Anaerobic Degradation Of Polycyclic Aromatic Hydrocarbons Research Articles

Occurrence and Sources of Polycyclic Aromatic Hydrocarbons and Factors Influencing Their Accumulation in Surface Sediment of a Deep-Sea Depression, Namely, the Tatar Trough (Tatar Strait, the Sea of Japan)

The concentrations of 14 polycyclic aromatic hydrocarbons (PAHs) in the sediment of the Tatar Trough were studied. Despite the increase in PAH concentrations over recent decades, which is likely the result of the handling and transportation of fossil fuels, PAH levels and ecological risk were found to be low. The spatial pattern revealed that higher PAH concentrations were mainly in the deeper water sites, suggesting that trough slope failure transported the PAHs to the deeper part of the basin. There was no correlation between the PAHs and grain size or the PAHs and organic carbon that is related to the PAH input from a variety of sources and the heterogeneity of organic matter. The PAH composition, isomer ratio, and PCA identified two areas with different PAH sources. The most northern part of the Tatar Trough received petrogenic PAHs that are probably transported downslope from the northern Tatar Strait where fossil fuels are handled in some ports. Another trough part was polluted by the PAHs from the combustion of coal and biomass and the exhaust of marine vehicles. The minor presence of genes responsible for aerobic PAH destruction can be explained by the anaerobic degradation of PAHs or the spontaneous creation of favorable conditions that promote bacterial PAH oxidation.

Nitrogen transformation promotes the anaerobic degradation of PAHs in water level fluctuation zone of the Three Gorges Reservoir in Yangtze River, China: Evidences derived from in-situ experiment

Polycyclic aromatic hydrocarbons (PAHs) pose a great threat to human health and ecological system safety. The interception of nitrogen is common found in the riparian zone. However, there is no evidence on how nitrogen addition affects the anaerobic degradation of PAHs in soil of the water-level-fluctuation zone (WLFZ) of the Three Gorges Reservoir (TGR) in Yangtze River, China. Here, we investigated the PAHs degradation rate, the variation of key functional genes and microbial communities after nitrogen addition in soil that experienced a flooding period of water-level-fluctuation. The results revealed that the ∑16PAHs were decreased 16.19 %–36.65 % and more 3–5-rings PAHs were biodegraded with nitrogen addition in WLFZ. The most genes involved in PAHs-anaerobic degradation and denitrification were up-regulated by nitrate addition, and phyla Firmicutes, Actinobacteria and Proteobacteria were more advantages in nitrogen addition groups. The Tax4Fun based genome function analysis revealed that the microbial activity of PAHs-degradation increased with nitrate addition. The co-occurrence network analysis indicated that nitrogen addition accelerated the metabolism of nitrogen and PAHs. It is the first time to provide the direct experimental evidences that nitrogen transformation in the WLFZ soil promotes anaerobic PAHs degradation. This work is of importance to understand the effect of nitrogen intercepted in the WLFZ soil of TGR in Yangtze River, China.

The combined enhancement of RL, nZVI and AQDS on the microbial anaerobic-aerobic degradation of PAHs in soil

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous persistent organic pollutants in soil, which have carcinogenic, teratogenic and mutagenic hazards. The effects of rhamnolipid (RL), nano zero-valent iron (nZVI), and anthraquinone-2,6-disulfonic acid (AQDS) on the degradation of PAHs in soil were studied. It was found that the treatment of 5 mg·kg−1RL + 1% nZVI +0.2 mmol·kg−1AQDS had the highest degradation rate. The degradation rate of total PAHs and HMW-PAHs was 72.81% and 79.47% respectively after 90 days. High-throughput sequencing showed that in RL + nZVI + AQDS enhanced soil, Clostridium, Geobacter, Anaeromyxobacter and Sphingomonas were the dominant species for anaerobic degradation of PAHs. Rhodococcus, Nocardioides, and Microvirga are the dominant species for aerobic degradation of PAHs. The activities of methyltransferase, dehydrogenase and catechol 1,2-dioxygenase in the anaerobic-aerobic degradation process of PAHs were consistent with the degradation process of PAHs, indicating the role of these enzymes in the degradation of PAHs. RL, nZVI, and AQDS combined enhanced microbial anaerobic-aerobic degradation has great application potential in remediation of PAHs-contaminated soil.

Simultaneously enhanced removal of PAHs and nitrogen driven by Fe2+/Fe3+ cycle in constructed wetland through automatic tidal operation

The lack of dissolved oxygen and weak substrate removal capacity in constructed wetlands (CW) leads to terrible removal of nitrogen and polycyclic aromatic hydrocarbons (PAHs). In this study, automatic tidal flow CW microcosms were constructed by improving the oxygen environment (siphon and air-duct) and substrate (magnetite) to enhance purification performance and the mechanism was explored. The results showed that the addition of air-duct could improve the oxygen collection and thus improved the NH4+ removal efficiency. Additionally, nitrogen removal was improved greatly due to the simultaneous nitrification and denitrification in aerobic layer with the addition of magnetite. Mass balance indicated the microbial degradation dominated (32–62%) the removal of PAHs. Metagenomic analysis proved the existence of magnetite enhanced the number of PAHs-degrading bacteria, functional groups and metabolic pathways and thus greatly improved the microbial degradation of PAHs. Furthermore, Fe2+/Fe3+ cycle played an important role in promoting the anaerobic degradation of PAHs, which might be served as an electron conduit to establish the direct interspecies electron transfer between iron-reducing bacteria (e.g. Deltaproteobacteria bacterium) and Anaerolineae bacterium to degrade PAHs efficiently. This study provided better understanding of the simultaneous removal of PAHs and nitrogen in tidal flow CWs.

Removal of polycyclic aromatic hydrocarbons during anaerobic biostimulation of marine sediments

This study proposes the supplementation of digestate, fresh organic fraction of municipal solid waste (OFMSW) and a nutrient solution during the anaerobic biostimulation of marine sediments contaminated by polycyclic aromatic hydrocarbons (PAHs). The experimental activity was conducted with four PAHs (i.e. phenanthrene, anthracene, fluoranthene and pyrene) under controlled mesophilic conditions (37 ± 1 °C) in 100 mL serum bottles maintained at 130 rpm. After 120 days of incubation, the highest total PAH degradation of 53 and 55% was observed in the experiments with digestate + nutrients and OFMSW + nutrients, respectively. Phenanthrene was the most degraded PAH and the highest removal of 69% was achieved with OFMSW + nutrients. The anaerobic PAH degradation proceeded through the accumulation of volatile fatty acids and the production of hydrogen and methane as biogas constituents. The highest cumulative biohydrogen production of 80 mL H2·g VS−1 was obtained when OFMSW was used as the sole amendment, whereas the highest biomethane yield of 140 mL CH4·g VS−1 was obtained with OFMSW + nutrients. The evolution of PAH removal during anaerobic digestion revealed a higher impact of the methanogenic phase rather than acidogenic phase on PAH degradation.

Polycyclic Aromatic Hydrocarbon-Induced Changes in Bacterial Community Structure under Anoxic Nitrate Reducing Conditions.

Although bacterial anaerobic degradation of mono-aromatic compounds has been characterized in depth, the degradation of polycyclic aromatic hydrocarbons (PAHs) such as naphthalene has only started to be understood in sulfate reducing bacteria, and little is known about the anaerobic degradation of PAHs in nitrate reducing bacteria. Starting from a series of environments which had suffered different degrees of hydrocarbon pollution, we used most probable number (MPN) enumeration to detect and quantify the presence of bacterial communities able to degrade several PAHs using nitrate as electron acceptor. We detected the presence of a substantial nitrate reducing community able to degrade naphthalene, 2-methylnaphthalene (2MN), and anthracene in some of the sites. With the aim of isolating strains able to degrade PAHs under denitrifying conditions, we set up a series of enrichment cultures with nitrate as terminal electron acceptor and PAHs as the only carbon source and followed the changes in the bacterial communities throughout the process. Results evidenced changes attributable to the imposed nitrate respiration regime, which in several samples were exacerbated in the presence of the PAHs. The presence of naphthalene or 2MN enriched the community in groups of uncultured and poorly characterized organisms, and notably in the Acidobacteria uncultured group iii1-8, which in some cases was only a minor component of the initial samples. Other phylotypes selected by PAHs in these conditions included Bacilli, which were enriched in naphthalene enrichments. Several nitrate reducing strains showing the capacity to grow on PAHs could be isolated on solid media, although the phenotype could not be reproduced in liquid cultures. Analysis of known PAH anaerobic degradation genes in the original samples and enrichment cultures did not reveal the presence of PAH-related nmsA-like sequences but confirmed the presence of bssA-like genes related to anaerobic toluene degradation. Altogether, our results suggest that PAH degradation by nitrate reducing bacteria may require the contribution of different strains, under culture conditions that still need to be defined.

Of mothballs and old yellow enzymes.

Polycyclic aromatic hydrocarbons (PAHs) are persistent and toxic environmental pollutants that accumulate in anoxic habitats. With the exception of naphthalene, nothing is known about the microbial degradation of PAH in these environments. The challenge that must be met in anaerobic PAH degradation is the destabilization of the resonance energy of the aromatic ring system, which requires electrons with very negative redox potentials. Estelmann et al. (2014) identify two enzymes from sulphate-reducing bacteria that perform successive 2-electron reductions of a coenzyme A thioester derivative of naphthalene. The first reduces 2-naphthoyl-CoA to 5,6-dihydro-2-naphthoyl-CoA and the second generates 5,6,7,8-tetrahydro-2-naphthoyl-CoA. Surprisingly, both enzymes are members of the 'old yellow enzyme' (OYE) family of flavoproteins. Neither uses adenosine triphosphate to achieve reduction of the aromatic ring. Typically, OYEs have flavin mononucleotide as cofactor and use nicotinamide adenine dinucleotide (phosphate) as reductant. Both ring reductases have flavin adenine dinucleotide and an iron-sulphur cluster as additional cofactors. Evidence also suggests that in the sulphate-reducing bacteria, these enzymes form a complex, allowing substrate channeling. The findings of this superb study represent unprecedented biochemistry. This work sheds light on how microbes meet the thermodynamic challenges of life at the redox limit.

A PCR-based assay for the detection of anaerobic naphthalene degradation

Polycyclic aromatic hydrocarbons (PAH) are widespread environmental pollutants of considerable risk to human health. The aerobic degradation of PAH via oxygenase reactions has been studied for several decades. In contrast, it was not until very recent that the first key enzyme involved in anaerobic PAH degradation, the dearomatizing 2-naphthoyl-CoA reductase, was isolated and characterized. In this work, a PCR-based functional assay was developed to detect microorganisms that have the ability to anaerobically degrade naphthalene, as a model for larger PAH. The degenerative oligonucleotide probes introduced here amplified a highly conserved region of the gene encoding 2-naphthoyl-CoA reductase (Ncr) in numerous sulfate-reducing pure cultures and environmental enrichments. The assay provides the first molecular tool for monitoring the anaerobic degradation of a model PAH.

Identification and characterization of 2‐naphthoyl‐coenzyme A reductase, the prototype of a novel class of dearomatizing reductases

The enzymatic dearomatization of aromatic ring systems by reduction represents a highly challenging redox reaction in biology and plays a key role in the degradation of aromatic compounds under anoxic conditions. In anaerobic bacteria, most monocyclic aromatic growth substrates are converted to benzoyl-coenzyme A (CoA), which is then dearomatized to a conjugated dienoyl-CoA by ATP-dependent or -independent benzoyl-CoA reductases. It was unresolved whether or not related enzymes are involved in the anaerobic degradation of environmentally relevant polycyclic aromatic hydrocarbons (PAHs). In this work, a previously unknown dearomatizing 2-naphthoyl-CoA reductase was purified from extracts of the naphthalene-degrading, sulphidogenic enrichment culture N47. The oxygen-tolerant enzyme dearomatized the non-activated ring of 2-naphthoyl-CoA by a four-electron reduction to 5,6,7,8-tetrahydro-2-naphthoyl-CoA. The dimeric 150 kDa enzyme complex was composed of a 72 kDa subunit showing sequence similarity to members of the flavin-containing 'old yellow enzyme' family. NCR contained FAD, FMN, and an iron-sulphur cluster as cofactors. Extracts of Escherichia coli expressing the encoding gene catalysed 2-naphthoyl-CoA reduction. The identified NCR is a prototypical enzyme of a previously unknown class of dearomatizing arylcarboxyl-CoA reductases that are involved in anaerobic PAH degradation; it fundamentally differs from known benzoyl-CoA reductases.

Anaerobic degradation of phenanthrene by a newly isolated humus-reducing bacterium, Pseudomonas aeruginosa strain PAH-1

Purpose The anaerobic degradation of polycyclic aromatic hydrocarbons (PAHs) has great significance to PAHs’ natural attenuation in contaminated sites. Previous studies mainly focused on anaerobic PAH degradation by mixed cultures with nitrate, sulfate, or Fe(III) oxides as electron acceptors, and the roles of pure cultures in the process was rarely reported. The aim of this paper is to isolate a pure culture that is capable of degrading phenanthrene anaerobically with anthraquinone-2,6-disulphonate (AQDS) as the sole electron acceptor and evaluate its environmental functions. Materials and methods A strain of pure culture was isolated from anodic solution of a microbial fuel cell via enrichment procedure with phenanthrene and AQDS under anaerobic conditions. Using the single colony isolation technique, a distinct colony was obtained and identified by the phenotypic and phylogenetic analysis. Its ability to reduce AQDS and degrade phenanthrene was conducted in serum bottles by standard anaerobic techniques (purged with80%N2−20% CO2). The concentration of AH2QDS and fructose was quantified by UV-vis spectrophotometer at 450 and 210 nm, respectively. Cells number was determined by direct plate counting on aerobic LB agar medium. The concentration of phenanthrene was determined using HPLC. Results and discussion Strain PAH-1 was identified as Pseudomonas aeruginosa. It could oxidize fructose or glucose to reduce AQDS and can support microbial growth by conserving energy from fructose to AQDS. It has the ability to degrade phenanthrene directly with AQDS as the sole electron acceptor (46.5% removal), and the microbial process may be AQDS dependent. The addition of small organic substances (fructose) could enhance the anaerobic biodegradation of phenanthrene from 46.5% to 56.7%. The anaerobic degradation of phenanthrene fits the pseudo-firstorder kinetics, giving the rate constants of 0.0233/day (R 2 =0.934) and 0.0328/day (R 2 =0.933) for non-fructose set and fructose set, respectively. Conclusions We successfully isolated a facultative anaerobe, P. aeruginosa strain PAH-1. This study was the first paper reporting that a pure culture (strain PAH-1) has the ability to anaerobically degrade phenanthrene with AQDS as the sole electronacceptor.Thefinding alsoexplorestheenvironmental significance of the Pseudomonas genus.

Genomic insights into the metabolic potential of the polycyclic aromatic hydrocarbon degrading sulfate‐reducing Deltaproteobacterium N47

Anaerobic degradation of polycyclic aromatic hydrocarbons (PAHs) is an important process during natural attenuation of aromatic hydrocarbon spills. However, knowledge about metabolic potential and physiology of organisms involved in anaerobic degradation of PAHs is scarce. Therefore, we introduce the first genome of the sulfate-reducing Deltaproteobacterium N47 able to catabolize naphthalene, 2-methylnaphthalene, or 2-naphthoic acid as sole carbon source. Based on proteomics, we analysed metabolic pathways during growth on PAHs to gain physiological insights on anaerobic PAH degradation. The genomic assembly and taxonomic binning resulted in 17 contigs covering most of the sulfate reducer N47 genome according to general cluster of orthologous groups (COGs) analyses. According to the genes present, the Deltaproteobacterium N47 can potentially grow with the following sugars including d-mannose, d-fructose, d-galactose, α-d-glucose-1P, starch, glycogen, peptidoglycan and possesses the prerequisites for butanoic acid fermentation. Despite the inability for culture N47 to utilize NO(3) (-) as terminal electron acceptor, genes for nitrate ammonification are present. Furthermore, it is the first sequenced genome containing a complete TCA cycle along with the carbon monoxide dehydrogenase pathway. The genome contained a significant percentage of repetitive sequences and transposase-related protein domains enhancing the ability of genome evolution. Likewise, the sulfate reducer N47 genome contained many unique putative genes with unknown function, which are candidates for yet-unknown metabolic pathways.

Anaerobic degradation of PAHs in soil: Impacts of concentration and amendment stability on the PAHs degradation and biogas production

In this study, the bioremediation of polycyclic aromatic hydrocarbons (PAHs)-contaminated soil under strict anaerobic-methanogenic conditions was systematically studied applying the central composite design approach. The effect of PAHs concentration and the stability of the compost as an organic amendment for anaerobic digestion were examined. In all assays, the used methanogenic consortium was able to degrade the PAHs although some inhibition effects were observed during the initial stage in some cases. The degradation rates varied between 31.4 and 90.6% during 50 days incubation period. The study demonstrated that the PAHs concentration influences the degradation rate where more degradation was observed by increasing the concentration of PAHs. However, the biogas production as a result of the digestion process was more influenced by the compost stability which also has its effect on the degradation rates as more degradation occurred with more stable compost, but more biogas was produced with less stable compost, which indicates that the biogas is mainly produced by the anaerobic digestion of the amended compost. Finally, it seems that compost addition is required to improve the process in some cases but in other circumstances it does not greatly improve the bioremediation of PAHs.

Anaerobic Degradation of Phenanthrene and Pyrene in Mangrove Sediment

This study investigated the anaerobic degradation of the polycyclic aromatic hydrocarbons (PAHs) phenanthrene and pyrene in mangrove sediment from Taiwan. The anaerobic degradation of PAH was enhanced by the addition of acetate, lactate, pyruvate, sodium chloride, cellulose, or zero-valent iron. However, it was inhibited by the addition of humic acid, di-(2-ethylhexyl) phthalate (DEHP), nonylphenol, or heavy metals. Of the microorganism strains isolated from the sediment samples, we found that strain MSA3 (Clostridium pascui), expressed the best ability to biodegrade PAH. The inoculation of sediment with the strain MSA3 could enhance PAH degradation.

Anaerobic PAH degradation in soil by a mixed bacterial consortium under denitrifying conditions

Polycyclic aromatic hydrocarbons (PAHs) are one of the main classes of contaminants in the terrestrial environment. Concentrations of biphenyl, fluorene, phenanthrene and pyrene were added to soil samples in order to investigate the anaerobic degradation potential of PAHs under denitrifying conditions. A mixed population of microorganisms obtained from a paddy soil was incubated for 20 days in anaerobic conditions in the presence of soil alone or with nitrate, adding, as electron donors, PAHs and, in some samples, glucose or acetate. At regular time intervals oxidation–reduction potential, PAHs concentration, microbial ATP and nitrate concentration into the solution were measured. Degradation trends for each hydrocarbon are similar under all conditions, indicating that the molecular conformation prevails over other parameters in controlling the degradation. Poor degradation results were obtained when PAHs were the only organic matter available for the inoculum, thus confirming the recalcitrance to degradation of these compounds. Biodegradation was influenced by the addition of other carbon sources. As better degradation results were generally obtained when acetate or glucose were added, the hypothesis of a co-metabolic enhancement of PAH biodegradation seems likely. Thus, anaerobic biodegradation of PAHs studied, biphenyl, fluorene, phenanthrene and pyrene, seems to be possible both through fermentative and respiratory metabolism, provided that low molecular weight co-metabolites and suitable electron acceptors (nitrate) are present.

Removal of polycyclic aromatic hydrocarbons (PAHs) from sewage sludge by anaerobic degradation

Due to the hydrophobic nature of the polyaromatic hydrocarbons (PAHs) they are mostly bound to the sludge and escape aerobic treatment in a wastewater treatment plant. They therefore proceed directly to the anaerobic post treatment, terminate in the sludge, and can be released to the environment if land spreading is used. PAH degradation in anaerobic methanogenic systems has only recently been shown to occur. In this study we have assessed several factors of anaerobic PAH degradation by evaluating thermodynamic feasibility of degradation, assessing degradation at different temperatures, and investigating the enriched cultures using fluorescent in-situ hybridization (FISH). Thermodynamic calculations indicated that PAH degradation was possible under methanogenic conditions, in the presence of hydrogen utilizing methanogens. Removal of naphthalene and 1-methyl naphthalene depended both on temperature and the initial inoculum. Inocula sourced from contaminated land sites were the most effective. The enrichments were all a mixture of Bacteria, and Archaea, and the Archaea were generally identified as Methanobacteriales, using an order-specific probe. The bacteria were not specifically identified. The results indicate a syntrophic culture, with the bacteria oxidizing the naphthalene, and the Archaea converting the hydrogen produced by oxidation, to methane.

Anaerobic degradation of polycyclic aromatic hydrocarbons

Polycyclic aromatic hydrocarbons (PAHs) are among the most important contaminants of groundwater. The 2- and 3-ring PAHs are of particular concern because they are water soluble in the 1–200 μg/l range and are transported with the groundwater over significant distances. Anaerobic degradation of PAH has been demonstrated in several microcosm studies with nitrate, ferric iron, or sulfate as electron acceptors and under methanogenic conditions. The biochemical degradation pathways were studied with naphthalene-degrading pure and enrichment cultures and revealed that 2-naphthoic acid is a central metabolite. Naphthalene is activated by addition of a C 1-unit to generate 2-naphthoic acid, whereas methylnaphthalene is activated by addition of fumarate to the methyl group and further degraded to 2-naphthoic acid. In the central 2-naphthoic acid degradation pathway the ring system is reduced prior to ring cleavage generating e.g. 5,6,7,8-tetrahydro-2-naphthoic acid. The ring cleavage produces metabolites such as 2-carboxycyclohexylacetic acid indicating that further degradation goes via cyclohexane derivatives and not via aromatic compounds. Anaerobic degradation of PAH has also been demonstrated in situ in contaminated aquifers by identification of compound specific metabolites and using stable isotope fraction studies. Detection of specific metabolites of anaerobic PAH degradation such as naphthyl-2-methylsuccinate indicated anaerobic degradation of 2-methylnaphthalene in situ whereas 2-naphthoic acid was indicative of naphthalene and 2-methylnaphthalene degradation. Other carboxylic acids that were detected in groundwater indicated anaerobic degradation of a wide range of PAH and heterocyclic compounds. Degradation of naphthalenes in contaminated aquifers could also be confirmed by carbon stable isotope shifts in the residual substrate fraction.