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Identification of a bacterium that specifically catalyzes the reductive dechlorination of polychlorinated biphenyls with doubly flanked chlorines.

ABSTRACT: A microorganism whose growth is linked to the dechlorination of polychlorinated biphenyls (PCBs) with doubly flanked chlorines was identified. Identification was made by reductive analysis of community 16S ribosomal DNA (rDNA) sequences from a culture enriched in the presence of 2,3,4,5-tetrachlorobiphenyl (2,3,4,5-CB), which was dechlorinated at the para position. Denaturing gradient gel electrophoresis (DGGE) analysis of total 16S rDNA extracted from the culture led to identification of three operational taxonomic units (OTUs 1, 2, and 3). OTU 1 was always detected when 2,3,4,5-CB or other congeners with doubly flanked chlorines were present and dechlorinated. Only OTUs 2 and 3 were detected in the absence of PCBs and when other PCBs (i.e., PCBs lacking doubly flanked chlorines) were not dechlorinated. Partial sequences of OTUs 2 and 3 exhibited 98.2% similarity to the sequence of "Desulfovibrio caledoniensis" (accession no. DCU53465). A sulfate-reducing vibrio isolated from the culture generated OTUs 2 and 3. This organism could not dechlorinate 2,3,4,5-CB. From these results we concluded that OTU 1 represents the dechlorinating bacterium growing in a coculture with a Desulfovibrio sp. The 16S rDNA sequence of OTU 1 is most similar to the 16S rDNA sequence of bacterium o-17 (89% similarity), an ortho-PCB-dechlorinating bacterium. The PCB dechlorinator, designated bacterium DF-1, reductively dechlorinates congeners with doubly flanked chlorines when it is supplied with formate or H(2)-CO(2) (80:20).

SUBMITTER: Wu Q

PROVIDER: S-EPMC126686 | biostudies-literature | 2002 Feb

REPOSITORIES: biostudies-literature

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Identification of a bacterium that specifically catalyzes the reductive dechlorination of polychlorinated biphenyls with doubly flanked chlorines.

Wu Qingzhong Q Watts Joy E M JE Sowers Kevin R KR May Harold D HD

Applied and environmental microbiology 20020201 2

A microorganism whose growth is linked to the dechlorination of polychlorinated biphenyls (PCBs) with doubly flanked chlorines was identified. Identification was made by reductive analysis of community 16S ribosomal DNA (rDNA) sequences from a culture enriched in the presence of 2,3,4,5-tetrachlorobiphenyl (2,3,4,5-CB), which was dechlorinated at the para position. Denaturing gradient gel electrophoresis (DGGE) analysis of total 16S rDNA extracted from the culture led to identification of three ...[more]

PMID: 11823222

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Project description:Reductive dehalogenation mediated by organohalide-respiring bacteria plays a critical role in the global cycling of organohalides. Nonetheless, information on the dehalogenation enantioselectivity of organohalide-respiring bacteria remains limited. In this study, we report the enantioselective dechlorination of chiral polychlorinated biphenyls (PCBs) by Dehalococcoides mccartyi CG1. CG1 preferentially removed halogens from the (-)-enantiomers of the three major environmentally relevant chiral PCBs (PCB174, PCB149, and PCB132), and the enantiomer compositions of the dechlorination products depended on their parent organohalides. The in vitro assays with crude cell extracts or concentrated whole cells and the in vivo experiments with living cells showed similar enantioselectivities, in contrast with the distinct enantiomeric enrichment factors (εER) of the substrate chiral PCBs. Additionally, these results suggest that concentrated whole cells might be an alternative to crude cell extracts in in vitro tests of reductive dehalogenation activities. The enantioselective dechlorination of other chiral PCBs that we resolved via gas chromatography further confirmed the preference of CG1 for the (-)-enantiomers.IMPORTANCE A variety of agrochemicals and pharmaceuticals are chiral. Due to the enantioselectivity in biological processes, enantiomers of chiral compounds may have different environmental occurrences, fates, and ecotoxicologies. Many chiral organohalides exist in anaerobic or anoxic soils and sediments, and organohalide-respiring bacteria play a major role in the environmental attenuation and global cycling of these chiral organohalides. Therefore, it is important to investigate the dehalogenation enantioselectivity of organohalide-respiring bacteria. This study reports the discovery of enantioselective dechlorination of chiral PCBs by Dehalococcoides mccartyi CG1, which provides insights into the dehalogenation enantioselectivity of Dehalococcoides and may shed light on future PCB bioremediation efforts to prevent enantioselective biological side effects.

Project description:Microbial reductive dechlorination of polychlorinated biphenyls (PCBs) has been observed in many PCB-impacted sediments. However, this biodegradation is relatively site-specific and can be affected by PCB compositions and sediment geochemical conditions. To better understand the influence of a common competing electron acceptor, ferric oxyhydroxide (FeOOH), on dechlorination, two sediments (Hudson River and Grasse River sediments), and two PCB mixtures (PCB 5/12, 64/71, 105/114, and 149/153/170 in Mixture 1 and PCB 5/12, 64/71, 82/97/99, 144/170 in Mixture 2) were used for this microcosm study. The addition of 40 mmole/kg FeOOH completely inhibited PCB dechlorination in the Hudson sediment, but only moderately inhibited PCB dechlorination in the Grasse sediment with a 3-week longer lag time. The inhibitory effect in the Grasse sediment was mainly due to the loss of unflanked para dechlorination activity. Fe(II) analysis showed that dechlorination started prior to the consumption of Fe(III), which indicates PCB reduction and Fe(III) reduction were able to take place concurrently. Dehalococcoides 16S rRNA genes increased with the commencement of dechlorination in the Grasse sediment, but not in the completely inhibited Hudson sediment. Rare ortho dechlorination pathways were identified in FeOOH-amended Grasse sediment microcosms, dominated by transformations of PCB 25(24-3-CB) to PCB 13(3-4-CB) and PCB 28(24-4-CB) to PCB 15(4-4-CB). The addition of carbon sources (acetate or a fatty acid mixture with acetate, propionate, and butyrate) after 27 weeks of incubation reinitiated dechlorination in FeOOH-amended Hudson sediment microcosms. Also, the addition of carbon sources greatly enhanced ortho dechlorination in FeOOH-amended Grasse microcosms, indicating the utilization of acetate and/or the fatty acid mixture for ortho dechlorination-related microorganisms. A dechlorination pathway analysis approach revealed that para-flanked meta dechlorination was primarily preferred followed by ortho-/double-flanked meta dechlorination and single-/double-flanked para dechlorination in the Grasse sediment.

Dataset Information

Identification of a bacterium that specifically catalyzes the reductive dechlorination of polychlorinated biphenyls with doubly flanked chlorines.

Publications

Identification of a bacterium that specifically catalyzes the reductive dechlorination of polychlorinated biphenyls with doubly flanked chlorines.

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