• Korean Journal of Microbiology
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• v.40 no.4
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• pp.313-319
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• 2004

The distribution and characteristics of acidotolerant heterotrophic and naphthalene-degrading bacteria were investigated in two forest areas, one near Ulsan petrochemical industrial complex (Sunam) and the other in countryside (Daeam). Average values of soil pH at Sunam and Daeam were 3.8 and 4.6, respectively. When het­erotrophic and naphthalene-degrading bacteria were enumerated by most probable number (MPN) procedures at Sunam, the median values of heterotrophs growing at pH 7.0 and pH 4.0 were $5.3{\times}10^7\;and\;3.3{times}10^7$ MPN/g, whereas those of naphthalene-degraders were $5.6{\times}10^4\;and\;4.0{times}10^5$ MPN/g, respectively. While the medians of heterotrophs at Daeam were larger than those at Sunam, the concentrations of naphthalene-degraders were higher at Sunam compared to those at Daeam. From the MPN tubes and enrichment cultures, we obtained 17 isolates of naphthalene-degraders which were identified as Sphingomonas paucimobilis, Brevundimonas vesic­ularis, Burkholderia cepacia, Ralstonia pickettii, Pseudomanas fluorescens, and Chryseomonas luteola. Among them, 6 isolates showed higher naphthalene-degrading activity on minimal media of pH 4 compared to pH 7, whereas the extent of growth was not greater at pH 4 than at pH 7 when they were inoculated on nutrient-rich media. It is plausible that the pH may affect naphthalene-degrading activity of the isolates by changing fatty acid composition of bacterial membrane.

• Journal of Microbiology and Biotechnology
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• v.16 no.11
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• pp.1819-1825
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• 2006

Four naphthalene-degrading bacteria (Pseudomonas sp. strains O1, W1, As1, and G1) were isolated feom pollutant-contaminated sites. Examination of their substrate utilization and analyses of key naphthalene-catabolic regulatory genes revealed that the pathway and regulation of naphthalene-degradation in all four strains resemble those of NAH7 from P. putida G7. Superoxide anion production, superoxide dismutase activity, and catalase activity during their growth on naphthalene-amended medium increased significantly, compared with those with glucose-amended medium. Addition of ascorbate, an antioxidant, or ferrous iron ($Fe^{2+}$) increased the growth rates of all tested microorganisms on naphthalene. Northern blot and HPLC analyses showed that both nahA gene expression and naphthalene degradation increased under those conditions. Our data suggest that naphthalene degradation can impose severe oxidative stress, and defenses against oxidative stress would play an important role in the metabolism of naphthalene.

• Microbiology and Biotechnology Letters
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• v.16 no.3
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• pp.199-204
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• 1988

A strain capable of growth on naphthalene minimal medium was isolated from soil by selective enrichment culture and identified as Pseudomonas putida N3 according to its morphological and physiological characteristics. The optimum pH and temperature for growth of the isolate were 7.0 and 3$0^{\circ}C$, respectively. This strain was resistant to ampicillin, chloramphenicol, kanamycin and streptomycin but. sensitive to tetracycline and rifampicin. Of the naphthalene related compounds, 1, 5-dihydroxynaphthalene was more easily utilized than naphthalene due to its solubility. And catechol was degraded through meta-cleavage pathway. A 110 Kb plasmid which encodes for a single set of enzymes responsible for the degradation of naphthalene was obtained.

• Advances in environmental research
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• v.1 no.3
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• pp.201-210
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• 2012

The bioavailability of sorbed organic contaminants is one of the most important factors used to determine their fate in the environment. This study was conducted to evaluate the bioavailability of slow-desorbable naphthalene in soils. An air sparging system was utilized to remove dissolved (or desorbed) naphthalene continuously and to limit the bacterial utilization of dissolved naphthalene. A biological air sparging system (air sparging system with bacteria) was developed to evaluate the bioavailability of the slow-desorption fraction in soils. Three different strains (Pseudomonas putida G7, Pseudomonas sp. CZ6 and Burkholderia sp. KM1) and two soils were used. Slow-desorbable naphthalene continuously decreased under air sparging; however, a greater decrease was observed in response to the biological air sparging system. Enhanced bioavailability was not observed in the Jangseong soil. Overall, the results of this study suggests that the removal rate of slow-desorbable contaminants may be enhanced by inoculation of degrading bacteria into an air sparging system during the remediation of contaminated soils. However, the enhanced bioavailability was found to depend more on the soil properties than the bacterial characteristics.

• Journal of Microbiology
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• v.43 no.6
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• pp.572-576
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• 2005

This study focused on detecting catabolic genes for polycyclic aromatic hydrocarbons (PAHs) distributed in the reed rhizosphere of Sunchon Bay, Korea. These marsh and mud environments were severely affected by human activities, including agriculture and fisheries. Our previous study on microbial roles in natural decontamination displayed the possibility that PAH-degrading bacteria, such as Achromobacter sp., Alcaligenes sp., Burkholderia sp. and Pseudomonas sp. play an important decontamination role in a reed rhizosphere. In order to gain further fundamental knowledge on the natural decontamination process, catabolic genes for PAH metabolism were investigated through PCR amplification of dioxygenase genes using soil genomic DNA and sequencing. Comparative analysis of predicted amino acid sequences from 50 randomly selected dioxygenase clones capable of hydroxylating inactivated aromatic nuclei indicated that these were divided into three groups, two of which might be originated from PAH-degrading bacteria. Amino acid sequences of each dioxygenase clone were a part of the genes encoding enzymes for initial catabolism of naphthalene, phenanthrene, or pyrene that might be originated from bacteria in the reed rhizosphere of Sunchon Bay.

• Korean Journal of Microbiology
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• v.24 no.1
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• pp.67-72
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• 1986

Aromatic hydrocarbon degrading bacteria were isolated from industrial waste by using an agar plate method. The isolate DY-1 was identified as Acinetobacter sp. and found to utilize phenanthrene as tis sole carbon source. THe bacteria were proved to produce salicylic acid as an intermediate from phenanthrene through naphthalene pathway, when the products in the culture were wxamined by thin-layer chromatography. THe $Phn^+$ genes were found to be involved in two plasmids of about 4 and 40kb which were lost and not detected in the DNA samples prepared from the mitomycin C-cured cells by a gel electrophoretic analysis.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.89-91
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• 2000

Polynuclear aromatic hydrocarbon (PAH) compounds are highly carcinogenic chemicals and common groundwater contaminants that are observed to persist in soils. The adherence and slow release of PAHs in soil is an obstacle to remediation and complicates the assessment of cleanup standards and risks. Biological degradation of PAHs in soil has been an area of active research because biological treatment may be less costly than conventional pumping technologies or excavation and thermal treatment. Biological degradation also offers the advantage to transform PAHs into non-toxic products such as biomass and carbon dioxide. Ample evidence exists for aerobic biodegradation of PAHs and many bacteria capable of degrading PAHs have been isolated and characterized. However, the microbial degradation of PAHs in sediments is impaired due to the anaerobic conditions that result from the typically high oxygen demand of the organic material present in the soil, the low solubility of oxygen in water, and the slow mass transfer of oxygen from overlying water to the soil environment. For these reasons, anaerobic microbial degradation technologies could help alleviate sediment PAH contamination and offer significant advantages for cost-efficient in-situ treatment. But very little is known about the potential for anaerobic degradation of PAHs in field soils. The objectives of this research were to assess: (1) the potential for biodegradation of PAH in field aged soils under denitrification conditions, (2) to assess the potential for biodegradation of naphthalene in soil microcosms under denitrifying conditions, and (3) to assess for the existence of microorganisms in field sediments capable of degrading naphthalene via denitrification. Two kinds of soils were used in this research: Harbor Point sediment (HPS-2) and Milwaukee Harbor sediment (MHS). Results presented in this seminar indicate possible degradation of PAHs in soil under denitrifying conditions. During the two months of anaerobic degradation, total PAH removal was modest probably due to both the low availability of the PAHs and competition with other more easily degradable sources of carbon in the sediments. For both Harbor Point sediment (HPS-2) and Milwaukee Harbor sediment (MHS), PAH reduction was confined to 3- and 4-ring PAHs. Comparing PAH reductions during two months of aerobic and anaerobic biotreatment of MHS, it was found that extent of PAHreduction for anaerobic treatment was compatible with that for aerobic treatment. Interestingly, removal of PAHs from sediment particle classes (by size and density) followed similar trends for aerobic and anaerobic treatment of MHS. The majority of the PAHs removed during biotreatment came from the clay/silt fraction. In an earlier study it was shown that PAHs associated with the clay/silt fraction in MHS were more available than PAHs associated with coal-derived fraction. Therefore, although total PAH reductions were small, the removal of PAHs from the more easily available sediment fraction (clay/silt) may result in a significant environmental benefit owing to a reduction in total PAH bioavailability. By using naphthalene as a model PAH compound, biodegradation of naphthalene under denitrifying condition was assessed in microcosms containing MHS. Naphthalene spiked into MHS was degraded below detection limit within 20 days with the accompanying reduction of nitrate. With repeated addition of naphthalene and nitrate, naphthalene degradation under nitrate reducing conditions was stable over one month. Nitrite, one of the intermediates of denitrification was detected during the incubation. Also the denitrification activity of the enrichment culture from MHS slurries was verified by monitoring the production of nitrogen gas in solid fluorescence denitrification medium. Microorganisms capable of degrading naphthalene via denitrification were isolated from this enrichment culture.

• Microbiology and Biotechnology Letters
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• v.41 no.1
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• pp.79-87
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• 2013

An unstable yet efficient phenanthrene-degrading bacterium strain Ph-3 was isolated from a petroleum-contaminated site at the Mathura Oil Refinery, India. The strain was identified as Pseudomonas sp. using a polyphasic approach. An analysis of the intermediates and assays of the degradative enzymes from a crude extract of phenanthrene-grown cells showed a novel and previously unreported pattern of 1, 2-dihydroxy naphthalene and salicylic acid production. While strain Ph-3 lost its phenanthrene- degrading potential during successive transfers on a rich medium, it maintained this trait in oligotrophic soil conditions under the stress of the pollutant and degraded phenanthrene efficiently in soil microcosms. Although the maintenance and in vitro study of unstable phenotypes are difficult and such strains are often missed during isolation, purification, and screening, these bacteria constitute a substantial fraction of the microbial community at contaminated sites and play an important role in pollutant degradation during biostimulation or monitored natural attenuation.

• Journal of Soil and Groundwater Environment
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• v.14 no.6
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• pp.45-52
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• 2009

Naphthalene-degrading bacteria Pseudomonas aeruginosa CZ6 isolated from contaminated soil can adhere to crystal naphthalene and produce extracellular polymeric substance. LB, YM and MSM medium were used as culture mediums to investigate the formation of biofilm. Biofilm was developed the most in LB medium by Pseudomonas aeruginosa CZ6. In the culture, strain CZ6 growth was rarely affected by naphthalene concentration. Optimal culture condition was $30^{\circ}C$ and pH 7 at 0.10% substrate and 150 rpm shaking. The effect of culture medium on naphthalene degradation in the two soil slurry system was evaluated. The initial degradation rate of naphthalene was highest in the MSM medium of soil slurry. However, the sorbed naphthalene was rapidly degraded at the LB medium when naphthalene availability in liquid was limited. The results of this study suggest that biofilm formation and extracellular polymeric substance production increased bioavailability of soil sorbed naphthalene.

• Journal of Soil and Groundwater Environment
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• v.12 no.6
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• pp.70-77
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• 2007

Pseudomonas sp. KM1 was separated from soil contaminated by petroleum and identified. The isolated strain is Gram-positive, rod-shaped and immotile. In batch culture, the optimum cultivation temperature and pH was $35^{\circ}C$ and 7, respectively. Biodegradation of PAHs experiment with soil slurry system was performed using Pseudomonas sp. KM1. Pseudomonas sp. KM1 could degrade 7 PAHs including naphthalene, acenaphthylene, acenaphthene, fluorene, phenanthrene, pyrene, and fluoranthene. These mixed PAHs was easily degraded within one day except fluoranthene, which was degraded much slowly, taking several days by this isolated bacteria. Pseudomonas sp. KM1 is good candidate for bioremediation of PAHs contaminated soils. Biodegradation rates of naphthalene, phenanthrene and pyrene in soils were different at each soil, and the rates were decreased as sorption capacity increased.