D. Y. Sorokin

The microbial sulfur cycle at extremely haloalkaline conditions of soda lakes

Soda lakes represent a unique ecosystem with extremely high pH (up to 11) and salinity (up to saturation) due to the presence of high concentrations of sodium carbonate in brines. Despite these double extreme conditions, most of the lakes are highly productive and contain a fully functional microbial system. The microbial sulfur cycle is among the most active in soda lakes. One of the explanations for that is high-energy efficiency of dissimilatory conversions of inorganic sulfur compounds, both oxidative and reductive, sufficient to cope with costly life at double extreme conditions. The oxidative part of the sulfur cycle is driven by chemolithoautotrophic haloalkaliphilic sulfur-oxidizing bacteria (SOB), which are unique for soda lakes. The haloalkaliphilic SOB are present in the surface sediment layer of various soda lakes at high numbers of up to 106 viable cells/cm3. The culturable forms are so far represented by four novel genera within the Gammaproteobacteria, including the genera Thioalkalivibrio, Thioalkalimicrobium, Thioalkalispira, and Thioalkalibacter. The latter two were only found occasionally and each includes a single species, while the former two are widely distributed in various soda lakes over the world. The genus Thioalkalivibrio is the most physiologically diverse and covers the whole spectrum of salt/pH conditions present in soda lakes. Most importantly, the dominant subgroup of this genus is able to grow in saturated soda brines containing 4 M total Na+ – a so far unique property for any known aerobic chemolithoautotroph. Furthermore, some species can use thiocyanate as a sole energy source and three out of nine species can grow anaerobically with nitrogen oxides as electron acceptor. The reductive part of the sulfur cycle is active in the anoxic layers of the sediments of soda lakes. The in situ measurements of sulfate reduction rates and laboratory experiments with sediment slurries using sulfate, thiosulfate, or elemental sulfur as electron acceptors demonstrated relatively high sulfate reduction rates only hampered by salt-saturated conditions. However, the highest rates of sulfidogenesis were observed not with sulfate, but with elemental sulfur followed by thiosulfate. Formate, but not hydrogen, was the most efficient electron donor with all three sulfur electron acceptors, while acetate was only utilized as an electron donor under sulfur-reducing conditions. The native sulfidogenic populations of soda lakes showed a typical obligately alkaliphilic pH response, which corresponded well to the in situ pH conditions. Microbiological analysis indicated a domination of three groups of haloalkaliphilic autotrophic sulfate-reducing bacteria belonging to the order Desulfovibrionales (genera Desulfonatronovibrio, Desulfonatronum, and Desulfonatronospira) with a clear tendency to grow by thiosulfate disproportionation in the absence of external electron donor even at salt-saturating conditions. Few novel representatives of the order Desulfobacterales capable of heterotrophic growth with volatile fatty acids and alcohols at high pH and moderate salinity have also been found, while acetate oxidation was a function of a specialized group of haloalkaliphilic sulfur-reducing bacteria, which belong to the phylum Chrysiogenetes.

Application of bacteria involved in the biological sulfur cycle for paper mill effluent purification

In anaerobic wastewater treatment, the occurrence of biological sulfate reduction results in the formation of unwanted hydrogen sulfide, which is odorous, corrosive and toxic. In this paper, the role and application of bacteria in anaerobic and aerobic sulfur transformations are described and exemplified for the treatment of a paper mill wastewater. The sulfate containing wastewater first passes an anaerobic UASB reactor for bulk COD removal which is accompanied by the formation of biogas and hydrogen sulfide. In an aeration pond, the residual CODorganic and the formed dissolved hydrogen sulfide are removed. The biogas, consisting of CH4 (80-90 vol.%), CO2 (10-20 vol.%) and H2S (0.8-1.2 vol.%), is desulfurised prior to its combustion in a power generator thereby using a new biological process for H2S removal. This process will be described in more detail in this paper. Biomass from the anaerobic bioreactor has a compact granular structure and contains a diverse microbial community. Therefore, other anaerobic bioreactors throughout the world are inoculated with biomass from this UASB reactor. The sludge was also successfully used in investigation on sulfate reduction with carbon monoxide as the electron donor and the conversion of methanethiol. This shows the biotechnological potential of this complex reactor biomass.

Thioalkalivibrio thiocyanoxidans sp. nov. and Thioalkalivibrio paradoxus sp. nov., novel alkaliphilic, obligately autotrophic, sulfur- oxidizing bacteria capable of growth on thiocyanate, from soda lakes

Nine strains of haloalkaliphilic, obligately autotrophic, sulfur-oxidizing bacteria able to grow with thiocyanate (SCN-) as the sole energy and nitrogen source were isolated from soda lakes in South-East Siberia, Kenya and Egypt after enrichment on sodium carbonate minerals buffered at pH 10 with thiocyanate as the substrate. The isolates fell into two groups that were substantially different in terms of cell morphology, growth parameters and the ability to oxidize carbon disulfide. The bacteria were able to oxidize sulfide, polysulfide, sulfur and tetrathionate, as well as thiocyanate. Two isolates belonged to an extremely halotolerant type growing in the presence of up to 4 M Na+. Cyanate (CNO-) was the main nitrogen-containing intermediate during thiocyanate degradation in both groups. According to DNA-DNA hybridization data and phylogenetic analysis, both groups of isolates belong to a recently described genus of haloalkaliphilic sulfur-oxidizing bacteria, i.e. Thioalkalivibrio, belonging to the gamma-Proteobacteria, in which where they represent two new species. The species name Thioalkalivibrio paradoxus (type strain ARh 1T = DSM 13531T = JCM 11367T) is proposed for the group with barrel-shaped cells, and the species name Thioalkalivibrio thiocyanoxidans (type strain ARh 2T, DSM 13532T = JCM 11368T) is proposed for the group with vibrio-shaped cells. The diagnosis of the genus Thioalkalivibrio is amended according to the new data.

Physiological and genomic features of highly alkaliphilic hydrogen-utilizing Betaproteobacteria from a continental serpentinizing site.

Serpentinization, or the aqueous alteration of ultramafic rocks, results in challenging environments for life in continental sites due to the combination of extremely high pH, low salinity and lack of obvious electron acceptors and carbon sources. Nevertheless, certain Betaproteobacteria have been frequently observed in such environments. Here we describe physiological and genomic features of three related Betaproteobacterial strains isolated from highly alkaline (pH 11.6) serpentinizing springs at The Cedars, California. All three strains are obligate alkaliphiles with an optimum for growth at pH 11 and are capable of autotrophic growth with hydrogen, calcium carbonate and oxygen. The three strains exhibit differences, however, regarding the utilization of organic carbon and electron acceptors. Their global distribution and physiological, genomic and transcriptomic characteristics indicate that the strains are adapted to the alkaline and calcium-rich environments represented by the terrestrial serpentinizing ecosystems. We propose placing these strains in a new genus ‘Serpentinomonas’.

Plasticicumulans acidivorans gen. nov., sp nov., a polyhydroxyalkanoate-accumulating gammaproteobacterium from a sequencing-batch bioreactor

Here, we describe a novel bacterium, strain TUD-YJ37(T), which can accumulate polyhydroxybutyrate (PHB) to more than 85 % (w/w) dry cell weight. The bacterium was isolated from a mixed-culture bioreactor by using a feast-famine regime and its properties were characterized. Phylogenetic analysis based on full 16S rRNA gene sequences revealed that the isolate is a member of the Gammaproteobacteria, forming an independent, deep phylogenetic lineage. It is most closely related to members of the genera Methylocaldum, Methylococcus and Natronocella, with sequence similarities below 91 %. Strain TUD-YJ37(T) was an obligately aerobic, ovoid, Gram-negative bacterium, motile by means of a polar flagellum. It utilized C₂-C₁₀ fatty acids as carbon and energy sources. The temperature range for growth was 20-35 °C, with an optimum of 30 °C; the pH range was 6.0-8.0, without a clear optimum. The major respiratory quinone was Q-8. Polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, three unidentified phospholipids, an unidentified aminolipid and another unidentified lipid. The predominant fatty acids in the membrane polar lipids were C₁₆:₁ω7c, C₁₆:₀ and C₁₈:₁ω7c. The G+C content of the genomic DNA was 67.4 mol%. On the basis of phenotypic, chemotaxonomic and molecular data, the isolate is proposed to represent a novel genus and species, for which the name Plasticicumulans acidivorans gen. nov., sp. nov. is proposed. The type strain of Plasticicumulans acidivorans is TUD-YJ37(T) ( = DSM 23606(T)  = CBS 122990(T)).

Halophilic and haloalkaliphilic sulfur-oxidizing bacteria

Chemotrophic sulfur-oxidizing bacteria (SOB) represent an important functional group of microorganisms responsible for the dark oxidation of reduced sulfur compounds generated by sulfidogens. Until recently, only a single genus of halophilic SOB (Halothiobacillus) has been described, and nothing was known about the ability of this group to grow at high pH. Investigation of soda lakes, unique extremely alkaline and saline habitats, led to the discovery of a novel ecotype of natronophilic SOB. In contrast to the previously known neutrophilic ecotype, this group cannot grow at neutral pH, but grows optimally in soda brines at pH values around 10. They were the first chemolithoautotrophs among the described alkaliphiles. The group, so far, includes four novel genera within the Gammaproteobacteria. The genera Thioalkalimicrobium and Thioalkalispira represent low salt-tolerant alkaliphiles tolerating up to 1.5 M Na+. The genus Thioalkalibacter is a high salt-tolerant facultative alkaliphile. The genus Thioalkalivibrio is the most diverse and includes aerobic extremely salt-tolerant members and moderately salt-tolerant thiocyanate-utilizing and facultatively anaerobic denitrifying strains. The genome sequence of two Thioalkalivibrio strains revealed the presence of a truncated Sox system lacking the SoxCD component which is typical for gammaproteobacterial SOB. Bioenergetic studies of high salt-tolerant Thioalkalivibrio strains showed an obligate sodium dependence for respiratory activity implying the presence of sodium-dependent elements. Investigation of hypersaline inland chloride-sulfate lakes and hypersaline brines of marine origin with neutral pH revealed an unexpectedly high culturable diversity of halophilic obligately chemolithoautotrophic SOB comprising seven different groups within the Gammaproteobacteria. Two groups of strictly aerobic moderate halophiles were represented by the known genera Halothiobacillus and Thiomicrospira. Under denitrifying conditions and with thiocyanate as e-donor, three novel groups of moderately halophilic SOB were represented by the genera Thiohalomonas, Thiohalophilus, and Thiohalobacter. At 4 M NaCl, two groups of extremely halophilic SOB (a type not known before among the SOB) had been discovered. The obligately aerobic extreme halophiles comprised a novel genus Thiohalospira, and the facultatively anaerobic nitrate-reducing extreme halophiles—a novel deep-lineage genus Thiohalorhabdus. Overall, the investigation of hypersaline and (halo)alkaline habitats uncovered a novel and diverse world of extremophilic SOB with properties previously unknown for chemolithoautotrophic bacteria.

Nitrolancea hollandica gen. nov., sp. nov., a chemolithoautotrophic nitrite-oxidizing bacterium isolated from a bioreactor belonging to the phylum Chloroflexi

A novel nitrite-oxidizing bacterium (NOB), strain Lb(T), was isolated from a nitrifying bioreactor with a high loading of ammonium bicarbonate in a mineral medium with nitrite as the energy source. The cells were oval (lancet-shaped) rods with pointed edges, non-motile, Gram-positive (by staining and from the cell wall structure) and non-spore-forming. Strain Lb(T) was an obligately aerobic, chemolitoautotrophic NOB, utilizing nitrite or formate as the energy source and CO2 as the carbon source. Ammonium served as the only source of assimilated nitrogen. Growth with nitrite was optimal at pH 6.8-7.5 and at 40 °C (maximum 46 °C). The membrane lipids consisted of C20 alkyl 1,2-diols with the dominant fatty acids being 10MeC18 and C(18 : 1)ω9. The peptidoglycan lacked meso-DAP but contained ornithine and lysine. The dominant lipoquinone was MK-8. Phylogenetic analyses of the 16s rRNA gene sequence placed strain Lb(T) into the class Thermomicrobia of the phylum Chloroflexi with Sphaerobacter thermophilus as the closest relative. On the basis of physiological and phylogenetic data, it is proposed that strain Lb(T) represents a novel species of a new genus, with the suggested name Nitrolancea hollandica gen. nov., sp. nov. The type strain of the type species is Lb(T) ( = DSM 23161(T) = UNIQEM U798(T)).

Erratum to: Bacterial chitin utilization at halophilic conditions

In the original publication of this paper the DSM number of strain HCh-An1 was incorrectly given as DSM26639. The correct number is DSM26670.