• 한국지하수토양환경학회지:지하수토양환경
• /
• 제21권1호
• /
• pp.72-79
• /
• 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
• /
• 제15권6호
• /
• pp.1170-1177
• /
• 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
• /
• 한국생물공학회 2001년도 추계학술발표대회
• /
• pp.578-581
• /
• 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
• /
• 제10권3호
• /
• pp.333-337
• /
• 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.

• 미생물학회지
• /
• 제46권1호
• /
• pp.109-111
• /
• 2010

다환 방향족 탄화수소(PAH; polycyclic aromatic hydrocarbon)는 발암성, 돌연변이 유발성, 유전독성을 지니기 때문에 인체위해성이 큰 물질로 알려져 있다. 기존 PAH 분해 미생물 탐색 방법중 독성이 강한 유기용매에 PAH를 용해시켜 미생물에 직접 분무하는 분무법, 미생물과 직접 혼합하는 한천중층법은 미생물 생장에 영향을 줄 뿐만 아니라, 특히 많이 쓰이는 분무법의 경우 분무되는 PAH의 양을 조절하기가 어렵고 멸균상태를 유지하기가 힘들다는 단점이 있다. 그래서 본 논문에서는 이와 같은 단점을 보완한 방법으로 승화법(Alley, Jeremy F. and Lewis R. Brown. 2000. Use of sublimation to prepare solid microbial media with water-insoluble substrates. Appl. Environ. Microbiol. 66, 439-442)을 도입하여 적용하였다. 그 결과 상용휘발유 및 태안유류유출지 시료로부터 분리한 350분리균주 중에서 7균주가 단일 PAH 또는 복수의 PAH 분해에 관여했다. 특히 Corynebacterium sp. SK20, Rhodococcus sp. TA24, Streptomyces sp. TA27은 시험한 pyene, phenanthrene, naphthalene에, Gordonia sp. H37는 pyrene, naphthalene에, Arthrobacter sp. S49는, naphthalene, phenanthrene에 활성이 있었다.

• 미생물학회지
• /
• 제34권3호
• /
• pp.101-107
• /
• 1998

지속성영양염제(Slow Release Fertilizer, SRF)와 유류분해미생물을 이용한 생물학적 방법으로 오염된 자갈환경으로부터 유류를 제거하는 방법을 연구하기 위해 microcosm 실험을 수행하였다. Microcosm은 직경 40 mm 이하의 자갈을 직경 10 cm 높이 20 cm의 glass column에 충진하여 준비하였으며, 전 실험구에 유류 약 2.4%(w/v)를 첨가하였다. 실험구 I은 대조구로서 아무것도 첨가되지 않았으며, 실험구 II에는 Inipol EAP-22(Elf Aquitaine, France)와 미생물제제, 실험구 III에는 SRF와 미생물제제, 실험구 IV에는 Inipol EAP-22가 첨가되었다. 유류의 생물학적 분해도를 $C_{17}$/Pristane과 $C_{18}$/Phytane 비율료 나타내었을 때, 실험구 III에서 유류분해도가 가장 높을 것으로 나타났다. 종속영양미생물 및 유유분해미생물의 전체적인 변화와 전자전달계 활성도는 유류분해도의 변화와 유사한 양상을 보이며 변화하는 것으로 나타났다. 무기영양염류의 분석 결과 실험구 III 에서는 전 실험기간에 걸쳐 질소와 인이 제한요인으로 작용하지 않은 것으로 나타났으며, 이는 SRF가 지용성영양염제인 Inipol EAP-22에 비해 지속적으로 영양염류를 제공함으로써 자갈환경에서 유류분해가 증가되었다고 추정할 수 있다. 따라서 본 실험결과로부터 SRF와 유류분해 미생물제제를 사용하여 해안 자갈지역의 유류오염을 효율적으로 제거할 수 있는 가능성이 확인되었다.

• Journal of Microbiology
• /
• 제37권3호
• /
• pp.128-135
• /
• 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
• /
• 제17권10호
• /
• pp.1700-1703
• /
• 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.

• 생명과학회지
• /
• 제24권6호
• /
• pp.694-699
• /
• 2014

토착 미생물은 친환경적 정화에 중요한 역할을 한다. 원유(crude oil)를 분해하는 80균주를 인천 연안에서 분리하고 oil film collapsing방법을 이용하여 유화능이 있는 12균주를 선별하였다. 이들 균주에 대해 ${\rho}$-nitrophenyl butylrate를 기질로 이용하여 리파아제(lipase)활성과 n-hexanedecane을 기질로 이용하여 유화(emulsification)활성을 측정하여 원유 분해 활성이 좋은 Incheon9를 선별하고 gas chromatography (FID)로 paraffine계 탄화수소를 감소시키는 것을 확인하였다. 이 균주의 16s rRNA유전자 분석을 통해 Acinetobacter sp.로 동정하고 NCBI에 등록하여 accession code (KF548540)를 부여 받았다. Acinetobacter sp. Incheon9의 성장과 유화능이 최적 배양 조건은 $20^{\circ}C$, pH 7, 1% NaCl였으며 대수증식기 기간에 가장 높은 유화능을 보였으며 탄화수소가 짧은 trybutyrin에서 분해능력이 좋았다. 이번 연구결과는 환경오염에 활용 가능한 미생물자원군의 확보를 위한 연구였으며 추후 활용을 위해서는 실제 환경에서 동일한 활성을 가지는지 여부에 대한 연구가 추가로 진행되어야 할 것이다.

• Environmental Sciences Bulletin of The Korean Environmental Sciences Society
• /
• 제4권4호
• /
• pp.227-230
• /
• 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.