Thiobacillus sedimenti sp. nov., a chemolithoautotrophic sulphur-oxidizing bacterium isolated from freshwater sediment.

A sulphur-oxidizing bacterium, designated strain SCUT-2T, was isolated from freshwater sediment collected from the Pearl River in Guangzhou, PR China. This strain was an obligate chemolithoautotroph, utilizing reduced sulphur compounds (elemental sulphur, thiosulphate, tetrathionate and sulphite) as the electron donor. Growth of strain SCUT-2T was observed at 20-40 â„ƒ (optimum at 30 °C), pH 5.0-9.0 (optimum at 6.0), and NaCl concentration range of 0-9 g L-1 (optimum at 1 g L-1). The major cellular fatty acids were C16:0 ω7c and cyclo-C17:0. The DNA G + C content of the complete genome sequence was 66.8 mol%. Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain SCUT-2T formed a lineage within the genus Thiobacillus, showing gene sequence identity of 98.0% with its closest relative Thiobacillus thioparus THI 115. The genome of strain SCUT-2T contains multiple genes encoding sulphur-oxidizing enzymes that catalyse the oxidation of reduced sulphur compounds, partial genes that are necessary for denitrification, and the genes encoding cbb3-type cytochrome c oxidase, aa3-type cytochrome c oxidase and bd-type quinol oxidase. Facultative anaerobic growth occurs when using nitrate as the electron acceptor and thiosulphate as the electron donor. On the basis of phenotypic, chemotaxonomic, genotypic and phylogenetic analysis, strain SCUT-2T (= GDMCC 1.4108T = JCM 39443T) is deemed to represent a novel Thiobacillus species, for which we propose the name Thiobacillus sedimenti sp. nov.

Improvement of the yield-related response of mycorrhized Lallemantia iberica to salinity through sulfur-oxidizing bacteria.

BACKGROUND: To investigate the effects of salinity as a serious environmental limiter of productivity on the yield-related traits of Lallemantia iberica, a split-plot experiment was performed for 2 years (2017-2018) based on a randomized complete block design with three replications at Urmia University (37°33'09″N, 45°05'53″E). The main plots included salinity stress at two levels (6.72 dS m-1 , and 0.91 dS m-1 as control), and subplots were inoculants at four levels (Funneliformis mosseae, Thiobacillus sp., F. mosseae + Thiobacillus sp., and no inoculation). RESULTS: In the saline condition, serious reductions in yield and yield components (numbers of capsules per plant, seeds per capsule, and seeds per plant, 1000-seed weight, seed and biological yields), concentrations of leaf phosphorus and potassium, and relative mycorrhizal dependency were observed, but against the harvest index the leaf sulfur and sodium contents were increased. Moreover, all morphological traits (plant height, number of branches and leaves, leaf weight, stem weight, and ratio of leaf weight to stem weight) were decreased under salinity conditions. Mycorrhizal inoculation enhanced the salinity-induced reduction of yield and morphological traits to some extent. Inoculation with Thiobacillus had superiority in some of the yield and morphological characteristics compared with those in the non-inoculated plants. CONCLUSION: Salinity stress can significantly affect the yield, morphological characteristics, nutrients content, and mycorrhizal dependency of L. iberica plants. This study exhibited the significant effects of single and simultaneous applications of F. mosseae and Thiobacillus on plant growth and yield in saline soils. © 2020 Society of Chemical Industry.

Mn3O4 Nanozyme Coating Accelerates Nitrate Reduction and Decreases N2O Emission during Photoelectrotrophic Denitrification by Thiobacillus denitrificans-CdS.

Biosemiconductors are highly efficient systems for converting solar energy into chemical energy. However, the inevitable presence of reactive oxygen species (ROS) seriously deteriorates the biosemiconductor performance. This work successfully constructed a Mn3O4 nanozyme-coated biosemiconductor, Thiobacillus denitrificans-cadmium sulfide (T. denitrificans-CdS@Mn3O4), via a simple, fast, and economic method. After Mn3O4 coating, the ROS were greatly eliminated; the concentrations of hydroxyl radicals, superoxide radicals, and hydrogen peroxide were reduced by 90%, 77.6%, and 26%, respectively, during photoelectrotrophic denitrification (PEDeN). T. denitrificans-CdS@Mn3O4 showed a 28% higher rate of nitrate reduction and 78% lower emission of nitrous oxide (at 68 h) than that of T. denitrificans-CdS. Moreover, the Mn3O4 coating effectively maintained the microbial viability and photochemical activity of CdS in the biosemiconductor. Importantly, no lag period was observed during PEDeN, suggesting that the Mn3O4 coating does not affect the metabolism of T. denitrificans-CdS. Immediate decomposition and physical separation are the two possible ways to protect a biosemiconductor from ROS damage by Mn3O4. This study provides a simple method for protecting biosemiconductors from the toxicity of inevitably generated ROS and will help develop more stable and efficient biosemiconductors in the future.

Bioaugmentation with Thiobacillus sp. H1 in an autotrophic denitrification desulfurization microbial reactor: Microbial community changes and relationship.

Thiobacillus sp. H1 was isolated and made into solid bacterial agent. The Thiobacillus sp. H1 agent was dosed into two reactor (all the agent dosed one-time, and multi-dosing bacteria evenly) and run for 40 days, a start-up with no microbial agent bioreactor as control. We found that the operational performance of multi-dosing inoculum reactor was stable, and the amount of elemental sulfur produced remained stable at 143.2-152.3 mg/L. The amount of elemental sulfur generated in the reactor without the addition of the inoculum was gradually increased, and the amount of elemental sulfur generated in the reactor with the inoculum added at one-time was decreased. Two kinds of Thiobacillus gen. and unclassified betaproteobacteria that coordinated the overall community function in the autotrophic denitrification desulfurization system with high-throughput sequencing. The trend of FccAB gene in each bioreactor was similar with the trend of elemental sulfur in the effluent. On the 5th day, the copy number of FccAB in bioreactor II was the highest among the three bioreactors, reaching 11.8 log copies L/g. This study explores the possibility of artificially synthesized denitrifying desulfurization flora in the future.

Mycorrhizal Fungi and Thiobacillus Co-inoculation Improve the Physiological Indices of Lallemantia iberica Under Salinity Stress.

Salinity, a serious environmental pressure on crop production, might be counteracted by free-living and symbiotic inoculants entailing positive synergistic effects. Enhancement in nutrient uptake and/or production of antioxidants under the stress condition, can improve plant growth and yield. In this study, inoculation of Lallemantia iberica with Funneliformis mosseae and the sulfur solubilizing bacterium (Thiobacillus sp. T95 and T40) was evaluated under two salinity levels (6.72 dS/m and 0.91 dS/m as control). The root colonization, spore density, seed and biological yield, total soluble sugars, and nutrients were reduced by salt stress. Antioxidant enzyme activity (catalase, superoxide dismutase, peroxidase and ascorbate peroxidase), proline, contents of sodium and sulfur have increased under salt stress. The enzyme activities as well as the concentrations of nitrogen, phosphorus, potassium, sodium, and sulfur were dropped at the flowering stage (75 days after sowing). Seed and biological yield, antioxidant enzymes activity, proline content, and nutrients were significantly improved in mycorrhizal treatments. Inoculation of Thiobacillus exhibited the positive effect on root colonization, spore density, enzymes activity, and nutrients. Bacterial treatments (dual and single) significantly increased the sulfur and total soluble sugars. Totally, the mycorrhizal plants accumulated more enzymatically produced antioxidants, osmolytes, and showed improved nutrient uptake. Our results provide new insights into the relationship among arbuscular mycorrhizal fungi (AMF), biosulfur bacteria, and plant growth under saline conditions. In conclusion, the Lallemantia iberica inoculation with mycorrhizal fungi, either alone, or in combination with Thiobacillus, is indicated for optimum plant yield through alleviation of the salinity stress.

Enhancement of tribromophenol removal in a sequencing batch reactor via submicron magnetite.

Conductive magnetite (Fe3O4) has been applied into some anaerobic bioprocesses to accelerate direct interspecies electron transfer (DIET), however, Fe3O4 is usually dissolved by iron-reducing bacteria under anaerobic conditions, resulting in the loss of magnetite. Therefore, submicron magnetite particles were added to the sequencing batch reactor (SBR) to build a Fe3O4/SBR system, which could alleviate magnetite dissolution and simultaneously remove tribromophenol (TBP) effectively. The average removal efficiencies of chemical oxygen demand (COD) and TBP in Fe3O4/SBR system were 81% and 91%, respectively, which were 51% and 18% higher than those of the control group without Fe3O4 (SBR system). The enhanced TBP biodegradation was likely related to potential DIET, which was supported by the scanning electron microscopy (SEM) analysis, the increase of dehydrogenase and heme c (fivefold and 1.7-fold), and the enrichment of iron-redoxing bacteria (Geobacter and Thiobacillus). Furthermore, magnetite mainly remained intact in structure as indicated by X-ray diffraction (XRD), which might be ascribed to in situ iron redox cycle and magnetite biosynthesis via Magnetospirillum. Notably, the content of hydrogen peroxide (H2O2) and hydroxyl radical (⋅OH) in Fe3O4/SBR system was 4-5 times higher than that of SBR system. These findings could provide insights into the development of cost-effective strategy for the removal of refractory organic pollutants.

Structural characterization of the bacterial proteasome homolog BPH reveals a tetradecameric double-ring complex with unique inner cavity properties.

Eukaryotic and archaeal proteasomes are paradigms for self-compartmentalizing proteases. To a large extent, their function requires interplay with hexameric ATPases associated with diverse cellular activities (AAA+) that act as substrate unfoldases. Bacteria have various types of self-compartmentalizing proteases; in addition to the proteasome itself, these include the proteasome homolog HslV, which functions together with the AAA+ HslU; the ClpP protease with its partner AAA+ ClpX; and Anbu, a recently characterized ancestral proteasome variant. Previous bioinformatic analysis has revealed a novel bacterial member of the proteasome family Betaproteobacteria proteasome homolog (BPH). Using cluster analysis, we here affirmed that BPH evolutionarily descends from HslV. Crystal structures of the Thiobacillus denitrificans and Cupriavidus metallidurans BPHs disclosed a homo-oligomeric double-ring architecture in which the active sites face the interior of the cylinder. Using small-angle X-ray scattering (SAXS) and electron microscopy averaging, we found that BPH forms tetradecamers in solution, unlike the dodecamers seen in HslV. Although the highly acidic inner surface of BPH was in striking contrast to the cavity characteristics of the proteasome and HslV, a classical proteasomal reaction mechanism could be inferred from the covalent binding of the proteasome-specific inhibitor epoxomicin to BPH. A ligand-bound structure implied that the elongated BPH inner pore loop may be involved in substrate recognition. The apparent lack of a partner unfoldase and other unique features, such as Ser replacing Thr as the catalytic residue in certain BPH subfamilies, suggest a proteolytic function for BPH distinct from those of known bacterial self-compartmentalizing proteases.

Change in the microbial community of saline geothermal fluids amended with a scaling inhibitor: effects of heat extraction and nitrate dosage.

Geothermal plants are often affected by corrosion caused by microbial metabolites such as H2S. In the Bad Blumau (Austria) geothermal system, an increase in microbially produced H2S was observed in the hot (107 °C) and scaling inhibitor-amended saline fluids and in fluids that had cooled down (45 °C). Genetic fingerprinting and quantification revealed the dominance, increasing abundance and diversity of sulfate reducers such as Desulfotomaculum spp. that accompanied the cooling and processing of the geothermal fluids. In addition, a δ34S isotopic signature showed the microbial origin of the H2S that has been produced either chemolithotrophically or chemoorganotrophically. A nitrate addition test in a test pipe as a countermeasure against the microbial H2S formation caused a shift from a biocenosis dominated by bacteria of the phylum Firmicutes to a community of Firmicutes and Proteobacteria. Nitrate supported the growth of nitrate-reducing sulfur-oxidizing Thiobacillus thioparus, which incompletely reduced nitrate to nitrite. The addition of nitrate led to a change in the composition of the sulfate-reducing community. As a result, representatives of nitrate- and nitrite-reducing SRB, such as Desulfovibrio and Desulfonatronum, emerged as additional community members. The interaction of sulfate-reducing bacteria and nitrate-reducing sulfur-oxidizing bacteria (NR-SOB) led to the removal of H2S, but increased the corrosion rate in the test pipe.

An efficient way to enhance the total nitrogen removal efficiency of the Anammox process by S0-based short-cut autotrophic denitrification.

In order to reduce the amount of NO3--N generated by the Anammox process, and alleviate the competition between denitrification and Anammox for NO2--N in a single reactor, the preference of S0 for reacting with coexisting NO2--N and NO3--N in the sulfur autotrophic denitrifying (SADN) process and the coupling effect of short-cut SADN and the Anammox process were studied. The results showed that S0 preferentially reacted with NO3- to produce NO2--N, and then reacted with NO2--N when NO3--N was insufficient, which could effectively alleviate the competition between SADN bacteria (SADNB) and Anammox bacteria (AnAOB) for NO2--N. After 170 days of operation, coupling between short-cut S0-SADN and the Anammox process was first successfully achieved. SADNB converted the NO3--N generated by the Anammox process into NO2--N, which was once again available to AnAOB. The total nitrogen removal efficiency eventually stabilized at over 95%, and the effluent NO3--N was controlled within 10 mg/L, when high NH4+-N wastewater was treated by the Anammox process. Microbial community analysis further showed that Candidatus Brocadia and Thiobacillus were the functional microorganisms for AnAOB and SADNB.

Identification of Thiobacillus bacteria in agricultural soil in Iran using the 16S rRNA gene.

Thiobacillus, as useful soil bacteria, plays an important role in sulfur cycling. The purpose of this study was to identify the species Thiobacillus thioparus, Thiobacillus novellas and Thiobacillus denitrificans in rainfed and irrigated lands soil in Ajabshir, Ilam, Qorveh, Rojintaak, Sonqor, Kermanshah and Research Farm of Razi University in Iran. Sampling was performed as randomized completely with three replications at depth of 0-30 cm. The Thiobacillus species were determined via 16S rRNA characteristics. The results of agarose gel electrophoresis indicated that T. thioparus was the highest amount in the irrigated land in Research Farm and its lowest amount was in the Rojintaak rainfed land. These species not found in four locations and conditions including the Ajabshir irrigated, Qorveh rainfed, Research Farm rainfed and Rojintaak irrigated lands. The results of the T. novellas indicated that this was found in Ilam irrigated, Qorveh rainfed, Research Farm irrigated, Rojintaak irrigated and Rojintaak rainfed lands. The highest and lowest amount of T. novellas was indicated in the Rojintaak and Ilam irrigated lands respectively. The T. denitrificans gene showed that this bacterium was observed only in both samples of Ajabshir. Our study showed that Thiobacillus was not detected in all of the soils. If sulfur fertilizer is given to the soil without this bacterium, it is necessary to use sulfur fertilizer with Thiobacillus bacteria inoculation for better sulfur oxidation.

A comparative study of eubacterial communities by PCR-DGGE fingerprints in anoxic and aerobic biotrickling filters used for biogas desulfurization.

Biological desulfurization has proven to be a process that is technically and economically feasible on using biotrickling filters that can be performed under aerobic and anoxic conditions. However, microbial communities are different mainly due to the use of different final electron acceptors. The analysis of microbial communities in these systems has not been addressed with regard to the anoxic process. The aim of the work reported here was to analyse the eubacterial community in the two types of bioreactor along the packed bed and during the operation time. The analysis was carried out using the 16S PCR-DGGE molecular fingerprint technique. The microbial profile analysis in the aerobic bioreactor revealed that the community was more diverse and stratified compared to those obtained in the two anoxic bioreactors, influenced by environmental factors. The main OTU involved in this process is genus Thiobacillus, although different species were detected depending on each operational condition.

Reclassification of Thiobacillus aquaesulis (Wood & Kelly, 1995) as Annwoodia aquaesulis gen. nov., comb. nov., transfer of Thiobacillus (Beijerinck, 1904) from the Hydrogenophilales to the Nitrosomonadales, proposal of Hydrogenophilalia class. nov. within the 'Proteobacteria', and four new families within the orders Nitrosomonadales and Rhodocyclales.

The genus Thiobacillus comprises four species with validly published names, of which Thiobacillus aquaesulis DSM 4255T (=ATCC 43788T) is the only species that can grow heterotrophically or mixotrophically - the rest being obligate autotrophs - and has a significant metabolic difference in not producing tetrathionate during the oxidation of thiosulfate during autotrophic growth. On the basis of this and differential chemotaxonomic properties and a 16S rRNA gene sequence similarity of 93.4 % to the type species Thiobacillus thioparus DSM 505T, we propose that it is moved to a novel genus, Annwoodia gen. nov., for which the type species is Annwoodia aquaesulis gen. nov., comb. nov. We confirm that the position of the genus Thiobacillus in the Betaproteobacteria falls within the Nitrosomonadales rather than the Hydrogenophilales as previously proposed. Within the Nitrosomonadales we propose the circumscription of genera to form the Thiobacilliaceae fam. nov. and the Sterolibacteriaceae fam. nov. We propose the merging of the family Methylophilaceae into the Nitrosomonadales, and that the Sulfuricellaceae be merged into the Gallionellaceae, leaving the orders Methylophilales and Sulfuricellales defunct. In the Rhodocyclales we propose the Azonexaceae fam. nov. and the Zoogloeaceae fam. nov. We also reject the Hydrogenophilales from the Betaproteobacteria on the basis of a very low 16S rRNA gene sequence similarity with the class-proper as well as physiological properties, forming the Hydrogenophilalia class. nov. in the 'Proteobacteria'. We provide emended descriptions of Thiobacillus, Hydrogenophilales, Hydrogenophilaceae, Nitrosomonadales, Gallionellaceae, Rhodocyclaceae and the Betaproteobacteria.

Genome-Resolved Meta-Omics Ties Microbial Dynamics to Process Performance in Biotechnology for Thiocyanate Degradation.

Remediation of industrial wastewater is important for preventing environmental contamination and enabling water reuse. Biological treatment for one industrial contaminant, thiocyanate (SCN-), relies upon microbial hydrolysis, but this process is sensitive to high loadings. To examine the activity and stability of a microbial community over increasing SCN- loadings, we established and operated a continuous-flow bioreactor fed increasing loadings of SCN-. A second reactor was fed ammonium sulfate to mimic breakdown products of SCN-. Biomass was sampled from both reactors for metagenomics and metaproteomics, yielding a set of genomes for 144 bacteria and one rotifer that constituted the abundant community in both reactors. We analyzed the metabolic potential and temporal dynamics of these organisms across the increasing loadings. In the SCN- reactor, Thiobacillus strains capable of SCN- degradation were highly abundant, whereas the ammonium sulfate reactor contained nitrifiers and heterotrophs capable of nitrate reduction. Key organisms in the SCN- reactor expressed proteins involved in SCN- degradation, sulfur oxidation, carbon fixation, and nitrogen removal. Lower performance at higher loadings was linked to changes in microbial community composition. This work provides an example of how meta-omics can increase our understanding of industrial wastewater treatment and inform iterative process design and development.

Development of PCR primer systems for amplification of 16S-rDNA to detect of Thiobacillus spp.

Thiobacillus is a genus of Gram-negative, rod-shaped and autotrophic Betaproteobacteria. They catalyze the dissimilatory oxidation of elemental sulfur and reduced inorganic sulfur compounds. Whereas more than 30 species have been known in this genus, most were never reliably or effectively published. The rest were either reclassified into Thiomonas, Paracoccus, Starkeya, Sulfuriferula, Halothiobacillus, Thermithiobacillus or Acidithiobacillus, were lost from culture. Most of Thiobacillus species are obligate autotrophs via elementary sulfur, thiosulfate or polythionates as energy sources. Based on 16S ribosomal RNA sequence analysis, many members of Thiobacillus have been reclassified. A system was developed for the detection of Thiobacillus bacteria by the amplification of specific 16S ribosomal RNA sequence gene (16S rDNA) fragments with PCR. Primer sequences were designed for the amplification of fragments of 16S rDNA.

Spatial variation in bacterial community in natural wetland-river-sea ecosystems.

Wetland-estuarine-marine environments are typical oxic/anoxic transition zones and have complex water flow-paths within the zone of mixing where freshwater interacts with ocean water. Little is known about the impact of this interaction on bacterial community structures or the relationship between bacterial community and geochemical factors in such transitional mixing environments. Hence, we investigated the distribution patterns and diversity in bacterial communities in the Yellow River estuary-coastal wetland-Bohai Sea transition zone by analyzing 39 samples from 13 ordered sites. High-throughput sequencing of the 16S rRNA gene revealed significant shifts in diversity and distribution of bacterial community in sediments from the Yellow River estuary to the Bohai Sea. Yellow River sediment was dominated by hydrogen-, nitrogen-, and iron-cycling bacteria, such as Hydrogenophaga, Nitrospira, Pseudomonas, and Thiobacillus. The coastal wetland had a haloduric community associated with different functions, such as Planctomyces, Marinobacter, Halomonas, Salinivibrio, and Salinibacter. The Bohai Sea sediment had a higher relative abundance of Lutimonas, Desulfococcus, Photobacterium, Propionigenium, and Vibrio. Spatial variation in bacterial community was correlated with pH, salinity and sulfate (SO42-) concentration in such coastal environments. The major bacterial taxa were significantly different across the wetland, estuary, and coastal marine ecosystems, indicating substantial spatial heterogeneity among the three ecosystems. Statistical analysis revealed strong links between variation in bacterial community structure and ecosystem type. Our results demonstrate the importance of geographic and geochemical factors in structuring the bacterial community in natural environments.

Thiosulfate oxidation by Thiomicrospira thermophila: metabolic flexibility in response to ambient geochemistry.

Previous studies of the stoichiometry of thiosulfate oxidation by colorless sulfur bacteria have failed to demonstrate mass balance of sulfur, indicating that unidentified oxidized products must be present. Here the reaction stoichiometry and kinetics under variable pH conditions during the growth of Thiomicrospira thermophila strain EPR85, isolated from diffuse hydrothermal fluids at the East Pacific Rise, is presented. At pH 8.0, thiosulfate was stoichiometrically converted to sulfate. At lower pH, the products of thiosulfate oxidation were extracellular elemental sulfur and sulfate. We were able to replicate previous experiments and identify the missing sulfur as tetrathionate, consistent with previous reports of the activity of thiosulfate dehydrogenase. Tetrathionate was formed under slightly acidic conditions. Genomic DNA from T. thermophila strain EPR85 contains genes homologous to those in the Sox pathway (soxAXYZBCDL), as well as rhodanese and thiosulfate dehydrogenase. No other sulfur oxidizing bacteria containing sox(CD)2 genes have been reported to produce extracellular elemental sulfur. If the apparent modified Sox pathway we observed in T. thermophila is present in marine Thiobacillus and Thiomicrospira species, production of extracellular elemental sulfur may be biogeochemically important in marine sulfur cycling.

Biogas desulfurization using autotrophic denitrification process.

The aim of this study was to evaluate the performance of an autotrophic denitrification process for desulfurization of biogas produced from a chicken manure digester. A laboratory scale upflow fixed bed reactor (UFBR) was operated for 105 days and fed with sodium sulfide or H2S scrubbed from the biogas and nitrate as electron donor and acceptor, respectively. The S/N ratio (2.5 mol/mol) of the feed solution was kept constant throughout the study. When the UFBR was fed with sodium sulfide solution with an influent pH of 7.7, about 95 % sulfide and 90 % nitrate removal efficiencies were achieved. However, the inlet of the UFBR was clogged several times due to the accumulation of biologically produced elemental sulfur particles and the clogging resulted in operational problems. When the UFBR was fed with the H2S absorbed from the biogas and operated with an influent pH of 8-9, around 98 % sulfide and 97 % nitrate removal efficiencies were obtained. In this way, above 95 % of the H2S in the biogas was removed as elemental sulfur and the reactor effluent was reused as scrubbing liquid without any clogging problem.

Structural insights into the efficient CO2-reducing activity of an NAD-dependent formate dehydrogenase from Thiobacillus sp. KNK65MA.

CO2 fixation is thought to be one of the key factors in mitigating global warming. Of the various methods for removing CO2, the NAD-dependent formate dehydrogenase from Candida boidinii (CbFDH) has been widely used in various biological CO2-reduction systems; however, practical applications of CbFDH have often been impeded owing to its low CO2-reducing activity. It has recently been demonstrated that the NAD-dependent formate dehydrogenase from Thiobacillus sp. KNK65MA (TsFDH) has a higher CO2-reducing activity compared with CbFDH. The crystal structure of TsFDH revealed that the biological unit in the asymmetric unit has two conformations, i.e. open (NAD(+)-unbound) and closed (NAD(+)-bound) forms. Three major differences are observed in the crystal structures of TsFDH and CbFDH. Firstly, hole 2 in TsFDH is blocked by helix α20, whereas it is not blocked in CbFDH. Secondly, the sizes of holes 1 and 2 are larger in TsFDH than in CbFDH. Thirdly, Lys287 in TsFDH, which is crucial for the capture of formate and its subsequent delivery to the active site, is an alanine in CbFDH. A computational simulation suggested that the higher CO2-reducing activity of TsFDH is owing to its lower free-energy barrier to CO2 reduction than in CbFDH.

Bioreactor microbial ecosystems for thiocyanate and cyanide degradation unravelled with genome-resolved metagenomics.

Gold ore processing uses cyanide (CN(-) ), which often results in large volumes of thiocyanate- (SCN(-) ) contaminated wastewater requiring treatment. Microbial communities can degrade SCN(-) and CN(-) , but little is known about their membership and metabolic potential. Microbial-based remediation strategies will benefit from an ecological understanding of organisms involved in the breakdown of SCN(-) and CN(-) into sulfur, carbon and nitrogen compounds. We performed metagenomic analysis of samples from two laboratory-scale bioreactors used to study SCN(-) and CN(-) degradation. Community analysis revealed the dominance of Thiobacillus spp., whose genomes harbour a previously unreported operon for SCN(-) degradation. Genome-based metabolic predictions suggest that a large portion of each bioreactor community is autotrophic, relying not on molasses in reactor feed but using energy gained from oxidation of sulfur compounds produced during SCN(-) degradation. Heterotrophs, including a bacterium from a previously uncharacterized phylum, compose a smaller portion of the reactor community. Predation by phage and eukaryotes is predicted to affect community dynamics. Genes for ammonium oxidation and denitrification were detected, indicating the potential for nitrogen removal, as required for complete remediation of wastewater. These findings suggest optimization strategies for reactor design, such as improved aerobic/anaerobic partitioning and elimination of organic carbon from reactor feed.

Aerobic and Anaerobic Thiosulfate Oxidation by a Cold-Adapted, Subglacial Chemoautotroph.

Geochemical data indicate that protons released during pyrite (FeS2) oxidation are important drivers of mineral weathering in oxic and anoxic zones of many aquatic environments, including those beneath glaciers. Oxidation of FeS2 under oxic, circumneutral conditions proceeds through the metastable intermediate thiosulfate (S2O3 (2-)), which represents an electron donor capable of supporting microbial metabolism. Subglacial meltwaters sampled from Robertson Glacier (RG), Canada, over a seasonal melt cycle revealed concentrations of S2O3 (2-) that were typically below the limit of detection, despite the presence of available pyrite and concentrations of the FeS2 oxidation product sulfate (SO4 (2-)) several orders of magnitude higher than those of S2O3 (2-). Here we report on the physiological and genomic characterization of the chemolithoautotrophic facultative anaerobe Thiobacillus sp. strain RG5 isolated from the subglacial environment at RG. The RG5 genome encodes genes involved with pathways for the complete oxidation of S2O3 (2-), CO2 fixation, and aerobic and anaerobic respiration with nitrite or nitrate. Growth experiments indicated that the energy required to synthesize a cell under oxygen- or nitrate-reducing conditions with S2O3 (2-) as the electron donor was lower at 5.1°C than 14.4°C, indicating that this organism is cold adapted. RG sediment-associated transcripts of soxB, which encodes a component of the S2O3 (2-)-oxidizing complex, were closely affiliated with soxB from RG5. Collectively, these results suggest an active sulfur cycle in the subglacial environment at RG mediated in part by populations closely affiliated with RG5. The consumption of S2O3 (2-) by RG5-like populations may accelerate abiotic FeS2 oxidation, thereby enhancing mineral weathering in the subglacial environment.