Heavy metal ions removed from imitating acid mine drainages with a thermoacidophilic archaea: Acidianus manzaensis YN25.

Metals in acid mine drainages (AMD) have posed a great threat to environment, and in situ economic environment-friendly remediation technologies need to be developed. Moreover, the effects of acidophiles on biosorption and migrating behaviors of metals in AMD have not been previously reported. In this study, the extremely thermoacidophilic Archaea, Acidianus manzaensis YN25 (A. manzaensis YN25) was used as a bio-adsorbent to adsorb metals (Cu2+, Ni2+, Cd2+ and Zn2+) from acidic solutions which were taken to imitate AMD. The values of their maximum biosorption capacities at both high (1 mM) and low (0.1 mM) metal concentrations followed the order: Cu2+ > Ni2+ > Cd2+ > Zn2+. With the elevations of temperature and pH value, the adsorption amounts of metals increased. The results also indicated that A. manzaensis YN25 had the highest adsorption affinity to Cu2+ in coexisting system of quaternary metals. Acid-base titration data revealed that carboxyl and phosphoryl groups provided adsorption sites for metals via deprotonation. Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS) further corroborated that amino played an important role in the biosorption process. The fitted Langmuir model illustrated monolayer adsorption occurring on cell surface. The possible adsorption mechanism of A. manzaensis YN25 mainly involved in electrostatic attraction and complexes formation. This study gives a profound insight into the biosorption behavior of heavy metals in acidic solution by thermoacidophilic Archaea and provides a probable novel strategy for in situ remediation of heavy metals pollution in AMD.

Biofilm formation and interspecies interactions in mixed cultures of thermo-acidophilic archaea Acidianus spp. and Sulfolobus metallicus.

The understanding of biofilm formation by bioleaching microorganisms is of great importance for influencing mineral dissolution rates and to prevent acid mine drainage (AMD). Thermo-acidophilic archaea such as Acidianus, Sulfolobus and Metallosphaera are of special interest due to their ability to perform leaching at high temperatures, thereby enhancing leaching rates. In this work, leaching experiments and visualization by microscopy of cell attachment and biofilm formation patterns of the crenarchaeotes Sulfolobus metallicus DSM 6482(T) and the Acidianus isolates DSM 29038 and DSM 29099 in pure and mixed cultures on sulfur or pyrite were studied. Confocal laser scanning microscopy (CLSM) combined with fluorescent dyes as well as fluorescently labeled lectins were used to visualize different components (e.g. DNA, proteins or glycoconjugates) of the aforementioned species. The data indicate that cell attachment and the subsequently formed biofilms were species- and substrate-dependent. Pyrite leaching experiments coupled with pre-colonization and further inoculation with a second species suggest that both species may negatively influence each other during pyrite leaching with respect to initial attachment and pyrite dissolution rates. In addition, the investigation of binary biofilms on pyrite showed that both species were heterogeneously distributed on pyrite surfaces in the form of individual cells or microcolonies. Physical contact between the two species seems to occur, as revealed by specific lectins able to specifically bind single species within mixed cultures.

[Adaptation of Acidianus hospitalis W1 to oligotrophic and acidic hot spring environments].

OBJECTIVE: To study the adaptation of A. hospitalis W1 to oligotrophic and acidic hot spring environments at the whole genome level. METHODS: We annotated the gene functions and constructed metabolic pathways of strain W1 by using different databases, such as NCBI non-redundant database (NRDB), UniProt, Sulfolobus protein database and Kyoto Encyclopedia of Genes and Genomes (KEGG). The metabolic pathways were polished according to the results of comparative genomics. RESULTS: Strain W1 grew autotrophically by fixing CO2 as carbon source through 3-hydroxypropionate/4-hydroxybutyrate or dicarboxylate-4-hydroxybutyrate cycle, and gained energy for growth by oxidation of reduced inorganic sulfur compounds (RISCs). Strain W1 differenced from A. ambivalens because its genome did not possess sulfur-metabolizing genes encoding sulfite: acceptor oxidoreductase, adenosine phosphosulfate reductase, sulfate adenylyl transferase and phosphoadenosine phosphosulfate reductase. Glucose was metabolized by strain W1 through non- phosphorylated Entner-Doudoroff pathway and tricarboxylic acid cycle. In addition, the sugar and amino acids transporters, as well as related hydrolysis enzymes were identified in the genome. These results suggest that strain W1 could also grow facultative autotrophically. Strain W1 cannot use H2 as electron donor due to lack of hydrogenase encoding genes. CONCLUSION: The versatile metabolic patterns afforded A. hospitalis W1 the ability to adapt to oligotrophic and acidic hot spring environments. Furthermore, the unique metabolic features of strain W1 will help to better understand the metabolic diversities of Acidianus.

Physiologic versatility and growth flexibility as the main characteristics of a novel thermoacidophilic Acidianus strain isolated from Copahue geothermal area in Argentina.

A novel thermoacidophilic archaeal strain has been isolated from three geothermal acidic hot springs in Copahue, Argentina. One of the most striking characteristic of ALE1 isolate is its metabolic versatility. It grows on sulphur, tetrathionate, iron (II) and sucrose under aerobic conditions, but it can also develop under anaerobic conditions using iron (III) or sulphur as electron acceptors and sulphur or hydrogen as electron donors autotrophically. A temperature of 75 °C and a pH between 2.5 and 3.0 are strain ALE1 optimal growth conditions, but it is able to oxidise iron (II) even at pH 1.0. Cells are irregular cocci surrounded by a regularly arrayed glycoprotein layer (S-layer). Phylogenetic analysis shows that strain ALE1 belongs to the family Sulfolobaceae in the class Thermoprotei, within the phylum Crenarchaeota. Based on 16S rRNA gene sequence similarity on NCBI database, ALE1 does not have closely related relatives, neither in culture nor uncultured, which is more surprising. Its closest related species are strains of Acidianus hospitalis (91 % of sequence similarity), Acidianus infernus (90 %), Acidianus ambivalens (90 %) and Acidianus manzanensis (90 %). Its DNA base composition of 34.5 % mol C + G is higher than that reported for other Acidianus species. Considering physiological and phylogenetic characteristics of strain ALE1, we considered it to represent a novel species of the genus Acidianus (candidatus "Acidianus copahuensis"). The aim of this study is to physiologically characterise this novel archaea in order to understand its role in iron and sulphur geochemical cycles in the Copahue geothermal area and to evaluate its potential applications in bioleaching and biooxidation.

X-ray Structure of a self-compartmentalizing sulfur cycle metalloenzyme.

Numerous microorganisms oxidize sulfur for energy conservation and contribute to the global biogeochemical sulfur cycle. We have determined the 1.7 angstrom-resolution structure of the sulfur oxygenase reductase from the thermoacidophilic archaeon Acidianus ambivalens, which catalyzes an oxygen-dependent disproportionation of elemental sulfur. Twenty-four monomers form a large hollow sphere enclosing a positively charged nanocompartment. Apolar channels provide access for linear sulfur species. A cysteine persulfide and a low-potential mononuclear non-heme iron site ligated by a 2-His-1-carboxylate facial triad in a pocket of each subunit constitute the active sites, accessible from the inside of the sphere. The iron is likely the site of both sulfur oxidation and sulfur reduction.

Acidianus tengchongensis sp. nov., a new species of acidothermophilic archaeon isolated from an acidothermal spring.

A new thermoacidophilic, obligately chemolithotrophic, facultatively aerobic archaeon Acidianus S5(T), was isolated from a Tengchong acidothermal spring in southwestern China. It is a Gram-negative, nonmotile, irregular coccoid organism with a cell diameter of 1.2 microm. The optimal pH and temperature for growth are 2.5 and 70 degrees C, respectively. Under anaerobic conditions, the organism reduces elemental sulfur with molecular hydrogen, producing hydrogen sulfide. Under aerobic conditions, it oxidizes elemental sulfur and produces sulfuric acid. No growth occurs when it is cultivated in an iron medium, indicating that ferrous iron cannot serve as an energy source. The G+C content is 38% (mol/mol), which is much different from that of other Acidianus species (31%-32.7%). The phylogenetic distances, based on 16S rDNA sequences, to A. brierleyi, A. infernus, and A. ambivalens were 0.2, 2.6, and 2.5%, respectively. DNA-DNA hybridization rates of strain S5(T) to A. brierleyi, A. infernus, and A. ambivalens are 44, 22, and 23%, respectively. Thus, a new name, Acidianus tengchongensis sp. nov., is proposed for this strain S5(T).