• 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.

• 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
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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.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.

• 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.

• 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 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 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.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.435-438
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• 2000

Crude oil-degrading microorganisms, Acinetobacter sp. A132, Pseudomonas aeruginosa F722, and Acinetobacter calcoaceticus OM1 were isolated from soil and sea. The optimal temperature of strain A132 and strain F722 on growth isolated from soil was $35^{\circ}C$ both, and also their growth were optimized at pH 8 and 9, respectively. The growth of the strains, A132 and F722, showed that crude oil of 2% (w/v) in culture broth in which crude oil was used as carbon and energy sources appeared to be an optimum. Optimal culture conditions of strain OM1 were different from those of the soil microorganisms except for temperature. The growth of strain OM1 was optimized at pH 7 and crude oil of 3.0% (w/v). The degradability to crude oil by strain A132 showed maximum $5.49g/\;l\;{\cdot}\;day$ under the conditions of $25^{\circ}C$, NaCl of 1.0% (w/v), and crude oil of 2.0% (w/v). The highest degradability of strain F722 to crude oil was $1.19g/\;l\;{\cdot}\;day$ under the culture conditions at $35^{\circ}C$, NaCl 1.0% (w/v), and crude oil of 2.0% (w/v). The degradation characteristics of kerosene $(nC_9-nC_{20})$ and diesel $(nC_9-nC_{28})$ by strain OM1, and F722 were analyzed by gas chromatography. Strain OM1 degraded more than 95% of kerosene and 75% of diesel for 7 days cultivation. Strain F722 showed degradation of more than 80% to kerosene in 10 days.

• Journal of Microbiology and Biotechnology
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• v.22 no.9
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• pp.1185-1192
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• 2012

This study investigated the hydrocarbonoclastic microbial community present on weathered crude oil and their ability to degrade weathered oil in seawater obtained from the Gulf St. Vincent (SA, Australia). Examination of the native seawater communities capable of utilizing hydrocarbon as the sole carbon source identified a maximum recovery of just $6.6{\times}10^1\;CFU/ml$, with these values dramatically increased in the weathered oil, reaching $4.1{\times}10^4\;CFU/ml$. The weathered oil (dominated by > $C_{30}$ fractions; $750,000{\pm}150,000mg/l$) was subject to an 8 week laboratory-based degradation microcosm study. By day 56, the natural inoculums degraded the soluble hydrocarbons (initial concentrations $3,400{\pm}700mg/l$ and $1,700{\pm}340mg/l$ for the control and seawater, respectively) to below detectable levels, and biodegradation of the residual oil reached 62% ($254,000{\pm}40,000mg/l$) and 66% ($285,000{\pm}45,000mg/l$) in the control and seawater sources, respectively. In addition, the residual oil gas chromatogram profiles changed with the presence of short and intermediate hydrocarbon chains. 16S rDNA DGGE sequence analysis revealed species affiliated with the genera Roseobacter, Alteromonas, Yeosuana aromativorans, and Pseudomonas, renowned oil-degrading organisms previously thought to be associated with the environment where the oil contaminated rather than also being present in the contaminating oil. This study highlights the importance of microbiological techniques for isolation and characterisation, coupled with molecular techniques for identification, in understanding the role and function of native oil communities.