• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.2
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• pp.145-150
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• 2001

A marine bacterium having a high oil-degrading activity was isolated from the coastal water of Yosu, Korea, identified as Pseudomonas sp. and named Pseudomonas sp. BCK-1. The optimal temperature, pH and NaCl concentration for cell growth was $30^{\circ}C$, 7.0 and $3\%$ (w/v), respectively. After cultivation at $30^{\circ}C$, 180 rpm in 250 mL erlenmeyer flask for 72 and 168 hours, $2\%$ (w/v) arabian light crude oil (ACO) and bunker C oil (BCO) which are considered to be hardly biodegradable compounds were degraded $92\%$ (w/w) and $72\%$ (w/w), respectively.

• 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.

• Microbiology and Biotechnology Letters
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• v.25 no.5
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• pp.520-526
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• 1997

Bacterial strains which degrade Arabian Light crude oil were isolated by enrichment culture from oil-spilled soil. The strain A54 was finally selected after testing emulsifying activity and oil conversion rate. Strain A54 was identified as a Acinetobacter sp. based on the morphological, biochemical and physiological characteristics. It appears to be highly specialized for growth on Arabian Light crude oil in minimal salts medium since it showed preference for oil or degradation products as substrates for growth. It was found that it could grow on at least fifteen different hydrocarbons. The optimum cultural and environmental conditions were as follows; 25$\circ$C for temperature, 7,5 for pH, 2.0% for NaCl concentration and 2.0% for crude oil concentration. Additionally, the optimal concentration of NH$_{4}$NO$_{3}$, and K$_{2}$HPO$_{4}$, were 12.5 mM and 0.057 mM, respectively. Cell growth and emulsifying activity as a function of time were also determined. Crude oil degradation and the reduction of product peaks were identified by the analysis of remnant oil by gas chromatography. Approximately 63% of crude oil were converted into a form no longer extractable by mixed organic solvents.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 2001.04a
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• pp.57-61
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• 2001

일반적으로 Asphaltene은 원유뿐만 아니라 화석연로원(Fessil Fuel Sources)에도 포함되어 있는 것으로 알려져 있으며 난분해성으로 탄화수소(Hydrocarbon)와 Heterocyclic 또는 N-, S-, 산소를 포함하는 화합물이다. 그리고 금속성 물질이 Asphaltene Fraction에 농축되어 있는 것으로 알려져 있다. 한편 이러한 metallo-complex는 Asphaltene의 구조화합물중 방향족계 sheet 사이에 Self-association 상태로 존재한다. Asphaltene의 분해는 일반 TPH나 BTEX계 유류의 분해 기작과는 매우 상이하며 현재까지 Asphaltene으로 오염된 토양의 복원 기술을 매우 미미한 것으로 알려져 있다. 따라서 본 연구는 Asphalteme으로 오염된 토양의 복원 기숙을 개발하기 위한 전단계로 Asphaletene의 구성물질과 저분자화 작용기작을 구명하여 효과적 복원기술 개발을 위하여 Arabian Light Oil에서 Oil 자체와 Oil에서 분리해낸 Asphaltene을 사용하여 용매조건과 처리제별로 저분자화 과정을 조사하였다.

• Microbiology and Biotechnology Letters
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• v.31 no.4
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• pp.408-412
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• 2003

The purpose of this study was to evaluate the Influence of oil concentration and inoculum size on petroleum biodegradation in soil by Nocardia sp. H17-1, isolated from oil-contaminated soil. To investigate the effect of initial oil concentration on total petroleum hydrocarbon (TPH) degradation, the soil was artificially contaminated with 10, 50 or 100 g of Arabian light oil per kg of soil, respectively. After 50 days, Nocardia sp. H17-1 degraded 78,94 and 53% of the each initial TPH concentration, respectively. Also, it produced 1.35, 4.21, and 5.91 mmol of $CO_2$ per g of soil, respectively. The degradation rate constant (k) of TPH was decreased in proportion to the initial oil concentrations while $CO_2$ production was increased with the concentration. The growth of Nocardia sp. H17-1 was remarkably inhibited when it was inoculated into soil containing 100 g of oil per kg of soil. To evaluate the effect of the inoculum size, the soil was artificially contaminated with 50 g of Arabian light oil per kg of soil, and inoculated with $3${\times}$10^{6}$ , $5${\times}$10^{7}$ , $2${\times}$10^{8}$ cells per g of soil, respectively. After 50 days, the degradation of TPH was remained with similar in all treatment but degradation rate constant (k) and evolved $CO_2$ was increased with increasing the inoculum size.

• KSBB Journal
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• v.13 no.5
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• pp.606-614
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• 1998

In this study, biodegradation rate of Arabian light crude oil by mixed cultures of selected petroleum-degraders was determined. Their modes of hydrocarbon uptake were then observed to determine whether there are differences in biodegradation rate by the mixed cultures. By the mixed cultures of petroleum-degraders having same modes of hydrocarbon uptake, such as strain US1 and K1 (using pseudo-solubilized hydrocarbons by a biosurfactants), K2-2 and P1(using hydrocarbons by direct contact), CL 180 and IC-10 (mixed type of uptake modes), the biodegradation rates of aliphatics in the crude oil were increased more than those by their pure cultures, about 40%, 25% and 20%, respectively. Biodegradation rate of strain KH3-2 (using only water- dissolved hydrocarbons) was increased by mixed cultures with strain K1, CL180 or IC-10 possessing high emulsifying activity. However, the biodegradation rate of the crude oil was decreased about 20%-40% by the mixed cultures of petroleum-degraders having different mode of hydrocarbon uptake, such as addition of strain US1 or K1 in the cultures of K2-2 or P1. Biosurfactants produced by US1 or K1 seems to enhance the emulsification of crude oil in aqueous phase but inhibit the attachment of K2-2 or P1 to crude oil. As same phenomena, the addition to Triton X-100 into the culture of strain US1, K1, CL180, IC-10 or KH3-2 increased the biodegradation rate, but the addition in the culture of strain K2-2 or P1 decreased the biodegradation rate. The mixed culture made of CL180, IC-10 and KH3-2 degraded 61.5% of aliphatics and 69% of aromatics in 3% (v/v) of Arabian light crude oil added.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 1996.11a
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• pp.114-117
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• 1996

Biodegradation potential of crude oil has been studied in liquid and soil slurry culture. Studies were performed to optimize the factors affecting metabolic activity. Arabian Light(sulfur content 1%) was used as a representative crude oil and Nocardia sp. was selected as an oil degrading microorganism based on its ability to degrade and emulsify Effects of various nutritional and environmental conditions as well as emulsification and surface tension were observed. Tentative optimization of environmental and nutritional condition were as follow; pH 8, sodium nitrate as inorganic nitrogen source, yeast extract 0.05%, phosphate concentration 0.25% and glucose addition of 1.0% (w/v basis), extent of degradation to 78 %.

• 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.8 no.6
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• pp.645-649
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• 1998

A glycolipid type of biosurfactants was obtained from a strain which had been isolated from soil. The cell was identified as Pseudomonas aeruginosa from taxonomic characteristics and was designated as YPJ80. Thin layer chromatography and deoxyhexose detection tests were done to verify the type of biosurfactant. Critical micelle concentration (CMC) of the surfactant was observed to be 50 ppm and the minimum surface tension was 30.1 mN/m. As an emulsifier, YPJ80 biosurfactant was superior to emulsan in the emulsification of crude Arabian light oil.

• KSBB Journal
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• v.14 no.2
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• pp.192-197
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• 1999

marine bacterium Pseudomonas sp. CHCS-2 produced the biosurfactant in the culture broth which contained 2%(w/v) arabian light crude oil and the productivity of biosurfactant was increased with the addition of glucose. The crude oil in the culture broth was degraded by this strain and carbon chain of $_nC_{12}~_nC_{22}$ was completely degradaded during the incubation for 196 h. The crude biosurfactant was purified by Amberlite XAD-7, Sepharose CL-4B and DEAE-Sepharose CL-6B column chromatography. Therefore, 0.21g/L of the purified biosurfactnat was obtained. The purified biosurfactant was a type of lipoprotein and the molecular weight was estimated as 67kDa by SDS-PAGE. The lipid composition was identified as octadecanoic acid by gas chromatography/mass spectrometry. And then, the N-terminal amino acid sequence of the protein was determined as Ser-Val-lle-Asn-Thr-lle-X-Met-lle-Gly-Gln-Gln- and the sequence did not show homology to any other known lipoprotein. Therefore, the purified lopoprotein was predicted novel biosurfactant.