Biodegradation and sorption of polyaromatic hydrocarbons by Phanerochaete chrysosporium

The ability of the white-rot fungus Phanerochaete chrysosporium (INA-12) to degrade various polynuclear aromatic hydrocarbons (PAH) was investigated. Under static, non-nitrogen-limiting conditions, P. chrysosporium mineralized both phenanthrene and benzo[a]pyrene. Total mineralization, based on radioactive tracing, was limited to 1.8%–3% for phenanthrene and benzo[a]pyrene respectively. In both cases the pattern of mineralization did not correlate temporally with the production of lignin peroxidase activity. Sorption of radiolabelled material to the biomass was very significant with 22% and 40% of the total radioactivity being sorbed for benzo[a]pyrene and phenanthrene respectively. A number of models were examined to predict the sorption isotherms, the best performance being obtained with a three-parameter empirical model. It is apparent that lignin peroxidase is not necessarily involved in the biodegradation of all PAH and that a significant factor in PAH biodegradation and/or disappearance in cultures with the intact fungus may be attributed to sorption phenomena.     

Microbial biodegradation of polyaromatic hydrocarbons

Polycyclic aromatic hydrocarbons (PAHs) are widespread in various ecosystems and are pollutants of great concern due to their potential toxicity, mutagenicity and carcinogenicity. Because of their hydrophobic nature, most PAHs bind to particulates in soil and sediments, rendering them less available for biological uptake. Microbial degradation represents the major mechanism responsible for the ecological recovery of PAH-contaminated sites. The goal of this review is to provide an outline of the current knowledge of microbial PAH catabolism. In the past decade, the genetic regulation of the pathway involved in naphthalene degradation by different gram-negative and gram-positive bacteria was studied in great detail. Based on both genomic and proteomic data, a deeper understanding of some high-molecular-weight PAH degradation pathways in bacteria was provided. The ability of nonligninolytic and ligninolytic fungi to transform or metabolize PAH pollutants has received considerable attention, and the biochemical principles underlying the degradation of PAHs were examined. In addition, this review summarizes the information known about the biochemical processes that determine the fate of the individual components of PAH mixtures in polluted ecosystems. A deeper understanding of the microorganism-mediated mechanisms of catalysis of PAHs will facilitate the development of new methods to enhance the bioremediation of PAH-contaminated sites.     

Biodegrader metabolic expansion during polyaromatic hydrocarbons rhizoremediation

Root-microbe interactions are considered to be the primary process of polyaromatic hydrocarbon (PAH) phytoremediation, since bacterial degradation has been shown to be the dominant pathway for environmental PAH dissipation. However, the precise mechanisms driving PAH rhizostimulation symbiosis remain largely unresolved. In this study, we assessed PAH degrading bacterial abundance in contaminated soils planted with 18 different native Michigan plant species. Phenanthrene metabolism assays suggested that each plant species differentially influenced the relative abundance of PAH biodegraders, though they generally were observed to increase heterotrophic and biodegradative cell numbers relative to unplanted soils. Further study of >1800 phenanthrene degrading isolates indicated that most of the tested plant species stimulated biodegradation of a broader range of PAH compounds relative to the unplanted soil bacterial consortia. These observations suggest that a principal contribution of planted systems for PAH bioremediation may be via expanded metabolic range of the rhizosphere bacterial community.     

Solubilization of polyaromatic hydrocarbons by recombinant bioemulsifier AlnA

AlnA is the protein responsible for the emulsifying and solubilizing activity of the Acinetobacter radioresistens KA53 bioemulsifier alasan. AlnA was produced in Escherichia coli, purified to homogeneity and then used to measure the enhanced solubility of 12 polyaromatic hydrocarbons (PAHs). The amount of PAH solubilized was directly proportional to AlnA concentration. The ratio of PAH solubilized by 40 μg/ml AlnA compared to that soluble in the aqueous buffer varied greatly, from 4 (fluorene) to 81 (hexylbenzylcyclosilane). Calculations of moles PAH solubilized per mole AlnA yielded values from 4.3 (hexylphenylbenzene) to 55.8 (1,10-phenanthrolene). There was no obvious relationship between the amount of PAH solubilized and its molecular weight or intrinsic solubility. Native gel electrophoresis indicated that AlnA formed hexamers in the presence of PAHs. With molar ratios of fluorene to AlnA of 0.75 or less, only the monomer was observed, whereas at ratios of 7.5 or higher, only the hexamer was detected. At an intermediate molar ratio of 2.6, both monomer and hexamer appeared. The data indicate that PAHs are initially solubilized by binding to the monomeric form of AlnA, and as the amount bound increases above one molecule PAH per AlnA, the protein aggregates to form a specific oligomer of 5–8 monomers which allows for the binding and solubilization of more PAH. Electronic Publication     

Biodegradation of polycyclic aromatic hydrocarbons

The intent of this review is to provide an outline of the microbial degradation of polycyclic aromatic hydrocarbons. A catabolically diverse microbial community, consisting of bacteria, fungi and algae, metabolizes aromatic compounds. Molecular oxygen is essential for the initial hydroxylation of polycyclic aromatic hydrocarbons by microorganisms. In contrast to bacteria, filamentous fungi use hydroxylation as a prelude to detoxification rather than to catabolism and assimilation. The biochemical principles underlying the degradation of polycyclic aromatic hydrocarbons are examined in some detail. The pathways of polycyclic aromatic hydrocarbon catabolism are discussed. Studies are presented on the relationship between the chemical structure of the polycyclic aromatic hydrocarbon and the rate of polycyclic aromatic hydrocarbon biodegradation in aquatic and terrestrial ecosystems.     

Degradation of polyaromatic hydrocarbons by Burkholderia cepacia 2A-12

A new strain of bacterium degrading polyaromatic hydrocarbons (PAHs), Burkholderia cepacia 2A-12, was isolated from oil-contaminated soil. Of three PAHs, the isolated strain could utilize naphthalene (Nap) and phenanthrene (Phe) as a sole carbon source but not pyrene (Pyr). However, the strain could degrade Pyr when a cosubstrate such as yeast extract (YE) was supplemented. The PAH degradation rate of the strain was enhanced by the addition of other organic materials such as YE, peptone, glucose, and sucrose. YE was a particularly effective additive in stimulating cell growth as well as PAH degradation. When 1 g YE l^–1, an optimum concentration, was supplemented into the basal salt medium (BSM) with 215 mg Phe l^–1, the specific growth rate (0.30 h^–1) and Phe-degrading rate (29.6 mol l^–1 h^–1) were enhanced approximately ten and three times more than those obtained in the BSM with 215 mg Phe l^–1, respectively. Both cell growth and PAH degradation rates were increased with increasing Phe and Pyr concentrations, and B. cepacia 2A-12 had a tolerance against Phe and Pyr toxicity at the high concentration of 730–760 mg l^–1. Through kinetic analysis, the maximum specific growth rate ( _max) and PAH degrading rate ( _max) for Phe were obtained as 0.39 h^–1 and 300 mol l^–1 h^–1, respectively. Also, _max and _max for Pyr were 0.27 h^–1 and 52 mol l^–1 h^–1, respectively. B. cepacia 2A-12 could simultaneously degrade crude oil as well as PAHs, indicating that this bacterium is very useful for the removal of oils and PAHs contaminants.     

Biodegradation of hydrocarbons in the presence of cyclodextrins

Aliphatic hydrocarbons are one of the main components of oil contamination. Bioremediation is considered to be a cost-effective treatment option among the conventional treatment methods with bioavailability being the limitation. Chemical surfactants could be used to increase the bioavailability of the hydrocarbons but they showed marked toxicity and environmental pollution. Cyclodextrins are cyclic oligosaccharides which can alter the solubility of the hydrocarbons by incorporating suitably sized hydrophobic molecules into their hydrophobic cavities. This paper focuses on studying the degradation of hydrocarbons by Pseudomonas like species named as Vid1 isolated previously from bilge oil contaminated waters in the presence of cyclodextrins. Among the three cyclodextrins (α, β and γ) tested at different concentrations, 2.5 mM of β-cyclodextrin showed higher amount of biodegradation when n-hexadecane was used as a model hydrocarbon compound. The percentage of residual hexadecane remaining in the 2.5 mM β-cyclodextrin supplied medium at 120 h was found to be 15% in comparison with the biotic control which was 43%. In the next experimental setup, degradation of mixture of hydrocarbons (tetradecane, hexadecane and octadecane) by Vid1 (Pseudomonas like species) was studied at a concentration of 2.5 mM β-cyclodextrin. The residual percentage of tetradecane, hexadecane and octadecane at 120 h was found to be 32, 43 and 61% in comparison with the biotic control 50, 58 and 67%, respectively. Our studies show that among a mixture of hydrocarbons (tetradecane, hexadecane and octadecane) in the presence of β-cyclodextrin, the highest concentration of hydrocarbon degradation was found in tetradecane, hexadecane and octadecane, respectively.     

Biodegradation and bioremediation of hydrocarbons in extreme environments

Many hydrocarbon-contaminated environments are characterized by low or elevated temperatures, acidic or alkaline pH, high salt concentrations, or high pressure, Hydrocarbon-degrading microorganisms, adapted to grow and thrive in these environments, play an important role in the biological treatment of polluted extreme habitats. The biodegradation (transformation or mineralization) of a wide range of hydrocarbons, including aliphatic, aromatic, halogenated and nitrated compounds, has been shown to occur in various extreme habitats. The biodegradation of many components of petroleum hydrocarbons has been reported in a variety of terrestrial and marine cold ecosystems. Cold-adapted hydrocarbon degraders are also useful for wastewater treatment. The use of thermophiles for biodegradation of hydrocarbons with low water solubility is of interest, as solubility and thus bioavailability, are enhanced at elevated temperatures. Thermophiles, predominantly bacilli, possess a substantial potential for the degradation of environmental pollutants, including all major classes. Indigenous thermophilic hydrocarbon degraders are of special significance for the bioremediation of oil-polluted desert soil. Some studies have investigated composting as a bioremediation process. Hydrocarbon biodegradation in the presence of high salt concentrations is of interest for the bioremediation of oil-polluted salt marshes and industrial wastewaters, contaminated with aromatic hydrocarbons or with chlorinated hydrocarbons. Our knowledge of the biodegradation potential of acidophilic, alkaliphilic, or barophilic microorganisms is limited.     

Measurement of various polyaromatic hydrocarbons in the urban area of Naples]

In order to investigate the organic compound fraction of the Naples aerosol a chromatographic method was used for the separation and analysis of the polycyclic aromatic (PAH). As a first step a suitable one-step thin-layer chromatography (TLC) separation of the cyclohexane extractable material from airborne particulate was sought. After the TLC separation the concentrated samples were analyzed by reverse-phase liquid chromatography with fluorescence detection. We obtained chromatographic separation of five PAH on the EPA Priority Pollutant List and we determined the concentration of these PAHs present in atmospheric matter.     

Biodegradation of polycyclic hydrocarbons by Phanerochaete chrysosporium

The ability of the white rot fungus Phanerochaete chrysosporium to degrade polycyclic aromatic hydrocarbons (PAHs) that are present in anthracene oil (a distillation product obtained from coal tar) was demonstrated. Analysis by capillary gas chromatography and high-performance liquid chromatography showed that at least 22 PAHs, including all of the most abundant PAH components present in anthracene oil, underwent 70 to 100% disappearance during 27 days of incubation with nutrient nitrogen-limited cultures of this fungus. Because phenanthrene is the most abundant PAH present in anthracene oil, this PAH was selected for further study. In experiments in which [14C]phenanthrene was incubated with cultures of P. chrysosporium containing anthracene oil for 27 days, it was shown that 7.7% of the recovered radiolabeled carbon originally present in [14C]phenanthrene was metabolized to 14CO2 and 25.2% was recovered from the aqueous fraction, while 56.1 and 11.0% were recovered from the methylene chloride and particulate fractions, respectively. High-performance liquid chromatography of the 14C-labeled material present in the methylene chloride fraction revealed that most (91.9%) of this material was composed of polar metabolites of [14C]phenanthrene. These results suggest that this microorganism may be useful for the decontamination of sites in the environment contaminated with PAHs.     

Characterization of selected actinomycetes degrading polyaromatic hydrocarbons in liquid culture and spiked soil

Summary Five strains of the Rhodococcus and Gordonia genera were evaluated for their potential use in bioremediation of polycyclic aromatic hydrocarbons (PAH) with or without another substrate (co-substrate). Their ability to produce biosurfactants or to degrade phenanthrene when growing on glucose, hexadecane and rapeseed oil was tested in liquid medium at 30 °C. All strains showed biosurfactant activity. The highest reduction in surface tension was recorded in whole cultures of Rhodococcus sp. DSM 44126 (23.1%) and R. erythropolis DSM 1069 (21.1%) grown on hexadecane and Gordonia sp. APB (20.4%) and R. erythropolis TA57 (18.2%) grown on rapeseed oil. Cultures of Gordonia sp. APB and G. rubripertincta formed emulsions when grown on rapeseed oil. After 14 days of incubation, Rhodococcus sp. DSM 44126 degraded phenanthrene (initial concentration 100 μg ml⁻¹) as sole carbon source (79.4%) and in the presence of hexadecane (80.6%), rapeseed oil (96.8%) and glucose (below the limit of detection). The other strains degraded less than 20%, and then with a co-substrate only. Rhodococcus sp. DSM 44126 was selected and its performance evaluated in soil spiked with a mixture of PAH (200 mg kg⁻¹). The effect of the addition of 0, 0.1 and 1% rapeseed oil as co-substrate was also tested. Inoculation enhanced the degradation of phenanthrene (55.7% and 95.2% with 0.1% oil and without oil respectively) and of anthracene (29.2% with 0.1% oil). Approximately 96% of anthracene and 62% of benzo(a)pyrene disappeared from the soil (inoculated and control) after 14 days and anthraquinone was detected as a metabolite. Rhodococcus sp. DSM 44126 was identified as Rhodococcus wratislaviensis by 16S rRNA sequencing and was able to degrade anthracene as sole carbon source in liquid culture.     

Biodegradation of petroleum hydrocarbons in soil inoculated with yeasts

Yeast species belonging to the Candida genus were added to the greyish-brown soil of the Apsheron Peninsula under laboratory conditions. The rate of CO2 production was used to estimate the degradation of crude oil, paraffin, cycloparaffin and aromatic hydrocarbons as well as their oxidized products. The rate of hydrocarbon degradation in the soil inoculated with yeast cells was shown to drop down gradually. The effective action on the process of hydrocarbon degradation depended on the special properties of an inoculated population and on the structure of a hydrocarbon. Some yeast species stimulated the degradation of various aromatic hydrocarbons and their oxidized products. Aromatic hydrocarbons were decomposed at a lower rate comparing to their oxidized products. When the soil was inoculated with C. guilliermondii populations, n-hexadecane added to the soil at a concentration of 1% was decomposed within 250-300 days. Field experiments confirmed that crude oil biodegradation was more intensive in the soil inoculated with yeast cells.     

Biodegradation of hydrocarbons by an extremely halophilic archaebacterium

An archaebacterium (strain EH4) able to biodegrade saturated and aromatic hydrocarbons has been isolated from a sail-marsh. Maximum growth on eicosane (62% of biodegradation, 10 h generation time) was reached in a medium prepared with a natural hypersaline water collected from a salt-marsh (3.5 mol/1 NaCl concentration). No growth on hydrocarbons was observed for NaCl concentration lower than 1.8 mol/1.     

Biodegradation of polycyclic hydrocarbons by Phanerochaete chrysosporium.

The ability of the white rot fungus Phanerochaete chrysosporium to degrade polycyclic aromatic hydrocarbons (PAHs) that are present in anthracene oil (a distillation product obtained from coal tar) was demonstrated. Analysis by capillary gas chromatography and high-performance liquid chromatography showed that at least 22 PAHs, including all of the most abundant PAH components present in anthracene oil, underwent 70 to 100% disappearance during 27 days of incubation with nutrient nitrogen-limited cultures of this fungus. Because phenanthrene is the most abundant PAH present in anthracene oil, this PAH was selected for further study. In experiments in which [14C]phenanthrene was incubated with cultures of P. chrysosporium containing anthracene oil for 27 days, it was shown that 7.7% of the recovered radiolabeled carbon originally present in [14C]phenanthrene was metabolized to 14CO2 and 25.2% was recovered from the aqueous fraction, while 56.1 and 11.0% were recovered from the methylene chloride and particulate fractions, respectively. High-performance liquid chromatography of the 14C-labeled material present in the methylene chloride fraction revealed that most (91.9%) of this material was composed of polar metabolites of [14C]phenanthrene. These results suggest that this microorganism may be useful for the decontamination of sites in the environment contaminated with PAHs.     

Biodegradation of polycyclic aromatic hydrocarbons by Pichia anomala

Pichia anomala 2.2540, isolated from soil contaminated by crude oil, degraded naphthalene, dibenzothiophene, phenanthrene and chrysene, both singly and in combination. The yeast degraded 4.5 mg naphthalene l(-1) within 24 h. Phenanthrene was degraded after a lag of 24 h. When a mixture of all four polycyclic aromatic hydrocarbons was treated at either 0.1-1.6 mg l(-1) or 3.1-5.3 mg l(-1), naphthalene was completely degraded first within 24 h, followed by phenanthrene and dibenzothiophene after 48 h. Chrysene, which remained in the mixture even after 96 h, could be degraded along with naphthalene. Chrysene at 0.7 and 1 mg l(-1), in the presence of 4.3 and 65 mg naphthalene l(-1), respectively, was removed within 96 h.     

Biodegradation of low-molecular-weight halogenated hydrocarbons by methanotrophic bacteria

Low-molecular-weight halogenated hydrocarbons are susceptible to degradation by anaerobic and aerobic bacteria. The methanotrophic bacterium Methylosinus trichosporium 0B3b degrades trichloroethylene more rapidly than other bacteria examined to date. Expression of soluble methane monooxygenase (MMO) is correlated with high rates of biodegradation. An analysis of 16 S rRNA sequences of 11 ribosomal RNAs from type I, type II and type X methanotrophs and methanol-utilizing bacteria have revealed four clusters of phylogenetically related methylotrophs. This information may be useful for the identification and enumeration of methylotrophs in bioreactors and other environments during remediation of contaminated waters.     

Biodegradation of petroleum hydrocarbons under tropical estuarine conditions

The physico-chemical parameters of water samples collected from three points in the Lagos lagoon were studied for 12 months. Safinity varied seasonally but the temperature pH, dissolved O₂, conductivity, NO₃ ^– and HPO₄ ^2– concentrations were relatively constant. There was a direct proportionality between the population density of hydro-carbon-utilizing bacteria and the oil content of water samples. Twelve hydrocarbon-utilizing bacteria were isolated by selective enrichment and characterized as species ofPseudomonas, Alcaligenes, Acinetobacter andBacilius. The organisms grew mainly on long-chain aliphatic hydrocarbons. Laboratory and field biodegradation studies showed both quantitative and qualitative changes in the hydrocarbon content of crude oil due to microbial degradative activities and a faster rate of oil depletion from the Lagos lagoon during the rainy season. The results obtained could offer a predictive model for estimating the rate of disappearance of petroleum hydrocarbons from the tropical estuarine environment.

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Résumé On a étudié les paramètres physicochimiques d'échantillons d'eau récoltés en trols endroits du lagon de Lagos pendant 12 mols. La salinite a varié avec les saisons mais tant la température que le pH, l'oxygène dissous, la conductivité et les teneurs en nitrales et en phosphates sont restés relativement constants. II y avait une proportionnalité directe entre la densité de population des bactéries utilisant les hydrocarbures et le contenu en pétrole des échantilions d'eau. Douze souches de bactéries utilisant les hydrocarbures ont été isolées par enrichissement sélectif et caractérisées au niveau de l'espèce comme desPseudomonas, Alcaligenes, Acinetobacter etBacillus. Ces organismes croissent principalement sur des hydrocarbures aliphatiques à longue chaîne. Des études de biodégradation au laboratoire et sur le terrain ont montré des changements tant quantitatifs que qualitatifs dans le contenu en hydrocarbures du pétroie brut, dus aux activités de dégradation microbienne ainsi qu'un appauvrissement plus rapide en pétrole du lagon de Lagos pendant la saison des pluies. Les résultats obtenus pourraient offrir un modèle prédictif pour l'estimation de la vitesse de disparition des hydrocarbures du pétrole dans l'environnement d'estuaires tropicaux.

     

Biodegradation of low-molecular-weight halogenated hydrocarbons by methanotrophic bacteria

Abstract Low-molecular-weight halogenated hydrocarbons are susceptible to degradation by anaerobic and aerobic bacteria. The methanotrophic bacterium Methylosinus trichosporium 0B3b degrades trichloroethylene more rapidly than other bacteria examined to date. Expression of soluble methane monooxygenase (MMO) is correlated with high rates of biodegradation.
An analysis of 16 S rRNA sequences of 11 ribosomal RNAs from type I, type II and type X methanotrophs and methanol-utilizing bacteria have revealed four clusters of phytogenetically related methylotrophs. This information may be useful for the identification and enumeration of methylotrophs in bioreactors and other environments during remediation of contaminated waters.     

Prediction of electrocatalytic activity of some nitrogen-doped polyaromatic hydrocarbons by molecular modelling

Density function theory calculations of minimum energy structure of an oxygen molecule and oxygen atom bonded to the two dimensional molecules, C₂₃NH₁₂, coronene and C₂₁NH₁₄, pentacene doped with nitrogen, indicate the structures are at a minimum on the potential energy surface having no imaginary frequencies. Calculation of the bond dissociation energy (BDE) to remove an oxygen atom from nitrogen-doped pentacene to which is bonded O₂, (C₂₁NH₁₄O₂) shows it is less than that to dissociate O₂. However, this is not the case for nitrogen-doped coronene. This suggests that nitrogen-doped pentacene could be an effective catalyst for the oxygen reduction reaction in fuel cells assuming it is O₂ dissociation. It is also shown that O₂H can bond to nitrogen-doped coronene and pentacene and that the BDE to remove OH is less than that to remove OH from O₂H indicating that N-doped coronene and pentacene could also catalyse this reaction. The calculated adsorption energy for O₂ and O₂H on these molecules is negative indicating they can bond to N-doped coronene and pentacene.