conventional bioleaching techniques.

A large scale, multiple genome comparison of acidophilic Archaea (pH ≤ 5.0) extends our understanding of oxidative stress responses in polyextreme environments

Acidophilic Archaea thrive in anaerobic and aerobic low pH environments (<pH 5) rich in dissolved heavy metals that exacerbate stress caused by the production of reactive oxygen species (ROS) such as hydrogen peroxide (H2O2), hydroxyl radical (·OH) and superoxide (O2·−). ROS react with lipids, proteins and nucleic acids causing oxidative stress and damage that can lead to cell death. Herein, genes and mechanisms potentially involved in ROS mitigation are predicted in over 200 genomes of acidophilic Archaea with sequenced genomes. These organisms can be subjected to simultaneous multiple stresses such as high temperature, high salinity, low pH and high heavy metal loads. Some of the topics addressed include: (1) the phylogenomic distribution of these genes and what can this tell us about the evolution of these mechanisms in acidophilic Archaea; (2) key differences in genes and mechanisms used by acidophilic versus non-acidophilic Archaea and between acidophilic Archaea and acidophilic Bacteria and (3) how comparative genomic analysis predicts novel genes or pathways involved in oxidative stress responses in Archaea and possible Horizontal Gene Transfer (HGT) events.

A large-scale genome-based survey of acidophilic Bacteria suggests that genome streamlining is an adaption for life at low pH

Genome streamlining theory suggests that reduction of microbial genome size optimizes energy utilization in stressful environments. Although this hypothesis has been explored in several cases of low nutrient (oligotrophic) and high temperature environments, little work has been carried out on microorganisms from low pH environments and what has been reported is inconclusive. In this study, we performed a large-scale comparative genomics investigation of more than 260 bacterial high-quality genome sequences of acidophiles, together with genomes of their closest phylogenetic relatives that live at circum-neutral pH. A statistically supported correlation is reported between reduction of genome size and decreasing pH that we demonstrate is due to gene loss and reduced gene sizes. This trend is independent from other genome size constraints such as temperature and G+C content. Genome streamlining in the evolution of acidophilic Bacteria is thus supported by our results. Analyses of predicted COG categories and subcellular location predictions indicate that acidophiles have a lower representation of genes encoding extra-cellular proteins, signal transduction mechanisms and proteins with unknown function, but are enriched in inner membrane proteins, chaperones, basic metabolism, and core cellular functions. Contrary to other reports for genome streamlining, there was no significant change in paralog frequencies across pH. However, a detailed analysis of COG categories revealed a higher proportion of genes in acidophiles in the following categories: 'Replication and repair', 'Amino acid transport' and 'Intracellular trafficking'. This study brings increasing clarity regarding genomic adaptations of acidophiles to life at low pH while putting elements such as the reduction of average gene size under the spotlight of streamlining theory.

Biogenic Platinum Nanoparticles’ Production by Extremely Acidophilic Fe(III)-Reducing Bacteria

Platinum nanoparticles (Pt(0)NPs) are expected to play a vital role in future technologies as high-performance catalysts. The microbiological route for Pt(0)NPs’ production is considered a greener and simpler alternative to conventional methods. In order to explore the potential utility of extreme acidophiles, Fe(III)-reducing acidophilic bacteria, Acidocella aromatica and Acidiphilium crytpum, were tested for the production of bio-Pt(0)NPs from an acidic solution. Bio-Pt(0)NPs were successfully formed via a simple one-step reaction with the difference in the size and location between the two strains. Intact enzymatic activity was essential to exhibit the site for Pt(0) crystal nucleation, which enables the formation of well-dispersed, fine bio-Pt(0)NPs. Active Ac. aromatica cells produced the finest bio-Pt(0)NPs of mean and median size of 16.1 and 8.5 nm, respectively. The catalytic activity of bio-Pt(0)NPs was assessed using the Cr(VI) reduction reaction, which was shown to be in a negative linear correlation with the mean particle size under the conditions tested. This is the first study reporting the recruitment of acidophilic extremophiles for the production of Pt(0)NPs. Acidophilic extremophiles often inhabit metal-rich acidic liquors in nature and are expected to become the promising tool for metal nanotechnology.

Glutathione synthetase overexpression in Acidithiobacillus ferrooxidans improves halotolerance of iron oxidation

Acidithiobacillus ferrooxidans are well-studied iron- and sulfur-oxidizing acidophilic chemolithoautotrophs that are exploited for their ability to participate in the bioleaching of metal sulfides. Here, we overexpressed the endogenous glutamate–cysteine ligase and glutathione synthetase genes in separate strains and found that glutathione synthetase overexpression increased intracellular glutathione levels. We explored the impact of pH on the halotolerance of iron oxidation in wild type and engineered cultures. The increase in glutathione allowed the modified cells to grow under salt concentrations and pH conditions that are fully inhibitory to wild type cells. Furthermore, we found that improved iron oxidation ability in the presence of chloride also resulted in higher levels of intracellular ROS in the strain. These results indicate that glutathione overexpression can be used to increase halotolerance in A. ferrooxidans and would likely be a useful strategy on other acidophilic bacteria. Importance The use of acidophilic bacteria in the hydrometallurgical processing of sulfide ores can enable many benefits including the potential reduction of environmental impacts. The cells involved in bioleaching tend to have limited halotolerance, and increased halotolerance could enable several benefits, including a reduction in the need for the use of fresh water resources. We show that the genetic modification of A. ferrooxidans for the overproduction of glutathione is a promising strategy to enable cells to resist the oxidative stress that can occur during growth in the presence of salt.

Bioleaching of Transition Metals From Limonitic Laterite Deposits and Reassessment of the Multiple Roles of Sulfur-Oxidizing Acidophiles in the Process

Using acidophilic bacteria to catalyze the reductive dissolution of oxidized minerals is an innovative process that facilitates the extraction of valuable base metals (principally cobalt and nickel) from limonites, which are otherwise often regarded as waste products of laterite mining. The most appropriate conditions required to optimize reductive mineral dissolution are unresolved, and the current work has reassessed the roles of Acidithiobacillus spp. in this process and identified novel facets. Aerobic bio-oxidation of zero-valent sulfur (ZVS) can generate sufficient acidity to counterbalance that consumed by the dissolution of oxidized iron and manganese minerals but precludes the development of low redox potentials that accelerate the reductive process, and although anaerobic oxidation of sulfur by iron-reducing species can achieve this, less acid is generated. Limited reduction of soluble iron (III) occurs in pure cultures of Acidithiobacillus spp. (Acidithiobacillus thiooxidans and Acidithiobacillus caldus) that do not grow by iron respiration. This phenomenon (“latent iron reduction”) was observed in aerated cultures and bioreactors and was independent of electron donor used (ZVS or hydrogen). Sufficient ferrous iron was generated in the presence of sterilized hydrophilic sulfur (bio-ZVS) to promote the effective reductive dissolution of Mn (IV) minerals in limonite and the solubilization of cobalt in the absence of viable acidophiles.