scholarly journals Genetic Identification of a Putative Vinyl Chloride Reductase in Dehalococcoides sp. Strain BAV1

2004 ◽  
Vol 70 (10) ◽  
pp. 6347-6351 ◽  
Author(s):  
Rosa Krajmalnik-Brown ◽  
Tina Hölscher ◽  
Ivy N. Thomson ◽  
F. Michael Saunders ◽  
Kirsti M. Ritalahti ◽  
...  
Keyword(s):  
Vinyl Chloride ◽  
Reductive Dechlorination ◽  
Complete Sequence ◽  
Mixed Cultures ◽  
Genetic Identification ◽  
Degenerate Primers ◽  
Transcription Analysis ◽  
Reductive Dehalogenase ◽  
Conserved Regions ◽  
Rdase Genes

ABSTRACT Dehalococcoides sp. strain BAV1 couples growth with the reductive dechlorination of vinyl chloride (VC) to ethene. Degenerate primers targeting conserved regions in reductive dehalogenase (RDase) genes were designed and used to PCR amplify putative RDase genes from strain BAV1. Seven unique RDase gene fragments were identified. Transcription analysis of VC-grown BAV1 cultures suggested that bvcA was involved in VC reductive dechlorination, and the complete sequence of bvcA was obtained. bvcA was absent in Dehalococcoides isolates that failed to respire VC, yet was detected in four of eight VC-respiring mixed cultures.

scholarly journals Multiple Reductive-Dehalogenase-Homologous Genes Are Simultaneously Transcribed during Dechlorination by Dehalococcoides-Containing Cultures

2005 ◽  
Vol 71 (12) ◽  
pp. 8257-8264 ◽  
Author(s):  
Alison S. Waller ◽  
Rosa Krajmalnik-Brown ◽  
Frank E. Löffler ◽  
Elizabeth A. Edwards
Keyword(s):  
Vinyl Chloride ◽  
Electron Acceptor ◽  
Reductive Dechlorination ◽  
Amino Acid Identity ◽  
Degenerate Primers ◽  
Acid Identity ◽  
Reductive Dehalogenase ◽  
Homologous Genes ◽  
Differential Transcription ◽  
Transcribed Sequences

ABSTRACT Degenerate primers were used to amplify 14 distinct reductive-dehalogenase-homologous (RDH) genes from the Dehalococcoides-containing mixed culture KB1. Most of the corresponding predicted proteins were highly similar (97 to >99% amino acid identity) to previously reported Dehalococcoides reductive dehalogenases. To examine the differential transcription of these RDH genes, KB1 was split into five subcultures amended with either trichloroethene, cis-1,2-dichloroethene, vinyl chloride, 1,2-dichlorethane, or no chlorinated electron acceptor. Total RNA was extracted following the onset of reductive dechlorination, and RDH transcripts were reverse transcribed and amplified using degenerate primers. The results indicate that the transcription of RDH genes requires the presence of a chlorinated electron acceptor, and for all treatments, multiple RDH genes were simultaneously transcribed, with transcripts of two of the genes being present under all four electron-accepting conditions. Two of the transcribed sequences were highly similar to reported vinyl chloride reductase genes, namely, vcrA from Dehalococcoides sp. strain VS and bvcA from Dehalococcoides sp. strain BAV1. These findings suggest that multiple RDH genes are induced by a single chlorinated substrate and that multiple reductive dehalogenases contribute to chloroethene degradation in KB1.

scholarly journals A comparative genomics and reductive dehalogenase gene transcription study of two chloroethene-respiring bacteria, Dehalococcoides mccartyi strains MB and 11a

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Adrian Low ◽  
Zhiyong Shen ◽  
Dan Cheng ◽  
Matthew J. Rogers ◽  
Patrick K. H. Lee ◽  
...  
Keyword(s):  
Draft Genome ◽  
Type I ◽  
Quantitative Real Time Pcr ◽  
Transcription Analysis ◽  
Reductive Dehalogenase ◽  
Dehalococcoides Mccartyi ◽  
Steady Level ◽  
Gene Transcripts ◽  
Large Genome Size ◽  
Rdase Genes

Abstract Genomes of two trichloroethene (TCE)-respiring Dehalococcoides (Dhc) mccartyi, strains MB and 11a, were sequenced to identify reductive dehalogenases (RDase) responsible for oraganohalide respiration. Transcription analyses were conducted to verify the roles of RDase subunit A genes (rdhA) in chloroethene respiration. Some interesting features of the strain MB draft genome include a large genome size, two CRISPR-cas type I systems and 38 rdhA genes. Strain 11a has a stream-lined genome with 11 rdhA genes, of which nine are distinct. Quantitative real-time PCR transcription analysis of RDase gene transcripts showed that a single RDase gene, designated mbrA, was up-regulated upon exposure to TCE and no other RDase genes were considerably expressed in strain MB. A single RDase gene, designated vcrA, was up-regulated upon exposure to TCE and expressed at a steady level until all chloroethenes were completely dechlorinated to ethene at 147 h in strain 11a. Overall, this study reports the genomes of two distinct Dhc strains; both contain numerous uncharacterized RDase genes, but in each strain only one such gene was expressed highly during organohalide respiration.

scholarly journals Temporal Expression of Respiratory Genes in an Enrichment Culture Containing Dehalococcoides ethenogenes

2006 ◽  
Vol 72 (8) ◽  
pp. 5486-5491 ◽  
Author(s):  
Brian G. Rahm ◽  
Robert M. Morris ◽  
Ruth E. Richardson
Keyword(s):  
Temporal Variability ◽  
Enrichment Culture ◽  
Transcript Level ◽  
Mixed Cultures ◽  
Temporal Expression ◽  
Expression Levels ◽  
Reductive Dehalogenase ◽  
Expression Of Genes ◽  
Dehalococcoides Ethenogenes ◽  
Rdase Genes

ABSTRACT Multiple reductive dehalogenase (RDase), hydrogenase (H2ase), and other respiration-associated (RA) oxidoreductase genes have been identified in cultured representatives of Dehalococcoides. Although their products are likely to play key roles in the environmentally important process of reductive dechlorination, very little information is available about their regulation and specific functions. Here we show increased expression and temporal variability in the expression of five RDase genes and in the expression of genes for a putative formate dehydrogenase (Fdh) and two H2ases, including a periplasmic [Ni/Fe] H2ase (Hup) and a cytoplasmic [Fe] H2ase (Vhu). mRNA transcripts extracted from tetrachloroethene-dechlorinating mixed cultures corresponding to Fdh, the H2ase Hup, and the RDase targets TceA and DET0162 were expressed most highly, with average levels 34 (± 7.5)-, 23 (± 6.7)-, 16 (± 3.3)-, and 13 (± 3.3)-fold higher, respectively, than that for RNA polymerase (RpoB). H2ase and RA transcripts reached their respective expression maxima within the first 2 h after feeding. RDase transcripts, however, were most highly expressed after 3 h and exhibited greater temporal variability than other transcripts. Comparison with D. ethenogenes strain 195 pure culture expression levels indicated that RDase DET1545 was more highly expressed in mixed cultures, where, on average, its transcript level was sixfold higher than that of RpoB. While the specific functions of several of these gene products remain elusive, the high expression levels and temporal variability reported here suggest that these groups of enzymes are metabolically important for the respiration of chlorinated ethenes in mixed cultures containing Dehalococcoides.

scholarly journals Fraction of Electrons Consumed in Electron Acceptor Reduction and Hydrogen Thresholds as Indicators of Halorespiratory Physiology

1999 ◽  
Vol 65 (9) ◽  
pp. 4049-4056 ◽  
Author(s):  
Frank E. Löffler ◽  
James M. Tiedje ◽  
Robert A. Sanford
Keyword(s):  
Vinyl Chloride ◽  
Electron Acceptor ◽  
Reductive Dechlorination ◽  
Theoretical Calculations ◽  
Mixed Cultures ◽  
Lower Threshold ◽  
Chlorinated Compound ◽  
Hydrogen Consumption ◽  
Pure Cultures ◽  
Good Agreement

ABSTRACT Measurements of the hydrogen consumption threshold and the tracking of electrons transferred to the chlorinated electron acceptor (fe) reliably detected chlororespiratory physiology in both mixed cultures and pure cultures capable of using tetrachloroethene,cis-1,2-dichloroethene, vinyl chloride, 2-chlorophenol, 3-chlorobenzoate, 3-chloro-4-hydroxybenzoate, or 1,2-dichloropropane as an electron acceptor. Hydrogen was consumed to significantly lower threshold concentrations of less than 0.4 ppmv compared with the values obtained for the same cultures without a chlorinated compound as an electron acceptor. The fe values ranged from 0.63 to 0.7, values which are in good agreement with theoretical calculations based on the thermodynamics of reductive dechlorination as the terminal electron-accepting process. In contrast, a mixed methanogenic culture that cometabolized 3-chlorophenol exhibited a significantly lower fe value, 0.012.

Reductive Dechlorination of Chlorinated Ethenes and 1,2-Dichloroethane by “Dehalococcoides ethenogenes” 195

1999 ◽  
Vol 65 (7) ◽  
pp. 3108-3113 ◽  
Author(s):  
Xavier Maymó-Gatell ◽  
Timothy Anguish ◽  
Stephen H. Zinder
Keyword(s):  
Vinyl Chloride ◽  
Growth Medium ◽  
Electron Acceptor ◽  
Reductive Dechlorination ◽  
Electron Acceptors ◽  
Chlorinated Compounds ◽  
Reductive Dehalogenase ◽  
Dehalococcoides Ethenogenes ◽  
Over Time ◽  
Number Of Cells

ABSTRACT “Dehalococcoides ethenogenes” 195 can reductively dechlorinate tetrachloroethene (PCE) completely to ethene (ETH). When PCE-grown strain 195 was transferred (2% [vol/vol] inoculum) into growth medium amended with trichloroethene (TCE),cis-dichloroethene (DCE), 1,1-DCE, or 1,2-dichloroethane (DCA) as an electron acceptor, these chlorinated compounds were consumed at increasing rates over time, which indicated that growth occurred. Moreover, the number of cells increased when TCE, 1,1-DCE, or DCA was present. PCE, TCE, 1,1-DCE, and cis-DCE were converted mainly to vinyl chloride (VC) and then to ETH, while DCA was converted to ca. 99% ETH and 1% VC. cis-DCE was used at lower rates than PCE, TCE, 1,1-DCE, or DCA was used. When PCE-grown cultures were transferred to media containing VC ortrans-DCE, products accumulated slowly, and there was no increase in the rate, which indicated that these two compounds did not support growth. When the intermediates in PCE dechlorination by strain 195 were monitored, TCE was detected first, followed bycis-DCE. After a lag, VC, 1,1-DCE, andtrans-DCE accumulated, which is consistent with the hypothesis that cis-DCE is the precursor of these compounds. Both cis-DCE and 1,1-DCE were eventually consumed, and both of these compounds could be considered intermediates in PCE dechlorination, whereas the small amount oftrans-DCE that was produced persisted. Cultures grown on TCE, 1,1-DCE, or DCA could immediately dechlorinate PCE, which indicated that PCE reductive dehalogenase activity was constitutive when these electron acceptors were used.

scholarly journals Genome Sequence of “Candidatus Dehalogenimonas etheniformans” Strain GP, a Vinyl Chloride-Respiring Anaerobe

2020 ◽  
Vol 9 (50) ◽  
Author(s):  
Yi Yang ◽  
Jun Yan ◽  
Xiuying Li ◽  
Yan Lv ◽  
Yiru Cui ◽  
...  
Keyword(s):  
Vinyl Chloride ◽  
Genome Sequence ◽  
Reductive Dechlorination ◽  
Reductive Dehalogenase ◽  
Reductive Dehalogenase Genes

ABSTRACT “Candidatus Dehalogenimonas etheniformans” strain GP couples growth with the reductive dechlorination of vinyl chloride and several polychlorinated ethenes. The genome sequence comprises a circular 2.07-Mb chromosome with a G+C content of 51.9% and harbors 50 putative reductive dehalogenase genes.

scholarly journals Auxiliary Substrates for Elimination of Trichloroethene, Monochlorobenzene, and Benzene in a Sequential Anaerobic–Aerobic GAC Biobarrier

2007 ◽  
Vol 7 (1 & 2) ◽  
pp. 68
Author(s):  
M. Gozan ◽  
A. Mueller ◽  
A. Tiehm
Keyword(s):  
Vinyl Chloride ◽  
Reductive Dechlorination ◽  
Biological Degradation ◽  
Anaerobic Conditions ◽  
Batch Studies ◽  
Second Stage ◽  
Competitive Reactions ◽  
Novel Concept ◽  
Tce Degradation ◽  
Granulated Activated Carbon

Sequential anaerobic-aerobic barrier is a novel concept for groundwater bioremediation. Trichloroethene (TCE), monochlorobenzene (MCB), and benzene (BZ) were used as model contaminants representing contaminant cocktails frequently found in the contaminated subsurface. The autochthonous microflora from a contaminated field was inoculated to eliminate model contaminants in a set of sequential anaerobic–aerobic granulated activated carbon (GAC) columns and batch studies. In the anaerobic column, the TCE was reductively dechlorinated through cis-dichloroethene (cis-DCE), vinyl chloride (VC), and ethene (ETH). Ethanol and sucrose as auxiliary substrates were added to donate electrons. In the second stage, MCB, BZ, and the lower chlorinated metabolites of TCE degradation, i.e. cis-Dichloroethene (cisDCE) and vinyl chloride (VC), were oxidatively degraded with addition of hydrogen peroxide and nitrate. This paper examines the influence of auxiliary substrates on the biological degradation of model pollutants. In the anaerobic barrier, the auxiliary substrates supply should be maintained low but stoichiometrically adequate for supporting reductive dechlorination. Supplying higher amount of auxiliary substrates provoked competitive reactions in anaerobic conditions, such as sulfate reduction and methanogenesis. If the auxiliary substrates are not utilized completely in the anaerobic phase, the remaining compounds flow into the aerobic phase. This led to unwanted conditions, i.e. oxidation of auxiliary substrates instead of pollutant elimination, and a higher consumption of electron acceptors. In the aerobic barrier, in particular, ethene proved to be a suitable auxiliary substrate for cometabolic degradation of cisDCE.

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