Thioalkalivibrio Versutus Research Articles

Heterologous expression of sulfur: quinone oxidoreductase (Sqr) to improve Thioalkalivibrio versutus D301 desulfurization performance.

Heterologous expression of sulfur: quinone oxidoreductase (Sqr) from Halomonas mongoliensis JS01, which is responsible for oxidizing sulfide to elemental sulfur, in Thioalkalivibrio versutus (T. versutus) D301 improves desulfurization. We expressed sqr in T. versutus D301 by conjugative transfer and then assayed its desulfurization capacity in an airlift reactor and analyzed its transcriptome at -380mV ORP. Our findings demonstrate that the D301-sqr+ strain, utilizing sodium sulfide as a sulfur source under optimal ORP conditions (-380mV), achieved an elemental sulfur yield of 95%. This represents an 8% increase over the T. versutus D301. Moreover, the sodium sulfide utilization rate for D301-sqr+showed a marked improvement [0.741vs. 0.651mmol∙(l·h)-1], with a concurrent increase in the rate of elemental sulfur production when compared to the T. versutus D301 (0.716vs. 0.518mmol ∙(l·h)-1). Transcriptome analysis revealed that the flavocytochrome c (fcc) and the sox system were differentially transcriptionally down-regulated in D301-sqr+ compared with the T. versutus D301. Heterologous expression of the gene sqr altered the transcription of related genes in T. versutus D301 sulfur oxidation pathway, increasing the yield of elemental sulfur and the rate of sulfur oxidation, and making D301-sqr+ more potential for industrial applications.

Biological Sulfur Production Potential of Local Sulfur Oxidizing Bacteria Culture and Thioalkalivibrio versutus AL2 in Haloalkaliphilic Conditions

Biological Sulfur Production Potential of Local Sulfur Oxidizing Bacteria Culture and Thioalkalivibrio versutus AL2 in Haloalkaliphilic Conditions

CRISPR/Cas12a-Mediated Genome Editing in Thioalkalivibrio versutus

Haloalkaliphilic Thioalkalivibrio versutus, a dominant species for sulfide removal, has attracted increasing attention. However, research on T. versutus is limited by the lack of genetic manipulation tools. In this work, we developed a CRISPR/AsCas12a-mediated system in T. versutus for an efficient and implementable genome editing workflow. Compared to the CRISPR/Cas9-mediated system, the CRISPR/AsCas12a system exhibited enhanced editing efficiency. Additionally, as Cas12a is capable of processing the crRNA maturation independently, the CRISPR/AsCas12a system allowed multiplex gene editing and large-fragment DNA knockout by expressing more than one crRNA under the control of one promoter. Using the CRISPR/AsCas12a system, five key genes of the elemental sulfur oxidation pathway were knocked out. Simultaneous deletion of the rhd and tusA genes disrupted the ability of T. versutus to metabolize elemental sulfur, resulting in a 24.7% increase in elemental sulfur generation and a 15.2% reduction in sulfate production. This genome engineering strategy significantly improved our understanding of sulfur metabolism in Thioalkalivibrio spp.

Organic layer characteristics and microbial utilization of the biosulfur globules produced by haloalkaliphilic Thioalkalivibrio versutus D301 during biological desulfurization.

The haloalkaliphilic genus Thioalkalivibrio, widely used in bio-desulfurization, can oxidize H2S to So, which is excreted outside cells in the form of biosulfur globules. As by-product of bio-desulfurization, information on biosulfur globules is still very scant, which limits its high-value utilization. In this paper, the characteristics of biosulfur globules produced by Thioalkalivibrio versutus D301 and the possibility of cultivating sulfur-oxidizing bacteria as a high biological-activity sulfur source were studied. The sulfur element in the biosulfur globules existed in the form α-S8, which was similar to chemical sulfur. The biosulfur globule was wrapped with an organic layer composed of polysaccharides and proteins. The composition of this organic layer could change. In the formation stage of biosulfur globules, the organic layer was dominated by polysaccharides, and in later stage, proteins became the main component. We speculated that the organic layer was mainly formed by the passive adsorption of organic matter secreted by cells. The existence of organic layer endowed biosulfur with better bioavailability. Compared with those found using chemical sulfur, the growth rates of Acidithiobacillus thiooxidans ATCC 19377T, Thiomicrospira microaerophila BDL05 and Thioalkalibacter halophilus BDH06 using biosulfur increased several folds to an order of magnitude, indicating that biosulfur was a good sulfur source for cultivating sulfur-oxidizing bacteria.

High tolerance of chemolithoautotrophic sulphur oxidizing bacteria towards pulp and paper mill wastewaters and their organic constituents supporting sulphur recovery in alkaline conditions

This study reports the tolerance of chemolithoautotrophic biotransformation of sulphurous compounds towards pulp and paper (P&P) mill wastewaters (primary filtrate of bleaching (PFB) and composite wastewater (WW)) and their constituents under haloalkaline conditions. The effects of organic compounds (methanol, acetate, D(+)-xylose, phenol and benzene) that may be present in P&P wastewaters, and yeast extract, a complex organic compound on thiosulphate biotransformation by Thioalkalivibrio versutus were investigated. All experiments were carried out in batch bioassays at pH 10 and 13–23 g Na+/L. Phenol and benzene reduced thiosulphate biotransformation by 88 and 94% at 0.25 and 1 g/L, respectively in 10 days. 20 g/L methanol, 20 g/L yeast extract and 10 g/L xylose reduced the biotransformation by 90, 88 and 56%, respectively. No inhibition of biotransformation occurred with acetate at concentrations up to 20 g/L. The growth was also enhanced by 1 to 10 g/L yeast extract likely serving as additional nutrients. At pH (∼10), the studied organic acids remain mostly unprotonated and, thus control their access through the cell membrane. Therefore, the inaccessibility of these compounds to the cytosol is a likely mechanism for having non-inhibitory effects. The 87% (v/v) WW did not affect thiosulphate biotransformation efficiency while 87% (v/v) PFB reduced it by 36% by day 10. The resistance of T. versutus to common organics present in P&P wastewaters indicates its potential use for sulphur recovery from P&P mill wastewaters at haloalkaline conditions and thus, supports the circular economy approach.

Enhanced Biodesulfurization with a Microbubble Strategy in an Airlift Bioreactor with Haloalkaliphilic Bacterium Thioalkalivibrio versutus D306.

Biodesulfurization under haloalkaline conditions requires limiting oxygen and additional energy in the system to deliver high mixing quality control. This study considers biodesulfurization in an airlift bioreactor with uniform microbubbles generated by a fluidic oscillation aeration system to enhance the biological desulfurization process and its hydrodynamics. Fluidic oscillation aeration in an airlift bioreactor requires minimal energy input for microbubble generation. This aeration system produced 81.87% smaller average microbubble size than the direct aeration system in a bubble column bioreactor. The biodesulfurization phase achieved a yield of 94.94% biological sulfur, 84.91% biological sulfur selectivity, and 5.06% sulfur oxidation performance in the airlift bioreactor with the microbubble strategy. The biodesulfurization conditions of thiosulfate via Thioalkalivibrio versutus D306 are revealed in this study. The biodesulfurization conditions in the airlift bioreactor with the fluidic oscillation aeration system resulted in the complete conversion of thiosulfate with 27.64% less sulfate production and 10.34% more biological sulfur production than in the bubble column bioreactor. Therefore, pleasant hydrodynamics via an airlift bioreactor mechanism with microbubbles is favored for biodesulfurization under haloalkaline conditions.

Identification of the Biosynthetic Pathway of Glycine Betaine That Is Responsible for Salinity Tolerance in Halophilic Thioalkalivibrio versutus D301.

Thioalkalivibrio versutus D301 has been widely used in the biodesulfurization process, as it is capable of oxidizing hydrogen sulfide to elemental sulfur under strongly halo-alkaline conditions. Glycine betaine contributes to the increased tolerance to extreme environments in some of Thioalkalivibrio species. However, the biosynthetic pathway of glycine betaine in Thioalkalivibrio remained unknown. Here, we found that genes associated with nitrogen metabolism of T. versutus D301 were significantly upregulated under high-salt conditions, causing the enhanced production of glycine betaine that functions as a main compatible solute in response to the salinity stress. Glycine betaine was synthesized by glycine methylation pathway in T. versutus D301, with glycine N-methyltransferase (GMT) and sarcosine dimethylglycine N-methyltransferase (SDMT) as key enzymes in this pathway. Moreover, substrate specificities of GMT and SDMT were quite different from the well characterized enzymes for glycine methylation in halophilic Halorhodospira halochloris. Our results illustrate the glycine betaine biosynthetic pathway in the genus of Thioalkalivibrio for the first time, providing us with a better understanding of the biosynthesis of glycine betaine in haloalkaliphilic Thioalkalivibrio.

Investigating role of abiotic side and finding optimum abiotic condition for improving gas biodesulfurization using Thioalkalivibrio versutus

Hydrogen sulfide (H2S) is a super toxic substance that produces SOx gases when combusted. Therefore, it should be removed from gas streams. Biodesulfurization is one of the developing methods for removing sulfide. Gas biodesulfurization must be accelerated to be competitive with chemical processes. This process has two sides: biotic and abiotic sides. To increase the rate of sulfide removal, this substance should be given to the bacteria in the maximum amount (Max. − RHS B). Therefore, it is necessary to minimize the rate of adverse abiotic reactions of sulfide (Min. − RHS A). Minimizing the sulfide reaction with biosulfur and oxygen and thiosulfate generation (Min. − RHS thio2) was assessed in de-microbized medium. It was concluded that the pH should be kept as low as possible. The kinetics of thiosulfate formation from sulfide oxidation (− RHS thio1) are strongly dependent on the sulfide concentration, and to minimize this reaction rate, sulfide should be gently injected into the culture. To minimize sulfide reduction to hydrogen sulfide (Min. − RHS rev), the pH should be kept as high as possible. Using the Design Expert v.13, a model was driven for the abiotic side to obtain optimum condition. The pH value was found to be 8.2 and the sulfide concentration to 2.5E−05 M. Thioalkalivibrio versutus cultivation under identified abiotic conditions resulted in biological removal of sulfide up to 1.5 g/h. The culture was not able to remove 2 g/h input sulfide, and to increase this, the biotic side should be studied.

Sodium Energetic Cycle in the Natronophilic Bacterium Thioalkalivibrio versutus.

As inhabitants of soda lakes, Thioalkalivibrio versutus are halo- and alkaliphilic bacteria that have previously been shown to respire with the first demonstrated Na+-translocating cytochrome-c oxidase (CO). The enzyme generates a sodium-motive force (Δs) as high as −270 mV across the bacterial plasma membrane. However, in these bacteria, operation of the possible Δs consumers has not been proven. We obtained motile cells and used them to study the supposed Na+ energetic cycle in these bacteria. The resulting motility was activated in the presence of the protonophore 2-heptyl-4-hydroxyquinoline N-oxide (HQNO), in line with the same effect on cell respiration, and was fully blocked by amiloride—an inhibitor of Na+-motive flagella. In immotile starving bacteria, ascorbate triggered CO-mediated respiration and motility, both showing the same dependence on sodium concentration. We concluded that, in T. versutus, Na+-translocating CO and Na+-motive flagella operate in the Na+ energetic cycle mode. Our research may shed light on the energetic reason for how these bacteria are confined to a narrow chemocline zone and thrive in the extreme conditions of soda lakes.

Elemental sulphur production from thiosulphate under haloalkaline conditions in a Thioalkalivibrio versutus amended fluidized bed bioreactor

Concentrated sulphurous and saline streams, produced for example by pulp and paper and petrochemical industries, pose challenges for both environmental and processes management. In this study, the potential of biological recovery of S0 from haloalkaline thiosulphate solution in a Thioalkalivibrio versutus amended continuous-flow fluidized bed bioreactor (FBBR) was investigated using different (12−5 h) hydraulic retention times (HRT) as well as physico-chemical means to separate the S0 produced. S2O32− was biotransformed to SO42− and S0 with the highest biotransformation efficiency of 99.9 %. At 7 h HRT, the capacity of the FBBR was reached, seen as incomplete thiosulphate conversion. S0 production rate increased up to 6.3 ± 0.6 g S/l/d at HRT 7 h, whilst the average S0 yield was 27 ± 2 %. The presence of biologically produced S0 was visual and identified by scanning electron microscopy. Separation of S0 from the effluent by centrifugation at 3417 relative centrifugal force (rcf) resulted in 93 % separation, while among the four tested coagulants, FeCl2 at 0.5 g/l resulted in 40 % separation. Also, FeCl2 enhanced thiosulphate biotransformation rates. In summary, continuous biological S0 production followed by separation by centrifugation indicates potential for sulphur recovery from alkaline and saline industrial streams.

Molecular and Physiological Adaptations to Low Temperature in Thioalkalivibrio Strains Isolated from Soda Lakes with Different Temperature Regimes.

ABSTRACTThe genus Thioalkalivibrio comprises sulfur-oxidizing bacteria thriving in soda lakes at high pH and salinity. Depending on the geographical location and the season, these lakes can strongly vary in temperature. To obtain a comprehensive understanding of the molecular and physiological adaptations to low temperature, we compared the responses of two Thioalkalivibrio strains to low (10°C) and high (30°C) temperatures. For this, the strains were grown under controlled conditions in chemostats and analyzed for their gene expression (RNA sequencing [RNA-Seq]), membrane lipid composition, and glycine betaine content. The strain Thioalkalivibrio versutus AL2T originated from a soda lake in southeast Siberia that is exposed to strong seasonal temperature differences, including freezing winters, whereas Thioalkalivibrio nitratis ALJ2 was isolated from an East African Rift Valley soda lake with a constant warm temperature the year round. The strain AL2T grew faster than ALJ2 at 10°C, likely due to its 3-fold-higher concentration of the osmolyte glycine betaine. Moreover, significant changes in the membrane lipid composition were observed for both strains, leading to an increase in their unsaturated fatty acid content via the Fab pathway to avoid membrane stiffness. Genes for the transcriptional and translational machinery, as well as for counteracting cold-induced hampering of nucleotides and proteins, were upregulated. Oxidative stress was reduced by induction of vitamin B12 biosynthesis genes, and growth at 10°C provoked downregulation of genes involved in the second half of the sulfur oxidation pathway. Genes for intracellular signal transduction were differentially expressed, and interestingly, AL2T upregulated flagellin expression, whereas ALJ2 downregulated it.IMPORTANCE In addition to their haloalkaline conditions, soda lakes can also harbor a variety of other extreme parameters, to which their microbial communities need to adapt. However, for most of these supplementary stressors, it is not well known yet how haloalkaliphiles adapt and resist. Here, we studied the strategy for adaptation to low temperature in the haloalkaliphilic genus Thioalkalivibrio by using two strains isolated from soda lakes with different temperature regimes. Even though the strains showed a strong difference in growth rate at 10°C, they exhibited similar molecular and physiological adaptation responses. We hypothesize that they take advantage of resistance mechanisms against other stressors commonly found in soda lakes, which are therefore maintained in the bacteria living in the absence of low-temperature pressure. A major difference, however, was detected for their glycine betaine content at 10°C, highlighting the power of this osmolyte to also act as a key compound in cryoprotection.Author Video: An author video summary of this article is available.

Identification of Na+-Pumping Cytochrome Oxidase in the Membranes of Extremely Alkaliphilic Thioalkalivibrio Bacteria

For the first time, the functioning of the oxygen reductase Na+-pump (Na+-pumping cytochrome c oxidase of the cbb3-type) was demonstrated by examining the respiratory chain of the extremely alkaliphilic bacterium Thioalkalivibrio versutus [Muntyan, M. S., et al. (2015) Cytochrome cbb3 of Thioalkalivibrio is a Na+-pumping cytochrome oxidase, Proc. Natl. Acad. Sci. USA, 112, 7695-7700], a product of the ccoNOQP operon. In this study, we detected and identified this enzyme using rabbit polyclonal antibody against the predicted C-terminal amino acid sequence of its catalytic subunit. We found that this cbb3-type oxidase is synthesized in bacterial cells, where it is located in the membranes. The 48-kDa oxidase subunit (CcoN) is catalytic, while subunits CcoO and CcoP with molecular masses of 29 and 34 kDa, respectively, are cytochromes c. The theoretical pI values of the CcoN, CcoO, and CcoP subunits were determined. It was shown that parts of the CcoO and CcoP subunits exposed to the aqueous phase on the cytoplasmic membrane P-side are enriched with negatively charged amino acid residues, in contrast to the parts of the integral subunit CcoN adjacent to the aqueous phase. Thus, the Na+-pumping cytochrome c oxidase of T. versutus, both in function and in structure, demonstrates adaptation to extremely alkaline conditions.

Performance and characteristic of a haloalkaliphilic bio-desulfurizing system using Thioalkalivibrio verustus D301 for efficient removal of H2S

Haloalkaliphilic biological desulfurization is attracting people's attention due to its remarkable economy and good performance. In this study, a bio-desulfurizing system was developed based on a sulfur-oxidizing bacterium Thioalkalivibrio versutus D301 to treat H2S under haloalkaliphilic conditions. Results showed the optimum performance of the system was obtained under sulfide load of 3.27 g/L/d and oxidization-reduction potential (ORP) value of −395 mV. The selection of elemental sulfur and sulfate reached 86.19 % ± 3.02 % and 8.6 ± 0.28 % under these conditions. Then, the obvious linear relationships between sulfide load and ORP as well as aeration rate were revealed. The gradient variations of ORP and sulfurous compounds concentration with bioreactor depth were revealed, which proved the existence of spatial and temporal heterogeneity in the bioreactor. The morphology difference of sulfur particles from bioreactor and shake flask implied the key role of operating conditions in shaping particle morphology, and it was found that 56.3 % of total S° particles were harvested within 2 h by gravity settling. This study will provide a valuable reference for the design of bio-desulfurizing equipment and the optimization of operational conditions in the industrial process in the future.

Improving confirmed nanometric sulfur bioproduction using engineered Thioalkalivibrio versutus

Complicated production procedures and superior characteristics of nano-sized sulfur elevate its price to 25–40 fold higher than micrograde kind. Also, natural gas hydrogen sulfide levels are restricted because of its toxic environmental consequences. Thioalkalivibrio versutus is a polyextremophilic industrial autotroph with high natural gas desulfurization capability. Here, nanometric (>50 nm) sulfur bioproduction using T. versutus while desulfurizing natural gas was validated. Also, this production was enhanced by 166.7% via lowering sulfate production by 55.1%. A specially-developed CRISPR system, with 42% editing efficiency, simplified the genome editing workflow scheme for this challenging bacterium. In parallel, sulfur metabolism was uncovered using proteins mining and transcriptome studies for defining sulfate-producing key genes (heterodisulfide reductase-like complex, sulfur dioxygenase, sulfite dehydrogenase and sulfite oxidase). This study provided cost-effective nanometric sulfur production and improved this production using a novel CRISPR strategy, which could be suitable for industrial polyextremophiles, after uncovering sulfur pathways in T. versutus.

Kinetics and modelling of thiosulphate biotransformations by haloalkaliphilic Thioalkalivibrio versutus

Biotransformation of thiosulphate by Thioalkalivibrio versutus was studied under haloalkaline conditions (pH 10, 0.66–1.2 M Na+) using batch assays and modelling tools for possible sulphur recovery from haloalkaline industrial streams. The thiosulphate was fully biotransformed to sulphate or to sulphate and elemental sulphur at initial S2O32−-S concentrations of 25–550 mM within 10 days. The highest biotransformation rate of 2.66 mM [S2O32−-S] h−1 was obtained at initial S2O32−-S concentration of 550 mM with half saturation constant (Ks) of 54.5 mM [S2O32−-S]. At initial concentrations below 100 mM S2O32−-S, the main product was sulphate whilst at above 100 mM also elemental sulphur was produced with up to 29% efficiency. The model approach developed incorporated S2O32− biotransformation to SO42− and S0. The kinetic modelling results were compatible (R2 > 0.90) with the experimental data. The maximum growth rate (µm) was 0.048 h−1 (0.47 mM C5H7NO2 h−1) and the maximum growth yield 0.18 mM C5H7NO2/mM S2O32−-S (20 g cell/mol S2O32−-S). The high rate thiosulphate biotransformation and elemental sulphur recovery results together with the developed kinetic model can be used for bioprocess design and operation. The potential industrial applications would aim at sustainable resource recovery from industrial haloalkaline and sulphurous process and/or effluent streams.

Deep and high-efficiency removal of sulfate through a coupling system with sulfate-reducing and sulfur-oxidizing capacity under haloalkaliphilic condition.

Sulfide from anaerobic treatment of high-sulfate wastewater would always have some adverse effects on downstream processes. In this study, a coupling anaerobic/aerobic system was developed and operated under haloalkaliphilic condition to realize deep and high-efficiency removal of sulfate without production of sulfide. A haloalkaliphilic sulfur-oxidizing strain, Thioalkalivibrio versutus SOB306, was responsible for oxidation of sulfide. The anaerobic part was first operated at optimum condition based on a previous study. Then, its effluent with an average sulfide concentration of 674 ± 33mg·l-1 was further directly treated by a set of 1 l biofilter with SOB306 strain under aerobic condition. Finally, 100% removal rate of sulfide was achieved at aeration rate of 0.75l·l-1·min-1, ORP of -392mV and HRT of 4h. The average yield of elemental sulfur reached 79.1 ± 1.3% in the filter, and the CROS achieved a conversion rate of sulfate to sulfur beyond 54%. This study for the first time revealed the characteristics and performance of the haloalkaliphilic CROS in deep treatment of high-sulfate wastewater, whichpavedthewayforthedevelopmentandapplicationofthis method in the real world.

Enhanced growth-driven stepwise inducible expression system development in haloalkaliphilic desulfurizing Thioalkalivibrio versutus

Highly toxic and flammable H2S gas has become an environmental threat. Because of its ability to efficiently remove H2S by oxidation, Thioalkalivibrio versutus is gaining more attention. Haloalkaliphilic autotrophs, like the bio-desulfurizing T. versutus, grow weakly. Weak growth makes any trial for developing potent genetic tools required for genetic engineering far from achieved. In this study, the fed-batch strategy improved T. versutus growth by 1.6 fold in maximal growth rate, 9-fold in O.D600 values and about 3-fold in biomass and protein productions. The strategy also increased the favorable desulfurization product, sulfur, by 2.7 fold in percent yield and 1.5-fold in diameter. A tight iron-inducible expression system for T. versutus was successfully developed. The system was derived from fed-batch cultivation coupled with new design, build, test and validate (DPTV) approach. The inducible system was validated by toxin expression. Fed-batch cultivation coupled with DPTV approach could be applied to other autotrophs.

Investigation of transient forms of sulfur during biological treatment of spent caustic

ABSTRACTIn the present study, the production of various transient forms of sulfur during biological oxidation of sulfidic spent caustics under haloalkaline conditions in a stirred tank bioreactor is investigated. Also, the effects of abiotic aeration (chemical oxidation), dissolved oxygen (DO) concentration and sodium concentration on forms of sulfur during biological treatment are demonstrated. Thioalkalivibrio versutus strain was used for sulfide oxidation in spent caustic (SC). The aeration had an important effect on sulfide oxidation and its final products. At DO concentrations above 2 mg l−1, majority of sulfide was oxidized to sulfate. Maximum sulfide removal efficiency (%R) and yield of sulfate production was obtained in Na+ concentration ranging from 0.6 to 2 M. Abiotic aeration, which is the most important factor of production of thiosulfate, resulted in the formation of an undesired product-polysulfide. However, abiotic aeration can be used as a pretreatment to biological treatment. In the bioreactor the removal efficiency was obtained as 82.7% and various forms of sulfur such as polysulfide, biosulfur, thiosulfate and sulfate was observed during biological treatment of SC.

Study on Haloalkaliphilic Sulfur-Oxidizing Bacterium for Thiosulfate Removal in Treatment of Sulfidic Spent Caustic

Due to the disadvantages of physiochemical methods for sulfidic spent caustic treatment, attentions are drawn to the environmental-friendly biotreatments including sulfur-oxidizing halo-alkaliphiles. Thioalkalivibrio versutus DSM 13738 was grown at alkaline (pH10) autotrophic medium with sodium carbonate/bicarbonate as the sole source of carbon and amended with sodium thiosulfate as the electron and energy source. The effect of various parameters including temperature (25-40 °C), pH (8-11), NaCl concentration (0.5-5 % w/v) and sodium thiosulfate concentrations (100-750 mM) was evaluated on bacterial growth and thiosulfate removal. This strain could eliminate sodium thiosulfate at very high concentrations up to 750 mM. The results showed that the highest specific growth rate was pH 9.5 and thiosulfate removal of Thioalkalivibrio versutus occurred at pH 10.5. The optimum salt concentration for thiosulfate removal was 2.5 % w/v and 5 % NaCl and specific growth rate elevated 2.5% w/v. It was also specified that this strain thrives occurred in 37 °C and at 35 and 37 °C higher removal of thiosulfate. Following chemical oxidation of sulfide to thiosulfate, application of Thioalkalivibrio versutus could be promising for spent caustic treatment. Since thiosulfate is utilized as an energy source, highest removal efficiency occurred at marginally different conditions compared to optimal growth.

Cytochrome cbb3 of Thioalkalivibrio is a Na+-pumping cytochrome oxidase

Cytochrome c oxidases (Coxs) are the basic energy transducers in the respiratory chain of the majority of aerobic organisms. Coxs studied to date are redox-driven proton-pumping enzymes belonging to one of three subfamilies: A-, B-, and C-type oxidases. The C-type oxidases (cbb3 cytochromes), which are widespread among pathogenic bacteria, are the least understood. In particular, the proton-pumping machinery of these Coxs has not yet been elucidated despite the availability of X-ray structure information. Here, we report the discovery of the first (to our knowledge) sodium-pumping Cox (Scox), a cbb3 cytochrome from the extremely alkaliphilic bacterium Thioalkalivibrio versutus. This finding offers clues to the previously unknown structure of the ion-pumping channel in the C-type Coxs and provides insight into the functional properties of this enzyme.