The first complete genome of the biotechnologically important species Sulfobacillus thermotolerans has been sequenced. Its 3 317 203-bp chromosome contains an 83 269-bp plasmid-like region, which carries heavy metal resistance determinants and the rusticyanin gene. Plasmid-mediated metal resistance is unusual for acidophilic chemolithotrophs. Moreover, most of their plasmids are cryptic and do not contribute to the phenotype of the host cells. A polyphosphate-based mechanism of metal resistance, which has been previously unknown in the genus Sulfobacillus or other Gram-positive chemolithotrophs, potentially operates in two Sulfobacillus species. The methylcitrate cycle typical for pathogens and identified in the genus Sulfobacillus for the first time can fulfill the energy and/or protective function in S. thermotolerans Kr1 and two other Sulfobacillus species, which have incomplete glyoxylate cycles. It is notable that the TCA cycle, disrupted in all Sulfobacillus isolates under optimal growth conditions, proved to be complete in the cells enduring temperature stress. An efficient antioxidant defense system gives S. thermotolerans another competitive advantage in the microbial communities inhabiting acidic metal-rich environments. The genomic comparisons revealed 80 unique genes in the strain Kr1, including those involved in lactose/galactose catabolism. The results provide new insights into metabolism and resistance mechanisms in the Sulfobacillus genus and other acidophiles.

A two-step process, which involved ferric leaching with biologically generated solution and subsequent biooxidation with the microbial community, has been previously proposed for the processing of low-grade zinc sulfide concentrates. In this study, we carried out the process of complete biological oxidation of the product of ferric leaching of the zinc concentrate, which contained 9% of sphalerite, 5% of chalcopyrite, and 29.7% of elemental sulfur. After 21 days of biooxidation at 40 °C, sphalerite and chalcopyrite oxidation reached 99 and 69%, respectively, while the level of elemental sulfur oxidation was 97%. The biooxidation residue could be considered a waste product that is inert under aerobic conditions. The results of this study showed that zinc sulfide concentrate processing using a two-step treatment is efficient and promising. The microbial community, which developed during biooxidation, was dominated by Acidithiobacillus caldus, Leptospirillum ferriphilum, Ferroplasma acidiphilum, Sulfobacillus thermotolerans, S. thermosulfidooxidans, and Cuniculiplasma sp. At the same time, F. acidiphilum and A. caldus played crucial roles in the oxidation of sulfide minerals and elemental sulfur, respectively. The addition of L. ferriphilum to A. caldus during biooxidation of the ferric leach product proved to inhibit elemental sulfur oxidation.

Acidiphilium multivorum LMS is an acidophile isolated from industrial bioreactors during the processing of the gold-bearing pyrite-arsenopyrite concentrate at 38–42 °C. Most strains of this species are obligate organoheterotrophs that do not use ferrous iron or reduced sulfur compounds as energy sources. However, the LMS strain was identified as one of the predominant sulfur oxidizers in acidophilic microbial consortia. In addition to efficient growth under strictly heterotrophic conditions, the LMS strain proved to be an active sulfur oxidizer both in the presence or absence of organic compounds. Interestingly, Ac. multivorum LMS was able to succeed more common sulfur oxidizers in microbial populations, which indicated a previously underestimated role of this bacterium in industrial bioleaching operations. In this study, the first draft genome of the sulfur-oxidizing Ac. multivorum was sequenced and annotated. Based on the functional genome characterization, sulfur metabolism pathways were reconstructed. The LMS strain possessed a complicated multi-enzyme system to oxidize elemental sulfur, thiosulfate, sulfide, and sulfite to sulfate as the final product. Altogether, the phenotypic description and genome analysis unraveled a crucial role of Ac. multivorum in some biomining processes and revealed unique strain-specific characteristics, including the ars genes conferring arsenic resistance, which are similar to those of phylogenetically distinct microorganisms.

Microbial population performing biooxidation of flotation concentrate of gold bearing sulfide ore containing pyrite, arsenopyrite, and pyrrhotite was studied using cultural and molecular biological (metagenomics sequencing of V3-V4 fragments of 16S rRNA gene) approaches. The biooxidation of the concentrate was conducted at temperatures from 38 to 42°C. Strains of Acidithiobacillus thiooxidans, Acidiphilium multivorum, Leptospirillum ferriphilum, Sulfobacillus thermotolerans, Ferroplasma acidarmanus, and Ferroplasma acidiphilum were isolated from the samples of the pulp from biooxidation reactors. It was shown that optimum temperatures of isolated strains were from 38 to 40°C. Metagenomic analysis demonstrated predominance of the genera Acidiferrobacter, Acidithiobacillus, Acidiphilum, Leptospirillum, and Ferroplasma. According to results of molecular biological analysis, share of the genus Acidithiobacillus was of 0 to 25%, share of the genus Acidiferrobacter was of 7 to 56%, share of the genus Acidiphilum was of 0.03 to 36%, share of the genus Leptospirillum was of 0.7 to 7%, whereas share of the archaea of the genus Ferroplasma was of 33 to 94%. Thus, it was shown that representatives of the genus Ferroplasma can play significant role in bioleach process. Representatives of the genus Acidiferrobacter were previously detected in acid mine drainages, acid soils as well as in bioleach heaps and reactors, whereas data on predominance of the genus in tank bioleach processes have not been presented in the literature. In the present study, strains of the genus Acidiferrobacter were not isolated despite application of the nutrient media recommended for Acidiferrobacter and their properties were not studied. Nevertheless, results of the present study suggest that representatives of the genus Acidiferrobacter have a great impact on industrial bioleach processes.

A thermoacidophilic consortium of chemolithotrophic microorganisms oxidizing the concentrate of high-pyrrhotite pyrite−arsenopyrite ore at 38–40°C was isolated. The most active members of the...