BphK shows dechlorination activity against 4-chlorobenzoate,an end product of bph-promoted degradation of PCBs

A bphK gene encoding glutathione S-transferase (GST) activity is located in the bph operon in Burkholderia sp. strain LB400 but its role in polychlorinated biphenyl (PCB) metabolism is unknown. This gene was over-expressed in Escherichia coli and an in vivo assay based on growth of E. coli containing GST activity was used to identify potential novel substrates for this enzyme. Using this assay, 4-chlorobenzoate (4-CBA) was identified as a substrate for the BphK enzyme. High pressure liquid chromatography analysis and chloride ion detection showed removal of 4-CBA and an equivalent increase of chloride in cell extracts when incubated with this enzyme. These results would indicate that this BphK enzyme has dechlorination activity in relation to 4-CBA and may have a role in protection of other Bph enzymes against certain chlorinated metabolites of PCB degradation.     

Dehalogenation of chlorinated aromatic compounds using a hybrid bioinorganic catalyst on cells of <Emphasis Type="Italic">Desulfovibrio desulfuricans</Emphasis>

A novel bioinorganic catalyst was obtained via reduction of Pd(II) to Pd⁰ on to the surface of cells of Desulfovibrio desulfuricans at the expense of H₂. Palladised biomass, supplied with formate or H₂ as an electron donor, catalysed the dehalogenation of 2-chlorophenol and polychlorinated biphenyls. In the example of 2,3,4,5-tetrachlorobiphenyl, the bioinorganic catalyst promoted a rate of chloride release of 9.33 ± 0.17 nmol min^–1 mg ^–1and only ~5% of this value was obtained using chemically reduced or commercially available Pd ⁰. In the case of 2,2,4,4,6,6-hexachlorobiphenyl the rate was more than four orders of magnitude faster than the degradation reported using a sulfidogenic culture. Negligible chloride release occurred from any of the chloroaromatic compounds using biomass alone, or from palladised biomass challenged with hexane carrier solvent only. Analysis of the spent solution showed that in addition to catalysis of reductive dehalogenation the new material was able to remove very effectively the organic residua, with neither any PCB nor any breakdown products identifiable by GC/MS.Revisions requested 8 September 2004; Revisions received 21 October 2004;     

Fluorescence resonance energy transfer (FRET) based molecular detection of a genetically modified PCB degrader in soil

Genetic analysis of the location of a mini-Tn5 promoted insertion of the LB400 bph operon in the rhizosphere coloniser Pseudomonas fluorescens F113rifPCB, allowed the development of a specific PCR detection system based on the unique DNA sequence at this insertion site. Real time PCR using both SYBR green chemistry and Fluorescence Resonance Energy Transfer probes allowed the precise identification of the recombinant strain and its quantitative detection in soil microcosms over a (bacteria/g) range of five orders of magnitude. This new assay can detect the genetically modified microorganism from soil in less than 90 min and at levels below the detection limits of standard PCR or cultivable counts on selective media.     

长江口两种江豚亚种体脂内多氯联苯同分异构体含量的比较分析

为了弄清长江口江豚(Neophocaena asiaeorientalis)体脂内多氯联苯(PCBs)的累积情况,采用气相色谱质谱联用仪(GC-MS)对5头东亚亚种(N.asiaeorientalis sunameri)和3头长江亚种(N.asiaeorientalis asiaeorientalis)的10种多氯联苯同族物(PCBs)含量进行了测定。研究发现,长江口江豚体脂中多氯联苯(PCBs)的平均含量为3.38μg/g(湿重),其中,长江亚种的多氯联苯(PCBs)含量(3.40~12.67μg/g)要显著高于东亚亚种(0.33~2.21μg/g)。在10种多氯联苯(PCBs)中,一氯、二氯和三氯联苯在所有个体中均未测出(小于0.005μg/g),但四氯、五氯、六氯和七氯联苯在所有个体中都被测出,其中,六氯联苯的平均含量高达1.48μg/g,占总含量的43.79%。从含量的组成看,六氯联苯和七氯联苯均构成了长江口江豚体脂内多氯联苯(PCBs)的主要成分,但相比东亚亚种,长江亚种的四氯联苯和五氯联苯也占较高的比例。研究表明,低氯代组分的多氯联苯(PCBs)可以随水流的稀释和生物转化而逐渐降解,但高氯代组分的多氯联苯(PCBs)则不易降解,并且随江豚年龄的增长而快速积累,产生更持久的毒性效应。     

Transformation of polychlorinated biphenyls by a novel BphA variant through the meta-cleavage pathway

The transformation of 20 polychlorinated biphenyls (PCBs) through the meta-cleavage pathway by recombinant Escherichia coli cells expressing the bphEFGBC locus from Burkholderia cepacia LB400 and the bphA genes from different sources was compared. The analysis of PCB congeners for which hydroxylation was observed but no formation of the corresponding yellow meta-cleavage product demonstrated that only lightly chlorinated congeners including one tetrachlorobiphenyl (2,2',4,4'-CB) were transformed into their corresponding yellow meta-cleavage products. Although many other tetrachlorobiphenyls (2, 2',5,5'-CB, 2,2',3,5'-CB, 2,4,4',5-CB, 2,3',4',5-CB, 2,3',4,4'-CB) and one pentachlorobiphenyl (2,2',4,5,5'-CB) tested were depleted from resting cell suspensions, no yellow meta-cleavage products were observed. For most of these congeners, dihydrodiol compounds accumulated as the endproducts, indicating that the bphB-encoded biphenyl-2,3-dihydrodiol-2,3-dehydrogenase is a key limiting step for further degradation of highly chlorinated congeners. These results suggest that engineering the biphenyl dioxygenase alone is insufficient for an improved removal of PCB. Rather, improved degradation of PCBs is more likely to be achieved with recombinant strains containing metabolic pathways not only specifically engineered for expanding the initial dioxygenation but also for the mineralization of PCBs.     

Cometabolic degradation of polychlorinated biphenyls (PCBs) by axenic cultures of Ralstonia sp. strain SA-5 and Pseudomonas sp. strain SA-6 obtained from Nigerian contaminated soils

Substantial metabolism of 2,3,4,5-tetrachlorobiphenyl (2,3,4,5-tetraCB) and 2,3′,4′,5-tetraCB by axenic cultures of Ralstonia sp. SA-5 and Pseudomonas sp. SA-6 was observed in the presence of biphenyl supplementation, although, the strains were unable to utilize tetrachlorobiphenyls as growth substrate. The former was more amenable to aerobic degradation (∼70% degradation) than the latter (22–45% degradation). Recovery of 2,5-chlorobenzoic acid and chloride from 2,3′,4′,5-tetraCB assay is an indication of initial dioxygenase attack on the 3,4-dichlorophenyl ring. The PCB-degradative ability of both strains was also investigated by GC analysis of individual congeners in Aroclor 1242 (100 ppm) following 12-day incubation with washed benzoate-grown cells. Results revealed two different catabolic properties. Whereas strain SA-6 required biphenyl as inducer of the degradation activity, such induction was not required by strain SA-5. Nearly all the detectable congeners in the mixture were extensively degraded (% reduction in ECD area counts for individual congeners ranged from 50.0 to 100% and 14.2 to 100%, respectively, for SA-5 and SA-6). The two strains exhibited no noticeable specificity for congeners with varying numbers of chlorine substitution and positions. The degradative competence of these isolates most especially SA-5 makes them among the most versatile PCB-metabolizing organisms yet reported.     

植物修复多氯联苯研究进展

综述了植物修复持久性有机污染物多氯联苯（PCBs）的研究进展,重点阐述了植物对PCBs的去除作用和机理,植物在从环境中去除PCBs的过程中,不仅仅是作为微生物降解的支持者,而且还作为积极的参与者对PCBs进行代谢：一方面植物通过根系从环境中吸收和积累PCBs,并将吸收的PCBs转化为非毒性的代谢产物累积于植物组织中;另一方面植物释放促进PCBs降解的酶直接降解PCBs,或释放根系分泌物,增加根际微生物的数量,提高其活性间接降解PCBs.文中对植物修复PCBs的影响因素如植物组织培养的类型、生物量、PCBs的初始浓度以及PCBs的类型、理化性质等进行了讨论.     

Identification of hot spots within harbour sediments through a new cumulative hazard index. Case study: Port of Bari,Italy

While numerous quality guidelines unquestionably drive the management of harbour sediments, port managers increasingly require easy-to-use tools supporting them in assessing the sediment quality. In this work, a new hazard index, named cumulative Normalized and Weighted Average Concentration (c_NWAC), is proposed, considering the concentrations of main hazardous, toxic and bio-accumulative sediment contaminants. This index is an upgraded version of the previously introduced NWAC index, which considered only the metal concentrations. The c_NWAC values range from 0 to 10 scores, and their visualization on a colour base code map leads to an easy identification of hotspots. The applicability of the new index was verified using a dataset derived from the analyses of 42 samples collected at different depths of the seabed of the Port of Bari (Italy). The concentrations of 58 parameters were considered, namely 11 metals and metalloids, 17 congeners of polychlorinated biphenyls (PCBs), 16 congeners of polycyclic aromatic hydrocarbons (PAHs), further 11 organic micropollutants, total organic carbon (TOC) and nutrients (N_tot, P_tot). For the study area, the obtained c_NWAC values resulted well correlated to the corresponding values of the commonly used mean Effects Range Median quotient (mERMq), and similar hazard categories were developed for the new index, even if the latter do not necessarily explain the real sediment toxicity, because they were derived from a different sediment dataset and different test species.Moreover, the hazard predictive ability of the new index was verified (with satisfactory results) by testing the considered samples by eco-toxicological assays, using three different biological species, and comparing the obtained results to the corresponding hazard categories developed for c_NWAC.     

Dioxin-like Polychlorinated Biphenyls Contamination and Distribution in Soils from the Modern Yellow River Delta,China

The concentrations of 12 dioxin-like Polychlorinated Biphenyls (dl-PCBs) were determined from 37 soil samples collected from the modern Yellow River Delta (China) in March 2006. The PCB concentrations were determined by dual capillary GC-ECD. The total dl-PCBs concentrations were in the range of 108.7 to 2153 ng/kg with an average of 800.5 ng/kg. The toxic equivalency (TEQ) in this area varies from 2.3 × 10^{? 5} to 9.5 × 10^{? 5} ng-TEQ/kg, so the delta is slightly polluted by PCBs. PCB118, the main congener, accounted for 46.7% of the total dl-PCB. More than 60% dl-PCBs were penta-PCBs. The spatial distribution of the dl-PCBs is discussed in detail. The distribution of the PCBs suggests that water runoff is the main source of the total contaminant burden in the delta, and that atmospheric precipitation plays an important role.     

多氯联苯在模拟水生态系统中的分布、积累与迁移动态研究

利用微宇宙模拟水生态系统对多氯联苯在水体环境中的行为 ,包括多氯联苯在水体、沉积物、生物体 (皇冠草、河蚬、鲢 )不同分配相及生物体不同组织、器官中含量、分布以及迁移、富集等时间变化动态过程进行了研究。结果表明 ,水草对PCB有一定程度的吸收 ;河蚬对PCBs有明显的吸收积累 ,并且吸收达到平衡的时间较长。鱼体对PCB的吸收积累在不同组织中有明显的差别 ,在内脏和肌肉组织中积累较高 ,鳃组织中积累较低。