• 한국수산과학회지
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• 제34권2호
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• pp.145-150
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• 2001

한국 남해안 유류 오염지역인 전남 여수시 소리도 지역 해수로부터 유류 분해능이 우수한 균주를 선별하여 동정한 결과 Pseudomonas sp.로 동정되었으며, Pseudomonas sp. BCK-1으로 명명하였다. 균성장에 대한 최적배양온도, pH, NaCl 농도는 각각 $30^{\circ}C$, 7.0, $3\%$(w/v)였으며, $2\%$ (w/v) arabian light crude oil과 bunker C oil을 기질로 72시간 및 168시간 배양한 결과 arabian light crude oil는 $92\%$ (w/w), bunker, C oil은 $72\%$ (w/w)가 각각 생분해되었다.

• KSBB Journal
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• 제15권1호
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• pp.27-34
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• 2000

남해안 유류 오염지역으로부터 유류분해능이 우수한 균주를 선별하여 동정한 결과 Pseudomonas aeruginosa로 동정되었으며, Pseudomonas aeruginosa BYK-2로 명명하였다. 균 성장 및 생물 유화제 생산을 위한 최적배양온도, 시간, pH, NaCl 농도는 각각 $25^{\circ}C$, 48시간, 7.0 이었으며, NaCl은 첨가하지 않았을 때 가장 많은 양의 생물유화체가 생산되었다. 본 균주의 경우, 1%(w/v) arabian light 원유와 bunker C oil을 기질로 7일간 배양한 결과 92.1%(w/w)와 76.2%(w/w)가 생분해되었다. n-hexadccane 에 대한 세포부착능이 실험에서는 탄화수소화합물들(arabian light 원유, kuwait 원유, bunker C 유, n-paraffine, n-hexadecane, n-tetradecane)을 기질로 배양했을때는 모두 80% 이상의 높은 세포부착능을 나타냈으며, 지방산들(oleic acid, olive oil, lecithin)중에는 lecithin을 기질로 배양했을 때 91.5%로 가장 높은 세포부착능을 나타냈어다. 또한 당들(arabinose, trehalose, dextrose, galactose, lactose, fructose, maltose, sorbitol, sucrose)을 기질로 했을 경우 arabinose, galactose, sorbitol, sucrose를 제외하고는 대부분 70%이하의 세포부착능을 나타냈다.

• 한국미생물·생명공학회지
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• 제25권5호
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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.

• 한국지하수토양환경학회:학술대회논문집
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• 한국지하수토양환경학회 2001년도 총회 및 춘계학술발표회
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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.

• 한국미생물·생명공학회지
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• 제31권4호
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• pp.408-412
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• 2003

오염토양에 유류분해능을 가진 Nocardia sp. H17-1의 접종시 고려되어야 할 인자중 하나인 초기 오염농도에 의한 탄화수소분해능과 초기 접종농도에 의한 분해능 및 균주의 생육을 조사하였다. H17-1은 실험 50일 동안 초기 오염농도 10, 50, 100 g Arabian light oil/kg of soil에 대해 각각 78.5%, 94.3%, 53.2%의 탄화수소를 제거하였으며, 오염농도가 높을수록 분해속도 상수(k) 낮아졌다. $CO_2$의 생성량 또한 오염농도가 높을수록 증가하였으나, 100 g/kg-soil의 오염농도에서는 균의 생육이 저해를 받는 것으로 나타났다. H17-1의 초기 접종농도에 의한 영향은 균의 접종량에 따라 최종 남은 TPH의 양은 큰 차이를 나타내지 않았으나, 분해속도상수(k)는 균의 접종량이 늘어남에 따라 크게 증가되었으며, $CO_2$의 생성량 또한 균의 접종농도에 따라 증가하였다.

• KSBB Journal
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• 제13권5호
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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.

• 한국지하수토양환경학회:학술대회논문집
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• 한국지하수토양환경학회 1996년도 경북지부 결성 및 추계학술발표회 논문집
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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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• 제37권3호
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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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• 제8권6호
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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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• 제14권2호
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• pp.192-197
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• 1999

해양세균 Pseudomonas sp. CHCS-2는 배양과정 중에 포도당 첨가에 의해 생물유화제의생산이 증가되었으며, 196시간 배양시 원유에 포함되어 있는 paraffine계 탄화수소 중에서 $_nC_{12}~_nC_{22}$까지 범위의 carbonchain이 거의 대부분 분해됨을 알 수 있었다. chloroform : metahnol = 1 : 1(V/V)로 추출하여 얻은 8.5g/L의 crude 생물 유화제를, Amberlite XAD-7, Sepharose CL-4B, DEAE-sepharose CL-6B 수지를 사용하여 정제한 결과 최종적으로 0.21g/L의 정제된 생물유화제를 얻었다. 본 균주가 생산하는 생물유화제는 단백질과 octadecanoic acid가 결합된 분자량이 약 67kDa인 lipoprotein으로 확인되었으며, 정재된 생물유화제중 단백질성분만을 분리하여 N-말단 아미노산배열을 분석한 결과, Ser-Val-He-Asn-Thr-Asn-IIe-Thr-X-Met-Ile-Gly-Gin-Gln-의 배열을 가지는 것으로 확인되었다. 이는 기존에 보고된 lipoproten과 다른 형태의 것으로, 본 균주가 생산하는 생물유화제의 경우 구성성분, 분자량 등을 고려해 볼 때 새로운 형태의 lipoprotein 계열 생물유화제로 추정된다.