• Journal of Soil and Groundwater Environment
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• v.21 no.1
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• pp.72-79
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• 2016

Bioremediation is one of the most effective ways to remediate TPH-contaminated sites. However, under actual field conditions that are not at the optimum temperature, degradation of microorganisms is generally reduced, which is why the efficiency of biodegradation is known to be significantly affected by the soil temperature. Therefore, in this study, the labscale experiment was conducted using indigenous crude oil degrading microorganisms isolated from crude oil contaminated site to evaluate the remediation efficiency. Crude oil degrading microorganisms were isolated from crude oil contaminated soil and temperature, which is a significant factor affecting the remediation efficiency of land farming, was adjusted to evaluate the microbial crude oil degrading ability, degradation time, and remediation efficiency. In order to assess the field applicability, the remediation efficiency was evaluated using crude oil contaminated soil (average TPH concentration of 10,000 mg/kg or more) from the OO premises. Followed by the application of microorganisms at 30℃, the bioremediation process reduced its initial TPH concentration of 10,812 mg/kg down to 1,890 mg/kg in 56 days, which was about an 83% remediation efficiency. By analyzing the correlation among the total number of cells, the number of effective cells, and TPH concentration, it was found that the number of effective microorganisms drastically increased during the period from 10 to 20 days while there was a sharp decrease in TPH concentration. Therefore, we confirmed the applicability of land farming with isolated microorganisms consortium to crude oil contaminated site, which is also expected to be applicable to bioremediation of other recalcitrant materials.

• Journal of Microbiology and Biotechnology
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• v.15 no.6
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• pp.1170-1177
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• 2005

Terminal-restriction fragment length polymorphism (T-RFLP) analysis was used to monitor inoculated oil-degrading microorganisms during bioremedial treatability tests. A pair of universal primers, fluorescently labeled 521F and 1392R, was employed to amplify small subunit rDNA in order to simultaneously detect two bacterial strains, Corynebacterium sp. IC10 and Sphingomonas sp. KH3-2, and a yeast strain, Yarrowia lipolytica 180. Digestion of the 5'-end fluorescence/labeled PCR products with HhaI produced specific terminal-restriction fragments (T-RFs) of 185 and 442 bases, corresponding to Corynebacterium sp. IC10 and Y. lipolytica 180, respectively. The enzyme NruI produced a specific T-RF of 338 bases for Sphingomonas sp. KH3-2. The detection limit for oildegrading microorganisms that were inoculated into natural environments was determined to be $0.01\%$ of the total microbial count, regardless of the background environment. When three oil-degrading microorganisms were released into oil-contaminated sand microenvironments, strains IC10 and 180 survived for 35 days after inoculation, whereas strain KH3-2 was detected at 8 days, but not at 35 days. This result implies that T-RFLP could be a useful tool for monitoring the survival and relative abundance of specific microbial strains inoculated into contaminated environments.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.578-581
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• 2001

A treatability study was conducted using a hydrocarbon-contaminated soil for the oPtimization of bioremediation strategy best fit to a given set of contamination. The applicability of nutrients, biosurfactant, and oil-degrading microorganisms were examined by monitoring $CO_2$ evolution and oil degradation The addition of inorganic nutrients in the form of slow released fertilizer accelerated the initial rate of $CO_2$ evolution by a factor of 3. The application of oil-degrading microorganisms did not significantly increased $CO_2$ evolution or biodegradation efficiency. Application of a commercial biosurfactant was most effect in terms of the total $CO_2$ evolution and the oil degradation rate. The results indicate that $CO_2$ evolution measurement was found to be a simple and reliable countermeasure of crude oil hydrocarbon mineralization for the rapid determination of the best-fit bioremediation strategy.

• Journal of Microbiology and Biotechnology
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• v.10 no.3
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• pp.333-337
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• 2000

An oil-degrading yeast, Yarrowia lipolytica 180, exhibits interesting cell surface characteristics under the growth on hydrocarbons. An electron microscopic study revealed that the cells grown on crude oil showed protrusions on the cell surface, and thicker periplasmic space and cell wall than the cell surface, and thicker periplasmic space and cell wall than the cells grown on glucose. Y. lipolytica cells lost its cell hydrophobicity after pronase(0.1 mg/ml) treatment. The strain produced two types of emulsifying materials during the growth on hydrocarbons; one was water-soluble extracellular materials and the other was cell wall-associated materials. Both emulsifying materials at lower concentration (0.12%) enhanced the oil-degrading activity of Moraxella sp. K12-7, which had medium emulsifying activity and negative cell hydrophobicity; however, it inhibited the oil-degrading activity of Pseudomunas sp. K12-5, which had medium emulsifying activity and cell hydrophobicity. These results suggest that the oil-degrading activity of Y. lipolytica 180 is closely associated with cell surface structure, and that a finely controlled application of Y.lipolytica 180 in combination with other oil-degrading microorganisms showed a possible enhancing efficiency of oil degradation.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.109-111
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• 2010

Recent studies have described various microorganisms that can degrade PAH, however, there are currently limited methods available to screen for PAH-degrading microorganisms. To screen for PAH-degrading microorganisms, a sublimation method (Alley, Jeremy F. and Lewis R. Brown. 2000. Appl. Environ. Microbiol. 66, 439-442) was modified to produce a simple screening system. In our results, there were several bacterial species capable of pyrene degradation including genera, Coryenbacterium, Gordonia, Rhodococcus, and Streptomyces, which have been screened from 350 bacterial isolates of commercial gasoline and oil-spilled sediment by the sublimation method. The main advantage of this method is that it (i) safely deposits an even, thin and visible layer of PAH onto the agar surface without the use of solvents and (ii) the quantity of PAH sublimed onto the agar can be easily controlled. Overall, this sublimation method may be an effective and simple technique to screen for PAH-degrading microorganisms.

• Korean Journal of Microbiology
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• v.34 no.3
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• pp.101-107
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• 1998

Biological treatment of Arabian light crude oil-contaminated pebble was investigated in laboratory microcosms after supplementation with inorganic nutrients and oil-degrading microorganisms. Glass columns ($10cm{\times}20cm$) were used as microcosms and each microcosm was filled with pebbles of diameter less than 40 mm. After initial oil contamination of 2.4% (w/v), Inipol EAP-22 or slow release fertilizer (SRF) was added as inorganic nutrients and microorganisms were sprayed over pebbles. When $C_{17}$/pristane and $C_{18}$/phytane ratios were used as a marker for oil biodegradation, both ratios for microcosm supplemented with SRF and microorganisms were the lowest (below detectable range) after 92 days. Elimination of oil by abiotic processes, however, were minimal with decrease of $C_{17}$/pristane and $C_{18}$/phytane ratios from 3.55 and 2.41 to 3.06 and 1.50, respectively. The numbers of heterotrophic and oil-degrading microorganisms, and biological activity (dehydrogenase activity) corresponded to the course of biodegradation activities in all microcosms. During the whole experimental period, there was no significant nutrient deficiency only in the microcosm with SRF and microorganisms. It seemed that a continuous supply of inorganic nutrients using SRF was the most important factor for the successful performance of biological treatment in oil-contaminated pebbles.

• Journal of Microbiology
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• v.37 no.3
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• pp.128-135
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• 1999

Among oil-degrading microorganisms isolated from oil-polluted industrial areas, one yeast strain showed high degradation activity of aliphatic hydrocarbons. From the analyses of 18S rRNA sequences, fatty acid, coenzyme Q system, G+C content of DNA, and biochemical characteristics, the strain was identified as Yarrowia lipolytica 180. Y. lipolytica 180 degraded 94% of aliphatic hydrocarbons in minimal salts medium containing 0.2% (v/v) of Arabian light crude oil within 3 days at 25$^{\circ}C$. Optimal growth conditions for temperature, pH, NaCl concentration, and crude oil concentration were 30$^{\circ}C$, pH 5-7, 1%, and 2% (v/v), respectively. Y. lipolytica 180 reduced surface tension when cultured on hydrocarbon substrates (1%, v/v), and the measured values of the surface tension were in the range of 51 to 57 dynes/cm. Both the cell free culture broth and cell debris of Y. lipolytica 180 were capable of emulsifying 2% (v/v) crude oil by itself. They were also capable of degrading crude oil (2%). The strain showed a cell surface hydrophobicity higher than 90%, which did not require hydrocarbon substrates for its induction. These results suggest that Y. lipolytica has high oil-degrading activity through its high emulsifying activity and cell hydrophobicity, and further indicate that the cell surface is responsible for the metabolism of aliphatic hydrocarbons.

• Journal of Microbiology and Biotechnology
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• v.17 no.10
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• pp.1700-1703
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• 2007

Microorganisms capable of degrading crude oil were isolated and grown in soybean oil as a sole carbon source. The microbial cultures were used to control green peach aphids in vitro. Approximately 60% mortality of aphids was observed when the cultures were applied alone onto aphids. To examine the cultures as a pesticide formulation mixture, the cultures were combined with a low dose of the insecticide imidacloprid (one-fourth dose of recommended field-application rate) and applied onto aphids. The cultures enhanced significantly the insecticidal effectiveness of imidacloprid, which was higher than imidacloprid alone applied at the low dose. The isolated microorganisms exhibited high emulsifying index values and decreased surface tension values after being grown in soybean oil media. GC/MS analyses showed that microorganisms degraded soybean oil to fatty acids. The cultures were suggested to play the roles of wetting, spreading, and sticking agents to improve the effectiveness of imidacloprid. This is the first report on the control of aphids by using oil-degrading microbial cultures.

• Journal of Life Science
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• v.24 no.6
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• pp.694-699
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• 2014

Indigenous microorganisms play decisive roles in biodegradation. In this study, eighty strains of hydrocarbon-degrading microbes were isolated from Incheon Bay. Among them, 12 strains were selected by an oil film collapsing method. The bacterial strain 'Incheon9' was eventually selected based on its relatively higher lipase and emulsification activities, and was identified as Acinetobacter sp. (NCBI accession code: KF54854). The optimum condition for the growth and emulsification activity of Acinetobacter sp. Incheon9 was $20^{\circ}C$, pH 7, and 1% NaCl. The optimum time for the best production of biosurfactant was 72 hrs. The oil degradation ability of Acinetobacter sp. Incheon9 was investigated by measuring the residual oils in the culture medium by gas chromatography (FID). This research provides foundational data for eco-friendly environmental remediation by microorganisms.

• Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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• v.4 no.4
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• pp.227-230
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• 2000

Bunker-A oil-degrading microorganisms were isolated from a marine environment using an enrichment culture technique. The isolated strain EL-081K was identified as the genus Acinetobacter based on the results of morphological, culture, and biochemical tests. The optimal temperature and initial pH for bunker-A oil degradation were $25^{\circ}C$ and 7.0, respectively, including aeration. The optimal medium composition for the degradation of bunker-A oil by Acinetobacter sp. EL_O81K was 10 ml/l bunker-A oil as the carbon source and 0.1% (NH$_4$)$_2$SO$_4$as the nitrogen source. Under the above conditions, the biodegradability of bunker-A oil was 38% after 96 hours of incubation. The addition of detergent did not increase the bunker-A oil degradation.