• Journal of Korea Soil Environment Society
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• v.1 no.2
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• pp.35-42
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• 1996

Seawater samples were collected from P'ohang coastal area during April 1995 - January 1996. The distribution of total heterotrophic bacteria and crude oil-degrading bacteria (CDB) were studied. In addition, biodegradation of crude oil was investigated through mono and mixed culture. The heterotrophic bacterial distribution was in the range of 4.1 $\times$ $10^4$- 1.2 $\times$ $10^5$ CFU/$m\ell$, respectively. The percent of crude oil-degrading bacteria against total heterotrophic bacteria was 0.05-0.54% which was lower than other marine samples reported. Therefore it could be suggested that the distribution of crude oil-degrading bacteria in the seawater of P'ohang coastal area was highly affected by presence of petroleum hydrocarbon. Taxonomical characteristics of 26 isolates were investigated. The results of identification were showed 7 genera which were Acinetobacter spp., Bacillus spp., Citrobacter spp., Micrococcus spp., Moraxella spp., Rhodococcus spp., and Serratia spp. Appearance of Enterobacteriaceae indicated that the seawater was polluted with wastewater. Also genus of Bacillus had predominant in CDB on P'ohang coastal area. In flask culture, biodegradation of crude oil was enhanced by addition of mixed culture of CDB.

• Journal of Microbiology
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• v.34 no.4
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• pp.363-369
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• 1996

As basic study to evaluate the treatability of oil-contaminated environment with bacteria, isolation and characterization of crude oil-degrading bacterium were carried out. A bacterial strain SH-14 capable of degrading crude oil was isolated from contaminated soils by enrichment culture technique and identified as Acinetobacter sp. by morphological, cultural and biochemical characteristics, and so named Acinetobacter sp. SH-14. The optimal medium composition and cultural conditions for the growth and emulsification of crude oil by Acinetobacter sp. SH-14 used were crude oil of 2.0%, $KNO_3$ of 0.2%, $K_2HPO_4$ of 0.05%, and $MgSO_4\;{\cdot}\;7H_2O$ of 1.0%, along with initial pH 7.0 at $30^{\circ}C$. Acinetobacter sp. SH-14 showed to be resistant to chloramphenicol and utilized various hydrocarbons such as dodecane, hexadecane, isooctane, cyclo-hexane etc., as a sole carbon source. Acinetobacter sp. SH-14 harbored a single plasmid. By agarose gel electrophoresis and curing experiment it was found that the genes for crude oil components degradation were encoded on the plasmid.

• KSBB Journal
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• v.15 no.1
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• pp.27-34
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• 2000

A marine bacterium having a high oil-degrading activity was isolated form the oil-polluted southern sea of Korea, and was identified as Pseudomonas aeruginosa and was named Pseudomonas aeruginosa BYK-2. The optimal tmeperatur, culture time, pH and NaCl concentration for biosurfactant production and cell growth showed $25^{\circ}C$, 48h, 7.0 and 0%(w/v), respectively. After cultivation at $25^{\circ}C$, 180 rpm in 250 mL erlenmeyer flask for 7days, 1%(w/v) arabian light crude oil and bunker C oil which are considered to be hardly degradable compounds were degraded 92.1%(w/w) and 76%(w/w) respectively. And then, cell adherence was measured on various carbon sources. The cell adherence indicated over 80% on hydrocarbons(arabian light crude oil, kuwait curde oil, bunker C oil, n-paraffine, n-hexadecane, n-tetradecane) as carbon sources. Lecithin among fatty acids(oleic acid, olive oil, lecithin) showed highest cell adherence of 91.5%. The cell adherence of sugars(arabinose, trehalose, dextrose, galactose, lactose, fructose, maltose, sorbitol, sucrose) observed to be less than 70% except for arabinose, galactose, sorbitol and sucrose.

• Journal of Microbiology
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• v.44 no.3
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• pp.354-359
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• 2006

This study examined the capacity of immobilized bacteria to degrade petroleum hydrocarbons. A mixture of hydrocarbon-degrading bacterial strains was immobilized in alginate and incubated in crude oil-contaminated artificial seawater (ASW). Analysis of hydrocarbon residues following a 30-day incubation period demonstrated that the biodegradation capacity of the microorganisms was not compromised by the immobilization. Removal of n-alkanes was similar in immobilized cells and control cells. To test reusability, the immobilized bacteria were incubated for sequential increments of 30 days. No decline in biodegradation capacity of the immobilized consortium of bacterial cells was noted over its repeated use. We conclude that immobilized hydrocarbon-degrading bacteria represent a promising application in the bioremediation of hydrocarbon-contaminated areas.

• Microbiology and Biotechnology Letters
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• v.21 no.1
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• pp.74-81
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• 1993

To investigate the oil degrading properties of psychrotrophic bacterium Acinetobacter calcoaceticus Al-l the effects of environmental factors on this bacterium were studied. The optimal environmental conditions for cell growth rate and oil-emulsifying activity were as follows; temperature $15^{\circ}C$, pH 7.5, salt concentration 0- 3% and crude oil concentration 0.1%. Additionally the optimal concentration of Nand P source for cell growth rate and oil-emulsifying activity were 0.76 mM and 0.057 mM as $(NH_4)HS0_4$ and $K_2HP0_4$, respectively. Analysis of remnant oil by gas chromatography showed time dependent oil degradation pattern by A. calcoaceticus during cultivation; At lOoe and $15^{circ}$e, most alkane peaks were disappeared and it showed large quantities of crude oil were degraded. But at $25^{circ}$e alkane compounds in the crude oil were partially degraded even after 120 hours incubation.

• Journal of Life Science
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• v.21 no.11
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• pp.1599-1606
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• 2011

To evaluate the effects of microorganism agents on oil biodegradation, treatability and microcosm studies were conducted. Petroleum oil degrading bacteria were isolated from enriched cultures of oil-contaminated sediment samples using a mineral salts medium (MSM) containing 0.5% Arabian heavy crude oil as the sole carbon source. After a 5 day-incubation period using MSM, mixed microorganisms of three species (strains BS1, BS2 and BS4) degraded 48.4% of aliphatic hydrocarbons and 30.5% of aromatic hydrocarbons. Treatability and microcosm tests were performed in the three different treatment conditions (AO: Arabian heavy crude oil, AO+IN: Arabian heavy crude oil+inorganic nutrient, AO+IN+MM: Arabian heavy crude oil+inorganic nutrient+mixed microorganism agents). Among these, significantly enhanced biodegradation of aliphatic hydrocarbons were observed in AO+IN and AO+IN+MM conditions, without showing any different biodegradation rates in either condition. However, the degradation rates of aromatic hydrocarbons in an AO+IN+MM condition were increased by 50% in the treatability test and by 13% in the microcosm test compared to those in an AO+IN condition. Taken together, it can be concluded that mixed microorganism agents enhance the biodegradation of aliphatic and aromatic hydrocarbons in laboratory, a treatability test, and a microcosm test. This agent could especially be a useful tool in the application of bioremediation for removal of aromatic hydrocarbons.

• Journal of Korean Society of Environmental Engineers
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• v.22 no.10
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• pp.1851-1859
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• 2000

Crude oil-degrading bacteria were isolated from the sites contaminated by oil products. The isolates were identified as Acinetobacter sp. A132, Pseudomonas putida A422, Pseudomonas aeruginosa F721, F722, and Xanthomonas maltophilia B823. The results of investigation on the degradability of crude oil indicated that the strain A132 had the highest rate of $6.04g/L{\cdot}day$. Also, the strain A132 and F722 almost degraded each of n-alkane compounds between $nC_{10}$ and $nC_{32}$. The strain A422 degraded benzene and xylene but not n-alkane. The strain B823 grew somewhat in crude oil but did not entirely degrade other substrates used in this study. The results of the GC/FID analysis on the degradability of the mixed n-alkane compounds showed that the strain F722 could degrade 100% of the compounds with $nC_7{\sim}nC_{10}$ and more than 80% of those with $nC_{11}{\sim}nC_{24}$.

• 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.

• Microbiology and Biotechnology Letters
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• v.21 no.1
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• pp.66-73
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• 1993

Psychrotrophic bacterial strains utilizing crude oil as their sole carbon and energy sources were isolated from Antarctic soil and sea sediments. One of the strains named AI-I showed the hightest activity for emulsification of crude oil and the best growth. This strain was identified as Acinetobacter calcoaceticus. A. calcoaceticus AI-I strain contains a plasmid (OCT plasmid) which was related to the utilization of alkane compounds. The molecular weight of this plasmid was estimated to be about 110 Md by agarose gel electrophoresis. The cured strain of A. calcoaceticus AI-I strain (OCT ) was not able to utilize normal hydrocarbon compounds ($C_6C_{17}$) as carbon and energy sources. A. ca/coaceticus AI-1 was resistant to ampicillin and sensitive to streptomycin, kanamycin, chloramphenicol, tetracycline. The results suggested that this strain carries a plasmid (OCT) responsible for oil utilization which is quite stable and might be concerned with antibiotics resistancy.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.36 no.3
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• pp.230-238
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• 2003

Biosurfactant-producing bacteria, showing strong crude oil degrading activity, were isolated from the caverns of National Oil Storage Basement. From the results of biochemical and molecular biological tests, the isolate was identified as Pseudomonas fluorescens PD101. It grows well on liquid media at temperature range from $20^{\circ}C\;to\;37^{\circ}C,$ but it does not produce biosurfactant when grown at $37^{\circ}C$ or at higher temperature. The biosurfactant was stable at broad pH range from 5 to 11 and under heat treatment condition of $100^{\circ}C$ for 30 min. The biosurfactant produced dark blue halo around the colony when grown on SW agar plates, which could confirm the biosurfactant as one of rhamnolipid group. The 700 bp of PCR product could be amplified from DNA of P. flurorescens PD101 by using PCR primers designed from rh1A gene of P. aeruginosa, and it showed $99\%$ of sequence homology with rh1A gene of P. aeruginosa encoding rhamnosyltransferase 1.