Biodegradation Of Naphthalene Research Articles

Assessment of biomass material as valuable electrode for high energy performance in microbial fuel cell with biodegradation of organic pollutant

Microbial fuel cells (MFC) have becomea more popular bio-electrochemical approach due to their environmental friendliness and sustainability. The anode is a vital component of the MFC, and its unstable material remains one of the primary causes of poor electron transportation throughout the reactor operations. This study seeks to improve MFC electron transport and naphthalene biodegradation by employing modified palm kernel shell (PKS)-based anode electrodes. These anode fabrications were successfully developed through the combination of PKS-based rGO hybridized into metal oxide modifiers (ZnO and TiO2). The anode electrodes were subsequently assigned PKS-rGO, PKS-rGO/TiO2, and PKS-rGO/ZnO. The power density displayed for PKS-rGO was 35.1 mW/m2, whereas the power density for the PKS-rGO/TiO2 composite anode was found to be 38.2 mW/m2. On the other hand, PKS-rGO/ZnO had a maximum power density of 45.4 mW/m2. The maximum current density (CD) in the anode PKS-rGO/ZnO was 114.0 mA/m2, indicating a comparable pattern of power output. Bacillus sp. and Niallia sp. were identified as exo-electrogens in the microbial study. In the MFC operation, the highest biodegradation efficiency was 85.50 % in the PKS-rGO settings, 92.80 % in PKS-rGO/TiO2, and94.30 % in PKS-rGO/ZnO. SEM and microbiological studies revealed that our anodes were highly biocompatible with microbes. Furthermore, this result proposes future research directions for the mortality ofanodic microbial communitiesin MFC.

Biodegradation of naphthalene using Kocuria rosea isolated from a Sawmill in Ikenne, Southwestern Nigeria

Abstract Naphthalene, a common member of the polycyclic aromatic hydrocarbons (PAHs) found in various industrial applications, has garnered attention due to its genotoxic, mutagenic, and potentially carcinogenic effects on living organisms. Concerns have escalated regarding the widespread presence of naphthalene in the environment, its resistance to biodegradation, capacity for bioaccumulation, and associated adverse impacts. In response, numerous pilot treatment approaches have been explored to mitigate the economic consequences and prevent the degradation of soil and water quality resulting from naphthalene pollution. This study investigates the degradation of naphthalene using Kocuria rosea, a microorganism previously isolated from sawdust samples collected from a Sawmill in Ikenne, Ogun State, Southwestern Nigeria. Kocuria rosea was cultivated in nutrient broth (NB) for 24 h, and its growth was quantified through UV–Visible spectrophotometric analysis. The phytotoxicity of both untreated and treated naphthalene solutions was assessed by measuring their impact on the germination of Zea mays (corn) seeds. Furthermore, biodegradation of naphthalene was confirmed by analyzing FT-IR spectra. The results indicate that naphthalene exhibits phytotoxic effects on Z. mays germination, whereas treatment with the Kocuria rosea solution substantially increased the germination rate from 13.3 % to 46.67 %. FT-IR analysis reveals that Kocuria rosea effectively degrades naphthalene, as evidenced by the disappearance of characteristic peaks at 3049.56, 1593.25, 958.65, and 781.2. In conclusion, Kocuria rosea, isolated from a Sawmill in Ikenne, demonstrates significant potential for the bioremediation of naphthalene, offering promise as an environmentally friendly and cost-effective approach for mitigating naphthalene pollution.

Evaluation of surfactant-aided polycyclic aromatic hydrocarbon biodegradation by molecular docking and molecular dynamic simulation in the marine environment

Marine oil spills directly cause polycyclic aromatic hydrocarbons (PAHs) pollution and affect marine organisms due to their toxic property. Chemical and bio-based dispersants composed of surfactants and solvents are considered effective oil spill-treating agents. Dispersants enhance oil biodegradation in the marine environment by rapidly increasing their solubility in the water column. However, the effect of dispersants, especially surfactants, on PAHs degradation by enzymes produced by microorganisms has not been studied at the molecular level. The role of the cytochrome P450 (CYP) enzyme in converting contaminants into reactive metabolites during the biodegradation process has been evidenced, but the activity in the presence of surfactants is still ambiguous. Thus, this study focused on the evaluation of the impact of chemical and bio-surfactants (i.e., Tween 80 (TWE) and Surfactin (SUC)) on the biodegradation of naphthalene (NAP), chrysene (CHR), and pyrene (PYR), the representative components of PAHs, with CYP enzyme from microalgae Parachlorella kessleri using molecular docking and molecular dynamics (MD) simulation. The molecular docking analysis revealed that PAHs bound to residues at the CYP active site through hydrophobic interactions for biodegradation. The MD simulation showed that the surfactant addition changed the enzyme conformation in the CYP-PAH complexes to provide more interactions between the enzyme and PAHs. This led to an increase in the enzyme's capability to degrade PAHs. Binding free energy (ΔG‌‌Bind) calculations confirmed that surfactant treatment could enhance PAHs degradation by the enzyme. The SUC gave a better result on NAP and PYR biodegradation based on ΔG‌‌Bind, while TWE facilitated the biodegradation of CHR. The research outputs could greatly facilitate evaluating the behaviors of oil spill-treating agents and oil spill response operations in the marine environment.

Optimized approach for naphthalene wastewater biodegradation by Enterobacter ludwigii NS12 isolated from petroleum industry sludge: Bioreactor study and kinetic investigation

Polycyclic aromatic hydrocarbons (PAHs) pollution is a pressing concern in environments, primarily due to human activities. Limited research exists on the degradation of PAHs by indigenous bacterial strains compared to other degraders. This study focuses on naphthalene biodegradation by isolated bacterial strains from petroleum waste. Naphthalene poses significant threats to ecosystems, inducing genotoxic, mutagenic, and carcinogenic effects. Bioremediation offers a safe and cost-effective solution by employing microbes to degrade naphthalene into non-toxic by-products. In this study, Enterobacter ludwigii NS12 found as the most efficient strain for naphthalene biodegradation. In the subsequent investigation the effect of key process parameters, including pH and temperature, on the biodegradation of naphthalene were studied. Under optimal conditions (pH 7.0 and temperature of 30 °C), Enterobacter ludwigii NS12 exhibited an 89.2 % removal efficiency in the bioreactor for naphthalene wastewater at a concentration of 300 mg/L. The confirmations of naphthalene biodegradation were investigated through residual analysis, revealing dibutyl phthalate and 1-octanol, 2-butyl as metabolites. Additionally, growth kinetic parameters μmax and Ks were evaluated, resulting in values of 0.2380 per day and 158.57 mg/L for Enterobacter ludwigii NS12, respectively. The findings of this study hold significant promise for the bioremediation of naphthalene contamination in the environment.

Major facilitator superfamily transporter PhaT modulates the efflux and degradation of polycyclic aromatic hydrocarbons in Novosphingobium pentaromativorans US6-1

Polycyclic aromatic hydrocarbons (PAHs) are a class of persistent and toxic organic pollutants that are widely distributed in nature. Although catabolic pathways are essential for bacteria to effectively utilize PAHs, transporting PAHs and their intermediate metabolites across the bacterial cell envelope is a key factor in PAH catabolism and bacterial survival. In this study, we identified a major facilitator superfamily (MFS) transporter member, PhaT, in a PAH-degrading plasmid of Novosphingobium pentaromativorans US6–1. Evolutionary analyses indicate that PhaT analogs are widely distributed in PAH-degrading bacteria, including the genera Novosphingobium and Sphingobium. Interestingly, deletion of the phaT gene upregulated the levels of PAH-degrading enzymes and PAH-degrading gene expression, thereby accelerating phenanthrene biodegradation. Further studies demonstrated that a deficiency of PhaT impaired the efflux of phenanthrene and its benzene ring-containing intermediate metabolites. Furthermore, the deficiency of PhaT also promoted the biodegradation of naphthalene but not of benzo[a]pyrene. Molecular docking and site-directed mutagenesis revealed the essential roles of hydrophobic amino acids in the PhaT central cavity for binding to benzene ring-containing substrates. Collectively, these findings indicate that PhaT plays a pivotal role in facilitating the efflux of PAHs and their benzene ring-containing intermediate metabolites.

An integrated evaluation of bioenhanced in situ LNAPL dissolution

Performance evaluation of in situ bioremediation processes in the field is difficult due to uncertainty created by matrix and contaminant heterogeneity, inaccessibility to direct observation, expense of sampling, and limitations of some measurements. The goal of this research was to develop a strategy for evaluating in situ bioremediation of light nonaqueous-phase liquid (LNAPL) contamination and demonstrating the occurrence of bioenhanced LNAPL dissolution by: (1) integrating a suite of analyses into a rational evaluation strategy; and (2) demonstrating the strategy's application in intermediate-scale flow-cell (ISFC) experiments simulating an aquifer contaminated with a pool of LNAPL (naphthalene dissolved in dodecane). Two ISFCs were operated to evaluate how the monitored parameters changed between a “no bioremediation” scenario and an “intrinsic in situ bioremediation” scenario. Key was incorporating different measures of microbial activity and contaminant degradation relevant to bioremediation: contaminant loss; consumption of electron acceptors; and changes in total alkalinity, pH, dissolved total inorganic carbon, carbon-stable isotopes, microorganisms, and intermediate metabolites. These measurements were integrated via mass-flux modeling and mass-balance analyses to document that in situ biodegradation of naphthalene was strongly accelerated in the “intrinsic in situ bioremediation” scenario versus “no bioremediation.” Furthermore, the integrated strategy provided consistent evidence of bioenhancement of LNAPL dissolution through intrinsic bioremediation by a factor of approximately 2 due to the biodegradation of the naphthalene near the pool/water interface.

Biodegradation of naphthalene – Ecofriendly approach for soil pollution mitigation

Naphthalene (NPT), a widely used household pest repellent and insecticide obtained from crude oil, serves as a toxic pollutant to non-target living matter. The stable and resistant nature of NPT makes it difficult to degrade through the physiochemical processes. The present study investigated the bacterial degradation of NPT isolated from crude oil-contaminated soil. Initially, the potent bacteria, Bacillus sp. GN 3.4, were isolated by enrichment culture method and subsequently assessed for NPT biodegradation. The optimum conditions for NPT biodegradation were pH 7.0 at 37 °C, 80 mg/L (initial NPT), 3% v/v (inoculum dose), and 7 days of treatment which showed 100% biodegradation. Furthermore, GC–MS analysis revealed the presence of degradation metabolites, namely, salicylate and hydroquinone indicating potential metabolic pathways. Considering the water-solubility and non-toxic nature of these metabolites, the results imply that Bacillus sp. GN 3.4. could potentially play a role in bioremediation by aiding in eliminating NPT from the soil.

Simultaneous naphthalene degradation and electricity production in a biowaste-powered microbial fuel cell

Naphthalene is a very common and hazardous environmental pollutant, and its biodegradation has received serious attention. As demonstrated in this study, naphthalene-contaminated wastewater can be biodegraded using a microbial fuel cell (MFC). Furthermore, the potential of MFC for electricity generation appears to be a promising technology to meet energy demands other than those produced from fossil fuels. Nowadays, efforts are being made to improve the overall performance of MFC by integrating biowaste materials for anode fabrication. In this study, palm kernel shell waste was used to produce palm kernel shell-derived graphene oxide (PKS-GO) and palm kernel shell-derived reduced graphene oxide (PKS-rGO), which were then fabricated into anode electrodes to improve the system's electron mobilization and transport. The MFC configuration with the PKS-rGO anode demonstrated greater energy production potential, with a maximum power density of 35.11 mW/m2 and a current density of 101.76 mA/m2, compared to the PKS-GO anode, which achieved a maximum power density of 17.85 mW/m2 and a current density of 72.56 mA/m2. Furthermore, there is simultaneous naphthalene biodegradation with energy production, where the biodegradation efficiency of naphthalene with PKS-rGO and PKS-GO is 85.5%, and 79.7%, respectively. In addition, the specific capacitance determined from the cyclic voltammetry curve revealed a value for PKS-rGO of 2.23 × 10−4 F/g, which is also higher than the value for PKS-GO (1.57 × 10−4 F/g) on the last day of operation. Anodic microbial analysis shows that electrogens thrive in the MFC process. Finally, a comparison with previous literature and the future prospects of the study are also presented.

Biodegradation of Polycyclic Aromatic Hydrocarbon Compound by Bacterial Cultures

In the present study biodegradation of Polycyclic Aromatic Hydrocarbon (PAH) compound Naphthalene by four bacterial cultures Bacillus subtilis PD6, Bacillussp. PD9, Enterobactersp. PD11 and Bacillussp. PD14 has been targeted. Biodegradation of Naphthalene by these four selected bacterial cultures was analysed by HPLC (High Performance Liquid Chromatography) technique. HPLC analysis revealed biodegradation of naphthalene by all the four bacterial cultures within a span of six days. Highest biodegradation 78.1% has been shown by Bacillus subtilis PD6 while other bacterial cultures Bacillus sp. PD9 has shown 77.90%, Enterobacter sp. PD11 showed 74.4% and Bacillus sp. PD14 exhibited 73.5% biodegradation of naphthalene.

Tween-80 enhanced biodegradation of naphthalene by Klebsiella quasipneumoniae.

Accidental spillage of petroleum products and industrial activities result in various hydrocarbons in the environment. While the n-hydrocarbons are readily degraded, the polycyclic aromatic hydrocarbons (PAHs) are recalcitrant to natural degradation, toxic to aquatic life and are responsible for diverse health challenges in terrestrial animals; suggesting the need for faster and more eco-friendly ways of removing PAHs from the environment. In this study, the surfactant tween-80 was used to enhance a bacterium's intrinsic naphthalene biodegradation activity. Eight bacteria isolated from oil-contaminated soils were characterised using morphological and biochemical methods. The most effective strain was identified as Klebsiella quasipneumoniae using 16S rRNA gene analysis. High-Performance Liquid Chromatography (HPLC) analyses showed that the detectable concentration of naphthalene was decreased from 500 to 157.18μg/mL (67.4%) after 7 d in the absence of tween-80, while 99.4% removal was achieved in 3 d in the presence of tween-80 at 60μg/mL concentration. The peaks observed in the Fourier Transform Infra-Red Spectroscopy (FTIR) spectrum of control (naphthalene), which were absent in that of the metabolites, further established naphthalene degradation. Furthermore, Gas Chromatography-Mass Spectrometer (GCMS) revealed metabolites of single aromatic ring, such as 3,4-dihydroxybenzoic acid 4-hydroxylmethylphenol, which confirmed that the removal of naphthalene is by biodegradation. Tyrosinase induction and laccase activities suggested the involvement of these enzymes in naphthalene biodegradation by the bacterium. Conclusively, a strain of K. quasipneumoniae that can effectively remove naphthalene from contaminated environments has been isolated, and its biodegradation rate was doubled in the presence of non-ionic surfactant, tween-80.

Biodegradation of Naphthalene Using Biosurfactant Producing Fusarium proliferatum WC416 Isolated from Refinery Effluent.

Naphthalene is one of the priority pollutants in the environment which can effectively managed through bioremediation approach. Here fungal remediation of PAHs may be cost-effective technology. In present research study, biodegradation of naphthalene at varying concentrations in laboratory scale was investigated using Fusarium proliferatum WC416 isolated from refinery effluent. Degradation of naphthalene was computed by high-performance liquid chromatography (HPLC) and gas chromatography -mass spectroscopy (GC-MS). The results show that Fusarium proliferatum WC416 has effectively degraded the naphthalene to salicylamide and salicylic acid. The highest degradation achieved was 83% for the concentration of 100ppm followed 74% and 63% for the concentration of 250ppm and 500ppm respectively. The intermediates confirmed by GC-MS were salicylic acid (m/z identification 138) and salicylamide (m/z identification 137). During the study, fungal growth, COD, pH, and surface tension have been monitored, which show the continuous variation that indicates progressive biodegradation of naphthalene. The qualitative assessment of extracellular enzymes shows positive activity for lipase, manganese peroxidase, and laccase. The present study also assessed the biosurfactant production by fungus which was characterized as sophorolipid in nature. The study suggests that Fusarium proliferatum WC416 would be an efficient degrader of naphthalene for environmental clean-up.

Biodegradation of naphthalene by biocatalysts isolated from the contaminated environment under optimal conditions

Polycyclic aromatic hydrocarbons (PAHs) pollution occurs in freshwater and marine environment by anthropogenic activities. Moreover, analysis of the PAHs-degradation by the indigenous bacterial strains is limited, compared with other degraders. In this study, naphthalene (NAP) biodegrading bacteria were screened by enrichment culture method. Three bacterial strains were obtained for NAP degradation and identified as Bacillus cereus CK1, Pseudomonas aeruginosa KD4 and Enterobacter aerogenes SR6. The amount of hydrogen, carbon, sulphur and nitrogen of wastewater were analyzed. Total bacterial count increased at increasing incubation time (6–60 days) and moderately decreased at higher NAP concentrations. The bacterial population increased after 48 days at 250 ppm NAP (519 ± 15.3 MPM/mL) concentration and this level increased at 500 ppm NAP concentration (541 ± 12.5 MPM/mL). NAP was degraded by bacterial consortium within 36 h–99% at 30 °C. PAHs degrading bacteria were grown optimally at 4% inoculum concentrations. Bacterial consortium was able to degrade 98% NAP at pH 7.0 after 36 h incubation and degradation potential was improved (100%) after 34 h (pH 8.0). Also at pH 9.0, 100% biodegradation was registered after 36 h incubation. When the agitation speed enhanced from 50 ppm to 150 ppm, increased bacteria growth and increased NAP degradation within 42 h incubation. Among the nutrient sources, beef extract, peptone and glucose supplemented medium supported complete degradation of PAHs within 30 h, whereas peptone supported 94.3% degradation at this time. Glucose supplemented medium showed only 2.8% NAP degradation after 6 h incubation and reached maximum (100%) within 42 h incubation. Bacterial consortium can be used to reduce NAP under optimal process conditions and this method can be used for the removal of various hydrocarbon-compounds.

Biodegradation of Naphthalene and Anthracene by Aspergillus glaucus Strain Isolated from Antarctic Soil

Biotechnologies based on microbial species capable of destroying harmful pollutants are a successful way to solve some of the most important problems associated with a clean environment. The subject of investigation is the Antarctic fungal strain Aspergillus glaucus AL1. The culturing of the examined strain was performed with 70 mg of wet mycelium being inoculated in a Czapek Dox liquid medium containing naphthalene, anthracene, or phenanthrene (0.3 g/L) as the sole carbon source. Progressively decreasing naphthalene and anthracene concentrations were monitored in the culture medium until the 15th day of the cultivation of A. glaucus AL1. The degradation was determined through gas chromatography–mass spectrometry. Both decreased by 66% and 44%, respectively, for this period. The GC-MS analyses were applied to identify salicylic acid, catechol, and ketoadipic acid as intermediates in the naphthalene degradation. The intermediates identified in anthracene catabolism are 2-hydroxy-1-naphthoic acid, o-phthalic acid, and protocatechuic acid. The enzyme activities for phenol 2-monooxygenase (1.14.13.7) and catechol 1,2-dioxygenase (1.13.11.1) were established. A gene encoding an enzyme with catechol 1,2-dioxygenase activity was identified and sequenced (GeneBank Ac. No KM360483). The recent study provides original data on the potential of an ascomycete’s fungal strain A. glaucus strain AL 1 to degrade naphthalene and anthracene.

TREATMENT OF Pb – NAPHTHALENE CO – CONTAMINATION IN SOILS BY ZERO VALENT IRON NANOPARTICLES (NZVI) AND BIOREMEDIATION

The co-contamination of soils with heavy metals and organic contaminants is a problem that requires a different and innovative approach to remediate these kinds of sites. The different nature of these two contaminant groups makes the problem complicated. Recently, zero-valent iron nanoparticles (nZVI) application has been integrated with bioremediation to be used for the remediation of halogenated organics. However, the use of this process for the remediation of co-contaminated soils has not been investigated. In this study, nZVI application integrated with bioremediation approaches (bioattenuation and biostimulation) was applied to Pb and naphthalene spiked soils in laboratory-scale reactors. Soil Pb fractions and naphthalene concentrations were monitored for 90 days. The nZVI decreased the mobility and bioavailability of Pb significantly by reducing exchangeable and carbonate bound fractions from 66.2 % to 25.8 – 37.2 % range. The nZVI also caused naphthalene degradation in the range of 38.8 – 58.5 %. Significant biodegradation of naphthalene occurred at samples subjected to nZVI and biostimulation. The highest overall naphthalene degradation (76.7 %) was obtained from the nZVI application which was integrated with biostimulation.

Biodegradation of Naphthalene Using Glass Beads Roller Bioreactor: Application of Artificial Neural Network Modeling

A roller bioreactor containing inert glass beads was employed to enhance naphthalene biodegradation in an aqueous solution. Mixed culture of microorganisms was isolated from sewage waste sludge and adopted for naphthalene biodegradation. The dissolution rate of naphthalene in the aqueous phase was studied at different conditions in the roller reactor. The results indicate that the best conditions for the biodegradation treatment in the roller bioreactor were 40% and 50% loads with 6 mm and 5 mm sized glass beads, respectively, at a rotation speed of 50 rpm. The biodegradation of 300 mg/L naphthalene in the bioreactor in the absence of glass beads proceeded slowly until depletion after seven days. In the presence of glass beads, the biodegradation rate was faster, depleting after only four days. At a concentration of 500 mg/L naphthalene, complete depletion was attained after nine days in the bioreactor with no glass beads and five days in the presence of glass beads. Both the biodegradation rate \(({r}_{s}\)) and the specific growth rate\(\left(\mu \right)\) increased by utilizing the glass beads bioreactor compared with the bioreactor with no glass beads, thus confirming that the presence of glass beads enhances the mass transfer of naphthalene from the solid to the aqueous phase where it becomes available for utilization. An artificial neural network was used to model naphthalene dissolution and biodegradation. Correlation coefficients of 99.2% and 98.3% were obtained between the experimental and predicted output values for dissolution and biodegradation, respectively, indicating that the ANN model can efficiently predict the experimental data.

Biodegradation of hazardous naphthalene and cleaner production of rhamnolipids — Green approaches of pollution mitigation

Toxic and hazardous waste poses a serious threat to human health and the environment. Green remediation technologies are required to manage such waste materials, which is a demanding and difficult task. Here, effort was made to explore the role of Pseudomonas aeruginosa SR17 in alleviating naphthalene via catabolism and simultaneously producing biosurfactant. The results showed up to 89.2% naphthalene degradation at 35 °C and pH 7. The GC/MS analysis revealed the generation of naphthalene degradation intermediates. Biosurfactant production led to the reduction of surface tension of the culture medium to 34.5 mN/m. The biosurfactant was further characterized as rhamnolipids. LC-MS of the column purified biosurfactant revealed the presence of both mono and di rhamnolipid congeners. Rhamnolipid find tremendous application in medical field and as well as in detergent industry and since they are of biological origin, they can be used as favorable alternative against their chemical counterparts. The study demonstrated that catabolism of naphthalene and concurrent formation of rhamnolipid can result in a dual activity process, namely environmental cleanup and production of a valuable microbial metabolite. Additionally, the present-day application of rhamnolipids is highlighted.

Effects of Fe (Iii) and Magnetite on Naphthalene Biodegradation Under Sulfate Reducing Conditions

Effects of Fe (Iii) and Magnetite on Naphthalene Biodegradation Under Sulfate Reducing Conditions

Hydrocarbon biodegradation potential of microbial communities from high Arctic beaches in Canada's Northwest Passage

Sea ice loss is opening shipping routes in Canada's Northwest Passage, increasing the risk of an oil spill. Harnessing the capabilities of endemic microorganisms to degrade oil may be an effective remediation strategy for contaminated shorelines; however, limited data exists along Canada's Northwest Passage. In this study, hydrocarbon biodegradation potential of microbial communities from eight high Arctic beaches was assessed. Across high Arctic beaches, community composition was distinct, potential hydrocarbon-degrading genera were detected and microbial communities were able to degrade hydrocarbons (hexadecane, naphthalene, and alkanes) at low temperature (4 °C). Hexadecane and naphthalene biodegradation were stimulated by nutrients, but nutrients had little effect on Ultra Low Sulfur Fuel Oil biodegradation. Oiled microcosms showed a significant enrichment of Pseudomonas and Rhodococcus. Nutrient-amended microcosms showed increased abundances of key hydrocarbon biodegradation genes (alkB and CYP153). Ultimately, this work provides insight into hydrocarbon biodegradation on Arctic shorelines and oil-spill remediation in Canada's Northwest Passage.

Paenibacillus polymyxa A26 and Its Surfactant-Deficient Mutant Degradation of Polycyclic Aromatic Hydrocarbons

We compared the ability of two bacterial strains, Paenibacillus polymyxa A26 and P. polymyxa A26Sfp, for biodegradation of naphthalene (NAP). The studies were performed under simulated laboratory conditions, in liquid medium and soil with different carbon sources, pH and salt contents. Changes in the luminescence inhibition of Aliivibrio fischeri, as an indicator of the baseline toxicity, were observed in degradation mixtures during 7 days of incubation. While both strains expressed the best growth and NAP degradation ability in the minimal salt medium containing sucrose and 5% NaCl at pH 7 and 8, the mutant strain remained effective even under extreme conditions. A26Sfp was found to be an efficient and potentially industrially important polycyclic aromatic hydrocarbon degradation strain. Its extracellular polysaccharide production is 30%, and glucan production is twice that of the wild type A 26. The surface tension reduction ability was ascertained as 25–30% increased emulsification ability.

Unveiling metabolic characteristics of an uncultured Gammaproteobacterium responsible for in situ PAH biodegradation in petroleum polluted soil.

Exploring the metabolic characteristics of indigenous PAH degraders is critical to understanding the PAH bioremediation mechanism in the natural environment. While stable-isotopic probing (SIP) is a viable method to identify functional microorganisms in complex environments, the metabolic characteristics of uncultured degraders are still elusive. Here, we investigated the naphthalene (NAP) biodegradation of petroleum polluted soils by combining SIP, amplicon sequencing and metagenome binning. Based on the SIP and amplicon sequencing results, an uncultured Gammaproteobacterium sp. was identified as the key NAP degrader. Additionally, the assembled genome of this uncultured degrader was successfully obtained from the 13 C-DNA metagenomes by matching its 16S rRNA gene with the SIP identified OTU sequence. Meanwhile, a number of NAP degrading genes encoding naphthalene/PAH dioxygenases were identified in this genome, further confirming the direct involvement of this indigenous degrader in the NAP degradation. The degrader contained genes related to the metabolisms of several carbon sources, energy substances and vitamins, illuminating potential reasons for why microorganisms cannot be cultivated and finally realize their cultivation. Our findings provide novel information on the mechanisms of in situ PAH biodegradation and add to our current knowledge on the cultivation of non-culturable microorganisms by combining both SIP and metagenome binning.