• 미생물학회지
• /
• 제40권4호
• /
• pp.313-319
• /
• 2004

울산석유화학공단과 인접한 지역(선암)및 농촌지역(대암)의 산림토양에서 내산성 종속영양세균 및 나프탈렌분해세균의 분포와 특성을 조사하였다. 토양 pH의 평균은 선암과 대암에서 각각 3.8과 4.6으로 측정되었다. 종속영양세균과 나프탈렌분해세균을 최확수치(MPN)법으로 계수하였으며, 선암의 경우 pH 7.0과 pH 4.0에서 생장하는 종속영양세균 수의 중앙간은 각각 $5.3{\times}10^7\;3.3{\times}10^7$ MPN/dried Soil g이었고, 나프탈렌분해세균 수의 중앙간은 pH 7.0과 4.0에서 각각 $5.6{\times}10^4$ 와 $4.0{\times}10^5$ MPN/dried soil g이었다. 대암에서 측정한 종속영양세균수의 중앙간은 두 pH 모두 선암에서보다 많았으나 나프탈렌분해세균의 농도는 선암이 대암보다 높았다. MPN시험관과 농화배양으로부터 17개의 나프탈렌분해세균을 분리하였으며, 이들은 Sphingomonas paucimobilis, Brevundimonas vesicularis, Burkholderia cepacia, Ralstonia pickettii, Pseudomonas fluorescens, Chryseomonas luteola 등으로 동정되었다. 분리한 17개 균주 중 6개는, 최소배지에서의 나프탈렌분해 활성이 PH 7.0에 비해 PH 4.0에서 더 높았으나 영양배지에 접종하였을 때의 생장정도는 pH 4.0이 pH 7.0보다 더 크지 않았다. 배지의 pH가 분리한 세균의 세포막 지방산 조성을 변화시켜 나프탈렌분해 활성에 영향을 주는 것으로 생각된다.

• Journal of Microbiology and Biotechnology
• /
• 제16권11호
• /
• pp.1819-1825
• /
• 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.

• 한국미생물·생명공학회지
• /
• 제16권3호
• /
• pp.199-204
• /
• 1988

Naphthalene을 유일한 탄소원으로 이용하는 균을 분식 배양과 연속식 배양에 의해서 토양과 폐수로부터 분리하였다. 이 균은 Pseudomonas putida로 동정되었으며, 최적 pH와 온도는 각각 7.0과 3$0^{\circ}C$ 이었다. 분리된 균은 1,5-dihydroxynaphthalene을 naphthalene보다 더욱 잘 이용하였으며 benzoate와 salicylate도 이용하였다. 또한 catechol dl meta-분해경로를 통해서 분해되었으며, ampicillin, chloramphenicol, kanamycin, streptomycin에 대해서 강한 저항성을 지니고 있었으며, naphthalene의 분해에 관여하는 약 110kb 크기의 plasmid를 1개 지니고 있었다.

• Advances in environmental research
• /
• 제1권3호
• /
• pp.201-210
• /
• 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
• /
• 제43권6호
• /
• pp.572-576
• /
• 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.

• 미생물학회지
• /
• 제24권1호
• /
• pp.67-72
• /
• 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
• /
• 한국생물공학회 2000년도 춘계학술발표대회
• /
• pp.89-91
• /
• 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.

• 한국미생물·생명공학회지
• /
• 제41권1호
• /
• pp.79-87
• /
• 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.

• 한국지하수토양환경학회지:지하수토양환경
• /
• 제14권6호
• /
• pp.45-52
• /
• 2009

나프탈렌 분해균주인 Pseudomonas aeruginosa CZ6을 오염된 토양에서 분리하였으며 분리된 균주는 결정상태의 나프탈렌에 부착하고 그 주변에 extracellular polymeric substance를 분비하는 특성을 가졌다. LB, YM과 MSM 배지를 사용하여 배지의 종류에 따른 biofilm 생성량을 측정한 결과, LB 배지에서 biofilm이 가장 많이 생성되는 것으로 나타났다. 나프탈렌을 기질로 한 배양조건에서 균주는 기질의 농도 영향을 크게 받지 않고 0.10% 기질, 150 rpm 조건에서 최적 배양조건은 $30^{\circ}C$, pH 7로 나타났다. 두 가지 토양에서 배지의 종류에 따른 나프탈렌의 분해특성을 관찰한 결과 초기에는 MSM 배지에서 나프탈렌이 가장 많이 분해가 되었다. 그러나 생물이용성이 제한을 받는 조건에서 LB 배지의 나프탈렌이 가장 빨리 제거가 되었다. 이런 결과는 biofilm의 형성과 extracellular polymeric substance 생성이 토양에 흡착된 잔류 나프탈렌의 생물학적 이용성을 향상시키기 때문인 것으로 사료된다.

• 한국지하수토양환경학회지:지하수토양환경
• /
• 제12권6호
• /
• pp.70-77
• /
• 2007

토양에 존재하는 다핵방향족탄화수소(Polycyclic Aromatic Hydrocarbons, PAHs)의 처리를 위하여 자연계로부터 분리된 균주는 Pseudomonas sp.로 동정되었으며, 이 균주를 KM1으로 명명하였다. 균주의 최적 성장조건은 회분식 배양에서 $35^{\circ}C$, pH 7로 나타났다. 분리균주 Pseudomonas sp. KM1에 의한 7-PAHs(naphthalene, acenaphthylene, acenaphthene, fluorene, phenanthrene, fluoranthene and pyrene)의 분해실험결과 배양 1일 만에 fluoranthene을 제외한 naphthalene, acenaphthylene, acenaphthene, fluorene, phenanthrene and pyrene 이 분해됨을 확인할 수 있었다. 그리고 토양유무에 따른 PAHs 분해실험 결과, 흡착분배계수와 유기물함량(%)이 큰 신동방이 경방이나 봉동보다 분리균주에 의한 생분해율(%)이 낮았다. 토양에 오염된 유기화합물의 분배특성과 토양 내 유기물함량(%)이 오염된 토양의 생물학적 처리효과에 영향을 미치는 중요한 인자인 것으로 나타났다.