• Korean Journal of Environmental Agriculture
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• v.34 no.3
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• pp.244-251
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• 2015

BACKGROUND: Recent studies have described the importance of bacteria that can degrade polycyclic aromatic hydrocarbons (PAHs). Here we screened bacterial isolates from commercial gasoline for PAH degraders and characterized their ability to degrade PAHs, lipids and proteins as well as their enantioselective epoxide hydrolase activity, salt tolerance, and seawater survival. METHODS AND RESULTS: One hundred two bacteria isolates from commercial gasoline were screened for PAH degraders by adding selected PAHs on to the surface of agar plates by the sublimation method. A clear zone was found only around the colonies of PAH degraders, which accounted for 13 isolates. These were identified as belonging to Bacillus sp., Brevibacterium sp., Micrococcus sp., Corynebacterium sp., Arthrobacter sp., and Gordonia sp. based on 16S rRNA sequences. Six isolates belonging to Corynebacterium sp., 3 of Micrococcus sp., Arthrobacter sp. S49, and Gordonia sp. H37 were lipid degraders. Arthrobacter sp. S49 was the only isolate showing high proteolytic activity. Among the PAH-degrading bacteria, Arthrobacter sp. S49, Brevibacterium sp. S47, Corynebacterium sp. SK20, and Gordonia sp. H37 showed enantioselective epoxide hydrolase activity with biocatalytic resolution of racemic styrene oxide. Among these, highest enantioselective hydrolysis activity was seen in Gordonia sp. H37. An intrinsic resistance to kanamycin was observed in most of the isolates and Corynebacterium sp. SK20 showed resistance to additional antibiotics such as tetracycline, ampicillin, and penicillin. CONCLUSION: Of the 13 PAH-degraders isolated from commercial gasoline, Arthrobacter sp. S49 showed the highest lipid and protein degrading activity along with high active epoxide hydrolase activity, which was the highest in Gordonia sp. H37. Our results suggest that bacteria from commercial gasoline may have the potential to degrade PAHs, lipids, and proteins, and may possess enantioselective epoxide hydrolase activity, high salt tolerance, and growth potential in seawater.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 2002.04a
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• pp.22-25
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• 2002

The effect of in-situ chemical oxidation on the indigenous soil microorganisms (total microbes and PAH-degrading microbes) and contaminant removal were investigated. Field soil contaminated with diesel in gas station was collected and the soil was treated from 0 to 900 minutes by in-situ ozonation as chemical remediation. The treated soil samples were incubated with supplying oxygen during the 9 weeks to understand the characteristics of microbes regrowth, damaged by ozone. The sharp decrease of aromatic fraction and TPH was observed within 60 minutes of ozone application and aromatic fraction and TPH then slowly decreased. The phenanthren-degrading bacteria were the most sensitive to ozonation, because 1 hour of ozonation reduced the microbes from 10$^{6}$ CFU/g-soil to below detection limits.

• 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.

• Proceedings of the Zoological Society Korea Conference
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• 1996.10a
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• pp.95.2-95
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• 1996

No Abstract, See Full Text

• Proceedings of the Zoological Society Korea Conference
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• 1997.10b
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• pp.130.2-130
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• 1997

No Abstract, See Full Text

• 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.

• Korean Journal of Microbiology
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• v.39 no.2
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• pp.108-113
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• 2003

Diesel oil-degrading bacterial strains were isolated from diesel oil contaminated soil and called HS series (HS1, HS2 and HS3). These strains were identified as Acinetobacter sp. (HS1) and Pseudomonas sp. (HS2 and HS3) based on Biolog test, cellular fatty acid composition, and 16S rDNA sequence analysis. These strains were coltivated in liquid minimal media containing 2% diesel oil, and diesel oil-degrading activity was measured. As result, all strains degraded over 70% of total diesel oil. But PAH (polycyclic aromatic hydrocarbon)- and pris- tane-degrading rate of these strain was below 20％ of total PAH and pristane. The HS 1 strain showed highest hydrophobicity and low emulsifying activity among the experimental strains and high diesel oil-degrading activity. From the above-mentioned result, microcosm experiment was performed with the HS1 strain. The HS1 strain showed a degrading activity of over 80% of total diesel oil in microcosm test. And microbial activity was correlated to diesel oil-degrading activity. Therefore, it is suggested that the HS1 strains could be effectively used for the bioremediation for diesel oil.

• Journal of Microbiology and Biotechnology
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• v.13 no.6
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• pp.942-948
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• 2003

Soil samples were obtained from 5 sites contaminated with polycyclic aromatic hydrocarbons (PAHs). These soil samples were cultured in using phenanthrene as a sole carbon and energy source, and 36 strains of phenanthrene-degrading bacteria were isolated from 3 sites. Most of them degraded 500 ppm of phenanthrene within 8 to 10 days, and these isolates could degrade a few other PAHs other than phenanthrene. Their genotypes were determined by restriction digests of the l6S rRNA genes [amplified ribosomal DNA restriction analysis (ARDRA)]. It was found that all the phenanthrene degrading isolates were included in 4 ARDRA types, and they showed a strict site endemism. l6S rDNAs of 12 strains selected from different sites were sequenced, and they were all confirmed as Sphingomonas strains. Their l6S rDNA sequences were compared for phylogenetic analysis; their sequence showed a similar result to ARDRA typing, thus indicating that these heterotrophic soil bacteria are not regionally mixed. In addition, it was found that the microbial diversity among sampling sites could be monitored by l6S rDNA PCR-RFLP pattern alone, which is simpler and easier to perform, without l6S rDNA sequence analysis.

• Korean Journal of Microbiology
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• v.41 no.3
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• pp.208-215
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• 2005

This study was accomplished in order to collect fundamental data on microbial roles in recycling process of reed rhizosphere. Sunchon bay, which is considered as one of the marsh and mud environments severely affected by human activities such agriculture and fisheries, was selected as a model place. In our initial efforts, two bacterial consortia were obtained by enrichment culture using PAH mixtures containing anthracene, naphthalene, phenanthrene and pyrene as the sources of carbon and energy, and four pure bacteria capable of rapid degradation of PAH were isolated from them. Four strains designated as SCB1, SCB2, SCB6, and SCB7 revealed by morphological, physiological and molecular analyses were identified as Burkholderia anthina, Alcaligenes sp., Achromobacter xylosoxidans., and Pseudomonas putida, respectively with over $99{\%}$ confidence. Notably, Burkholderia anthina SCB1 and Alcaligenes sp. SCB2 were found to utilize anthracene and pyrene more quickly than naphthalene and phenanthrene, whereas Achromobacter xylosoxidans SCB6 and Pseudomonas putida SCB7 exhibited similar growth and degradation patterns except for pyrene. These facts suggest that the rhizosphere microorganisms capable of PAH degradation might be used to clean up the contamination sites with polycyclic aromatic hydrocarbons.

• Journal of Life Science
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• v.21 no.11
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• pp.1511-1517
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• 2011

We isolated a potent phenanthrene-degrading bacterium from oil-contaminated soils of Suzhou, China, and assessed the potential use of these bacteria for bioremediation of soils contaminated by polycyclic aromatic hydrocarbons (PAHs) in a microcosm. Based on 16S rDNA sequencing, we identified this bacteria as Sphigobacterium sp. SW-09. By PCR amplification, we also identified catechol 2,3-dioxygenase genes (nahH genes) mediating PAH degradation. Staphylococcus sp. KW-07, which has been identified in our previous study, showed potential for use in bioremediation of oil-contaminated soils. In this experiment, we compared the rate of phenanthrene-degradation between Staphylococcus sp. KW-07 and Sphingobacterium sp. SW-09 in a microcosm condition. Newly isolated Sphingobacterium sp. SW-09 showed a higher phenanthrene-degradation rate than that of Staphylococcus sp. KW-07 in soil microcosms. Together, our results suggest that the Sphingobacterim sp. SW-09 strain isolated from the Suzhou area may also be useful in bioremediation of PAH-contaminated soils.