Reductive dechlorination pathways for substituted benzenes: a correlation with electronic properties

Electronic properties were correlated with observed reductive dechlorination pathways by unacclimated consortia for chlorinated phenols, dihydroxybenzenes, benzoic acids, and anilines. Molecular structures and properties were calculated using the semi-empirical Modified Neglect of Differential Overlap method at the Cornell Supercomputing Facility. Observed preferential positions for reductive dechlorination by unacclimated consortia correlate well with the largest negative value for the carbon-chlorine bond charge. Of 16 dechlorination pathways observed for unacclimated bacteria, the most negative carbon-chlorine bond charge correlated with 15 pathways.This correlation between the observed dechlorination position and the parent compound's electronic properties is consistent with the observed reductive dechlorination of chlorophenols and chlorinated dihydroxybenzenes at the ortho position, and the meta dechlorination of chlorobenzoic acids. Net carbonchlorine bond charges also correlate with the preferred dechlorination position for two of three known chloroaniline pathways, suggesting preferential removal of chlorines from the ortho position of chloroanilines.Abbreviations CA chloroaniline - CBz chlorobenzoic acid - CC chlorocatechol - CP chlorophenol - DCA dichloroaniline - DCBz dichlorobenzoic acid - DCC dichlorocatechol - DCH dichlorohydroquinone - DCP dichlorophenol - DCR dichlororesorcinol - PCP pentachlorophenol - TCA trichloroaniline - TCBz trichlorobenzoic acid - TCC trichlorocatechol - TCH trichlorohydroquinone - TCP trichlorophenol - TCR trichlororesorcinol - TeCA tetrachloroaniline - TeCBz tetrachlorobenzoic acid - TeCC tetrachlorocatechol - TeCH tetrachlorohydroquinone - TeCP tetrachlorophenol - TeCR tetrachlororesorcinol     

Redox Potential as a Parameter To Predict the Reductive Dechlorination Pathway of Chloroanilines in Anaerobic Environments

Abstract The hypothesis that the microbially catalyzed pathway proceeds with a step that would yield the highest energy was examined for the reductive dechlorination of chloroanilines (CAs) under anaerobic conditions. The Gibbs free energy of formation was estimated with Benson's method, then the redox potentials were determined using an H⁺/H₂ couple as the reference system. The observed pathways were compared using the redox potential of the reaction, and the results showed that the redox potential correctly predicts the pathway that yields the highest energy for the dechlorination step. Received: 5 April 1996; Accepted: 9 August 1996     

Regiospecificity of chlorophenol reductive dechlorination by vitamin b(12s)

Vitamin B(12), reduced by titanium (III) citrate to vitamin B(12s), catalyzes the reductive dechlorination of chlorophenols. Reductive dechlorination of pentachlorophenol and of all tetrachlorophenol and trichlorophenol isomers was observed. Reaction of various chlorophenols with vitamin B(12) favored reductive dechlorination at positions adjacent to another chlorinated carbon, but chlorines ortho to the hydroxyl group of a phenol were particularly resistant to reductive dechlorination, even if they were also ortho to a chlorine. This resulted in a reductive dechlorination pattern favoring removal of para and meta chlorines, which differs substantially from the pattern exhibited by anaerobic microbial consortia.     

Modeling Natural Attenuation of Chlorinated Ethenes Under Spatially Varying Redox Conditions

A three-dimensional model for the transport and reductive dechlorination of chlorinated ethenes in ground-water systems with variable redox conditions is demonstrated and applied to a pilot test for accelerated natural attenuation of trichloroethene (TCE). The rate and extent of biotransformation of TCE and chlorinated progeny is controlled by the dominant terminal electron accepting process (TEAP) that is simulated over space and time. The solute transport code, Sequential Electron Acceptor Model, 3D-transport, (SEAM3D) which simulates aerobic and sequential anaerobic biodegradation of organic carbon, is modified to implement the equations. Results of a generic model for TCE transport in ground-water systems with different redox conditions demonstrate that the degree of chlorinated ethene attenuation is influenced by background concentrations of aqueous- and solid-phase electron acceptors, but that model results are sensitive to other input parameters (inhibition coefficients, maximum rate of reductive dechlorination, biomass concentrations, and ground-water velocity). Simulation results of enhanced in situ bioremediation using dissolved organic carbon as a reducing agent show that spatial and temporal changes in the dominant TEAP and the subsequent rate of reductive dechlorination are adequately represented with the model. Initial concentrations of Fe(III) and the dechlorinating microbial population influence the simulated time lag observed during the pilot test.     

Regiospecificity of Chlorophenol Reductive Dechlorination by Vitamin B12s

Vitamin B₁₂, reduced by titanium (III) citrate to vitamin B_12s, catalyzes the reductive dechlorination of chlorophenols. Reductive dechlorination of pentachlorophenol and of all tetrachlorophenol and trichlorophenol isomers was observed. Reaction of various chlorophenols with vitamin B₁₂ favored reductive dechlorination at positions adjacent to another chlorinated carbon, but chlorines ortho to the hydroxyl group of a phenol were particularly resistant to reductive dechlorination, even if they were also ortho to a chlorine. This resulted in a reductive dechlorination pattern favoring removal of para and meta chlorines, which differs substantially from the pattern exhibited by anaerobic microbial consortia.     

Microbial reductive dehalogenation of polychlorinated biphenyls

Under anaerobic conditions, microbial reductive dechlorination of polychlorinated biphenyls (PCBs) occurs in soils and aquatic sediments. In contrast to dechlorination of supplemented single congeners for which frequently ortho dechlorination has been observed, reductive dechlorination mainly attacks meta and/or para chlorines of PCB mixtures in contaminated sediments, although in a few instances ortho dechlorination of PCBs has been observed. Different microorganisms appear to be responsible for different dechlorination activities and the occurrence of various dehalogenation routes. No axenic cultures of an anaerobic microorganism have been obtained so far. Most probable number determinations indicate that the addition of PCB congeners, as potential electron acceptors, stimulates the growth of PCB-dechlorinating microorganisms. A few PCB-dechlorinating enrichment cultures have been obtained and partially characterized. Temperature, pH, availability of naturally occurring or of supplemented carbon sources, and the presence or absence of H(2) or other electron donors and competing electron acceptors influence the dechlorination rate, extent and route of PCB dechlorination. We conclude from the sum of the experimental data that these factors influence apparently the composition of the active microbial community and thus the routes, the rates and the extent of the dehalogenation. The observed effects are due to the specificity of the dehalogenating bacteria which become active as well as changing interactions between the dehalogenating and non-dehalogenating bacteria. Important interactions include the induced changes in the formation and utilization of H(2) by non-dechlorinating and dechlorinating bacteria, competition for substrates and other electron donors and acceptors, and changes in the formation of acidic fermentation products by heterotrophic and autotrophic acidogenic bacteria leading to changes in the pH of the sediments.     

Enhanced dechlorination of 2,3-chlorophenol in an anaerobic bioreactor by addition of auxiliary carbon sources

Addition of an auxiliary carbon source (sodium acetate, sodium lactate, or yeast extract) to microbial consortia from an anaerobic bioreactor stimulated reductive dechlorination of 2,3-chlorophenol (CP) compared to unamended consortia or consortia amended with sodium formate or sodium propionate. However, no significant effects on dechlorination of 3-CP were observed by addition of auxiliary carbon sources. In a continuous-flow, anaerobic bioreactor, addition of sodium lactate resulted in a 2 to 3-fold increase in the rate of dechlorination of 2,3-CP compared to addition of sodium formate. Enhanced transformation of the dechlorination product (3-CP) was also evident. © Rapid Science Ltd. 1998     

Transformation of carbon tetrachloride under sulfate reducing conditions

The removal of carbon tetrachloride under sulfate reducing conditions was studied in an an aerobic packed-bed reactor. Carbon tetrachloride, up to a concentration of 30 μM, was completely converted. Chloroform and dichloromethane were the main transformation products, but part of the carbon tetrachloride was also completely dechlorinated to unknown products. Gram-positive sulfate-reducing bacteria were involved in the reductive dechlorination of carbon tetrachloride to chloroform and dichloromethane since both molybdate, an inhibitor of sulfate reduction, and vancomycin, an inhibitor of gram-positive bacteria completely inhibited carbon tetrachloride transformation. Carbon tetrachloride transformation by these bacteria was a cometabolic process and depended on the input of an electron donor and electron acceptor (sulfate). The rate of carbon tetrachloride transformation by sulfate reducing bacteria depended on the type of electron donor present. A transformation rate of 5.1 nmol·ml^-1·h^-1 was found with ethanol as electron donor. At carbon tetrachloride concentrations higher than18 μM, sulfate reduction and reductive dechlorination of carbon tetrachloride decreased and complete inhibition was observed at a carbon tetrachloride concentration of 56.6 μM. It is not clear what type of microorganisms were involved in the observed partial complete dechlorination of carbon tetrachloride. Sulfate reducing bacteria probably did not play a role since inhibition of these bacteria with molybdate had no effect on the complete dechlorination of carbon tetrachloride. This revised version was published online in August 2006 with corrections to the Cover Date.     

Transition Metal Catalyst-Assisted Reductive Dechlorination of Perchloroethylene by Anaerobic Aquifer Enrichments

Bioremediation of groundwater contaminated with chlorinated solvents, such as perchloroethylene (PCE) or carbon tetrachloride, can be accomplished by adding nutrients to stimulate a microbial community capable of reductive dechlorination. However, biotransformation of these solvents, especially PCE, typically occurs very slowly or not at all. Experiments were conducted to evaluate whether the addition of transition metal tetrapyrrole catalysts would increase the reductive transformation of PCE to trichloroethylene (TCE) by sulfate-reducing enrichment cultures. Batch assays were used to test vitamin B₁₂ and two synthetic sulfonatophenyl porphine catalysts for the stimulation of reductive dechlorination of PCE by sulfate-reducing bacteria (SRB) enriched from aquifer sediments from two locations at Dover Air Force Base. Cells from the enrichments were concentrated and added to batch assay vials. Vials containing SRB cells amended with vitamin B12 exhibited enhanced transformation of PCE to TCE compared with reactors amended with either synthetic catalysts or reactors containing cells alone. Methane production was observed in reactors that exhibited maximum levels of dechlorination. Storage of aquifer sediments between enrichments led to decreased levels of PCE dechlorination in subsequent assays.     

Sequential anaerobic/aerobic biodegradation of chloroethenes--aspects of field application

Because of a range of different industrial activities, sites contaminated with chloroethenes are a world-wide problem. Chloroethenes can be biodegraded by reductive dechlorination under anaerobic conditions as well as by oxidation under aerobic conditions. The tendency of chloroethenes to undergo reductive dechlorination decreases with a decreasing number of chlorine substituents, whereas with less chlorine substituents chloroethenes more easily undergo oxidative degradation. There is currently a growing interest in aerobic metabolic degradation of chloroethenes, which demonstrates advantages compared to cometabolic degradation pathways. Sequential anaerobic/aerobic biodegradation can overcome the disadvantages of reductive dechlorination and leads to complete mineralization of the chlorinated pollutants. This approach shows promise for site remediation in natural settings and in engineered systems.     

Synchronous anaerobic and aerobic degradation of DDT by an immobilized mixed culture system

Summary For the investigation of a mixed anaerobic and aerobic degradation of xenobiotics the reductive dechlorination of 1,1,1-trichloro-2,2-bis (4-chlorophenyl)ethane (DDT) to 1,1-dichloro-2,2-bis (4-chlorophenyl)ethane (DDD) and the oxidative degradation of the DDT-conversion product 4,4-dichlorodiphenylmethane (DDM) were studied. Enrichments from digested sewage sludge led to the isolation of an Enterobacter cloacae-strain which is able to reductive dechlorination of DDT during the fermentation of lactose. From fresh sewage sludge 11 bacterial strains were isolated in batch-culture and in continuous culture utilizing diphenylmethane, a non chlorinated structural analogon of DDM, as sole source of carbon and energy. One of these isolates, Alcaliaenes sp. cometabolizes DDM during the aerobic growth with diphenylmethane. By coimmobilization of Alcaligenes sp. and Enterobacter cloacae in Ca-alginate a system could be established, in which the reductive dechlorination of DDT and the oxidative degradation of DDM and diphenylmethane proceeds simultaneously in one reactor vessel.     

Influence of Substituents on Reductive Dehalogenation of 3-Chlorobenzoate Analogs

The biochemical effects of aryl substituents on the reductive dechlorination of 3-chlorobenzoate analogs were quantified with (i) a stable 3-chlorobenzoate-grown methanogenic sludge enrichment, (ii) Desulfomonile tiedjei DCB-1, isolated from this enrichment and able to catalyze the reductive dechlorination of 3-chlorobenzoate, and (iii) a defined 3-chlorobenzoate-degrading methanogenic consortium with D. tiedjei as the key dechlorinating organism. The addition of hydrogen stimulated the dechlorination rate in the consortium. The extent of this stimulation depended on the substituent. The data were evaluated with various sets of substituent constants compiled for the Hammett equation. None of the sets yielded a satisfactory correlation between experimental values and theoretical constants. This suggests that the microbially catalyzed reductive dechlorination of 3-chlorobenzoate cannot be described simply as either a nucleophilic or an electrophilic substitution reaction. Nevertheless, observations that the presence of a para-amino or -hydroxy group inhibited the rate of dechlorination suggest that the rate-limiting step in the reductive dechlorination of 3-chlorobenzoate is a nucleophilic attack on the negatively charged π electron cloud around the benzene nucleus.     

Formulation of the CBC-Model for Modelling the Contaminants and Footprints in Natural Attenuation of BTEX

The Refuse Hideaway Landfill (23-acre) received municipal, commercial, and industrial waste between 1974 and 1988. It was designed as a "natural attenuation" landfill and no provision was made to collect and treat contaminated water. Natural biological degradation through sequential reductive dechlorination had been an important mechanism for natural attenuation at the site. We used the concentration of hydrogen to forecast whether reductive dechlorination would continue over time at particular locations in the plume. Based on published literature, reductive dechlorination and natural attenuation of PCE, TCE, and cis-DCE can be expected in the aquifer if the concentration of molecular hydrogen in monitoring wells are adequate (> 1 nanomolar). Reductive dechlorination can be expected to continue as the ground water moves down gradient. Natural attenuation through reductive dechlorination is not expected in flow paths that originate at down gradient monitoring wells with low concentrations of molecular hydrogen (< 1 nanomolar). In three monitoring wells at the margin of the landfill and in five monitoring wells down gradient of the landfill, ground water maintained a molecular hydrogen concentration, ranging from 1.30 to 9.17 nanomolar, that is adequate for reductive dechlorination. In three of the monitoring wells far down gradient of the landfill, the concentration of molecular hydrogen (0.33 to 0.83 nanomolar) was not adequate to support reductive dechlorination. In wells with adequate concentrations of hydrogen, the concentrations of chlorinated volatile organic compounds were attenuated over time, or concentrations of chlorinated volatile organics were below the detection limit. In wells with inadequate concentrations of hydrogen, the concentrations of chlorinated organic compounds attenuated at a slower rate over time. In wells with adequate hydrogen the first order rate of attenuation of PCE, TCE, cis-DCE and total chlorinated volatile organic compounds varies from 0.38 to 0.18 per year. In wells without adequate hydrogen the rate varies from 0.015 to 0.006 per year.     

Use of Silicate Minerals for pH Control during Reductive Dechlorination of Chloroethenes in Batch Cultures of Different Microbial Consortia

In chloroethene-contaminated sites undergoing in situ bioremediation, groundwater acidification is a frequent problem in the source zone, and buffering strategies have to be implemented to maintain the pH in the neutral range. An alternative to conventional soluble buffers is silicate mineral particles as a long-term source of alkalinity. In previous studies, the buffering potentials of these minerals have been evaluated based on abiotic dissolution tests and geochemical modeling. In the present study, the buffering potentials of four silicate minerals (andradite, diopside, fayalite, and forsterite) were tested in batch cultures amended with tetrachloroethene (PCE) and inoculated with different organohalide-respiring consortia. Another objective of this study was to determine the influence of pH on the different steps of PCE dechlorination. The consortia showed significant differences in sensitivities toward acidic pH for the different dechlorination steps. Molecular analysis indicated that Dehalococcoides spp. that were present in all consortia were the most pH-sensitive organohalide-respiring guild members compared to Sulfurospirillum spp. and Dehalobacter spp. In batch cultures with silicate mineral particles as pH-buffering agents, all four minerals tested were able to maintain the pH in the appropriate range for reductive dechlorination of chloroethenes. However, complete dechlorination to ethene was observed only with forsterite, diopside, and fayalite. Dissolution of andradite increased the redox potential and did not allow dechlorination. With forsterite, diopside, and fayalite, dechlorination to ethene was observed but at much lower rates for the last two dechlorination steps than with the positive control. This indicated an inhibition effect of silicate minerals and/or their dissolution products on reductive dechlorination of cis-dichloroethene and vinyl chloride. Hence, despite the proven pH-buffering potential of silicate minerals, compatibility with the bacterial community involved in in situ bioremediation has to be carefully evaluated prior to their use for pH control at a specific site.     

Isolation and characterization of Desulfovibrio dechloracetivorans sp. nov., a marine dechlorinating bacterium growing by coupling the oxidation of acetate to the reductive dechlorination of 2-chlorophenol

Strain SF3, a gram-negative, anaerobic, motile, short curved rod that grows by coupling the reductive dechlorination of 2-chlorophenol (2-CP) to the oxidation of acetate, was isolated from San Francisco Bay sediment. Strain SF3 grew at concentrations of NaCl ranging from 0.16 to 2.5%, but concentrations of KCl above 0. 32% inhibited growth. The isolate used acetate, fumarate, lactate, propionate, pyruvate, alanine, and ethanol as electron donors for growth coupled to reductive dechlorination. Among the halogenated aromatic compounds tested, only the ortho position of chlorophenols was reductively dechlorinated, and additional chlorines at other positions blocked ortho dechlorination. Sulfate, sulfite, thiosulfate, and nitrate were also used as electron acceptors for growth. The optimal temperature for growth was 30 degrees C, and no growth or dechlorination activity was observed at 37 degrees C. Growth by reductive dechlorination was revealed by a growth yield of about 1 g of protein per mol of 2-CP dechlorinated, and about 2.7 g of protein per mole of 2,6-dichlorophenol dechlorinated. The physiological features and 16S ribosomal DNA sequence suggest that the organism is a novel species of the genus Desulfovibrio and which we have designated Desulfovibrio dechloracetivorans. The unusual physiological feature of this strain is that it uses acetate as an electron donor and carbon source for growth with 2-CP but not with sulfate.     

Isolation and Characterization of Desulfovibrio dechloracetivorans sp. nov., a Marine Dechlorinating Bacterium Growing by Coupling the Oxidation of Acetate to the Reductive Dechlorination of 2-Chlorophenol

Strain SF3, a gram-negative, anaerobic, motile, short curved rod that grows by coupling the reductive dechlorination of 2-chlorophenol (2-CP) to the oxidation of acetate, was isolated from San Francisco Bay sediment. Strain SF3 grew at concentrations of NaCl ranging from 0.16 to 2.5%, but concentrations of KCl above 0.32% inhibited growth. The isolate used acetate, fumarate, lactate, propionate, pyruvate, alanine, and ethanol as electron donors for growth coupled to reductive dechlorination. Among the halogenated aromatic compounds tested, only the ortho position of chlorophenols was reductively dechlorinated, and additional chlorines at other positions blocked ortho dechlorination. Sulfate, sulfite, thiosulfate, and nitrate were also used as electron acceptors for growth. The optimal temperature for growth was 30°C, and no growth or dechlorination activity was observed at 37°C. Growth by reductive dechlorination was revealed by a growth yield of about 1 g of protein per mol of 2-CP dechlorinated, and about 2.7 g of protein per mole of 2,6-dichlorophenol dechlorinated. The physiological features and 16S ribosomal DNA sequence suggest that the organism is a novel species of the genus Desulfovibrio and which we have designated Desulfovibrio dechloracetivorans. The unusual physiological feature of this strain is that it uses acetate as an electron donor and carbon source for growth with 2-CP but not with sulfate.     

Factors controlling the carbon isotope fractionation of tetra- and trichloroethene during reductive dechlorination by Sulfurospirillum ssp. and Desulfitobacterium sp. strain PCE-S

Carbon stable isotope fractionation of tetrachloroethene (PCE) and trichloroethene (TCE) was investigated during reductive dechlorination. Growing cells of Sulfurospirillum multivorans, Sulfurospirillum halorespirans, or Desulfitobacterium sp. strain PCE-S, the respective crude extracts and the abiotic reaction with cyanocobalamin (vitamin B(12)) were used. Fractionation of TCE (alphaC=1.0132-1.0187) by S. multivorans was more than one order of magnitude higher than values previously observed for tetrachloroethene (PCE) (alphaC=1.00042-1.0017). Similar differences in fractionation were observed during reductive dehalogenation by the close relative S. halorespirans with alphaC=1.0046-1.032 and alphaC=1.0187-1.0229 for PCE and TCE respectively. TCE carbon isotope fractionation (alphaC=1.0150) by the purified PCE-reductive dehalogenase from S. multivorans was more than one order of magnitude higher than fractionation of PCE (alphaC=1.0017). Carbon isotope fractionation of TCE by Desulfitobacterium sp. strain PCE-S (alphaC=1.0109-1.0122) as well as during the abiotic reaction with cyanocobalamin (alphaC=1.0154) was in a similar range to previously reported values for fractionation by mixed microbial cultures. In contrast with previous results with PCE, no effects due to rate limitations, uptake or transport of the substrate to the reactive site could be observed during TCE dechlorination. Our results show that prior to a mechanistic interpretation of stable isotope fractionation factors it has to be carefully verified how other factors such as uptake or transport affect the isotope fractionation during degradation experiments with microbial cultures.     

零价金属降解多氯联苯(PCBs)

多氯联苯(polychlorinated biphenyls,简称PCBs)是一类对环境有不利影响的有毒有机物,它在环境中广泛而大量分布。许多科学家都在致力于有效处理PCBs污染介质(包括水、油、沉积物和土壤)的修复技术的研究。本文综述了国内外在零价金属还原脱氯降解PCBs领域的研究状况。在高温等特殊条件下或有钯、铂、镍和铜等催化剂存在的条件下,零价金属能有效促进PCBs还原脱氯。讨论了零价铁还原脱氯的3个可能的途径:金属直接反应,将零价铁表面的电子转移到有机氯化物使之脱氯;铁腐蚀的直接产物Fe2 具有还原能力,它可使得一部分氯代烃脱氯;铁反应产生的氢气可使有机氯化物还原。评述了零价金属还原脱氯PCBs具有有效、廉价和易得的特点。展望了零价金属还原脱氯降解PCBs研究领域的发展前景。     

Successive rapid reductive dehalogenation and mineralization of pentachlorophenol by the indigenous microflora of farmyard manure compost

AIMS: To determine whether composting with animal manure can be used to effectively remediate soil from a pentachlorophenol (PCP)-contaminated site, and to establish the fate of the degraded xenobiotic. METHODS AND RESULTS: Contaminated soil from a sawmill site was mixed with farm animal manure and composted in a 0.5 m3 silo under fully aerobic conditions. The disappearance and fate of PCP was monitored by gas chromatography (GC-ECD) and extensive mineralization confirmed in experiments with 14C-radiolabelled PCP. The disappearance of PCP was rapid and virtually complete within 6 days, prior to the onset of thermophilic conditions. Dechlorination of the PCP was found to be both reductive and sequential. CONCLUSIONS: PCP removal from contaminated soil by aerobic composting with animal manure is efficient and proceeds via reductive dechlorination to virtually complete mineralization. This contrasts with other chlorophenol composting regimes in which mineralization is achieved but dechlorination intermediates do not accumulate to detectable levels. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study demonstrate that anaerobic reductive dechlorination can proceed in an aerobic composting environment and contribute to efficient pentachlorophenol removal. Farmyard manure composts may represent a rapid, low-cost, low-technology option for treatment of chlorophenol-contaminated soils.