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1.
Gut Microbes ; 13(1): 1-20, 2021.
Article in English | MEDLINE | ID: mdl-34125646

ABSTRACT

The emerging evidence supports the use of prebiotics like herb-derived polysaccharides for treating nonalcoholic fatty liver disease (NAFLD) by modulating gut microbiome. The present study was initiated on the microbiota-dependent anti-NAFLD effect of Astragalus polysaccharides (APS) extracted from Astragalus mongholicus Bunge in high-fat diet (HFD)-fed mice. However, the exact mechanisms underlying the beneficial effects of APS on NAFLD formation remain poorly understood.Co-housing experiment was used to assess the microbiota dependent anti-NAFLD effect of APS. Then, targeted metabolomics and metagenomics were adopted for determining short-chain fatty acids (SCFAs) and bacteria that were specifically enriched by APS. Further in vitro experiment was carried out to test the capacity of SCFAs-producing of identified bacterium. Finally, the anti-NAFLD efficacy of identified bacterium was tested in HFD-fed mice.Our results first demonstrated the anti-NAFLD effect of APS in HFD-fed mice and the contribution of gut microbiota. Moreover, our results indicated that SCFAs, predominantly acetic acid were elevated in APS-supplemented mice and ex vivo experiment. Metagenomics revealed that D. vulgaris from Desulfovibrio genus was not only enriched by APS, but also a potent generator of acetic acid, which showed significant anti-NAFLD effects in HFD-fed mice. In addition, D. vulgaris modulated the hepatic gene expression pattern of lipids metabolism, particularly suppressed hepatic fatty acid synthase (FASN) and CD36 protein expression.Our results demonstrate that APS enriched D. vulgaris is effective on attenuating hepatic steatosis possibly through producing acetic acid, and modulation on hepatic lipids metabolism in mice. Further studies are warranted to explore the long-term impacts of D. vulgaris on host metabolism and the underlying mechanism.


Subject(s)
Acetic Acid/metabolism , Desulfovibrio vulgaris/metabolism , Non-alcoholic Fatty Liver Disease/drug therapy , Probiotics/administration & dosage , Animals , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Bacteria/metabolism , CD36 Antigens/genetics , CD36 Antigens/metabolism , Desulfovibrio vulgaris/growth & development , Diet, High-Fat/adverse effects , Fatty Acid Synthases/genetics , Fatty Acid Synthases/metabolism , Fatty Acids, Volatile/metabolism , Gastrointestinal Microbiome , Humans , Liver/enzymology , Liver/metabolism , Male , Mice , Mice, Inbred C57BL , Non-alcoholic Fatty Liver Disease/etiology , Non-alcoholic Fatty Liver Disease/genetics , Non-alcoholic Fatty Liver Disease/microbiology
2.
Appl Environ Microbiol ; 86(1)2019 12 13.
Article in English | MEDLINE | ID: mdl-31628147

ABSTRACT

Sulfate-reducing bacteria (SRB) are key contributors to microbe-induced corrosion (MIC), which can lead to serious economic and environmental impact. The presence of a biofilm significantly increases the MIC rate. Inhibition of the quorum-sensing (QS) system is a promising alternative approach to prevent biofilm formation in various industrial settings, especially considering the significant ecological impact of conventional chemical-based mitigation strategies. In this study, the effect of the QS stimulation and inhibition on Desulfovibrio vulgaris is described in terms of anaerobic respiration, cell activity, biofilm formation, and biocorrosion of carbon steel. All these traits were repressed when bacteria were in contact with QS inhibitors but enhanced upon exposure to QS signal molecules compared to the control. The difference in the treatments was confirmed by transcriptomic analysis performed at different time points after treatment application. Genes related to lactate and pyruvate metabolism, sulfate reduction, electron transfer, and biofilm formation were downregulated upon QS inhibition. In contrast, QS stimulation led to an upregulation of the above-mentioned genes compared to the control. In summary, these results reveal the impact of QS on the activity of D. vulgaris, paving the way toward the prevention of corrosive SRB biofilm formation via QS inhibition.IMPORTANCE Sulfate-reducing bacteria (SRB) are considered key contributors to biocorrosion, particularly in saline environments. Biocorrosion imposes tremendous economic costs, and common approaches to mitigate this problem involve the use of toxic and hazardous chemicals (e.g., chlorine), which raise health and environmental safety concerns. Quorum-sensing inhibitors (QSIs) can be used as an alternative approach to inhibit biofilm formation and biocorrosion. However, this approach would only be effective if SRB rely on QS for the pathways associated with biocorrosion. These pathways would include biofilm formation, electron transfer, and metabolism. This study demonstrates the role of QS in Desulfovibrio vulgaris on the above-mentioned pathways through both phenotypic measurements and transcriptomic approach. The results of this study suggest that QSIs can be used to mitigate SRB-induced corrosion problems in ecologically sensitive areas.


Subject(s)
Biofilms/drug effects , Desulfovibrio vulgaris/growth & development , Quorum Sensing/drug effects , Acyl-Butyrolactones/pharmacology , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Biofilms/growth & development , Carbon/metabolism , Corrosion , Desulfovibrio vulgaris/genetics , Desulfovibrio vulgaris/metabolism , Energy Metabolism/genetics , Gene Expression Regulation , Genes, Bacterial , Lactic Acid/metabolism , Plankton/microbiology , Pyruvic Acid/metabolism , Seawater/chemistry , Steel , Sulfates/metabolism , Transcription Factors/genetics , Transcriptome
3.
Environ Microbiol ; 21(1): 360-373, 2019 01.
Article in English | MEDLINE | ID: mdl-30394641

ABSTRACT

Desulfovibrio species are representatives of microorganisms at the boundary between anaerobic and aerobic lifestyles, since they contain the enzymatic systems required for both sulfate and oxygen reduction. However, the latter has been shown to be solely a protective mechanism. By implementing the oxygen-driven experimental evolution of Desulfovibrio vulgaris Hildenborough, we have obtained strains that have evolved to grow with energy derived from oxidative phosphorylation linked to oxygen reduction. We show that a few mutations are sufficient for the emergence of this phenotype and reveal two routes of evolution primarily involving either inactivation or overexpression of the gene encoding heterodisulfide reductase. We propose that the oxygen respiration for energy conservation that sustains the growth of the O2 -evolved strains is associated with a rearrangement of metabolite fluxes, especially NAD+ /NADH, leading to an optimized O2 reduction. These evolved strains are the first sulfate-reducing bacteria that exhibit a demonstrated oxygen respiratory process that enables growth.


Subject(s)
Desulfovibrio vulgaris/growth & development , Desulfovibrio vulgaris/metabolism , Energy Metabolism/physiology , Oxygen/metabolism , Sulfates/metabolism , Anaerobiosis , Desulfovibrio vulgaris/genetics , NAD/metabolism , Oxidation-Reduction , Oxidative Phosphorylation , Oxidoreductases/genetics , Oxidoreductases/metabolism
4.
Biotechnol Bioeng ; 115(6): 1465-1474, 2018 06.
Article in English | MEDLINE | ID: mdl-29476629

ABSTRACT

In situ bioreduction of soluble hexavalent uranium U(VI) to insoluble U(IV) (as UO2 ) has been proposed as a means of preventing U migration in the groundwater. This work focuses on the bioreduction of U(VI) and precipitation of U(IV). It uses anaerobic batch reactors with Desulfovibrio vulgaris, a well-known sulfate, iron, and U(VI) reducer, growing on lactate as the electron donor, in the absence of sulfate, and with a 30-mM bicarbonate buffering. In the absence of sulfate, D. vulgaris reduced >90% of the total soluble U(VI) (1 mM) to form U(IV) solids that were characterized by X-ray diffraction and confirmed to be nano-crystalline uraninite with crystallite size 2.8 ± 0.2 nm. pH values between 6 and 10 had minimal impact on bacterial growth and end-product distribution, supporting that the mono-nuclear, and poly-nuclear forms of U(VI) were equally bioavailable as electron acceptors. Electron balances support that H2 transiently accumulated, but was ultimately oxidized via U(VI) respiration. Thus, D. vulgaris utilized H2 as the electron carrier to drive respiration of U(VI). Rapid lactate utilization and biomass growth occurred only when U(VI) respiration began to draw down the sink of H2 and relieve thermodynamic inhibition of fermentation.


Subject(s)
Desulfovibrio vulgaris/growth & development , Desulfovibrio vulgaris/metabolism , Hydrogen/metabolism , Uranium/metabolism , Bioreactors/microbiology , Biotransformation , Culture Media/chemistry , Desulfovibrio vulgaris/drug effects , Hydrogen-Ion Concentration , Lactates/metabolism , Oxidation-Reduction
5.
Appl Microbiol Biotechnol ; 102(6): 2839-2850, 2018 Mar.
Article in English | MEDLINE | ID: mdl-29429007

ABSTRACT

Desulfovibrio spp. are capable of heavy metal reduction and are well-studied systems for understanding metal fate and transport in anaerobic environments. Desulfovibrio vulgaris Hildenborough was grown under environmentally relevant conditions (i.e., temperature, nutrient limitation) to elucidate the impacts on Cr(VI) reduction on cellular physiology. Growth at 20 °C was slower than 30 °C and the presence of 50 µM Cr(VI) caused extended lag times for all conditions, but once growth resumed the growth rate was similar to that without Cr(VI). Cr(VI) reduction rates were greatly diminished at 20 °C for both 50 and 100 µM Cr(VI), particularly for the electron acceptor limited (EAL) condition in which Cr(VI) reduction was much slower, the growth lag much longer (200 h), and viability decreased compared to balanced (BAL) and electron donor limited (EDL) conditions. When sulfate levels were increased in the presence of Cr(VI), cellular responses improved via a shorter lag time to growth. Similar results were observed between the different resource (donor/acceptor) ratio conditions when the sulfate levels were normalized (10 mM), and these results indicated that resource ratio (donor/acceptor) impacted D. vulgaris response to Cr(VI) and not merely sulfate limitation. The results suggest that temperature and resource ratios greatly impacted the extent of Cr(VI) toxicity, Cr(VI) reduction, and the subsequent cellular health via Cr(VI) influx and overall metabolic rate. The results also emphasized the need to perform experiments at lower temperatures with nutrient limitation to make accurate predictions of heavy metal reduction rates as well as physiological states in the environment.


Subject(s)
Carcinogens, Environmental/metabolism , Carcinogens, Environmental/toxicity , Chromium/metabolism , Chromium/toxicity , Desulfovibrio vulgaris/drug effects , Desulfovibrio vulgaris/metabolism , Anaerobiosis , Desulfovibrio vulgaris/growth & development , Microbial Viability/drug effects , Oxidation-Reduction , Sulfates/metabolism , Temperature
6.
Water Res ; 129: 163-171, 2018 02 01.
Article in English | MEDLINE | ID: mdl-29149671

ABSTRACT

The intensive use of silver nanoparticles (AgNPs) in cosmetics and textiles causes their release into sewer networks of urban water systems. Although a few studies have investigated antimicrobial activities of nanoparticles against environmental bacteria, little is known about potential impacts of the released AgNPs on sulfate reducing bacteria in sewers. Here, we investigated the effect of AgNPs on Desulfovibrio vulgaris Hidenborough (D. vulgaris), a typical sulfate-reducing bacterium (SRB) in sewer systems. We found AgNPs stimulated the proliferation of D. vulgaris, rather than exerting inhibitory or biocidal effects. Based on flow cytometer detections, both the cell growth rate and the viable cell ratio of D. vulgaris increased during exposure to AgNPs at concentrations of up to 100 mg/L. The growth stimulation was dependent on the AgNP concentration. These results imply that the presence of AgNPs in sewage may affect SRB abundance in sewer networks. Our findings also shed new lights on the interactions of nanoparticles and bacteria.


Subject(s)
Desulfovibrio vulgaris/drug effects , Metal Nanoparticles/toxicity , Silver/pharmacology , Bacteria/drug effects , Cell Proliferation/drug effects , Desulfovibrio vulgaris/growth & development , Desulfovibrio vulgaris/metabolism , Lactates/metabolism , Sewage/microbiology , Sulfates/metabolism
7.
Environ Microbiol ; 19(8): 3059-3069, 2017 08.
Article in English | MEDLINE | ID: mdl-28419704

ABSTRACT

Microbial populations can withstand, overcome and persist in the face of environmental fluctuation. Previously, we demonstrated how conditional gene regulation in a fluctuating environment drives dilution of condition-specific transcripts, causing a population of Desulfovibrio vulgaris Hildenborough (DvH) to collapse after repeatedly transitioning from sulfate respiration to syntrophic conditions with the methanogen Methanococcus maripaludis. Failure of the DvH to successfully transition contributed to the collapse of this model community. We investigated the mechanistic basis for loss of robustness by examining whether conditional gene regulation altered heterogeneity in gene expression across individual DvH cells. We discovered that robustness of a microbial population across environmental transitions was attributable to the retention of cells in two states that exhibited different condition-specific gene expression patterns. In our experiments, a population with disrupted conditional regulation successfully alternated between cell states. Meanwhile, a population with intact conditional regulation successfully switched between cell states initially, but collapsed after repeated transitions, possibly due to the high energy requirements of regulation. These results demonstrate that the survival of this entire model microbial community is dependent on the regulatory system's influence on the distribution of distinct cell states among individual cells within a clonal population.


Subject(s)
Desulfovibrio vulgaris/growth & development , Methanococcus/growth & development , Microbial Consortia/physiology , Microbial Interactions/physiology , Desulfovibrio vulgaris/genetics , Energy Metabolism/physiology , Oxidation-Reduction , Sulfates/metabolism
8.
Mol Syst Biol ; 13(3): 919, 2017 03 20.
Article in English | MEDLINE | ID: mdl-28320772

ABSTRACT

Managing trade-offs through gene regulation is believed to confer resilience to a microbial community in a fluctuating resource environment. To investigate this hypothesis, we imposed a fluctuating environment that required the sulfate-reducer Desulfovibrio vulgaris to undergo repeated ecologically relevant shifts between retaining metabolic independence (active capacity for sulfate respiration) and becoming metabolically specialized to a mutualistic association with the hydrogen-consuming Methanococcus maripaludis Strikingly, the microbial community became progressively less proficient at restoring the environmentally relevant physiological state after each perturbation and most cultures collapsed within 3-7 shifts. Counterintuitively, the collapse phenomenon was prevented by a single regulatory mutation. We have characterized the mechanism for collapse by conducting RNA-seq analysis, proteomics, microcalorimetry, and single-cell transcriptome analysis. We demonstrate that the collapse was caused by conditional gene regulation, which drove precipitous decline in intracellular abundance of essential transcripts and proteins, imposing greater energetic burden of regulation to restore function in a fluctuating environment.


Subject(s)
Desulfovibrio vulgaris/growth & development , Methanococcus/growth & development , Systems Biology/methods , Desulfovibrio vulgaris/genetics , Directed Molecular Evolution , Gene Expression Profiling , Methanococcus/genetics , Oxidation-Reduction , Phenotype , Proteomics , Sequence Analysis, RNA , Single-Cell Analysis , Sulfates/metabolism
9.
Chemosphere ; 168: 529-538, 2017 Feb.
Article in English | MEDLINE | ID: mdl-27852449

ABSTRACT

The iron-reducing bacterium Acidiphilium cryputum JF-5 and a sulfate reducing bacterium (SRB) collected and purified from the mine drainage of a copper mine in the northwest of Sichuan Province, China, were used to biologically synthesize nano-sized FeS-coated limestone to remove As(V) from solution. The adsorption efficiency of As(V) is improved from 6.64 µg/g with limestone alone to 187 µg/g with the FeS coated limestone in both batch and column experiments. The hydraulic conductivity of the columns are also improved by the presence of the nano-sized FeS coatings, but the solution neutralization performance of the limestone can be reduced by passivation by gypsum and Fe(III) precipitates. Calculations for FeS-coated limestone dissolution experiments show that the process can be described as nCa.sol = At1/2 - nCa,gyp. The results suggest that FeS-coated limestone may be an effective medium for remediating As(V)-bearing solutions such as acid mine drainage in systems such as Permeable Reactive Barriers.


Subject(s)
Arsenicals/isolation & purification , Calcium Carbonate/chemistry , Ferrous Compounds/chemistry , Mining , Nanoparticles/chemistry , Water Pollutants, Chemical/isolation & purification , Water Purification/methods , Acidiphilium/growth & development , Adsorption , Biodegradation, Environmental , China , Desulfovibrio vulgaris/growth & development , Groundwater/chemistry , Groundwater/microbiology , Oxidation-Reduction
10.
Appl Environ Microbiol ; 82(18): 5563-75, 2016 09 15.
Article in English | MEDLINE | ID: mdl-27371588

ABSTRACT

Hydrogen sulfide produced by sulfate-reducing bacteria (SRB) in sewers causes odor problems and asset deterioration due to the sulfide-induced concrete corrosion. Free nitrous acid (FNA) was recently demonstrated as a promising antimicrobial agent to alleviate hydrogen sulfide production in sewers. However, details of the antimicrobial mechanisms of FNA are largely unknown. Here, we report the multiple-targeted antimicrobial effects of FNA on the SRB Desulfovibrio vulgaris Hildenborough by determining the growth, physiological, and gene expression responses to FNA exposure. The activities of growth, respiration, and ATP generation were inhibited when exposed to FNA. These changes were reflected in the transcript levels detected during exposure. The removal of FNA was evident by nitrite reduction that likely involved nitrite reductase and the poorly characterized hybrid cluster protein, and the genes coding for these proteins were highly expressed. During FNA exposure, lowered ribosome activity and protein production were detected. Additionally, conditions within the cells were more oxidizing, and there was evidence of oxidative stress. Based on an interpretation of the measured responses, we present a model depicting the antimicrobial effects of FNA on D. vulgaris These findings provide new insight for understanding the responses of D. vulgaris to FNA and will provide a foundation for optimal application of this antimicrobial agent for improved control of sewer corrosion and odor management.IMPORTANCE Hydrogen sulfide produced by SRB in sewers causes odor problems and results in serious deterioration of sewer assets that requires very costly and demanding rehabilitation. Currently, there is successful application of the antimicrobial agent free nitrous acid (FNA), the protonated form of nitrite, for the control of sulfide levels in sewers (G. Jiang et al., Water Res 47:4331-4339, 2013, http://dx.doi.org/10.1016/j.watres.2013.05.024). However, the details of the antimicrobial mechanisms of FNA are largely unknown. In this study, we identified the key responses (decreased anaerobic respiration, reducing FNA, combating oxidative stress, and shutting down protein synthesis) of Desulfovibrio vulgaris Hildenborough, a model sewer corrosion bacterium, to FNA exposure by examining the growth, physiological, and gene expression changes. These findings provide new insight and underpinning knowledge for understanding the responses of D. vulgaris to FNA exposure, thereby benefiting the practical application of FNA for improved control of sewer corrosion and odor.


Subject(s)
Anti-Infective Agents/pharmacology , Desulfovibrio vulgaris/drug effects , Nitrous Acid/pharmacology , Adenosine Triphosphate/metabolism , Desulfovibrio vulgaris/genetics , Desulfovibrio vulgaris/growth & development , Desulfovibrio vulgaris/metabolism , Electron Transport/drug effects , Energy Metabolism/drug effects , Gene Expression Profiling , Protein Biosynthesis/drug effects , Ribosomes/metabolism , Transcription, Genetic
11.
Environ Sci Technol ; 49(24): 14655-62, 2015 Dec 15.
Article in English | MEDLINE | ID: mdl-26579558

ABSTRACT

Formate is recognized as a superior substrate for biological H2 production by several bacteria. However, the growth of a single organism coupled to this energetic pathway has not been shown in mesophilic conditions. In the present study, a bioreactor with gas sparging was used, where we observed for the first time that H2 production from formate can be coupled with growth of the model sulfate-reducing bacterium Desulfovibrio vulgaris in the absence of sulfate or a syntrophic partner. In these conditions, D. vulgaris had a maximum growth rate of 0.078 h(-1) and a doubling time of 9 h, and the ΔG of the reaction ranged between -21 and -18 kJ mol(-1). This is the first report of a single mesophilic organism that can grow while catalyzing the oxidation of formate to H2 and bicarbonate. Furthermore, high volumetric and specific H2 production rates (125 mL L(-1) h(-1) and 2500 mL gdcw(-1) h(-1)) were achieved in a new bioreactor designed and optimized for H2 production. This high H2 production demonstrates that the nonconventional H2-producing organism D. vulgaris is a good biocatalyst for converting formate to H2.


Subject(s)
Desulfovibrio vulgaris/growth & development , Desulfovibrio vulgaris/metabolism , Formates/metabolism , Hydrogen/metabolism , Bioreactors , Oxidation-Reduction , Sulfates/metabolism
12.
PLoS One ; 10(4): e0123455, 2015.
Article in English | MEDLINE | ID: mdl-25837676

ABSTRACT

Although obligate anaerobe, the sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough (DvH) exhibits high aerotolerance that involves several enzymatic systems, including two membrane-bound oxygen reductases, a bd-quinol oxidase and a cc(b/o)o3 cytochrome oxidase. Effect of constant low oxygen concentration on growth and morphology of the wild-type, single (Δbd, Δcox) and double deletion (Δcoxbd) mutant strains of the genes encoding these oxygen reductases was studied. When both wild-type and deletion mutant strains were cultured in lactate/sulfate medium under constant 0.02% O2 sparging, they were able to grow but the final biomasses and the growth yield were lower than that obtained under anaerobic conditions. At the end of the growth, lactate was not completely consumed and when conditions were then switched to anaerobic, growth resumed. Time-lapse microscopy revealed that a large majority of the cells were then able to divide (over 97%) but the time to recover a complete division event was longer for single deletion mutant Δbd than for the three other strains. Determination of the molar growth yields on lactate suggested that a part of the energy gained from lactate oxidation was derived toward cells protection/repairing against oxidative conditions rather than biosynthesis, and that this part was higher in the single deletion mutant Δbd and, to a lesser extent, Δcox strains. Our data show that when DvH encounters oxidative conditions, it is able to stop growing and to rapidly resume growing when conditions are switched to anaerobic, suggesting that it enters active dormancy sate under oxidative conditions. We propose that the pyruvate-ferredoxin oxidoreductase (PFOR) plays a central role in this phenomenon by reversibly switching from an oxidative-sensitive fully active state to an oxidative-insensitive inactive state. The oxygen reductases, and especially the bd-quinol oxidase, would have a crucial function by maintaining reducing conditions that permit PFOR to stay in its active state.


Subject(s)
Desulfovibrio vulgaris/growth & development , Desulfovibrio vulgaris/metabolism , Electron Transport Complex IV/metabolism , Oxidoreductases/metabolism , Oxygen/metabolism , Anaerobiosis , Biomass , Cell Proliferation/physiology , Desulfovibrio vulgaris/genetics , Lactic Acid/metabolism , Oxidation-Reduction , Pyruvate Synthase/metabolism
13.
Appl Environ Microbiol ; 81(8): 2676-89, 2015 Apr.
Article in English | MEDLINE | ID: mdl-25662968

ABSTRACT

Dissimilatory sulfate reduction is a microbial catabolic pathway that preferentially processes less massive sulfur isotopes relative to their heavier counterparts. This sulfur isotope fractionation is recorded in ancient sedimentary rocks and generally is considered to reflect a phenotypic response to environmental variations rather than to evolutionary adaptation. Modern sulfate-reducing microorganisms isolated from similar environments can exhibit a wide range of sulfur isotope fractionations, suggesting that adaptive processes influence the sulfur isotope phenotype. To date, the relationship between evolutionary adaptation and isotopic phenotypes has not been explored. We addressed this by studying the covariation of fitness, sulfur isotope fractionation, and growth characteristics in Desulfovibrio vulgaris Hildenborough in a microbial evolution experiment. After 560 generations, the mean fitness of the evolved lineages relative to the starting isogenic population had increased by ∼ 17%. After 927 generations, the mean fitness relative to the initial ancestral population had increased by ∼ 20%. Growth rate in exponential phase increased during the course of the experiment, suggesting that this was a primary influence behind the fitness increases. Consistent changes were observed within different selection intervals between fractionation and fitness. Fitness changes were associated with changes in exponential growth rate but changes in fractionation were not. Instead, they appeared to be a response to changes in the parameters that govern growth rate: yield and cell-specific sulfate respiration rate. We hypothesize that cell-specific sulfate respiration rate, in particular, provides a bridge that allows physiological controls on fractionation to cross over to the adaptive realm.


Subject(s)
Desulfovibrio vulgaris/physiology , Genetic Fitness , Sulfates/metabolism , Biological Evolution , Desulfovibrio vulgaris/genetics , Desulfovibrio vulgaris/growth & development , Oxidation-Reduction , Sulfur Isotopes/metabolism
14.
Anal Chem ; 87(2): 884-91, 2015 Jan 20.
Article in English | MEDLINE | ID: mdl-25496351

ABSTRACT

An autonomous metabolomic workflow combining mass spectrometry analysis with tandem mass spectrometry data acquisition was designed to allow for simultaneous data processing and metabolite characterization. Although previously tandem mass spectrometry data have been generated on the fly, the experiments described herein combine this technology with the bioinformatic resources of XCMS and METLIN. As a result of this unique integration, we can analyze large profiling datasets and simultaneously obtain structural identifications. Validation of the workflow on bacterial samples allowed the profiling on the order of a thousand metabolite features with simultaneous tandem mass spectra data acquisition. The tandem mass spectrometry data acquisition enabled automatic search and matching against the METLIN tandem mass spectrometry database, shortening the current workflow from days to hours. Overall, the autonomous approach to untargeted metabolomics provides an efficient means of metabolomic profiling, and will ultimately allow the more rapid integration of comparative analyses, metabolite identification, and data analysis at a systems biology level.


Subject(s)
Computational Biology , Desulfovibrio vulgaris/metabolism , Electronic Data Processing/methods , Metabolomics/methods , Chromatography, Liquid/methods , Databases, Factual , Desulfovibrio vulgaris/growth & development , Software , Tandem Mass Spectrometry/methods
15.
Sci Rep ; 4: 7478, 2014 Dec 15.
Article in English | MEDLINE | ID: mdl-25504148

ABSTRACT

Microbial syntrophic metabolism has been well accepted as the heart of how methanogenic and other anaerobic microbial communities function. In this work, we applied a single-cell RT-qPCR approach to reveal gene-expression heterogeneity in a model syntrophic system of Desulfovibrio vulgaris and Methanosarcina barkeri, as compared with the D. vulgaris monoculture. Using the optimized primers and single-cell analytical protocol, we quantitatively determine gene-expression levels of 6 selected target genes in each of the 120 single cells of D. vulgaris isolated from its monoculture and dual-culture with M. barkeri. The results demonstrated very significant cell-to-cell gene-expression heterogeneity for the selected D. vulgaris genes in both the monoculture and the syntrophic dual-culture. Interestingly, no obvious increase in gene-expression heterogeneity for the selected genes was observed for the syntrophic dual-culture when compared with its monoculture, although the community structure and cell-cell interactions have become more complicated in the syntrophic dual-culture. In addition, the single-cell RT-qPCR analysis also provided further evidence that the gene cluster (DVU0148-DVU0150) may be involved syntrophic metabolism between D. vulgaris and M. barkeri. Finally, the study validated that single-cell RT-qPCR analysis could be a valuable tool in deciphering gene functions and metabolism in mixed-cultured microbial communities.


Subject(s)
Bacterial Proteins/genetics , Biomarkers/metabolism , Desulfovibrio vulgaris/genetics , Gene Expression Profiling , Methanosarcina barkeri/genetics , Single-Cell Analysis/methods , Bacterial Proteins/metabolism , Cells, Cultured , Coculture Techniques , Desulfovibrio vulgaris/growth & development , Desulfovibrio vulgaris/metabolism , Gene Expression Regulation, Bacterial , Methanosarcina barkeri/growth & development , Methanosarcina barkeri/metabolism , Oligonucleotide Array Sequence Analysis , RNA, Messenger/genetics , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction
16.
Water Res ; 64: 255-264, 2014 Nov 01.
Article in English | MEDLINE | ID: mdl-25073000

ABSTRACT

We evaluated a hydrogen-based membrane biofilm reactor (MBfR) for its capacity to reduce and remove hexavalent uranium [U(VI)] from water. After a startup period that allowed slow-growing U(VI) reducers to form biofilms, the MBfR successfully achieved and maintained 94-95% U(VI) removal over 8 months when the U surface loading was 6-11 e(-) mEq/m(2)-day. The MBfR biofilm was capable of self-recovery after a disturbance due to oxygen exposure. Nanocrystalline UO2 aggregates and amorphous U precipitates were associated with vegetative cells and apparently mature spores that accumulated in the biofilm matrix. Despite inoculation with a concentrated suspension of Desulfovibrio vulgaris, this bacterium was not present in the U(VI)-reducing biofilm. Instead, the most abundant group in the biofilm community contained U(VI) reducers in the Rhodocyclaceae family when U(VI) was the only electron acceptor. When sulfate was present, the community dramatically shifted to the Clostridiaceae family, which included spores that were potentially involved in U(VI) reduction.


Subject(s)
Bacteria/isolation & purification , Biofilms , Uranium/isolation & purification , Water Pollutants, Radioactive/isolation & purification , Water Purification/methods , Bacteria/growth & development , Bacteria/metabolism , Clostridium/growth & development , Clostridium/isolation & purification , Clostridium/metabolism , Desulfovibrio vulgaris/growth & development , Hydrogen/chemistry , Membranes, Artificial , RNA, Ribosomal, 16S/analysis , Rhodocyclaceae/growth & development , Rhodocyclaceae/isolation & purification , Rhodocyclaceae/metabolism , Sulfates/metabolism , Uranium/metabolism
17.
Environ Sci Technol ; 48(12): 6928-37, 2014 Jun 17.
Article in English | MEDLINE | ID: mdl-24871825

ABSTRACT

The capacity of Desulfovibrio vulgaris to reduce U(VI) was studied previously with nongrowth conditions involving a high biomass concentration; thus, bacterial growth through respiration of U(VI) was not proven. In this study, we conducted a series of batch tests on U(VI) reduction by D. vulgaris at a low initial biomass (10 to 20 mg/L of protein) that could reveal biomass growth. D. vulgaris grew with U(VI) respiration alone, as well as with simultaneous sulfate reduction. Patterns of growth kinetics and solids production were affected by sulfate and Fe(2+). Biogenic sulfide nonenzymatically reduced 76% of the U(VI) and greatly enhanced the overall reduction rate in the absence of Fe(2+) but was rapidly scavenged by Fe(2+) to form FeS in the presence of Fe(2+). Biogenic U solids were uraninite (UO2) nanocrystallites associated with 20 mg/g biomass as protein. The crystallite thickness of UO2 was 4 to 5 nm without Fe(2+) but was <1.4 nm in the presence of Fe(2+), indicating poor crystallization inhibited by adsorbed Fe(2+) and other amorphous Fe solids, such as FeS or FeCO3. This work fills critical gaps in understanding the metabolic utilization of U by microorganisms and formation of UO2 solids in bioremediation sites.


Subject(s)
Desulfovibrio vulgaris/growth & development , Uranium/isolation & purification , Adsorption , Aerobiosis , Bacterial Proteins/analysis , Biodegradation, Environmental , Biomass , Carbonates/metabolism , Crystallization , Desulfovibrio vulgaris/metabolism , Ferric Compounds/metabolism , Hydrogen-Ion Concentration , Iron/metabolism , Kinetics , Lactates/analysis , Microscopy, Electron, Transmission , Oxidation-Reduction , Particle Size , Photoelectron Spectroscopy , Sulfates/metabolism , Sulfides/metabolism , Uranium Compounds/chemistry , X-Ray Absorption Spectroscopy , X-Ray Diffraction
18.
Biochim Biophys Acta ; 1844(7): 1238-47, 2014 Jul.
Article in English | MEDLINE | ID: mdl-24713144

ABSTRACT

An alternative route for haem b biosynthesis is operative in sulfate-reducing bacteria of the Desulfovibrio genus and in methanogenic Archaea. This pathway diverges from the canonical one at the level of uroporphyrinogen III and progresses via a distinct branch, where sirohaem acts as an intermediate precursor being converted into haem b by a set of novel enzymes, named the alternative haem biosynthetic proteins (Ahb). In this work, we report the biochemical characterisation of the Desulfovibrio vulgaris AhbD enzyme that catalyses the last step of the pathway. Mass spectrometry analysis showed that AhbD promotes the cleavage of S-adenosylmethionine (SAM) and converts iron-coproporphyrin III via two oxidative decarboxylations to yield haem b, methionine and the 5'-deoxyadenosyl radical. Electron paramagnetic resonance spectroscopy studies demonstrated that AhbD contains two [4Fe-4S](2+/1+) centres and that binding of the substrates S-adenosylmethionine and iron-coproporphyrin III induces conformational modifications in both centres. Amino acid sequence comparisons indicated that D. vulgaris AhbD belongs to the radical SAM protein superfamily, with a GGE-like motif and two cysteine-rich sequences typical for ligation of SAM molecules and iron-sulfur clusters, respectively. A structural model of D. vulgaris AhbD with putative binding pockets for the iron-sulfur centres and the substrates SAM and iron-coproporphyrin III is discussed.


Subject(s)
Bacterial Proteins/metabolism , Desulfovibrio vulgaris/enzymology , Heme/biosynthesis , Iron-Sulfur Proteins/metabolism , Amino Acid Sequence , Catalysis , Desulfovibrio vulgaris/growth & development , Electron Spin Resonance Spectroscopy , Molecular Sequence Data , Oxidation-Reduction , Protein Conformation , S-Adenosylmethionine/metabolism , Sequence Homology, Amino Acid , Spectrophotometry, Ultraviolet
19.
J Hazard Mater ; 272: 28-35, 2014 May 15.
Article in English | MEDLINE | ID: mdl-24675611

ABSTRACT

Sulfate-reducing bacteria (SRB) can produce iron sulfide (FeS) solids with mineralogical characteristics that may be beneficial for a variety of biogeochemical applications, such as long-term immobilization of uranium. In this study, the growth and metabolism of Desulfovibrio vulgaris, one of the best-studied SRB species, were comprehensively monitored in batch studies, and the biogenic FeS solids were characterized by X-ray diffraction. Controlling the pH by varying the initial pH, the iron-to-sulfate ratio, or the electron donor - affected the growth of D. vulgaris and strongly influenced the formation and growth of FeS solids. In particular, lower pH (from initial conditions or a decrease caused by less sulfate reduction, FeS precipitation, or using pyruvate as the electron donor) produced larger-sized mackinawite (Fe1+xS). Greater accumulation of free sulfide, from more sulfate reduction by D. vulgaris, also led to larger-sized mackinawite and particularly stimulated mackinawite transformation to greigite (Fe3S4) when the free sulfide concentration was 29.3mM. Furthermore, sufficient free Fe(2+) led to the additional formation of vivianite [Fe3(PO4)2·8(H2O)]. Thus, microbially relevant conditions (initial pH, choice of electron donor, and excess or deficiency of sulfide) are tools to generate biogenic FeS solids of different characteristics.


Subject(s)
Anti-Infective Agents/chemistry , Biodegradation, Environmental , Desulfovibrio vulgaris/metabolism , Ferrous Compounds/chemistry , Crystallization , Desulfovibrio vulgaris/growth & development , Electrons , Hydrogen-Ion Concentration , Sulfates/chemistry , Sulfides/chemistry , Thermodynamics , Water Pollutants, Chemical , Water Purification , X-Ray Diffraction
20.
Appl Environ Microbiol ; 80(7): 2133-41, 2014 Apr.
Article in English | MEDLINE | ID: mdl-24463969

ABSTRACT

Corrinoids are essential cofactors of reductive dehalogenases in Dehalococcoides mccartyi, an important bacterium in bioremediation, yet sequenced D. mccartyi strains do not possess the complete pathway for de novo corrinoid biosynthesis. Pelosinus sp. and Desulfovibrio sp. have been detected in dechlorinating communities enriched from contaminated groundwater without exogenous cobalamin corrinoid. To investigate the corrinoid-related interactions among key members of these communities, we constructed consortia by growing D. mccartyi strain 195 (Dhc195) in cobalamin-free, trichloroethene (TCE)- and lactate-amended medium in cocultures with Desulfovibrio vulgaris Hildenborough (DvH) or Pelosinus fermentans R7 (PfR7) and with both in tricultures. Only the triculture exhibited sustainable dechlorination and cell growth when a physiological level of 5,6-dimethylbenzimidazole (DMB), the lower ligand of cobalamin, was provided. In the triculture, DvH provided hydrogen while PfR7 provided corrinoids to Dhc195, and the initiation of dechlorination and Dhc195 cell growth was highly dependent on the growth of PfR7. Corrinoid analysis indicated that Dhc195 imported and remodeled the phenolic corrinoids produced by PfR7 into cobalamin in the presence of DMB. Transcriptomic analyses of Dhc195 showed the induction of the CbiZ-dependent corrinoid-remodeling pathway and BtuFCD corrinoid ABC transporter genes during corrinoid salvaging and remodeling. In contrast, another operon annotated to encode a putative iron/cobalamin ABC transporter (DET1174-DET1176) was induced when cobalamin was exogenously provided. Interestingly, a global upregulation of phage-related genes was observed when PfR7 was present. These findings provide insights into both the gene regulation of corrinoid salvaging and remodeling in Dhc195 when it is grown without exogenous cobalamin and microbe-to-microbe interactions in dechlorinating microbial communities.


Subject(s)
Chloroflexi/growth & development , Chloroflexi/metabolism , Corrinoids/metabolism , Lactic Acid/metabolism , Microbial Consortia , Benzimidazoles/metabolism , Chlorine/metabolism , Culture Media/chemistry , Desulfovibrio vulgaris/growth & development , Desulfovibrio vulgaris/metabolism , Fermentation , Hydrogen/metabolism , Transcriptome , Veillonellaceae/growth & development , Veillonellaceae/metabolism
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