Phenotype-driven assessment of the ancestral trajectory of sulfur biooxidation in the thermoacidophilic archaea Sulfolobaceae.

Certain members of the family Sulfolobaceae represent the only archaea known to oxidize elemental sulfur, and their evolutionary history provides a framework to understand the development of chemolithotrophic growth by sulfur oxidation. Here, we evaluate the sulfur oxidation phenotype of Sulfolobaceae species and leverage comparative genomic and transcriptomic analysis to identify the key genes linked to sulfur oxidation. Metabolic engineering of the obligate heterotroph Sulfolobus acidocaldarius revealed that the known cytoplasmic components of sulfur oxidation alone are not sufficient to drive prolific sulfur oxidation. Imaging analysis showed that Sulfolobaceae species maintain proximity to the sulfur surface but do not necessarily contact the substrate directly. This indicates that a soluble form of sulfur must be transported to initiate cytoplasmic sulfur oxidation. Conservation patterns and transcriptomic response implicate an extracellular tetrathionate hydrolase and putative thiosulfate transporter in a newly proposed mechanism of sulfur acquisition in the Sulfolobaceae.IMPORTANCESulfur is one of the most abundant elements on earth (2.9% by mass), so it makes sense that the earliest biology found a way to use sulfur to create and sustain life. However, beyond evolutionary significance, sulfur and the molecules it comprises have important technological significance, not only in chemicals such as sulfuric acid and in pyritic ores containing critical metals but also as a waste product from oil and gas production. The thermoacidophilic Sulfolobaceae are unique among the archaea as sulfur oxidizers. The trajectory for how sulfur biooxidation arose and evolved can be traced using experimental and bioinformatic analyses of the available genomic data set. Such analysis can also inform the process by which extracellular sulfur is acquired and transported by thermoacidophilic archaea, a phenomenon that is critical to these microorganisms but has yet to be elucidated.

Autotrophic denitrification by sulfur-based immobilized electron donor for enhanced nitrogen removal: Denitrification performance, microbial interspecific interaction and functional traits.

Sulfur-driven autotrophic denitrification (SdAD) is a promising nitrogen removing process, but its applications were generally constrained by conventional electron donors (i.e., thiosulfate (Na2S2O3)) with high valence and limited bioavailability. Herein, an immobilized electron donor by loading elemental sulfur on the surface of polyurethane foam (PFSF) was developed, and its feasibility for SdAD was investigated. The denitrification efficiency of PFSF was 97.3%, higher than that of Na2S2O3 (91.1%). Functional microorganisms (i.e., Thiobacillus and Sulfurimonas) and their metabolic activities (i.e., nir and nor) were substantially enhanced by PFSF. PFSF resulted in the enrichment of sulfate-reducing bacteria, which can reduce sulfate (SO42-). It attenuated the inhibitory effect of SO42-, whereas the generated product (hydrogen sulfide) also served as an electron donor for SdAD. According to the economic evaluation, PFSF exhibited strong market potential. This study proposes an efficient and low-cost immobilized electron donor for SdAD and provides theoretical support to its practical applications.

A review on sulfur transformation during anaerobic digestion of organic solid waste: Mechanisms, influencing factors and resource recovery.

Anaerobic digestion (AD) is an economical and environment-friendly technology for treating organic solid wastes (OSWs). OSWs with high sulfur can lead to the accumulation of toxic and harmful hydrogen sulfide (H2S) during AD, so a considerable amount of studies have focused on removing H2S emissions. However, current studies have found that sulfide induces phosphate release from the sludge containing iron­phosphorus compounds (FePs) and the feasibility of recovering elemental sulfur (S0) during AD. To tap the full potential of sulfur in OSWs resource recovery, deciphering the sulfur transformation pathway and its influencing factors is required. Therefore, in this review, the sulfur species and distributions in OSWs and the pathway of sulfur transformation during AD were systematically summarized. Then, the relationship between iron (ferric compounds and zero-valent iron), phosphorus (FePs) and sulfur were analyzed. It was found that the reaction of iron with sulfide during AD drove the conversion of sulfide to S0 and iron sulfide compounds (FeSx), and consequently iron was applied in sulfide abatement. In particular, ferric (hydr)oxide granules offer possibilities to improve the economic viability of hydrogen sulfide control by recovering S0. Sulfide is an interesting strategy to release phosphate from the sludge containing FePs for phosphorus recovery. Critical factors affecting sulfur transformation, including the carbon source, free ammonia and pretreatment methods, were summarized and discussed. Carbon source and free ammonia affected sulfur-related microbial diversity and enzyme activity and different sulfur transformation pathways in response to varying pretreatment methods. The study on S0 recovery, organic sulfur conversion, and phosphate release mechanism triggered by sulfur deserves further investigation. This review is expected to enrich our knowledge of the role of sulfur during AD and inspire new ideas for recovering phosphorus and sulfur resources from OSWs.

Realizing a high-rate sulfidogenic reactor driven by sulfur-reducing bacteria with organic substrate dosage minimization and cost-effectiveness maximization.

Biological sulfur reduction is an attractive sulfidogenic technology for the treatment of organics-deficient metal-laden wastewater, because it theoretically reduces the electron donor consumption by 75%, compared to sulfate reduction. However, reducing the external organic substrate dosage may lower the sulfur reduction rate. Supplying with a more biodegradable organic substrate could possibly enhance sulfidogenic activity but also increase the chemical cost. Therefore, the sulfide production performance of a sulfur-reducing bioreactor feeding with varied levels of organic supply, and different types of organic substrates were investigated. The results showed that high-rate sulfide production (12.30 mg S/L/h) in a sulfur-reducing bioreactor can be achieved at the minimal dosage of organic substrate as low as 39 mg C/L of organic carbon in the influent. Changing the type of organic substrate posed a significant effect on the sulfidogenic activity in the sulfur-reducing bioreactor. Sodium acetate was found to be the optimal substrate to achieve the highest sulfide production rate (28.20 mg S/L/h) by sulfur-reducing bacteria (S0RB), followed by ethanol, methanol, glycerol, pyruvic acid, acetic acid, glucose, sucrose, malic acid, sodium formate, formic acid, N-propanol, N-butanol, lactic acid, sodium lactate, propionic acid and sodium propionate (2.87 mg S/L/h as the lowest rate). However, the cost-effectiveness analysis showed that glucose was the most cost-effective organic substrate to realize the sulfur reduction process in high sulfide production rate (20.13 mg S/L/h) and low chemical cost (5.94 kg S/$). The utilization pathway of the different organic substrates in the sulfur-reducing bioreactor was also discussed.

QMEC: a tool for high-throughput quantitative assessment of microbial functional potential in C, N, P, and S biogeochemical cycling.

Microorganisms are major drivers of elemental cycling in the biosphere. Determining the abundance of microbial functional traits involved in the transformation of nutrients, including carbon (C), nitrogen (N), phosphorus (P) and sulfur (S), is critical for assessing microbial functionality in elemental cycling. We developed a high-throughput quantitative-PCR-based chip, Quantitative microbial element cycling (QMEC), for assessing and quantifying the genetic potential of microbiota to mineralize soil organic matter and to release C, N, P and S. QMEC contains 72 primer pairs targeting 64 microbial functional genes for C, N, P, S and methane metabolism. These primer pairs were characterized by high coverage (average of 18-20 phyla covered per gene) and sufficient specificity (>70% match rate) with a relatively low detection limit (7-102 copies per run). QMEC was successfully applied to soil and sediment samples, identifying significantly different structures, abundances and diversities of the functional genes (P<0.05). QMEC was also able to determine absolute gene abundance. QMEC enabled the simultaneous qualitative and quantitative determination of 72 genes from 72 samples in one run, which is promising for comprehensively investigating microbially mediated ecological processes and biogeochemical cycles in various environmental contexts including those of the current global change.

Inferring Methionine Sulfoxidation and serine Phosphorylation crosstalk from Phylogenetic analyses.

BACKGROUND: The sulfoxidation of methionine residues within the phosphorylation motif of protein kinase substrates, may provide a mechanism to couple oxidative signals to changes in protein phosphorylation. Herein, we hypothesize that if the residues within a pair of phosphorylatable-sulfoxidable sites are functionally linked, then they might have been coevolving. To test this hypothesis a number of site pairs previously detected on human stress-related proteins has been subjected to analysis using eukaryote ortholog sequences and a phylogenetic approach. RESULTS: Overall, the results support the conclusion that in the eIF2α protein, serine phosphorylation at position 218 and methionine oxidation at position 222, belong to the same functional network. First, the observed data were much better fitted by Markovian models that assumed coevolution of both sites, with respect to their counterparts assuming independent evolution (p-value = 0.003). Second, this conclusion was robust with respect to the methods used to reconstruct the phylogenetic relationship between the 233 eukaryotic species analyzed. Third, the co-distribution of phosphorylatable and sulfoxidable residues at these positions showed multiple origins throughout the evolution of eukaryotes, which further supports the view of an adaptive value for this co-occurrence. Fourth, the possibility that the coevolution of these two sites might be due to structure-driven compensatory mutations was evaluated. The results suggested that factors other than those merely structural were behind the observed coevolution. Finally, the relationship detected between other modifiable site pairs from ataxin-2 (S814-M815), ataxin-2-like (S211-M215) and Pumilio homolog 1 (S124-M125), reinforce the view of a role for phosphorylation-sulfoxidation crosstalk. CONCLUSIONS: For the four stress-related proteins analyzed herein, their respective pairs of PTM sites (phosphorylatable serine and sulfoxidable methionine) were found to be evolving in a correlated fashion, which suggests a relevant role for methionine sulfoxidation and serine phosphorylation crosstalk in the control of protein translation under stress conditions.

Binding of Copper and Cisplatin to Atox1 Is Mediated by Glutathione through the Formation of Metal-Sulfur Clusters.

Copper is an essential nutrient required for many biological processes involved in primary metabolism, but free copper is toxic due to its ability to catalyze formation of free radicals. To prevent toxic effects, in the cell copper is bound to proteins and low molecular weight compounds, such as glutathione, at all times. The widely used chemotherapy agent cisplatin is known to bind to copper-transporting proteins, including copper chaperone Atox1. Cisplatin interactions with Atox1 and other copper transporters are linked to cancer resistance to platinum-based chemotherapy. Here we analyze the binding of copper and cisplatin to Atox1 in the presence of glutathione under redox conditions that mimic intracellular environment. We show that copper(I) and glutathione form large polymers with a molecular mass of approximately 8 kDa, which can transfer copper to Atox1. Cisplatin also can form polymers with glutathione, albeit at a slower rate. Analysis of simultaneous binding of copper and cisplatin to Atox1 under physiological conditions shows that both metals are bound to the protein through copper-sulfur-platinum bridges.

Cost-effective enhanced iron bioavailability in rice grain grown on calcareous soil by sulfur mediation and its effect on heavy metals mineralization.

Calcareous soil, high pH, and low organic matter are the major factors that limit iron (Fe) availability to rice crop. The present study was planned with the aim to biofortified rice grain with Fe, by integrated use of chemical and organic amendments in pH-manipulated calcareous soil. The soil pH was reduced (pHL2) by using elemental sulfur (S) at the rate of 0.25 % (w/w). The organic amendments, biochar (BC) and poultry manure (PM) [1 % (w/w)], along with ferrous sulfate at the rate of 7.5 mg kg-1 soil were used. The incorporation of Fe with BC in soil at pHL2 significantly improved plant biomass, photosynthetic rate, and paddy yield up to 99, 97, and 36 %, respectively, compared to control. A significant increase in grain Fe (190 %), protein (58 %), and ferritin (400 %) contents was observed while anti-nutrients, i.e., polyphenols (37 %) and phytate (21 %) were significantly decreased by the addition of Fe and BC in soil at pHL2 relative to control. Among the organic amendments, PM significantly increased Cd, Pb, Ni, and Cr concentrations in rice grain relative to control but their concentration values were below as compared to the toxic limits of hazard quotients and hazard index (HQ and HI). Hence, this study implies that Fe applied with BC in the soil at pHL2 can be considered as an effective strategy to augment Fe bioavailability and to reduce non-essential heavy metal accumulation in rice grain.

Novel cost effective full scale mussel shell bioreactors for metal removal and acid neutralization.

Acid mine drainage (AMD) impacted waters are a worldwide concern for the mining industry and countries dealing with this issue; both active and passive technologies are employed for the treatment of such waters. Mussel shell bioreactors (MSB) represent a passive technology that utilizes waste from the shellfish industry as a novel substrate. The aim of this study is to provide insight into the biogeochemical dynamics of a novel full scale MSB for AMD treatment. A combination of water quality data, targeted geochemical extractions, and metagenomic analyses were used to evaluate MSB performance. The MSB raised the effluent pH from 3.4 to 8.3 while removing up to ∼99% of the dissolved Al, and Fe and >90% Ni, Tl, and Zn. A geochemical gradient was observed progressing from oxidized to reduced conditions with depth. The redox conditions helped define the microbial consortium that consists of a specialized niche of organisms that influence elemental cycling (i.e. complex Fe and S cycling). MSB technology represents an economic and effective means of full scale, passive AMD treatment that is an attractive alternative for developing economies due to its low cost and ease of implementation.

Pyrosequencing-Based Assessment of the Microbial Community Structure of Pastoruri Glacier Area (Huascarán National Park, Perú), a Natural Extreme Acidic Environment.

The exposure of fresh sulfide-rich lithologies by the retracement of the Nevado Pastoruri glacier (Central Andes, Perú) is increasing the presence of heavy metals in the water as well as decreasing the pH, producing an acid rock drainage (ARD) process in the area. We describe the microbial communities of an extreme ARD site in Huascarán National Park as well as their correlation with the water physicochemistry. Microbial biodiversity was analyzed by FLX 454 sequencing of the 16S rRNA gene. The suggested geomicrobiological model of the area distinguishes three different zones. The proglacial zone is located in the upper part of the valley, where the ARD process is not evident yet. Most of the OTUs detected in this area were related to sequences associated with cold environments (i.e., psychrotolerant species of Cyanobacteria or Bacteroidetes). After the proglacial area, an ARD-influenced zone appeared, characterized by the presence of phylotypes related to acidophiles (Acidiphilium) as well as other species related to acidic and cold environments (i.e., acidophilic species of Chloroflexi, Clostridium and Verrumicrobia). Sulfur- and iron-oxidizing acidophilic bacteria (Acidithiobacillus) were also identified. The post-ARD area was characterized by the presence of OTUs related to microorganisms detected in soils, permafrost, high mountain environments, and deglaciation areas (Sphingomonadales, Caulobacter or Comamonadaceae).

Jerusalem artichoke as low-cost fructose-rich feedstock for fossil fuels desulphurization by a fructophilic bacterium.

AIMS: Through biodesulphurization (BDS) is possible to remove the sulphur present in fossil fuels to carry out the very strict legislation. However, this biological process is limited by the cost of the culture medium, and thus, it is important to explore cheaper alternative carbon sources, such as Jerusalem artichoke (JA). These carbon sources usually contain sulphates which interfere with the BDS process. The goal of this work was to remove the sulphates from Jerusalem artichoke juice (JAJ) through BaCl2 precipitation viewing the optimization of dibenzothiophene (DBT) desulphurization by Gordonia alkanivorans strain 1B. METHODS AND RESULTS: Using a statistical design (Doehlert distribution), the effect of BaCl2 concentration (0.125-0.625%) and pH (5-9) was studied on sulphate concentration in hydrolysed JAJ. A validated surface response derived from data indicated that zero sulphates can be achieved with 0.5-0.55% (w/v) BaCl2 at pH 7; however, parallel BDS assays showed that the highest desulphurization was obtained with the juice treated with 0.5% (w/v) BaCl2 at pH 8.73. Further assays demonstrated that enhanced DBT desulphurization was achieved using hydrolysed JAJ treated in these optimal conditions. A total conversion of 400 µmol l(-1) DBT into 2-hydroxybiphenyl (2-HBP) in <90 h was observed, attaining a 2-HBP maximum production rate of 28.2 µmol l(-1) h(-1) and a specific production rate of 5.06 µmol(-1) g(-1) (DCW) h(-1) . CONCLUSIONS: These results highlight the efficacy of the treatment applied to JAJ in making this agromaterial a promising low-cost renewable feedstock for improved BDS by the fructophilic strain 1B. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is a fundamental step viewing BDS application at the industrial level as it accounts a cost-effective production of the biocatalysts, one of the main drawbacks for BDS scale-up.

Assessment of silver nanoparticle-induced physiological and molecular changes in Arabidopsis thaliana.

In this study, the effect of silver nanoparticles and silver ions on Arabidopsis thaliana was investigated at physiological and molecular levels. The seedlings were grown in sublethal concentrations of silver nanoparticles and silver ions (0.2, 0.5, and 1 mg/L) in 1/4 Hoagland's medium for 14 days under submerged hydroponic conditions. Significantly higher reduction in the total chlorophyll and increase in anthocyanin content were observed after exposure to 0.5 and 1 mg/L silver nanoparticles as compared to similar concentrations of silver ions. Lipid peroxidation increased significantly after exposure to 0.2, 0.5, and 1 mg/L of silver nanoparticles and 0.5 and 1 mg/L of silver ions. Qualitative analysis with dichloro-dihydro-fluorescein diacetate and rhodamine 123 fluorescence showed a dose-dependent increase in reactive oxygen species production and changes in mitochondrial membrane potential in the roots of seedlings exposed to different concentrations of silver nanoparticles. Real-time PCR analysis showed significant upregulation in the expression of sulfur assimilation, glutathione biosynthesis, glutathione S-transferase, and glutathione reductase genes upon exposure to silver nanoparticles as compared with silver ions. Overall, based on the physiological and molecular level responses, it was observed that exposure to silver nanoparticles exerted more toxic response than silver ions in A. thaliana.

A combined proteomic and transcriptomic analysis on sulfur metabolism pathways of Arabidopsis thaliana under simulated acid rain.

With rapid economic development, most regions in southern China have suffered acid rain (AR) pollution. In our study, we analyzed the changes in sulfur metabolism in Arabidopsis under simulated AR stress which provide one of the first case studies, in which the systematic responses in sulfur metabolism were characterized by high-throughput methods at different levels including proteomic, genomic and physiological approaches. Generally, we found that all of the processes related to sulfur metabolism responded to AR stress, including sulfur uptake, activation and also synthesis of sulfur-containing amino acid and other secondary metabolites. Finally, we provided a catalogue of the detected sulfur metabolic changes and reconstructed the coordinating network of their mutual influences. This study can help us to understand the mechanisms of plants to adapt to AR stress.

NAA-SLR and ICP-AES application in the assessment of mass fraction of 19 chemical elements in pediatric and young adult prostate glands.

The effect of age on the mass fraction of 19 chemical elements in the intact prostate of 50 apparently healthy 0-30-year-old males was investigated by neutron activation analysis with high-resolution spectrometry of short-lived radionuclides and inductively coupled plasma atomic emission spectrometry. Mean values (M ± standard error of the mean) for mass fraction (in milligrams per kilogram, on dry weight basis) of chemical elements were as follows: Al, 77 ± 17; B, 1.31 ± 0.29; Ba, 4.0 ± 1.2; Br, 37.7 ± 4.3; Ca, 1,536 ± 189; Cl, 13,414 ± 949; Cu, 12.3 ± 2.1; Fe, 132 ± 11; K, 11,547 ± 468; Li, 0.064 ± 0.009; Mg, 922 ± 89; Mn, 1.88 ± 0.16; Na, 9,834 ± 411; P, 6,741 ± 335; S, 8,034 ± 251; Si, 199 ± 34; Sr, 1.40 ± 0.19; and Zn, 277 ± 33. The upper limit of mean mass fraction of V was ≤0.24. This work revealed that there is significant tendency for the mass fractions of Ca, K, Mg, and Zn in the prostate tissue of healthy individuals to increase with age from the time of birth up to 30 years. It means that Ca, K, Mg, and Zn mass fractions in prostate tissue are the androgen-dependent parameters. Our finding of a positive correlation between the prostatic Zn and Ca, K, Mg, P, and S mass fractions indicates that there is a special relationship of Zn with some main electrolytes (Ca, K, and Mg) and with P- and S-containing compounds in the prostate. It was shown also that high levels of Al, B, Ba, Br, Cl, Li, Na, and Sr mass fraction in prostate tissue do not indicate a direct involvement of these elements in the reproductive function of the prostate.

Scalable economic extracellular synthesis of CdS nanostructured particles by a non-pathogenic thermophile.

We report microbially facilitated synthesis of cadmium sulfide (CdS) nanostructured particles (NP) using anaerobic, metal-reducing Thermoanaerobacter sp. The extracellular CdS crystallites were <10 nm in size with yields of ~3 g/L of growth medium/month with demonstrated reproducibility and scalability up to 24 L. During synthesis, Thermoanaerobacter cultures reduced thiosulfate and sulfite salts to H2S, which reacted with Cd²âº cations to produce thermodynamically favored NP in a single step at 65 °C with catalytic nucleation on the cell surfaces. Photoluminescence (PL) analysis of dry CdS NP revealed an exciton-dominated PL peak at 440 nm, having a narrow full width at half maximum of 10 nm. A PL spectrum of CdS NP produced by dissimilatory sulfur reducing bacteria was dominated by features associated with radiative exciton relaxation at the surface. High reproducibility of CdS NP PL features important for scale-up conditions was confirmed from test tubes to 24 L batches at a small fraction of the manufacturing cost associated with conventional inorganic NP production processes.

Elemental economy: microbial strategies for optimizing growth in the face of nutrient limitation.

Microorganisms play a dominant role in the biogeochemical cycling of nutrients. They are rightly praised for their facility for fixing both carbon and nitrogen into organic matter, and microbial driven processes have tangibly altered the chemical composition of the biosphere and its surrounding atmosphere. Despite their prodigious capacity for molecular transformations, microorganisms are powerless in the face of the immutability of the elements. Limitations for specific elements, either fleeting or persisting over eons, have left an indelible trace on microbial genomes, physiology, and their very atomic composition. We here review the impact of elemental limitation on microbes, with a focus on selected genetic model systems and representative microbes from the ocean ecosystem. Evolutionary adaptations that enhance growth in the face of persistent or recurrent elemental limitations are evident from genome and proteome analyses. These range from the extreme (such as dispensing with a requirement for a hard to obtain element) to the extremely subtle (changes in protein amino acid sequences that slightly, but significantly, reduce cellular carbon, nitrogen, or sulfur demand). One near-universal adaptation is the development of sophisticated acclimation programs by which cells adjust their chemical composition in response to a changing environment. When specific elements become limiting, acclimation typically begins with an increased commitment to acquisition and a concomitant mobilization of stored resources. If elemental limitation persists, the cell implements austerity measures including elemental sparing and elemental recycling. Insights into these fundamental cellular properties have emerged from studies at many different levels, including ecology, biological oceanography, biogeochemistry, molecular genetics, genomics, and microbial physiology. Here, we present a synthesis of these diverse studies and attempt to discern some overarching themes.

Sulfur transformations in pilot-scale constructed wetland treating high sulfate-containing contaminated groundwater: a stable isotope assessment.

Current understanding of the dynamics of sulfur compounds inside constructed wetlands is still insufficient to allow a full description of processes involved in sulfur cycling. Experiments in a pilot-scale horizontal subsurface flow constructed wetland treating high sulfate-containing contaminated groundwater were carried out. Application of stable isotope approach combined with hydro-chemical investigations was performed to evaluate the sulfur transformations. In general, under inflow concentration of about 283 mg/L sulfate sulfur, sulfate removal was found to be about 21% with a specific removal rate of 1.75 g/m(2)·d. The presence of sulfide and elemental sulfur in pore water about 17.3 mg/L and 8.5 mg/L, respectively, indicated simultaneously bacterial sulfate reduction and re-oxidation. 70% of the removed sulfate was calculated to be immobilized inside the wetland bed. The significant enrichment of (34)S and (18)O in dissolved sulfate (δ(34)S up to 16‰, compared to average of 5.9‰ in the inflow, and δ(18)O up to 13‰, compared to average of 6.9‰ in the inflow) was observed clearly correlated to the decrease of sulfate loads along the flow path through experimental wetland bed. This enrichment also demonstrated the occurrence of bacterial sulfate reduction as well as demonstrated by the presence of sulfide in the pore water. Moreover, the integral approach shows that bacterial sulfate reduction is not the sole process controlling the isotopic composition of dissolved sulfate in the pore water. The calculated apparent enrichment factor (ɛ = -22‰) for sulfur isotopes from the δ(34)S vs. sulfate mass loss was significantly smaller than required to produce the observed difference in δ(34)S between sulfate and sulfide. It indicated some potential processes superimposing bacterial sulfate reduction, such as direct re-oxidation of sulfide to sulfate by oxygen released from plant roots and/or bacterial disproportionation of elemental sulfur. Furthermore, 41% of residual sulfate was calculated to be from sulfide re-oxidation, which demonstrated that the application of stable isotope approach combined with the common hydro-chemical investigations is not only necessary for a general qualitative evaluation of sulfur transformations in constructed wetlands, but also leads to a quantitative description of intermediate processes.

Metagenomic assessment of a sulfur-oxidizing enrichment culture derived from marine sediment.

The biological oxidation of reduced sulfur compounds is a critically important process in global sulfur biogeochemistry. In this study, we enriched from marine sediments under denitrifying conditions, chemolithotrophic sulfur oxidizers that could oxidize a variety of reduced sulfur compounds: thiosulfate, tetrathionate, sulfide, and polysulfide. Two major phylotypes of 16S rRNA gene (>99% identity in each phylotype) were detected in this enrichment culture. In order to characterize sulfide oxidation, we sequenced and characterized one fosmid clone (43.6 kb) containing the group I sulfide-quinone reductase (sqr) gene. Interestingly, four putative rhodanese genes were found in this clone. Furthermore, comparative alignment with the closest genome of Thiomicrospira crunogena XCL2 revealed that three homologous genes were located within the vicinity of the sqr gene. Fosmid clones harboring carbon fixation (cbbL and cbbM) and denitrification (narG) genes were screened, and the phylogeny of the functional genes was analyzed. Along with the comparison between the sqr-containing fosmid clones and the relevant -proteobacteria, our phylogenetic study based on the 16S rRNA gene and carbon fixation genes suggest the prevalence of chemolithotrophic -proteobacteria in the denitrifying cultures. The findings of this study imply that a combination of cultivation and metagenomic approaches might provide us with a glimpse into the characteristics of sulfur oxidizers in marine sediments.

[pH regulation to continuous treatment system of high-sulfur antibiotic wastewater].

With the pH regulation to the batch-treatment system as a control, the effect of different pH levels, i.e., 7.0, 5.1, 4.1, on the treatment efficiencies and the contents of yeast and bacteria content in aerated column were investigated for the continuous micro-aerobic treatment system of the antibiotic wastewater. With the decrease in pH levels in aerated column, the COD removals reached 53%, 42% and 45%, respectively, BOD5 removals were 79%, 58% and 50%, respectively, and the DHA activities (TF/MLSS x t) of the sludge were 47,088, 10,506 and 14,390 mg/(g h), respectively, during the three experimental stages. The results on the basis of scanning electron microscope (SEM) and fluorescent in-situ hybridization (FISH)-flow cytometry (FCM) showed that different pH levels didn't affect the contents of yeast and bacteria in the aerated column content at three different pH levels, and bacteria were the dominant microbes with a content of above 98% throughout the experimental periods. In addition, the variation of pH levels had the same influence on the treatment efficiencies of both continuous treatment system and batch treatment system, although different changes of microbial ecologies happened in these two systems.

Biocatalytic desulfurization (BDS) of petrodiesel fuels.

Oil refineries are facing many challenges, including heavier crude oils, increased fuel quality standards, and a need to reduce air pollution emissions. Global society is stepping on the road to zero-sulfur fuel, with only differences in the starting point of sulfur level and rate reduction of sulfur content between different countries. Hydrodesulfurization (HDS) is the most common technology used by refineries to remove sulfur from intermediate streams. However, HDS has several disadvantages, in that it is energy intensive, costly to install and to operate, and does not work well on refractory organosulfur compounds. Recent research has therefore focused on improving HDS catalysts and processes and also on the development of alternative technologies. Among the new technologies one possible approach is biocatalytic desulfurization (BDS). The advantage of BDS is that it can be operated in conditions that require less energy and hydrogen. BDS operates at ambient temperature and pressure with high selectivity, resulting in decreased energy costs, low emission, and no generation of undesirable side products. Over the last two decades several research groups have attempted to isolate bacteria capable of efficient desulfurization of oil fractions. This review examines the developments in our knowledge of the application of bacteria in BDS processes, assesses the technical viability of this technology and examines its future challenges.