Isolation and transcriptional analysis of novel tetrachloroethene reductive dehalogenase gene from Desulfitobacterium sp. strain KBC1

Tsukagoshi, Norihiko; Ezaki, Satoshi; Uenaka, Tetsuya; Suzuki, Nobukazu; Kurane, Ryuichiro

doi:10.1007/s00253-005-0022-x

Cited by 33 publications

(14 citation statements)

References 41 publications

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“…Experimental support (transcription under organohalide compounds) has been reported for the following proteins: BvcA.AAT64888.1 (Krajmalnik-Brown et al, 2004). MbrA.ADF96893.1 (Chow et al, 2010), PrdA.BAE45338.1 (Tsukagoshi et al, 2006). (B) Residues of the rdhA -homologous genes in the sediment samples around the conserved regions defined in (A) .…”

Section: Resultsmentioning

confidence: 99%

High frequency of phylogenetically diverse reductive dehalogenase-homologous genes in deep subseafloor sedimentary metagenomes

et al. 2014

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Marine subsurface sediments on the Pacific margin harbor diverse microbial communities even at depths of several hundreds meters below the seafloor (mbsf) or more. Previous PCR-based molecular analysis showed the presence of diverse reductive dehalogenase gene (rdhA) homologs in marine subsurface sediment, suggesting that anaerobic respiration of organohalides is one of the possible energy-yielding pathways in the organic-rich sedimentary habitat. However, primer-independent molecular characterization of rdhA has remained to be demonstrated. Here, we studied the diversity and frequency of rdhA homologs by metagenomic analysis of five different depth horizons (0.8, 5.1, 18.6, 48.5, and 107.0 mbsf) at Site C9001 off the Shimokita Peninsula of Japan. From all metagenomic pools, remarkably diverse rdhA-homologous sequences, some of which are affiliated with novel clusters, were observed with high frequency. As a comparison, we also examined frequency of dissimilatory sulfite reductase genes (dsrAB), key functional genes for microbial sulfate reduction. The dsrAB were also widely observed in the metagenomic pools whereas the frequency of dsrAB genes was generally smaller than that of rdhA-homologous genes. The phylogenetic composition of rdhA-homologous genes was similar among the five depth horizons. Our metagenomic data revealed that subseafloor rdhA homologs are more diverse than previously identified from PCR-based molecular studies. Spatial distribution of similar rdhA homologs across wide depositional ages indicates that the heterotrophic metabolic processes mediated by the genes can be ecologically important, functioning in the organic-rich subseafloor sedimentary biosphere.

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Section: Resultsmentioning

confidence: 99%

High frequency of phylogenetically diverse reductive dehalogenase-homologous genes in deep subseafloor sedimentary metagenomes

et al. 2014

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“…Within the PCE dehalogenase operon in Desulfitobacterium sp. strain KBC1, a gene encoding a transcriptional regulator, PrdK, of the CAP family activators was detected (22). The PCE dehalogenase (PrdA) gene transcription of this organism is induced by addition of PCE.…”

Section: Discussionmentioning

confidence: 99%

“…Tsukagoshi and coworkers (22) reported the induction of PCE dehalogenase gene expression in Desulfitobacterium hafniense strain KCB1 as controlled by the presence of PCE.…”

mentioning

confidence: 99%

Retentive Memory of Bacteria: Long-Term Regulation of Dehalorespiration in Sulfurospirillum multivorans

et al. 2009

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The gram-negative, strictly anaerobic epsilonproteobacterium Sulfurospirillum multivorans is able to gain energy from dehalorespiration with tetrachloroethene (perchloroethylene [PCE]) as a terminal electron acceptor. The organism can also utilize fumarate as an electron acceptor. Prolonged subcultivation of S. multivorans in the absence of PCE with pyruvate as an electron donor and fumarate as an electron acceptor resulted in a decrease of PCE dehalogenase (PceA) activity. Concomitantly, the pceA transcript level equally decreased as shown by reverse transcriptase PCR. (19). The organism utilizes pyruvate, hydrogen, or formate as an electron donor. When grown with hydrogen or formate as an electron donor and PCE as an electron acceptor, ATP synthesis is coupled to reductive dechlorination via electron transport phosphorylation (dehalorespiration) (8). The key enzyme of PCE utilization that converts PCE via TCE to cis-1,2-dichloroethene (cDCE) is the PCE reductive dehalogenase (PceA). Mature PceA is a monomeric enzyme harboring one norpseudovitamin B 12 and two Fe/S clusters as cofactors (12,15). The cytoplasmic precursor of the PCE dehalogenase (pre-PceA) bears an N-terminal signal peptide of 37 amino acids, including the motif SRRXFXK, which identifies pre-PceA as a substrate of the Tat (twin arginine translocation) protein export pathway (2). Mature PceA is localized in the periplasm, as shown by freeze fracture immunogold labeling techniques (9).The pce operon comprises pceA, the PCE dehalogenase gene, and pceB, encoding a putative membrane integral protein (16). Genes for transcriptional regulation of the pceAB genes are not found in a 6-kb DNA fragment including the pce operon (GenBank accession number AF022812).The ability of reductive dechlorination by anaerobic bacteria was found in phylogenetically distant species (8). Transcriptional regulation of dehalogenase gene expression was described for the 3-chloro-4-hydroxy-phenylacetate (Cl-OHPA) dehalogenase (CprA) of the gram-positive Desulfitobacterium dehalogenans. In this organism, the enzyme was strictly regulated by the absence or presence of the substrate. When the cultures were grown with alternative electron acceptors, such as fumarate or nitrate, CprA transcription ceased within the first subcultivation and was induced within less than 30 min (21). A CRP/FNR-like transcriptional activator, CprK, encoded within the cpr operon, was shown to interact with the DNA upstream of the cpr promoter region only when Cl-OHPA was bound to the protein (18). A direct role of CprK in the induction of dehalogenase gene expression in D. dehalogenans was assumed, and a model for transcriptional regulation of cprA expression was developed (13).While detailed information is available on the control of dehalogenase gene expression in D. dehalogenans grown in the presence of chlorinated aromatic compounds, such as Cl-OHPA, a transcriptional regulation of the PCE dehalogenase genes of Desulfitobacterium species has not been described so far (23). Tsukagoshi and coworker...

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“…Desulfitobacterium sp. strains PCE1 and KBC1 dechlorinate PCE to yield TCE and also dechlorinate chlorophenols 28,29…”

Section: Dehalorespiring Bacteriamentioning

confidence: 99%

Biochemical and genetic bases of dehalorespiration

Futagami

Goto

Furukawa

2008

The Chemical Record

105

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Some anaerobic bacteria can effi ciently eliminate one or more halide atoms from halogenated compounds such as chlorophenols and chloroethenes through reductive dehalogenation. During this process, the bacteria utilize halogenated compounds as the terminal electron acceptors in their anaerobic respiration, called dehalorespiration, to yield energy for growth. Currently the genera of Desulfi tobacterium and Dehalococcoides occupy the major part of the dehalorespiring isolates. The former can acquire energy not only by dehalorespiration but also by other respirations utilizing organic compounds and metals. In sharp contrast, the latter is specialized in dehalorespiration and plays a crucial role in the detoxifi cation of chlorinated compounds in nature. From these bacteria, various reductive dehalogenases, which catalyze the dehalogenation reaction, were purifi ed and their corresponding genes were identifi ed. Most reductive dehalogenases exhibit similar features such as the presences of a Tat (twin arginine translocation) signal sequence, two Fe-S clusters, and a corrinoid cofactor. Some of dehalogenase-encoding genes are found to be fl anked by insertion sequences. Thus, dehalogenase genes act as a catabolic transposon, and genetic rearrangements mediated by transposable elements occur well in dehalorespirers. Moreover, the genome sequences of some dehalorespiring bacteria provide many insights into the mechanism of dehalorespiration and the evolution of a dehalogenase gene.

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Isolation and transcriptional analysis of novel tetrachloroethene reductive dehalogenase gene from Desulfitobacterium sp. strain KBC1

Cited by 33 publications

References 41 publications

High frequency of phylogenetically diverse reductive dehalogenase-homologous genes in deep subseafloor sedimentary metagenomes

High frequency of phylogenetically diverse reductive dehalogenase-homologous genes in deep subseafloor sedimentary metagenomes

Retentive Memory of Bacteria: Long-Term Regulation of Dehalorespiration in Sulfurospirillum multivorans

Biochemical and genetic bases of dehalorespiration

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