Purple Sulfur Research Articles

Identification of metal-sensitive structural changes in the Ca2+-binding photocomplex from Thermochromatium tepidum by isotope-edited vibrational spectroscopy.

Calcium ions play a dual role in expanding the spectral diversity and structural stability of photocomplexes from several Ca2+-requiring purple sulfur phototrophic bacteria. Here, metal-sensitive structural changes in the isotopically labeled light-harvesting 1 reaction center (LH1-RC) complexes from the thermophilic purple sulfur bacterium Thermochromatium (Tch.) tepidum were investigated by perfusion-induced attenuated total reflection (ATR) Fourier transform infrared (FTIR) spectroscopy. The ATR-FTIR difference spectra induced by exchanges between native Ca2+ and exogenous Ba2+ exhibited interconvertible structural and/or conformational changes in the metal binding sites at the LH1 C-terminal region. Most of the characteristic Ba2+/Ca2+ difference bands were detected even when only Ca ions were removed from the LH1-RC complexes, strongly indicating the pivotal roles of Ca2+ in maintaining the LH1-RC structure of Tch. tepidum. Upon 15N-, 13C- or 2H-labeling, the LH1-RC complexes exhibited characteristic 15N/14N-, 13C/12C-, or 2H/1H-isotopic shifts for the Ba2+/Ca2+ difference bands. Some of the 15N/14N or 13C/12C bands were also sensitive to further 2H-labelings. Given the band frequencies and their isotopic shifts along with the structural information of the Tch. tepidum LH1-RC complexes, metal-sensitive FTIR bands were tentatively identified to the vibrational modes of the polypeptide main chains and side chains comprising the metal binding sites. Furthermore, important new IR marker bands highly sensitive to the LH1 BChl a conformation in the Ca2+-bound states were revealed based on both ATR-FTIR and near-infrared Raman analyses. The present approach provides valuable insights concerning the dynamic equilibrium between the Ca2+- and Ba2+-bound states statically resolved by x-ray crystallography.

Genome Sequences of Marichromatium gracile HOL-1 and Its Purple Photosynthetic Coisolate, Afifella sp. H1R.

ABSTRACTWe sequenced the genomes of both the purple sulfur gammaproteobacterium Marichromatium gracile HOL-1 and another purple photosynthetic organism, strain H1R, that was originally isolated as an unidentified contaminant. Through genome sequencing, we have now identified organism H1R as a species of Afifella. A whole-genome-based phylogenetic analysis of both species is provided.

Abrupt changes in the physical and biological structure of endorheic upland lakes due to 8‐m lake‐level variation during the 20th century

AbstractClimate‐induced variation in lake level can affect physicochemical properties of endorheic lakes, but its consequences for phototrophic production and regime shifts are not well understood. Here, we quantified changes in the abundance and community composition of phototrophs in Kenosee and White Bear lakes, two endorheic basins in the parkland Moose Mountain uplands of southeastern Saskatchewan, Canada, which have experienced > 8 m declines in water level since ~ 1900. We hypothesized that lower water levels and warmer temperatures should manifest as increased abundance of phytoplankton, particularly cyanobacteria, and possibly trigger a regime shift to turbid conditions due to evaporative concentration of nutrients and solutes. High‐resolution analysis of sedimentary pigments revealed an increase in total phototrophic abundance (as β‐carotene) concurrent with lake‐level decline beginning ~ 1930, but demonstrated little directional change in cyanobacteria. Instead, significant increases in obligately anaerobic purple sulfur bacteria (as okenone) occurred in both lakes during ~ 1930–1950, coeval with alterations to light environments and declines in lake level. The presence of okenone suggests that climate‐induced increases in solute concentrations may have favored the formation of novel bacterial habitats where photic and anoxic zones overlapped. Generalized additive models showed that establishment of this unique habitat was likely preceded by increased temporal variance of sulfur bacteria, but not phytoplankton or cyanobacteria, suggesting that this abrupt change to physical lake structure was unique to deep‐water environments. Such climate‐induced shifts may become more frequent in the region due to hydrological stress on lake levels due to warming temperatures across the Northern Great Plains.

Genomic Comparison, Phylogeny and Taxonomic Reevaluation of the Ectothiorhodospiraceae and Description of Halorhodospiraceae fam. nov. and Halochlorospira gen. nov.

The Ectothiorhodospiraceae family represents purple sulfur bacteria of the Gammaproteobacteria found primarily in alkaline soda lakes of moderate to extremely high salinity. The main microscopically visible characteristic separating them from the Chromatiaceae is the excretion of the intermediate elemental sulfur formed during oxidation of sulfide prior to complete oxidation to sulfate rather than storing it in the periplasm. We present a comparative study of 38 genomes of all species of phototrophic Ectothiorhodospiraceae. We also include a comparison with those chemotrophic bacteria that have been assigned to the family previously and critically reevaluate this assignment. The data demonstrate the separation of Halorhodospira species in a major phylogenetic branch distant from other Ectothiorhodospiraceae and support their separation into a new family, for which the name Halorhodospiraceae fam. nov. is proposed. In addition, the green-colored, bacteriochlorophyll-containing species Halorhodospira halochloris and Halorhodospira abdelmalekii were transferred to the new genus Halochlorospira gen. nov. of this family. The data also enable classification of several so far unclassified isolates and support the separation of Ectothiorhodospira shaposhnikovii and Ect. vacuolata as well as Ect. mobilis and Ect. marismortui as distinct species.

Emissions of H2S from Hog Finisher Farm Anaerobic Manure Treatment Lagoons: Physical, Chemical and Biological Influence

Hydrogen sulfide (H2S) from hog operations contributes to noxious odors in the surrounding environment and can be life-threatening. There is, however, limited understanding of what influences H2S emissions from these farms. Emissions of H2S were measured periodically over the course of two years at hog finisher farms in humid mesothermal (North Carolina, NC, USA) and semi-arid (Oklahoma, OK, USA) climates. Emissions were determined using an inverse dispersion backward Lagrangian stochastic model in conjunction with line-sampled H2S concentrations and measured turbulence. Daily emissions at the two lagoons were characterized by low emissions on most days with occasional days of high emissions. Mean annual area-specific emissions were much lower for the NC lagoon (1.32 µg H2S m−2 s−1 ± 0.07 µg H2S m−2 s−1) than the OK lagoon (6.88 µg H2S m−2 s−1 ± 0.13 µg H2S m−2 s−1). Mean annual hog-specific emissions for the NC lagoon were 0.75 g H2S hd−1 d−1 while those for the OK lagoon were 1.92 g H2S hd−1 d−1. Emissions tended to be higher during the afternoon, likely due to higher mean winds. Daily H2S emissions from both lagoons were greatest during the first half of the year and decreased as the year progressed and a reddish color (indicating high populations of purple sulfur bacteria (PSB)) appeared in the lagoon. The generally low emissions at the NC lagoon and higher emissions at the OK lagoon were likely a result of the influence of wind on mixing the lagoon and not the presence of PSB.

Allochromatium tepidum, sp. nov., a hot spring species of purple sulfur bacteria.

We describe a new species of purple sulfur bacteria (Chromatiaceae, anoxygenic phototrophic bacteria) isolated from a microbial mat in the sulfidic geothermal outflow of a hot spring in Rotorua, New Zealand. This phototroph, designated as strain NZ, grew optimally near 45°C but did not show an absorption maximum at 915nm for the light-harvesting-reaction center core complex (LH1-RC) characteristic of other thermophilic purple sulfur bacteria. Strain NZ had a similar carotenoid composition as Thermochromatium tepidum, but unlike Tch. tepidum, grew photoheterotrophically on acetate in the absence of sulfide and metabolized thiosulfate. The genome of strain NZ was significantly larger than that of Tch. tepidum but slightly smaller than that of Allochromatium vinosum. Strain NZ was phylogenetically more closely related to mesophilic purple sulfur bacteria of the genus Allochromatium than to Tch. tepidum. This conclusion was reached from phylogenetic analyses of strain NZ genes encoding 16S rRNA and the photosynthetic functional gene pufM, from phylogenetic analyses of entire genomes, and from a phylogenetic tree constructed from the concatenated sequence of 1090 orthologous proteins. Moreover, average nucleotide identities and digital DNA:DNA hybridizations of the strain NZ genome against those of related species of Chromatiaceae supported the phylogenetic analyses. From this collection of properties, we describe strain NZ here as the first thermophilic species of the genus Allochromatium, Allochromatium tepidum NZT, sp. nov.

Single-cell genomics-based analysis reveals a vital ecological role of Thiocapsa sp. LSW in the meromictic Lake Shunet, Siberia.

Meromictic lakes usually harbour certain prevailing anoxygenic phototrophic bacteria in their anoxic zone, such as the purple sulfur bacterium (PSB) Thiocapsa sp. LSW (hereafter LSW) in Lake Shunet, Siberia. PSBs have been suggested to play a vital role in carbon, nitrogen and sulfur cycling at the oxic–anoxic interface of stratified lakes; however, the ecological significance of PSBs in the lake remains poorly understood. In this study, we explored the potential ecological role of LSW using a deep-sequencing analysis of single-cell genomics associated with flow cytometry. An approximately 2.7 Mb draft genome was obtained based on the co-assembly of five single-cell genomes. LSW might grow photolithoautotrophically and could play putative roles not only as a carbon fixer and diazotroph, but also as a sulfate reducer/oxidizer in the lake. This study provides insights into the potential ecological role of Thiocapsa sp. in meromictic lakes.

Cultivation of a vampire: 'Candidatus Absconditicoccus praedator'.

Halorhodospira halophila, one of the most-xerophilic halophiles, inhabits biophysically stressful and energetically expensive, salt-saturated alkaline brines. Here, we report an additional stress factor that is biotic: a diminutive Candidate-Phyla-Radiation bacterium, that we named 'Ca. Absconditicoccus praedator' M39-6, which predates H. halophila M39-5, an obligately photosynthetic, anaerobic purple-sulfur bacterium. We cultivated this association (isolated from the hypersaline alkaline Lake Hotontyn Nur, Mongolia) and characterized their biology. 'Ca. Absconditicoccus praedator' is the first stably cultivated species from the candidate class-level lineage Gracilibacteria (order-level lineage Absconditabacterales). Its closed-and-curated genome lacks genes for the glycolytic, pentose phosphate- and Entner-Doudoroff pathways which would generate energy/reducing equivalents and produce central carbon currencies. Therefore, 'Ca. Absconditicoccus praedator' is dependent on host-derived building blocks for nucleic acid-, protein-, and peptidoglycan synthesis. It shares traits with (the uncultured) 'Ca. Vampirococcus lugosii', which is also of the Gracilibacteria lineage. These are obligate parasitic lifestyle, feeding on photosynthetic anoxygenic Gammaproteobacteria, and absorption of host cytoplasm. Commonalities in their genomic composition and structure suggest that the entire Absconditabacterales lineage consists of predatory species which act to cull the populations of their respective host bacteria. Cultivation of vampire : host associations can shed light on unresolved aspects of their metabolism and ecosystem dynamics at life-limiting extremes.

Complete genome of the thermophilic purple sulfur Bacterium Thermochromatium tepidum compared to Allochromatium vinosum and other Chromatiaceae.

The complete genome sequence of the thermophilic purple sulfur bacterium Thermochromatium tepidum strain MCT (DSM 3771T) is described and contrasted with that of its mesophilic relative Allochromatium vinosum strain D (DSM 180T) and other Chromatiaceae. The Tch. tepidum genome is a single circular chromosome of 2,958,290 base pairs with no plasmids and is substantially smaller than the genome of Alc. vinosum. The Tch. tepidum genome encodes two forms of RuBisCO and contains nifHDK and several other genes encoding a molybdenum nitrogenase but lacks a gene encoding a protein that assembles the Fe-S cluster required to form a functional nitrogenase molybdenum-iron cofactor, leaving the phototroph phenotypically Nif-. Tch. tepidum contains genes necessary for oxidizing sulfide to sulfate as photosynthetic electron donor but is genetically unequipped to either oxidize thiosulfate as an electron donor or carry out assimilative sulfate reduction, both of which are physiological hallmarks of Alc. vinosum. Also unlike Alc. vinosum, Tch. tepidum is obligately phototrophic and unable to grow chemotrophically in darkness by respiration. Several genes present in the Alc. vinosum genome that are absent from the genome of Tch. tepidum likely contribute to the major physiological differences observed between these related purple sulfur bacteria that inhabit distinct ecological niches.

Relations between Hydrogen and Sulfur Metabolism in Purple Sulfur Bacteria

Relations between Hydrogen and Sulfur Metabolism in Purple Sulfur Bacteria

Photosynthetic Growth and Energy Conversion in an Engineered Phototroph Containing Thermochromatium tepidum Light-Harvesting Complex 1 and the Rhodobacter sphaeroides Reaction Center Complex.

Light-harvesting complex 1 (LH1) of the thermophilic purple sulfur bacterium Thermochromatium tepidum can be expressed in the purple non-sulfur bacterium Rhodobacter sphaeroides and forms a functional RC-LH1 complex with the native Rba. sphaeroides reaction center (Nagashima, K. V. P., et al. Proc. Natl. Acad. Sci. U. S. A. 2017, 114, 10906-10911). Although there is a large uphill energy gap between Tch. tepidum LH1 and the Rba. sphaeroides RC in this chimeric complex, it has been shown that light energy can be transferred, consistent with that seen in the native Rba. sphaeroides RC-LH1 complex. In this study, the contribution of this chimeric complex to growth and photosynthetic energy conversion in the hybrid organism was quantified. The mutant synthesizing this chimeric complex was grown phototrophically under 940 nm light-emitting diode (LED) light preferentially absorbed by Tch. tepidum LH1 and showed faster growth at low intensities of this wavelength than both a mutant strain of Rba. sphaeroides lacking LH2 and a mutant lacking all light-harvesting complexes. When grown with 850 nm LED light, the strain containing the native Rba. sphaeroides LH1-RC grew faster than the chimeric strain. Electron transfer from the RC to the membrane-integrated cytochrome bc1 complex was also estimated by flash-induced absorption changes in heme b. The rate of ubiquinone transport through the LH1 ring structure in the chimeric strain was virtually the same as that in native Rba. sphaeroides. We conclude that Tch. tepidum LH1 can perform the physiological functions of native LH1 in Rba. sphaeroides.

Purple sulfur bacteria fix N2 via molybdenum-nitrogenase in a low molybdenum Proterozoic ocean analogue

Biological N2 fixation was key to the expansion of life on early Earth. The N2-fixing microorganisms and the nitrogenase type used in the Proterozoic are unknown, although it has been proposed that the canonical molybdenum-nitrogenase was not used due to low molybdenum availability. We investigate N2 fixation in Lake Cadagno, an analogue system to the sulfidic Proterozoic continental margins, using a combination of biogeochemical, molecular and single cell techniques. In Lake Cadagno, purple sulfur bacteria (PSB) are responsible for high N2 fixation rates, to our knowledge providing the first direct evidence for PSB in situ N2 fixation. Surprisingly, no alternative nitrogenases are detectable, and N2 fixation is exclusively catalyzed by molybdenum-nitrogenase. Our results show that molybdenum-nitrogenase is functional at low molybdenum conditions in situ and that in contrast to previous beliefs, PSB may have driven N2 fixation in the Proterozoic ocean.

Carotenoid biomarkers in Namibian shelf sediments: Anoxygenic photosynthesis during sulfide eruptions in the Benguela Upwelling System

Aromatic carotenoid-derived hydrocarbon biomarkers are ubiquitous in ancient sediments and oils and are typically attributed to anoxygenic phototrophic green sulfur bacteria (GSB) and purple sulfur bacteria (PSB). These biomarkers serve as proxies for the environmental growth requirements of PSB and GSB, namely euxinic waters extending into the photic zone. Until now, prevailing models for environments supporting anoxygenic phototrophs include microbial mats, restricted basins and fjords with deep chemoclines, and meromictic lakes with shallow chemoclines. However, carotenoids have been reported in ancient open marine settings for which there currently are no known modern analogs that host GSB and PSB. The Benguela Upwelling System offshore Namibia, known for exceptionally high primary productivity, is prone to recurrent toxic gas eruptions whereupon hydrogen sulfide emanates from sediments into the overlying water column. These events, visible in satellite imagery as water masses clouded with elemental sulfur, suggest that the Benguela Upwelling System may be capable of supporting GSB and PSB. Here, we compare distributions of biomarkers in the free and sulfur-bound organic matter of Namibian shelf sediments. Numerous compounds-including acyclic isoprenoids, steranes, triterpanes, and carotenoids-were released from the polar lipid fractions upon Raney nickel desulfurization. The prevalence of isorenieratane and β-isorenieratane in sampling stations along the shelf verified anoxygenic photosynthesis by low-light-adapted, brown-colored GSB in this open marine setting. Renierapurpurane was also present in the sulfur-bound carotenoids and was typically accompanied by lower abundances of renieratane and β-renierapurpurane, thereby identifying cyanobacteria as an additional aromatic carotenoid source.

Lipid composition of the microbial mat from a hypersaline environment (Vermelha Lagoon, Rio de Janeiro, Brazil)

ABSTRACTThis study determines organic-matter (OM) composition in the different color layers of a stratified hypersaline microbial mat and verifies the hypothesis that each layer includes a distinct group of lipids. The relation of precursor lipids from the microbial mat to the hydrocarbon composition in fossil records was also evaluated. To that end, the composition was studied of glycolipids (GLs), phospholipids (PLs), and “neutral” lipids (NLs, including hydrocarbons, n-alkanols, sterols, hopanols, free fatty acids, and wax esters) in four different color layers (A–D; depth intervals: up to 0.5 cm, 0.5–1.0 cm, 1.5–3.0 cm, and 3.0–6.0 cm, respectively) of a stratified hypersaline mat from the Vermelha Lagoon, Rio de Janeiro, Brazil.Microscopic characterization revealed the presence of 16 cyanobacterial morphospecies, with predominance of Microcoleus chthonoplastes. The notable prevalence of saturated straight-chain fatty acids (FAs), n-16:0 and n-18:0 and their monounsaturated counterparts, n-16:1 and n-18:1 in all three lipid fractions (GLs, PLs, and NLs), associated with the domination of n-C17 alkane and n-C17:1 alkene among the hydrocarbons confirmed the main imprint of cyanobacteria. The composition of the studied lipid classes implies the contribution of sulfate-reducing bacteria such as Desulfomicrobium sp. strain, purple sulfur bacteria, as well as the possible input of Geobacter spp. and Desulfovibrio spp., particularly in the deeper layers.The notable decrease in total extractable lipids (TELs) yield from layers A to D indicates that lipid synthesis is far more intense by photosynthesizing cyanobacteria than by anaerobic microorganisms. The content of PLs was uniform and low (< 5%) in all layers, implying their extremely quick degradation. GLs, followed by NLs, were the most abundant in all layers indicating the medium, which is characterized by carbon source excess and limited nitrogen source, which regulates microorganism growth. The upper layers, A (green) and B (reddish-brown) differ from those lower, C (dark brown greenish) and D (brown) according to the NLs/GLs ratio, which is higher in the former.The lipid compositions reveal distinctions between the individual layers in the microbial mat. The observed layers clearly differ according to the amount of high-molecular-weight (C22–C31) n-alkanes and long-chain (C21–C30) n-alkanols, the content of phytol, bishomohopanol, tetrahymanol, C27–C29 sterols, the stanol/stenol ratio in the neutral lipid fraction, as well as the content of branched (iso and anteiso) FAs and w9/w7 FA ratio in the GLs fraction. The mentioned parameters imply a greater contribution of sulfate-reducing and purple sulfur bacteria to layer B, higher impact of photosynthetic red algae in upper layers A and B, the elevated contribution of marine ciliate species, feeding on bacteria to layers B and C, as well as the increment of anoxygenic phototrophic and heterotrophic bacteria to layer D. The greatest capability for the synthesis of hydrocarbons is observed in layer B.The composition of lipid classes in the microbial mat showed a significant relationship with the most important biomarkers' fingerprints in the source rocks extracts and petroleum derived from the carbonate hypersaline environments, including the distribution of n-alkanes, a high abundance of phytane and gammacerane, as well as a distribution of C27–C29 regular steranes. Therefore, these results offer an insight into the transformation of microbial OM during the sedimentation processes in a hypersaline environment and its contribution to the fossil record.

Holocene phototrophic community and anoxia dynamics in meromictic Lake Jaczno (NE Poland) using high-resolution hyperspectral imaging and HPLC data

Abstract. Global spread of hypoxia and less frequent mixing in lakes is a major growing environmental concern. Climate change and human impact are expected to increasingly deteriorate aquatic ecosystems. The study of processes and drivers of such changes in the past provides a great asset for prevention and remediation in the future. We used a multiproxy approach combining high-resolution bulk pigment data measured by hyperspectral imaging (HSI) with lower-resolution specific chlorophyll types and carotenoids measured by HPLC to examine Holocene trophic state changes and anoxia evolution in the meromictic Lake Jaczno, NE Poland. A redundancy analysis (RDA) including pollen-inferred vegetation cover, temperature and human impacts provides insight into specific conditions and drivers of changing trophic and redox states in the lake. Anoxic and sulfidic conditions were established in Lake Jaczno after initial basin infilling 9500 years ago. Until 6700 cal BP, lake trophy was relatively low, water turbidity was high and green sulfur bacteria (GSB) were abundant within the phototrophic community, suggesting a deep oxic–anoxic boundary and weak stratification. The period between 6700–500 cal BP is characterized by constantly increasing lake production and a gradual shift from GSB to purple sulfur bacteria (PSB), suggesting a shallower oxic–anoxic boundary and pronounced stratification. Yet, the presence of spheroidene and speroidenone in the sediments indicates intermittent anoxia. After 500 cal BP, increasing human impact, deforestation and intensive agriculture promoted lake eutrophication, with a shift to PSB dominance and establishment of permanent anoxia and meromixis. Our study unambiguously documents the legacy of human impact on processes determining eutrophication and anoxia.

Nitrite oxidation by phototrophic bacteria of Chlorobium, Thiocapsa and Lamprocystis genera under the influence of inorganic pollutants

Pollutants of inorganic nature (acids, alkalis, mineral salts of different composition, metals) change the course of biological processes of environmental purification, but their influence on the physiological properties of phototrophic sulfur bacteria has not been studied enough. The usage of nitrite ions as an electron donor of anoxygenic photosynthesis by cells of phototrophic green and purple sulfur bacteria Chlorobium limicola IMV K-8, Thiocapsa sp. Ya-2003 and Lamprocystis sp. Ya-2003, isolated from Yavorivske Lake, under the influence of the most widespread inorganic pollutants – hydro- and dihydrophosphates, sulfates, chlorides and chlorates, has been studied. It is shown that KH2PO4, K2HPO4, Na2SO4, NaCl and KClO3, present in the van Niel medium with 4.2 mM NaNO2 at concentrations that are 0.5, 1.0, 2.0, 3.0, 4.0 times different from the maximum permissible concentrations (MPC), influenced the biomass accumulation and nitrite ions oxidation by phototrophic green and purple sulfur bacteria. In media with hydro- and dihydrophosphate ions at concentrations 4.0 times higher than the MPC, inhibition of bacterial growth was up to 1.7 times lower than in the control. The biomass accumulation by bacteria in media with chloride and chlorate ions at concentrations 3.0–4.0 times higher than MPC was 2.0–2.8 times lower compared to the control. In the medium with Na2SO4 at concentrations 2.0–4.0 times higher than MPC, the biomass was 2.0–4.0 times lower than in the control. Nitrites’ oxidation by all strains in the media with the studied pollutants was slowed down. The residual content of nitrite ions in media with hydro- and dihydrophosphate, chloride and chlorate ions at their concentrations 4.0 times higher than MPC, exceeded the NO2– content in the control variants up to 1.7 times. If in the medium without pollutants the cells of C. limicola IMV K-8, Thiocapsa sp. Ya-2003 and Lamprocystis sp. Ya-2003 strains oxidized 72.7%, 72.2% and 71.4%, respectively, of nitrite ions present in the medium, then in the medium with sulfate ions at concentration 4.0 times higher than the MPC, bacteria oxidized nitrite ions only at 39.6%, 34.4% and 27.0%, respectively. Oxidation of a lower quantity of nitrites by phototrophic bacteria in the media with inorganic pollutants led to the production by them of a lower quantity of nitrates. The content of NO3– in the media with hydro-, dihydrophosphate and chlorate ions at all concentrations was up to 1.9 times lower than in the control. In media with sulfate ions at concentrations 2.0–4.0 times higher than MPC and chloride at concentration 4.0 times higher than MPC, the content of nitrate ions was 2.1–4.3 and 2.0 times, respectively, lower than in the control variants. Inorganic pollutants stimulated the synthesis of intracellular carbohydrates in C. limicola IMV K-8. If the content of intracellular glucose in cells grown in the medium without pollutants was 10.3 mg/g dry cell weight, then in cells grown in media with K2HPO4, KH2PO4, Na2SO4, NaCl and KClO3 at concentrations 4.0 times higher than MPC, its content increased by 12.2%, 10.7%, 51.6%, 17.1% and 35.9%, respectively. The glycogen content in the cells grown in the medium without pollutants was 45.1 mg/g dry cell weight. Hydro- and dihydrophosphate, chloride and chlorate ions at concentrations 4.0 times higher than MPC stimulated glycogen synthesis in cells by 47.5%, 57.6%, 67.4% and 74.6%, respectively. The glycogen content in cells grown in the medium with Na2SO4 at concentrations 3.0 and 4.0 times higher than MPC increased by 102.9% and 107.5%, respectively. Therefore, it is established that pollutants of inorganic nature affect the physiological properties of photosynthetic sulfur bacteria and thus change the course of biological processes of environment purification, in particular, from nitrite ions.

Anoxygenic photo- and chemo-synthesis of phototrophic sulfur bacteria from an alpine meromictic lake.

ABSTRACTMeromictic lakes are interesting ecosystems to study anaerobic microorganisms due their permanent stratification allowing the formation of a stable anoxic environment. The crenogenic meromictic Lake Cadagno harbors an important community of anoxygenic phototrophic sulfur bacteria responsible for almost half of its total productivity. Besides their ability to fix CO2 through photosynthesis, these microorganisms also showed high rates of dark carbon fixation via chemosyntesis. Here, we grew in pure cultures three populations of anoxygenic phototrophic sulfur bacteria previously isolated from the lake, accounting for 72.8% of the total microbial community and exibiting different phenotypes: (1) the motile, large-celled purple sulfur bacterium (PSB) Chromatium okenii, (2) the small-celled PSB Thiodictyon syntrophicum and (3) the green sulfur bacterium (GSB) Chlorobium phaeobacteroides. We measured their ability to fix CO2 through photo- and chemo-synthesis, both in situ in the lake and in laboratory under different incubation conditions. We also evaluated the efficiency and velocity of H2S photo-oxidation, an important reaction in the anoxygenic photosynthesis process. Our results confirm that phototrophic sulfur bacteria strongly fix CO2 in the presence of light and that oxygen increases chemosynthesis at night, in laboratory conditions. Moreover, substancial differences were displayed between the three selected populations in terms of activity and abundance.

Fifty years of limnology (1969-2019) at Mahoney Lake, British Columbia, Canada

Abstract Mahoney Lake is a small, meromictic saline lake in south-central British Columbia noted for its unique layer of purple sulfur bacteria. First examined in 1969, this lake has undergone physical, chemical, biological, and pre-historical research to generate an understanding of how the lake and its biota function have developed through time. Advances in understanding the sulfur transformations and bacterial nutrient cycling over the last fifty years have been prolific, resulting in the description of several new taxa. Mahoney Lake is exceptional in its limnological characteristics and is an ideal site for training future limnologists.

Insights into the catalytic mechanism of type VI sulfide:quinone oxidoreductases

Sulfide oxidation is catalyzed by ancient membrane-bound sulfide:quinone oxidoreductases (SQR) which are classified into six different types. For catalysis of sulfide oxidation, all SQRs require FAD cofactor and a redox-active centre in the active site, usually formed between conserved essential cysteines. SQRs of different types have variation in the number and position of cysteines, highlighting the potential for diverse catalytic mechanisms. The photosynthetic purple sulfur bacterium, Thiocapsa roseopersicina contains a type VI SQR enzyme (TrSqrF) having unusual catalytic parameters and four cysteines likely involved in the catalysis. Site-directed mutagenesis was applied to identify the role of cysteines in the catalytic process of TrSqrF. Based on biochemical and kinetic characterization of these TrSqrF variants, Cys121 is identified as crucial for enzyme activity. The cofactor is covalently bound via a heterodisulfide bridge between Cys121 and the C8M group of FAD. Mutation of another cysteine present in all SQRs (Cys332) causes remarkably decreased enzyme activity (14.6% of wild type enzyme) proving important, but non-essential role of this residue in enzyme catalysis. The sulfhydril-blocking agent, iodoacetamide can irreversibly inactivate TrSqrF but only if substrates are present and the enzyme is actively catalyzing its reaction. When the enzyme is inhibited by iodoacetamide, the FAD cofactor is released. The inhibition studies support a mechanism that entails opening and reforming of the heterodisulfide bridge during the catalytic cycle of TrSqrF. Our study thus reports the first detailed structure-function analysis of a type VI SQR enzyme which enables the proposal of a distinct mechanism of sulfide oxidation for this class.

Filamentous Anoxygenic Phototrophic Bacteria in Microbial Communities of the Kulunda Steppe Soda Lakes (Altai Krai, Russia)

Soda lakes are relic ecosystems inhabited by unique microorganisms, which are doubly extremophilic: both haloalkaliphilic and natronophilic. Two morphologically and physiologically similar monocultures of filamentous anoxygenic phototrophic bacteria (FAPB) were isolated from the water samples and biofilms on plants of the coastal zones of the steppe soda lakes Tanatar 6 and Gorchina 1 (Kulunda, Altai krai) during the season of their desalination. In their natural environments, FAPB coexisted with anoxygenic purple sulfur bacteria of the genera Chromatium, Thiocapsa, Ectothiorhodospira, and Thiorhodospira, as well as with oxygenic phototrophs (alkaliphilic cyanobacteria, euglenophytes, and diatoms). FAPB formed filaments surrounded by thin sheaths; their cells contained antenna structures (chlorosomes). Apart from small amounts of bacteriochlorophyll (BChl) a in the reaction center, both isolates contained also antenna BChl c and BChl d. Alpha- and beta-carotenes and their derivatives were also detected. FAPB grew well in the medium containing Na2S ∙ 9H2O (500 mg/L) at total mineralization of 15−30 g/L and pH 8−9.5. Identification of both cultures based on amino acid composition of the PufLM complex revealed that they belonged to the phylum Chloroflexi and were almost 100% identical both to each other and to the halo-alkaliphilic isolate “Candidatus Viridilinea mediisalina” Kir15-3F revealed previously in the Kiran soda lake (Eastern Siberia, Russia). It may be concluded that “Cand. Viridilinea mediisalina” is a typical component of Siberian soda lakes with moderate salinity.