scholarly journals The Complexome of Dehalococcoides mccartyi Reveals Its Organohalide Respiration-Complex Is Modular

2018 ◽  
Vol 9 ◽  
Author(s):  
Katja Seidel ◽  
Joana Kühnert ◽  
Lorenz Adrian
Keyword(s):  
Organohalide Respiration ◽  
Dehalococcoides Mccartyi

scholarly journals Dehalococcoides mccartyi Strain DCMB5 Respires a Broad Spectrum of Chlorinated Aromatic Compounds

2014 ◽  
Vol 81 (2) ◽  
pp. 587-596 ◽  
Author(s):  
Marlén Pöritz ◽  
Christian L. Schiffmann ◽  
Gerd Hause ◽  
Ulrike Heinemann ◽  
Jana Seifert ◽  
...  
Keyword(s):  
Electron Acceptor ◽  
Aromatic Compounds ◽  
Lag Phase ◽  
Organohalide Respiration ◽  
Content Type ◽  
Reductive Dehalogenase ◽  
Dehalococcoides Mccartyi ◽  
Type Iv ◽  
Transmission Electron ◽  
Almost All

ABSTRACTPolyhalogenated aromatic compounds are harmful environmental contaminants and tend to persist in anoxic soils and sediments.Dehalococcoides mccartyistrain DCMB5, a strain originating from dioxin-polluted river sediment, was examined for its capacity to dehalogenate diverse chloroaromatic compounds. Strain DCMB5 used hexachlorobenzenes, pentachlorobenzenes, all three tetrachlorobenzenes, and 1,2,3-trichlorobenzene as well as 1,2,3,4-tetra- and 1,2,4-trichlorodibenzo-p-dioxin as electron acceptors for organohalide respiration. In addition, 1,2,3-trichlorodibenzo-p-dioxin and 1,3-, 1,2-, and 1,4-dichlorodibenzo-p-dioxin were dechlorinated, the latter to the nonchlorinated congener with a remarkably short lag phase of 1 to 4 days following transfer. Strain DCMB5 also dechlorinated pentachlorophenol and almost all tetra- and trichlorophenols. Tetrachloroethene was dechlorinated to trichloroethene and served as an electron acceptor for growth. To relate selected dechlorination activities to the expression of specific reductive dehalogenase genes, the proteomes of 1,2,3-trichlorobenzene-, pentachlorobenzene-, and tetrachloroethene-dechlorinating cultures were analyzed. Dcmb_86, an ortholog of the chlorobenzene reductive dehalogenase CbrA, was the most abundant reductive dehalogenase during growth with each electron acceptor, suggesting its pivotal role in organohalide respiration of strain DCMB5. Dcmb_1041 was specifically induced, however, by both chlorobenzenes, whereas 3 putative reductive dehalogenases, Dcmb_1434, Dcmb_1339, and Dcmb_1383, were detected only in tetrachloroethene-grown cells. The proteomes also harbored a type IV pilus protein and the components for its assembly, disassembly, and secretion. In addition, transmission electron microscopy of DCMB5 revealed an irregular mode of cell division as well as the presence of pili, indicating that pilus formation is a feature ofD. mccartyiduring organohalide respiration.

scholarly journals The MarR-Type Regulator Rdh2R RegulatesrdhGene Transcription in Dehalococcoides mccartyi Strain CBDB1

2016 ◽  
Vol 198 (23) ◽  
pp. 3130-3141 ◽  
Author(s):  
Lydia Krasper ◽  
Hauke Lilie ◽  
Anja Kublik ◽  
Lorenz Adrian ◽  
Ralph Golbik ◽  
...  
Keyword(s):  
Heterologous Expression ◽  
Direct Repeat ◽  
Negative Regulator ◽  
Control Of Gene Expression ◽  
Organohalide Respiration ◽  
Content Type ◽  
Reductive Dehalogenase ◽  
Dehalococcoides Mccartyi ◽  
Transcriptional Start Sites

ABSTRACTReductive dehalogenases are essential enzymes in organohalide respiration and consist of a catalytic subunit A and a membrane protein B, encoded byrdhABgenes. Thirty-twordhABgenes exist in the genome ofDehalococcoides mccartyistrain CBDB1. To gain a first insight into the regulation ofrdhoperons, the control of gene expression of twordhABgenes (cbdbA1453/cbdbA1452 and cbdbA1455/cbdbA1454) by the MarR-type regulator Rdh2R (cbdbA1456) encoded directly upstream was studied using heterologous expression andin vitrostudies. Promoter-lacZreporter fusions were generated and integrated into the genome of theEscherichia colihost. ThelacZreporter activities of bothrdhApromoters decreased upon transformation of the cells with a plasmid carrying therdh2Rgene, suggesting that Rdh2R acts as repressor, whereas thelacZreporter activity of therdh2Rpromoter was not affected. The transcriptional start sites of bothrdhAgenes in strain CBDB1 and/or the heterologous host mapped to a conserved direct repeat with 11- to 13-bp half-sites. DNase I footprinting revealed binding of Rdh2R to a ∼30-bp sequence covering the complete direct repeat in both promoters, including the transcriptional start sites. Equilibrium sedimentation ultracentrifugation revealed that Rdh2R binds as tetramer to the direct-repeat motif of therdhA(cbdbA1455) promoter. Using electrophoretic mobility shift assays, a similar binding affinity was found for bothrdhApromoters. In the presence of only one half-site of the direct repeat, the interaction was strongly reduced, suggesting a positive cooperativity of binding, for which unusual short palindromes within the direct-repeat half-sites might play an important role.IMPORTANCEDehalococcoides mccartyistrains are obligate anaerobes that grow by organohalide respiration. They have an important bioremediation potential because they are capable of reducing a multitude of halogenated compounds to less toxic products. We are now beginning to understand how these organisms make use of this large catabolic potential, wherebyD. mccartyiexpresses dehalogenases in a compound-specific fashion. MarR-type regulators are often encoded in the vicinity of reductive dehalogenase genes. In this study, we made use of heterologous expression andin vitrostudies to demonstrate that the MarR-type transcription factor Rdh2R acts as a negative regulator. We identify its binding site on the DNA, which suggests a mechanism by which it controls the expression of two adjacent reductive dehalogenase operons.

scholarly journals Impact of Ammonium on Syntrophic Organohalide-Respiring and Fermenting Microbial Communities

mSphere ◽  
2016 ◽  
Vol 1 (2) ◽  
Author(s):  
Anca G. Delgado ◽  
Devyn Fajardo-Williams ◽  
Kylie L. Kegerreis ◽  
Prathap Parameswaran ◽  
Rosa Krajmalnik-Brown
Keyword(s):  
Chlorinated Solvents ◽  
Reductive Dehalogenation ◽  
Ammonium Concentration ◽  
Organohalide Respiration ◽  
Content Type ◽  
Dehalococcoides Mccartyi ◽  
Metabolic Capability ◽  
Subsurface Environments ◽  
Fermentative Bacteria ◽  
High Concentrations

ABSTRACT Contamination with ammonium and chlorinated solvents has been reported in numerous subsurface environments, and these chemicals bring significant challenges for in situ bioremediation. Dehalococcoides mccartyi is able to reduce the chlorinated solvent trichloroethene to the nontoxic end product ethene. Fermentative bacteria are of central importance for organohalide respiration and bioremediation to provide D. mccartyi with H2, their electron donor, acetate, their carbon source, and other micronutrients. In this study, we found that high concentrations of ammonium negatively correlated with rates of trichloroethene reductive dehalogenation and fermentation. However, detoxification of trichloroethene to nontoxic ethene occurred even at ammonium concentrations typical of those found in animal waste (up to 2 g liter−1 NH4 +-N). To date, hundreds of subsurface environments have been bioremediated through the unique metabolic capability of D. mccartyi. These findings extend our knowledge of D. mccartyi and provide insight for bioremediation of sites contaminated with chlorinated solvents and ammonium. Syntrophic interactions between organohalide-respiring and fermentative microorganisms are critical for effective bioremediation of halogenated compounds. This work investigated the effect of ammonium concentration (up to 4 g liter−1 NH4 +-N) on trichloroethene-reducing Dehalococcoides mccartyi and Geobacteraceae in microbial communities fed lactate and methanol. We found that production of ethene by D. mccartyi occurred in mineral medium containing ≤2 g liter−1 NH4 +-N and in landfill leachate. For the partial reduction of trichloroethene (TCE) to cis-dichloroethene (cis-DCE) at ≥1 g liter−1 NH4 +-N, organohalide-respiring dynamics shifted from D. mccartyi and Geobacteraceae to mainly D. mccartyi. An increasing concentration of ammonium was coupled to lower metabolic rates, longer lag times, and lower gene abundances for all microbial processes studied. The methanol fermentation pathway to acetate and H2 was conserved, regardless of the ammonium concentration provided. However, lactate fermentation shifted from propionic to acetogenic at concentrations of ≥2 g liter−1 NH4 +-N. Our study findings strongly support a tolerance of D. mccartyi to high ammonium concentrations, highlighting the feasibility of organohalide respiration in ammonium-contaminated subsurface environments. IMPORTANCE Contamination with ammonium and chlorinated solvents has been reported in numerous subsurface environments, and these chemicals bring significant challenges for in situ bioremediation. Dehalococcoides mccartyi is able to reduce the chlorinated solvent trichloroethene to the nontoxic end product ethene. Fermentative bacteria are of central importance for organohalide respiration and bioremediation to provide D. mccartyi with H2, their electron donor, acetate, their carbon source, and other micronutrients. In this study, we found that high concentrations of ammonium negatively correlated with rates of trichloroethene reductive dehalogenation and fermentation. However, detoxification of trichloroethene to nontoxic ethene occurred even at ammonium concentrations typical of those found in animal waste (up to 2 g liter−1 NH4 +-N). To date, hundreds of subsurface environments have been bioremediated through the unique metabolic capability of D. mccartyi. These findings extend our knowledge of D. mccartyi and provide insight for bioremediation of sites contaminated with chlorinated solvents and ammonium.

scholarly journals Changes of the Proteome and Acetylome during Transition into the Stationary Phase in the Organohalide-Respiring Dehalococcoides mccartyi Strain CBDB1

2021 ◽  
Vol 9 (2) ◽  
pp. 365
Author(s):  
Franziska Greiner-Haas ◽  
Martin von Bergen ◽  
Gary Sawers ◽  
Ute Lechner ◽  
Dominique Türkowsky
Keyword(s):  
Membrane Protein ◽  
Stationary Phase ◽  
Electron Acceptor ◽  
Acetate Metabolism ◽  
Strictly Anaerobic ◽  
Uptake Hydrogenase ◽  
Organohalide Respiration ◽  
Dehalococcoides Mccartyi ◽  
Protein Levels ◽  
Twin Arginine Translocation

The strictly anaerobic bactGIerium Dehalococcoides mccartyi obligatorily depends on organohalide respiration for energy conservation and growth. The bacterium also plays an important role in bioremediation. Since there is no guarantee of a continuous supply of halogenated substrates in its natural environment, the question arises of how D. mccartyi maintains the synthesis and activity of dehalogenating enzymes under these conditions. Acetylation is a means by which energy-restricted microorganisms can modulate and maintain protein levels and their functionality. Here, we analyzed the proteome and Nε-lysine acetylome of D. mccartyi strain CBDB1 during growth with 1,2,3-trichlorobenzene as an electron acceptor. The high abundance of the membrane-localized organohalide respiration complex, consisting of the reductive dehalogenases CbrA and CbdbA80, the uptake hydrogenase HupLS, and the organohalide respiration-associated molybdoenzyme OmeA, was shown throughout growth. In addition, the number of acetylated proteins increased from 5% to 11% during the transition from the exponential to the stationary phase. Acetylation of the key proteins of central acetate metabolism and of CbrA, CbdbA80, and TatA, a component of the twin-arginine translocation machinery, suggests that acetylation might contribute to maintenance of the organohalide-respiring capacity of the bacterium during the stationary phase, thus providing a means of ensuring membrane protein integrity and a proton gradient.

scholarly journals Guided cobalamin biosynthesis supports Dehalococcoides mccartyi reductive dechlorination activity

2013 ◽  
Vol 368 (1616) ◽  
pp. 20120320 ◽  
Author(s):  
Jun Yan ◽  
Jeongdae Im ◽  
Yi Yang ◽  
Frank E. Löffler
Keyword(s):  
Reductive Dechlorination ◽  
Methanosarcina Barkeri ◽  
Chlorinated Ethenes ◽  
Vitamin B ◽  
Organohalide Respiration ◽  
Dehalococcoides Mccartyi ◽  
Subsurface Environments ◽  
Cobalamin Biosynthesis ◽  
Axenic Cultures ◽  
Sporomusa Ovata

Dehalococcoides mccartyi strains are corrinoid-auxotrophic Bacteria and axenic cultures that require vitamin B 12 (CN-Cbl) to conserve energy via organohalide respiration. Cultures of D. mccartyi strains BAV1, GT and FL2 grown with limiting amounts of 1 µg l −1 CN-Cbl quickly depleted CN-Cbl, and reductive dechlorination of polychlorinated ethenes was incomplete leading to vinyl chloride (VC) accumulation. In contrast, the same cultures amended with 25 µg l −1 CN-Cbl exhibited up to 2.3-fold higher dechlorination rates, 2.8–9.1-fold increased growth yields, and completely consumed growth-supporting chlorinated ethenes. To explore whether known cobamide-producing microbes supply Dehalococcoides with the required corrinoid cofactor, co-culture experiments were performed with the methanogen Methanosarcina barkeri strain Fusaro and two acetogens, Sporomusa ovata and Sporomusa sp. strain KB-1, as Dehalococcoides partner populations. During growth with H 2 /CO 2 , M. barkeri axenic cultures produced 4.2 ± 0.1 µg l −1 extracellular cobamide (factor III), whereas the Sporomusa cultures produced phenolyl- and p -cresolyl-cobamides. Neither factor III nor the phenolic cobamides supported Dehalococcoides reductive dechlorination activity suggesting that M. barkeri and the Sporomusa sp. cannot fulfil Dehalococcoides ' nutritional requirements. Dehalococcoides dechlorination activity and growth occurred in M. barkeri and Sporomusa sp. co-cultures amended with 10 µM 5′,6′-dimethylbenzimidazole (DMB), indicating that a cobalamin is a preferred corrinoid cofactor of strains BAV1, GT and FL2 when grown with chlorinated ethenes as electron acceptors. Even though the methanogen and acetogen populations tested did not produce cobalamin, the addition of DMB enabled guided biosynthesis and generated a cobalamin that supported Dehalococcoides ' activity and growth. Guided cobalamin biosynthesis may offer opportunities to sustain and enhance Dehalococcoides activity in contaminated subsurface environments.

scholarly journals Effects of the Feeding Solution Composition on a Reductive/Oxidative Sequential Bioelectrochemical Process for Perchloroethylene Removal

Processes ◽  
2021 ◽  
Vol 9 (3) ◽  
pp. 405
Author(s):  
Edoardo Dell’Armi ◽  
Marco Zeppilli ◽  
Bruna Matturro ◽  
Simona Rossetti ◽  
Marco Petrangeli Papini ◽  
...  
Keyword(s):  
Reductive Dechlorination ◽  
Nitrate Reduction ◽  
Coulombic Efficiency ◽  
Reducing Power ◽  
Mineral Medium ◽  
Process Conditions ◽  
Microbial Electrolysis Cells ◽  
Dehalococcoides Mccartyi ◽  
In Series ◽  
Synthetic Groundwater

Chlorinated aliphatic hydrocarbons (CAHs) are common groundwater contaminants due to their improper use in several industrial activities. Specialized microorganisms are able to perform the reductive dechlorination (RD) of high-chlorinated CAHs such as perchloroethylene (PCE), while the low-chlorinated ethenes such as vinyl chloride (VC) are more susceptible to oxidative mechanisms performed by aerobic dechlorinating microorganisms. Bioelectrochemical systems can be used as an effective strategy for the stimulation of both anaerobic and aerobic microbial dechlorination, i.e., a biocathode can be used as an electron donor to perform the RD, while a bioanode can provide the oxygen necessary for the aerobic dechlorination reaction. In this study, a sequential bioelectrochemical process constituted by two membrane-less microbial electrolysis cells connected in series has been, for the first time, operated with synthetic groundwater, also containing sulphate and nitrate, to simulate more realistic process conditions due to the possible establishment of competitive processes for the reducing power, with respect to previous research made with a PCE-contaminated mineral medium (with neither sulphate nor nitrate). The shift from mineral medium to synthetic groundwater showed the establishment of sulphate and nitrate reduction and caused the temporary decrease of the PCE removal efficiency from 100% to 85%. The analysis of the RD biomarkers (i.e., Dehalococcoides mccartyi 16S rRNA and tceA, bvcA, vcrA genes) confirmed the decrement of reductive dechlorination performances after the introduction of the synthetic groundwater, also characterized by a lower ionic strength and nutrients content. On the other hand, the system self-adapted the flowing current to the increased demand for the sulphate and nitrate reduction, so that reducing power was not in defect for the RD, although RD coulombic efficiency was less.

scholarly journals A Microcosm Treatability Study for Evaluating Wood Mulch-Based Amendments as Electron Donors for Trichloroethene (TCE) Reductive Dechlorination

Water ◽  
2021 ◽  
Vol 13 (14) ◽  
pp. 1949
Author(s):  
Edoardo Masut ◽  
Alessandro Battaglia ◽  
Luca Ferioli ◽  
Anna Legnani ◽  
Carolina Cruz Viggi ◽  
...  
Keyword(s):  
Electron Donor ◽  
Environmental Sustainability ◽  
Reductive Dechlorination ◽  
Environmental Remediation ◽  
Low Cost ◽  
Chlorinated Solvents ◽  
Electron Donors ◽  
Chlorinated Ethenes ◽  
Dehalococcoides Mccartyi ◽  
Wood Mulch

In this study, wood mulch-based amendments were tested in a bench-scale microcosm experiment in order to assess the treatability of saturated soils and groundwater from an industrial site contaminated by chlorinated ethenes. Wood mulch was tested alone as the only electron donor in order to assess its potential for stimulating the biological reductive dechlorination. It was also tested in combination with millimetric iron filings in order to assess the ability of the additive to accelerate/improve the bioremediation process. The efficacy of the selected amendments was compared with that of unamended control microcosms. The results demonstrated that wood mulch is an effective natural and low-cost electron donor to stimulate the complete reductive dechlorination of chlorinated solvents to ethene. Being a side-product of the wood industry, mulch can be used in environmental remediation, an approach which perfectly fits the principles of circular economy and addresses the compelling needs of a sustainable and low environmental impact remediation. The efficacy of mulch was further improved by the co-presence of iron filings, which accelerated the conversion of vinyl chloride into the ethene by increasing the H2 availability rather than by catalyzing the direct abiotic dechlorination of contaminants. Chemical analyses were corroborated by biomolecular assays, which confirmed the stimulatory effect of the selected amendments on the abundance of Dehalococcoides mccartyi and related reductive dehalogenase genes. Overall, this paper further highlights the application potential and environmental sustainability of wood mulch-based amendments as low-cost electron donors for the biological treatment of chlorinated ethenes.

scholarly journals Substrate-dependent competition and cooperation relationships between Geobacter and Dehalococcoides for their organohalide respiration

2021 ◽  
Vol 1 (1) ◽  
Author(s):  
Yongyi Liang ◽  
Qihong Lu ◽  
Zhiwei Liang ◽  
Xiaokun Liu ◽  
Wenwen Fang ◽  
...  
Keyword(s):  
Electron Donors ◽  
Community Stability ◽  
Organohalide Respiration ◽  
Competition And Cooperation ◽  
Free Competition ◽  
Isolation And Characterization ◽  
Geochemical Cycling ◽  
New Strategy ◽  
Insight Into

AbstractObligate and non-obligate organohalide-respiring bacteria (OHRB) play central roles in the geochemical cycling and environmental bioremediation of organohalides. Their coexistence and interactions may provide functional redundancy and community stability to assure organohalide respiration efficiency but, at the same time, complicate isolation and characterization of specific OHRB. Here, we employed a growth rate/yield tradeoff strategy to enrich and isolate a rare non-obligate tetrachloroethene (PCE)-respiring Geobacter from a Dehalococcoides-predominant microcosm, providing experimental evidence for the rate/yield tradeoff theory in population selection. Surprisingly, further physiological and genomic characterizations, together with co-culture experiments, revealed three unique interactions (i.e., free competition, conditional competition and syntrophic cooperation) between Geobacter and Dehalococcoides for their respiration of PCE and polychlorinated biphenyls (PCBs), depending on both the feeding electron donors (acetate/H2 vs. propionate) and electron acceptors (PCE vs. PCBs). This study provides the first insight into substrate-dependent interactions between obligate and non-obligate OHRB, as well as a new strategy to isolate fastidious microorganisms, for better understanding of the geochemical cycling and bioremediation of organohalides.

Sign in / Sign up

Export Citation Format

Share Document