Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.4.16.2 (PCP)
 3,761 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A new membrane-associated 2,4,6-trichlorophenol reductive dehalogenase from Desulfitobacterium frappieri PCP-1 was isolated. Initial characterization of the crude preparation showed that the dechlorinating activity was sensitive to oxygen, and its optimum pH was 7.0. Its dechlorinating activity was not inhibited by sulphate, was completely inhibited by 1 mM sulphite, and partially inhibited by 5 mM sodium azide and by more than 5 mM nitrate. Several polychlorophenols were dechlorinated in the ortho position with respect to the hydroxy group. A dehalogenase was purified to apparent homogeneity. SDS gel electrophoresis revealed a single protein band with a molecular mass of 37 kDa. However, after two-dimensional gel electrophoresis, this band was composed of three isoforms. MS analyses showed that the three isoforms were from the same protein and the molecular mass of the most abundant isoform is 33800 Da. A mixture of iodopropane and titanium citrate caused a light-reversible inhibition of the dechlorinating activity, suggesting the involvement of a corrinoid cofactor. The apparent K(m) value for 2,4,6-trichlorophenol and pentachlorophenol were 18.3+/-2.8 microM and 26.8+/-2.9 microM respectively, at a methyl viologen concentration of 2 mM. The N-terminal amino acid sequence and an internal tryptic peptide sequence were determined. One open reading frame (ORF) was found in the Desulfitobacterium hafniense genome containing these peptides sequences. The corresponding ORF in D. frappieri PCP-1 was cloned and sequenced. This ORF, that we designated crdA, showed no homology with any known dehalogenase, suggesting a distinct reductive dehalogenase.
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PMID:Purification, cloning and sequencing of an enzyme mediating the reductive dechlorination of 2,4,6-trichlorophenol from Desulfitobacterium frappieri PCP-1. 1269 29

Desulfitobacterium hafniense PCP-1 (formerly frappieri PCP-1) has two reductive dehalogenases (RDases) that have been characterized. One is a membrane-associated 2,4,6-trichlorophenol RDase, which is encoded by crdA, and the other is a 3,5-dichlorophenol RDase encoded by cprA5. In this report, we determined the occurrence of these two RDase genes in seven other Desulfitobacterium strains. The presence or absence of these two RDases may explain the differences in the spectrum of halogenated compounds by these Desulfitobacterium strains. crdA gene sequences were found in all of the tested strains. It was expressed in strain PCP-1 regardless of the absence or presence of chlorophenols in the culture medium. crdA was also expressed in D. hafniense strains DCB-2 and TCE-1. cprA5 was detected only in D. hafniense strains PCP-1, TCP-A, and DCB-2. In these strains, cprA5 transcripts were detected only in the presence of chlorophenols. We also examined the expression of putative cprA RDases (cprA2, cprA3, and cprA4) that were shown to exist in the D. hafniense DCB-2 genome. RT-PCR experiments showed that cprA2, cprA3, and cprA4 were expressed in D. hafniense strains PCP-1, DCB-2, and TCP-A in the presence of chlorophenols. However, contrary to cprA5, these three genes were also expressed in the absence of halogenated compounds in the culture medium.
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PMID:Occurrence and expression of crdA and cprA5 encoding chloroaromatic reductive dehalogenases in Desulfitobacterium strains. 1654 Nov 58

Three chlorophenolic compounds (2-chlorophenol, 2,4,6-trichlorophenol, and pentachlorophenol) were tested to assess their effects on two soils with different properties: a granitic soil (Haplic Arenosol) and a calcareous one (Calcaric Regosol). Different concentrations of the pollutants (ranging from 0.001 to 10,000 mg kg(-1) soil, d.w.) were assayed for their effects on soil microbial activity and composition, using manometric respirometry and PCR-DGGE analysis, respectively. Other ecotoxicity tests such as Lactuca sativa seedling growth in the contaminated soils and algal growth inhibition (Pseudokirschneriella subcapitata) in their water extracts were done. The behaviour of the pollutants in the soils with respect to biodegradability and volatilization was also investigated. In the Haplic Arenosol, volatilization is the main process affecting 2-chlorophenol. Degradation and fixation of this compound in the soil matrix are favored in the Calcaric Regosol. This is the least toxic pollutant assayed. For 2,4,6-trichlorophenol, the soil pH is a critical parameter in the toxicity assays due to the neutral pKa of the compound. It is toxic in the soil microbial activity assay, but some recovery of the biotic processes can be observed, particularly in the Calcaric Regosol. This compound is more toxic in the Haplic Arenosol than in the Calcaric Regosol. Pentachlorophenol is ionized in both soils due to its low pKa, increasing its water solubility. It is highly toxic to the soil microbiota, thus inhibiting respiration, biodegradation and other biotic dissipation processes. Plant and alga tests, were more sensitive than soil microbial tests, except for PCP. The microbial populations tend to show changes at lower concentrations than the microbial activity. Some soil types (abundant in the Mediterranean area), with alkaline pH and fine textures could show higher level of ecotoxicity for ionizable organic pollutants than the soil type recommended by the OECD in ecotoxicity testing.
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PMID:Ecotoxicity of chlorophenolic compounds depending on soil characteristics. 2153 45

The binding interactions of lysozyme with 2-chlorophenol, 2,4-dichlorophenol, 2,4,6-trichlorophenol and pentachlorophenol were investigated by UV-vis absorption, CD, fluorescence, synchronous fluorescence, and three-dimensional fluorescence spectra techniques under physiological pH 7.40. The binding constants, quenching mechanism, and the number of binding sites were determined by the quenching of lysozyme fluorescence in presence of chlorophenols. H-bonds and hydrophobic interactions played major roles in stabilizing the chlorophenols-lysozyme complex. The distances r between chlorophenols and lysozyme were calculated to be 1.94nm, 2.75nm, 3.54nm, and 3.76nm for 2-CP, 2,4-DCP, 2,4,6-TCP, and PCP, respectively. The effects of chlorophenols on the conformation of lysozyme were analyzed using CD, synchronous fluorescence and three-dimensional fluorescence spectra.
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PMID:Investigation of the interaction between chlorophenols and lysozyme in solution. 2159 81

Relative to those of unexposed cultures, the transcript levels of the four CprA-type reductive dehalogenase genes (cprA2, cprA3, cprA4, and cprA5) in Desulfitobacterium hafniense PCP-1 were measured in cultures exposed to chlorophenols. In 2,4,6-trichlorophenol-amended cultures, cprA2 and cprA3 were upregulated, as was cprA5, but concomitantly with the appearance of 2,4-dichlorophenol (DCP). In 3,5-DCP-amended cultures, only cprA5 was upregulated. In pentachlorophenol-amended cultures grown for 12 h, cprA2 and cprA3 were upregulated but not cprA5. cprA4 was not upregulated significantly in cultures containing any tested chlorophenols.
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PMID:Quantitative analysis of the relative transcript levels of four chlorophenol reductive dehalogenase genes in Desulfitobacterium hafniense PCP-1 exposed to chlorophenols. 2174 10