Methyl-beta-cyclodextrin-enhanced solubilization and aerobic biodegradation of polychlorinated biphenyls in two aged-contaminated soils

The bioremediation of aged polychlorinated biphenyl (PCB)-contaminated soils is adversely affected by the low bioavailability of the pollutants. Randomly methylated-beta-cyclodextrins (RAMEB) were tested as a potential PCB-bioavailability-enhancing agent in the aerobic treatment of two aged-contaminated soils. The soils, contaminated by about 890 and 8500 mg/kg of Aroclor 1260 PCBs, were amended with biphenyl (4 g/kg), inorganic nutrients (to adjust their C:N ratio to 20:1), and variable amounts of RAMEB (0%, 0.5%, or 1.0% [w/w]) and treated in both aerobic 3-L solid-phase reactors and 1.5-L packed-bed loop reactors for 6 months. Notably, significant enhancement of the PCB biodegradation and dechlorination, along with a detectable depletion of the initial soil ecotoxicity, were generally observed in the RAMEB-treated reactors of both soils. RAMEB effects were different in the two soils, depending upon the treatment conditions employed, and generally increased proportionally with the concentration at which RAMEB was applied. RAMEB, which was slowly metabolized by the soil's aerobic microorganisms, was found to markedly enhance the occurrence of the indigenous aerobic, cultivable biphenyl-growing bacteria harboring genes homologous to those of two highly specialized PCB degraders (i.e., bphABC genes of Pseudomonas pseudoalcaligenes KF707 and bphA1A2A3A4BC1 genes of Rhodococcus globerulus P6) and chlorobenzoic acid-degrading bacteria as well as the occurrence of PCBs in the water phase of the soil reactors. These findings indicate that RAMEB enhanced the aerobic bioremediation of the two soils by increasing the bioavailability of PCBs and the occurrence of specialized bacteria in the soil reactors.     

Induction of glutathione-S-transferase in the Northern quahog Mercenaria mercenaria after exposure to the polychlorinated biphenyl (PCB) mixture Aroclor 1248

The glutathione-S-transferases (GSTs) from the Northern quahog (Mercenaria mercenaria) were examined after an injection of a polychlorinated biphenyl (PCB) mixture, Aroclor 1248, to a concentration of ~50 ppm. Enzymatic analysis indicated a fourfold increase in the GST activity of quahogs injected with PCBs compared with that of the control. An electrophoretic analysis of the GST from the PCB-exposed quahogs showed a 1.5-fold increase in the concentration over that of the control. Purification of the GST on a glutathione affinity column yielded a glutathione binding protein, in addition to the GSTs. However, the amount of the glutathione binding protein in the PCB-injected quahogs was found to decrease by ~50% in comparison to the glutathione binding protein in the control quahogs.     

Persistente chlororganische Verbindungen, Quecksilber und radioaktive Nuklide in Eiern von Rotmilanen (Milvus milvus) aus Sachsen-Anhalt

Zusammenfassung  Insgesamt wurden 60 Resteier des Rotmilans in drei Untersuchungsgebieten im südlichen Teil des Bundeslandes Sachsen-Anhalt zwischen 1987–1993 gesammelt und auf elf persistente Chlorkohlenwasserstoff-Verbindungen und Quecksilber sowie z. T. auf radioaktive Nuklide untersucht:

Effect of nutrient enrichment on the distribution and sedimentation of polychlorinated biphenyls (PCBs) in seawater

The effect of nutrient enrichment on the distribution of polychlorinated biphenyl's (PCBs) in the microbial food web and the residence time of PCBs in seawater was studied in an experimental mesocosm system. Two 5 m high temperature and light controlled mesocosm tubes (⊘ = 0,5 m) were filled with seawater from the northern Baltic Sea. Inorganic phosphorus and nitrogen were added daily to one mesocosm, while the other served as a control. Experiments were conducted at 5, 10 and 20°>C. Three ¹⁴C-labelled PCBs of different degree of chlorination were added to subsamples of the mesocosms: 4 chlorobiphenyl (MCB), IUPAC # 3; 2,2′,5,5′-tetrachlorobiphenyl (TCB), IUPAC # 52 and 2,2′, 4,4′,5,5′-hexachlorobiphenyl (HCB) IUPAC # 153. The biomasses and growth rates of the microorganisms as well as the sedimentation rate of particulate organic material increased with nutrient enrichment. The size distribution of the microorganisms changed with nutrient status, from dominance of picoplankton (< 2 μm) in the control towards increased importance of micro (> 10 μm) and nanoplankton (2– 10 μm) in nutrient enrichment. The specific growth rate of the bacterial community was found to be more temperature dependent than that of the phytoplankton community. The relative proportion of PCBs in the >2 μm fraction was observed to be in the order MCB < TCB < HCB, while the opposite distribution prevailed in the < 2 μm fraction. We hypothesize that this is due to the combined effect of the different Kow values of the PCBs and a different composition of the particulate organic carbon in the > 2 μm and < 2 μm fractions (e.g. different lipid composition). The residence time of the PCBs in the mesocosm generally decreased with nutrient enrichment, but was dependent on the degree of chlorination of the PCB. Our results indicate that the transport of organic pollutants up through the food web is more important in nutrient poor than in nutrient rich waters and that the importance of sedimentation is higher in eutrophic ecosystems. This revised version was published online in July 2006 with corrections to the Cover Date.     

A novel reductive graphene oxide‐based magnetic molecularly imprinted poly(ethylene‐co‐vinyl alcohol) polymers for the enrichment and determination of polychlorinated biphenyls in fish samples

The novel reductive graphene oxide‐based magnetic molecularly imprinted poly(ethylene‐co‐vinyl alcohol) polymers (rGO@m‐MIPs) were successfully synthesized as adsorbents for six kinds of polychlorinated biphenyls (PCBs) in fish samples. rGO@m‐MIPs was prepared by surface molecular imprinting technique. Besides, Fe₃O₄ nanoparticles (NPs) were employed as magnetic supporters, and rGO@Fe₃O₄ was in situ synthesis. Different from functional monomer and cross‐linker in traditional molecularly imprinted polymer, here, 3,4‐dichlorobenzidine was employed as dummy molecular and poly(ethylene‐co‐vinyl alcohol) was adopted as the imprinted polymers. After morphology and inner structure of the magnetic adsorbent were characterized, the adsorbent was employed for disperse solid phase extraction toward PCBs and exhibited great selectivity and high adsorption efficiency. This material was verified by determination of PCBs in fish samples combined with gas chromatography‐mass spectrometry (GC‐MS) method. According to the detection, the low detection limits (LODs) of PCBs were 0.0035–0.0070 µg l⁻¹ and spiked recoveries ranged between 79.90 and 94.23%. The prepared adsorbent can be renewable for at least 16 times and expected to be a new material for the enrichment and determination of PCBs from contaminated fish samples. Copyright © 2015 John Wiley & Sons, Ltd.     

Alginate beads as a storage, delivery and containment system for genetically modified PCB degrader and PCB biosensor derivatives of Pseudomonas fluorescens F113

Aims: Pseudomonas fluorescens F113Rifpcb is a genetically engineered rhizosphere bacterium with the potential to degrade polychlorinated biphenyls (PCBs). F113Rifpcbgfp and F113L::1180gfp are biosensor strains capable of detecting PCB bioavailability and biodegradation. The aim of this paper is to evaluate the use of alginate beads as a storage, delivery and containment system for use of these strains in PCB contaminated soils. Methods and Results: The survival and release of Ps. fluorescens F113Rifpcb from alginate beads were evaluated. Two Ps. fluorescens F113‐based biosensor strains were encapsulated, and their ability to detect 3‐chlorobenzoate (3‐CBA) and 3‐chlorobiphenyl (3‐CBP) degradation in soil was assessed. After 250 days of storage, 100% recovery of viable F113Rifpcb cells was possible. Amendments to the alginate formulation allowed for the timed release of the inoculant. Encapsulation of the F113Rifpcb cells provided a more targeted approach for the inoculation of plants and resulted in lower inoculum populations in the bulk soil, which may reduce the risk of unintentional spread of these genetically modified micro‐organisms in the environment. Encapsulation of the biosensor strains in alginate beads did not interfere with their ability to detect either 3‐CBA or 3‐CBP degradation. In fact, detection of 3‐CBP degradation was enhanced in encapsulated biosensors. Conclusions: Alginate beads are an effective storage and delivery system for PCB degrading inocula and biosensors. Significance and Impact of the Study: Pseudomonas fluorescens F113Rifpcb and the F113 derivative PCB biosensor strains have excellent potential for detecting and bioremediation of PCB contaminated soils. The alginate bead delivery system could facilitate the application of these strains as biosensors.     

Structural basis for preferential binding of non-ortho-substituted polychlorinated biphenyls by the monoclonal antibody S2B1

Polychlorinated biphenyls (PCBs) are a family of 209 isomers (congeners) with a wide range of toxic effects. In structural terms, they are of two types: those with and those without chlorines at the ortho positions (2, 2', 6 and 6'). Only 20 congeners have no ortho chlorines. Three of these are bound by the aryl hydrocarbon receptor and are one to four orders of magnitude more toxic than all others. A monoclonal antibody, S2B1, and its recombinant Fab have high selectivity and nanomolar binding affinities for two of the most toxic non-ortho-chlorinated PCBs, 3,4,3',4'-tetrachlorobiphenyl and 3,4,3',4',5'-pentachlorobiphenyl. To investigate the basis for these properties, we built a three-dimensional structure model of the S2B1 variable fragment (Fv) based on the high-resolution crystallographic structures of antibodies 48G7 and N1G9. Two plausible conformations for the complementarity-determining region (CDR) H3 loop led to two putative PCB-binding pockets with very different shapes (models A and B). Docking studies using molecular mechanics and potentials of mean force (PMF) indicated that model B was most consistent with the selectivity observed for S2B1 in competition ELISAs. The binding site in model B had a deep, narrow pocket between V(L) and V(H), with a slight constriction at the top that opened into a wider pocket between CDRs H1 and H3 on the antibody surface. This binding site resembles those of esterolytic antibodies that bind haptens with phenyl rings. One phenyl ring of the PCB fits into the deep pocket, and the other ring is bound in the shallower one. The bound PCB is surrounded by the side chains of TyrL91, TyrL96 and TrpH98, and it has a pi-cation interaction with ArgL46. The tight fit of the binding pocket around the ortho positions of the bound PCBs indicates that steric hindrance of ortho chlorines in the binding site, rather than induced conformational change of the PCBs, is responsible for the selectivity of S2B1.     

Structural and biochemical characterization of a novel aldehyde dehydrogenase encoded by the benzoate oxidation pathway in Burkholderia xenovorans LB400

The recently identified benzoate oxidation (box) pathway in Burkholderia xenovorans LB400 (LB400 hereinafter) assimilates benzoate through a unique mechanism where each intermediate is processed as a coenzyme A (CoA) thioester. A key step in this process is the conversion of 3,4-dehydroadipyl-CoA semialdehyde into its corresponding CoA acid by a novel aldehyde dehydrogenase (ALDH) (EC 1.2.1.x). The goal of this study is to characterize the biochemical and structural properties of the chromosomally encoded form of this new class of ALDHs from LB400 (ALDH_C) in order to better understand its role in benzoate degradation. To this end, we carried out kinetic studies with six structurally diverse aldehydes and nicotinamide adenine dinucleotide (phosphate) (NAD ⁺ and NADP ⁺). Our data definitively show that ALDH_C is more active in the presence of NADP ⁺ and selective for linear medium-chain to long-chain aldehydes. To elucidate the structural basis for these biochemical observations, we solved the 1.6-Å crystal structure of ALDH_C in complex with NADPH bound in the cofactor-binding pocket and an ordered fragment of a polyethylene glycol molecule bound in the substrate tunnel. These data show that cofactor selectivity is governed by a complex network of hydrogen bonds between the oxygen atoms of the 2′-phosphoryl moiety of NADP ⁺ and a threonine/lysine pair on ALDH_C. The catalytic preference of ALDH_C for linear longer-chain substrates is mediated by a deep narrow configuration of the substrate tunnel. Comparative analysis reveals that reorientation of an extended loop (Asn478-Pro490) in ALDH_C induces the constricted structure of the substrate tunnel, with the side chain of Asn478 imposing steric restrictions on branched-chain and aromatic aldehydes. Furthermore, a key glycine (Gly104) positioned at the mouth of the tunnel allows for maximum tunnel depth required to bind medium-chain to long-chain aldehydes. This study provides the first integrated biochemical and structural characterization of a box-pathway-encoded ALDH from any organism and offers insight into the catalytic role of ALDH_C in benzoate degradation.     

Microbial Degradation of Organic Pollutants in Soil in a Cold Climate

Cold-adapted microorganisms are potentially interesting for use in environmental biotechnology applications since a large part of the biosphere has low temperatures during at least parts of the year. Many studies have shown that both oil-contaminated and uncontaminated soils in the Arctic, the Antarctic and the Alps contain microbes that can degrade different hydrocarbons deriving from oils. A few studies have also been conducted on degradation of herbicides in soils at low temperatures. Furthermore, phenols and some polychlorinated biphenyl (PCB) congeners have proved to be degradable at low temperatures, using microorganisms isolated from sediments or soils. Additions of nitrogen and phosphorous to polluted soils have been shown to enhance the degradation of hydrocarbons in many cases. Bioaugmentation with hydrocarbon degrading cold-adapted microorganisms has given varying results. The inoculated microorganisms have probably been out-competed by the indigenous microorganisms in some cases. Different ways to increase the efficiency of microbial degradation of organic pollutants in soil in a cold climate is discussed.     

珠江三角洲池塘养殖中多氯联苯的环境归趋

对珠江三角洲4个不同地方的养殖池塘中水样,泥样和不同种类的鱼样分析,结果表明水产养殖中多氯联苯的分布和归趋在不同分配相中有很大差别,水相中含量较低(8.0～2 4 .0 3ng·L- 1 ) ,沉积相中较高(7.32～36 .2 3ng·g- 1 ,干重) ,特别是工业活动频繁的地方,而乡村相对较低。鱼类不同食性对多氯联苯的积累及其同分异构体的分布有很大影响,肉食性鱼类比草食性鱼类积累更多的多氯联苯.其中代表性同分异构体有IUPAC 118,138,81/ 87,15 3,180 ,5 2 ,4 9,99,4 4。珠江三角洲水产养殖中多氯联苯对生态环境的影响相对较低。