• Journal of Microbiology
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• 제35권1호
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• pp.53-60
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• 1997

Pseudomonas sp. P20 and Pseudomonas sp. DJ-12 isolated from the polluted environment are capable of degrading biphenyl and 4-chlorobiphenyl (4CB) to produce benzoic acid and 4-chlorobenzoic acid (4CBA) respectively, by pcbABCD-encoded enzymes. 4CBA can be further degraded by Pseudomonas sp. DJ-12, but not by Pseudomonas sp P20. However, the meta-cleavage activities of 2, 3-dihydroxybiphenyl (2, 3-DHBP) and 4-chloro-2, 3-DHBP dioxygenases (2, 3-DHBD) encoded by pcbC in Pseudomonas sp. P20 were stronger than Pseudomonas sp. DJ-12. In this study, the pcbC gene encoding 2, 3-DHBD was cloned from the genomic DNA of Pseudomonas sp. P20 by using pKT230. A hybrid plasmid pKK1 was constructed and E. coli KK1 transformant was selected by transforming the pKK1 hybrid plasmid carrying pcbC into E. coli XL1-Blue. By transferring the pKK1 plasmide of E. coli KK1 into Pseudomonas sp. DJ-12 by conjugation, a recombinant strain Pseudomonas sp. P20, Pseudomonas sp. DJ-12, and the recombinant cell assay methods. Pseudomonas sp. DJ12-C readily degraded 4CB and 2, 3-DHBP to produce 2-hydroxy-6-oxo-6-phenylhexa-2, 4-dienoic acid (HOPDA), and the resulting 4CBA and benzoic acid were continuously catabolized. Pseudomonas sp. DJ12-C degraded 1 mM 4CB completely after incubation for 20 h, but Pseudomonas sp. P20 and Pseudomonas sp. DJ-12 showed only 90% and Pseudomonas sp. DJ-12 had, but its degradation activity to 2, 3-DHBP, 3-methylcatechol, and catechol was improved.

• 한국미생물·생명공학회지
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• 제34권3호
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• pp.185-195
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• 2006

산업발달과 인구증가로 인하여 석유계 탄화수소의 사용량이 점차 증가함에 따라 많은 양의 석유계 탄화수소가 환경에 잔류하여 토양과 지하수에 심각한 오염을 야기시키고 있으며, 인체에도 피해를 주게 된다. 유류오염토양을 복원하는 방법 중 생물을 이용한 복원기술은 경제적이고 환경친화적인 기술로서, phytoremediation 방법은 유류오염물질을 분해할 수 있는 미생물과 토양 내의 미생물량을 증가시킬 수 있는 고등식물을 함께 이용함으로써 생물복원기술의 효율을 극대화할 수 있는 방법이다. 토양 내 유류오염물질은 중금속, polychlorinated biphenyl, trichloroethylene, perchloroethylene 등의 오염물질과 달리 식물에 의해 분해 될 수 있기 때문에 유류오염물질 정화효율이 높은 식물종을 선택하는 것이 무엇보다 중요하다. 본 연구에서는 phytoremediation 기법을 이용하여 유류오염토양을 정화하는 과정에서 식물과 근권 미생물을 역할을 밝히고, 이전에 보고된 연구결과를 바탕으로 유류오염토양복원에 효과적인 식물종과 근권미생물을 알아보았다. 토양 내의 유류오염물질은 식물과 근권 미생물에 의해 분해제거되는데, 식물과 근권 미생물은 유류오염물질을 직접 분해하기도 하며 서로의 분해작용을 촉진하는 간접적 역할을 하기도 한다. 유류오염 토양의 정화에 선호되는 식물종은 alfalfa, ryegrass, tall fescue, poplar, corn 등이었으며, 탄화수소를 분해할 수 있는 것으로 밝혀진 미생물종은 주로 Pseudomonas spp., Bacillus spp., Alcaligenes spp. 등이었다. Phytoremediation 방법을 통해 토양 내 유류오염물질의 정화효율을 높일 수 있다는 연구결과가 보고되고 있다. 따라서 phytoremediation 과정에서 식물과 근권 미생물의 역할과 상호작용을 정확히 이해한다면 보다 효과적인 토양복원을 기대할 수 있을 것이다.

• 한국환경과학회지
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• 제5권5호
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• pp.611-617
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• 1996

유일한 탄소원으로서 Biphenyl을 포함하고 있는 액체배지에서 Pseudomonas sp. P2의 생육에 미치는 영양원의 효과에 관해 연구하였다. Pseudomonas sp. P2의 특성을 조사하기 위한, 배양시간은 균의 생육 유선에서 대수성장기 말인 100시간으로 하였다. PCBs를 분해하는 Pseudomonas sp. P2의 생육을 위한 최적 조성은 1000 mg/L $NH_4NO_3$, 1000 mg/L KH_2PO_4$, 100 mg/L MgSO_4$.$7H_2O$, 30 mg/L $CaCl_2$.$2H_2O$, 200 mg/L NaCl, 10mg/L $FeSO_4$.$7H_2O$였다. Pseudomonas sp. P2에 의한 100시간 배양후 절연유 내에 있는 PCBs의 분해률은 500mg/L에서 59.3%, 1000mg/L에서 57.6%, 1500mg/L에서 51.4%, 2000mg/L에서는 48.775 였다.

• Journal of Microbiology
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• 제34권3호
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• pp.241-247
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• 1996

The stability of the genetically engineered microorganisms and their recombinant plasmids released in natural environments has been regarded as one of the molecular ecological topics. In this study, the recombinant plasmids pCU103 in which the pcbCD genes involved in biodegradation of biphenyl and 4-chlorobiphenyl were cloned in pBluescript SK(+) vector, were examined for their structural and functional stability in different waters at 15 $^{\circ}C$ by the methods of electrophoresis, Southern hybridization, quantification with fluorescent dye, and transformation. The recombinant plamids maintained their stabilities for about 30 days in sterilized distilled water (SDW), 15 days in autoclaved creek water (AW), 25 days in filtered and autoclaved non-sterile creek water (FAW), 4 days in Luria-Bertani (LB) broth, and less than one day in filtered non-sterile creek water (FW). The covalently closed circular (CCC) form of the plasmid was decreased and open circular (OC) form was increased as a function of incubation time, and then linear (L) form was produced to be ultimately degraded out. The degradation rates of the plasmid were proportionally correlated to trophic level of the water, and the biological factor such as DNases was found to be one of the most critical factors affecting structural and functional stability of the plasmid in non-sterile natural water.

• 임산에너지
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• 제17권1호
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• pp.56-70
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• 1998

This study was carried out to investigate the structural characteristics of kraft lignin and the wood degrading characteristics, the productivity of ligninolytic enzymes and the enzymatic degradation of kraft lignin by white-rot fungi. To purify kraft lignin, precipitation of kraft pulping black liquors of pitch pine meal was done by titration with lN $H_{2}SO_{4}$ reaching to pH 2, and isolation of the precipitates done by centrifugation. The isolated precipitates from pitch pine were redissloved in lN NaOH, reprecipitated by titration with lN $H_{2}SO_{4}$, washed with deionized water, and kept ofr analysis after freeze drying. Fractionation of the precipitates in solution by successive extraction with $CH_{2}Cl_{2}$ and MeOH, and the fractionates were named SwKL, SwKL I, SwKL II, and SwKL III for pitch pine kraft lignin. The more molecular weights of kraft lignin increased, the less phenolic hydroxyl groups and the more aliphatic hydroxyl groups. Because as the molecular weights increased, the ratio of etherified guaiayl/syringyl(G/S ratio) and the percentage were increased. The spectra obtained by 13C NMR and FTIR assigned by comparing the chemical shifts of various signals with shifts of signals from autherized ones reported. The optimal growth temperature and pH of white-rot fungi in medium were $28^{\circ}C$ and 4.5-5.0, respectively. Especially, in temperature and pH range, and mycelial growth, the best white-rot fungus selected was Phanerochaete chrysosporium for biodegradation. For the degradation pathways, the ligninolytic fungus jcultivated with stationary culture using medium of 1% kraft lignin as a substrate for 3 weeks at $28^{\circ}C$. The weight loss of pitch pine kraft lignin was 15.8%. The degraded products extracted successively methoanol, 90% dioxane and diethyl ether. The ether solubles were analyzed by HPLC. Kraft lignin degradation was initiated in $\beta$-O-4 bonds of lignin by the laccase from Phanerochaete chrysosporium and the degraded compounds were produced from the cleavage of $C\alpha$-$C\beta$ linkages at the side chains by oxidation process. After $C\alpha$-$C\beta$ cleavage, $C\alpha$-Carbon was oxidized and changed into aldehyde and acidic compounds such as syringic acid, syringic aldehyde and vanilline. And the other compound as quinonemethide, coumarin, was analyzed. The structural characteristics of kraft lignin were composed of guaiacyl group substituted functional OHs, methoxyl, and carbonyl at C-3, -4, and -5 and these groups were combinated with $\alpha$ aryl ether, $\beta$ aryl ether and biphenyl. Kraft lignin degradation pathways by Phanerochaete chrysosporium were initially accomplished cleavage of $C\alpha$-$C\beta$ linkages and $C\alpha$ oxidation at the propyl side chains and finally cleavage of aromatic ring and oxidation of OHs.

• 생명과학회지
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• 제19권5호
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• pp.659-663
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• 2009

장기간 경유로 오염된 지역의 토양으로부터 분리한 세균 3Y는 석유계 탄화수소를 구성하는 다양한 화합물을 유일 탄소원으로하여 성장하였다. Sphingomonas sp. 3Y는 지방족 화합물은 물론이고 방향족 화합물을 이용해서 성장할 수 있었다. 지방족 화합물로서는 hexane과 hexadecane을 이용하여 성장하였고, 한편 방향족 화합물로서는 BTEX는 물론이고 phenol, biphenyl, 또는 phenanthrene을 유일 탄소원으로 이용하여 성장하였다. 본 균주는 indole과 catechol을 이용한 실험결과 방향족 탄화수소의 생분해 과정에서 맨 첫 단계 반응에 관여하는 효소인 aromatic ring dioxygenase 활성과 benzene 환을 깨는 효소인 meta-cleavage dioxygenase 활성을 나타내었다. Sphingomonas sp. 3Y의 16S rRNA 유전자의 염기서열 분석과 계통수 작성 결과 본 균주는 ${\alpha}$-Proteobacteria인 Sphingomonas속에 해당하였으며 지금까지 잘 알려진 석유계 탄화수소를 분해하는 Sphingomonas sp. 균주들과는 다른 cluster를 형성하였다. 다양한 석유계 탄화수소 성분을 이용하여 성장하는 Sphingomonas sp. 3Y는 유류로 오염된 토양의 복원에 유용하게 사용될 것으로 여겨지며 이 균주의 최적 분해 조건을 조사한다면 그 결과는 이 균주가 분리된 오염지역의 생물학적 분해를 최적화하는데 기여할 것이다.

• 한국미생물·생명공학회지
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• 제33권3호
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• pp.215-221
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• 2005

Hexane을 유일 탄소원으로 첨가한 무기염 배지에서 hexane 생분해 속도와 비성장속도를 구하였고, $^{14}C-hexane$을 이용하여 무기화(mineralization) 정도를 측정하였다. 또한, 16S rDNA PCR-DGGE 분석기법을 활용하여 consortium의 미생물 군집 특성을 조사하였다. Consortium CH의 최대 비성장속도 (${\mu}_{max}$)값은 $0.2\;h^{-1}$이고, 최대 hexane 분해속도 ($V_{max}$)와 포화상수 ($K_{s}$)는 각각 460 ${\mu}mol{\cdot}g-DCW^{-1}{\cdot}h^{-1}$ 및 25.87mM 이었다. Consortium CH는 $^{14}C-hexane$의 약 $49.1\%$를 무기화하였고, $43.6{\%}$의 $^{14}C-hexane$는 biomass를 증가시키는데 사용하였다. DGGE band로부터 얻은 clone들은 유류, biphenyl, PCE 및 폐가스 등과 같은 오염물질 분해능을 가진 세균들과 유사성이 가장 높았다. 본 연구에서 얻은 hexane 생분해용 consortium은 향후 hexane 제거용 생물학적 시스템을 개발하는데 유용하게 활용될 수 있다.

• 한국전기전자재료학회:학술대회논문집
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• 한국전기전자재료학회 1998년도 추계학술대회 논문집
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• pp.125-128
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• 1998

Electroluminescent(EL) devices based on organic thin films have attracted lots of interests in large-area light-emitting display. One of the problems of such device is a lifetime, where a degradation of the cell is possibly due to an organic layer's thickness, morphology and interface with electrode. In this study, light-emitting organic electroluminescent devices were fabricated using Alq$_3$(8-hydroxyquinolinate aluminum) and TPD(N,N'-diphenyl-N,N'-bis(3-methylphenyl)-[1-1'-biphenyl]-4,4'-diamine).Where Alq$_3$ is an electron-transport and emissive layer, TPD is a hole-transport layer. The cell structure is ITO/TPD/Alq$_3$/Al and the cell is fabricated by vacuum evaporation method. In a measurement of current-voltage characteristics, we obtained a turn-on voltage at about 9 V. And we used other buffer layer of PPy(Polypyrrole) with ITO/PPy/TPD/Alq$_3$/Al structure. We observed a surface morphology by AFM(Atomic Force Microscopy), UV/visible absorption spectrum, and PL(Photoluminescence) spectrum. We obtained the UV/visible absorption peak at 358nm in TPD and at 359nm in Alq$_3$, and at 225nm and the PL peaks at 410nm in TPD and at 510nm in Alq$_3$ and at 350nm. We also studied EL spectrum in the cell structure of ITO/TPD/Alq$_3$/Al and ITO/PPy/TPD/Alq$_3$/Al and we observed the EL spectrum peak at 510nm from our cell

• The Korean Journal of Ecology
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• 제18권1호
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• pp.109-120
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• 1995

As the genetically modified microorganisms (GMMs) and their recombinant plasmid DNAs could be released into natural environments, their stabilities and impacts to indigenous microorganisls have become very importhant research subjects concerning with environmental and ecological aspects. In this study, the genetically modified E. coli CU103 and its recombinant pCU103 plasmid DNA, in which pcbCD genes involving in degradation of biphenyl and 4-chlorobiphenyl were cloned, were studied for their survival and stability in several different waters established under laboratory conditions. E. coli CU103 and its host E. coli XL1-Blue survived longer in sterile distilled water (SDW) and filtered autoclaved river water (FAW) than in filtered river water (FW). A lot of extracellular DNAs were released from E. coli CU103 by lytic action of phages in FW and the released DNAs were degraded by DNase dissolved in the water. Such effects of the factors in FW on stability of the recombinant pCU103 plasmid were also observed in the results of gel electrophoresis, quantitative analysis with bisbenzimide, and transformation assay. Therefore, the recombinant plasmids of pCU103 were found to be readily liberated from the genetically modified E. coli CU103 into waters by normal metabolic processes and lysis of cells. And the plasmid DNAs were quite stable in waters, but their stabilities could be affected by physicoKDICical and biological factors in non-sterile natural waters.

• 한국전기전자재료학회:학술대회논문집
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• 한국전기전자재료학회 1998년도 춘계학술대회 논문집
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• pp.399-402
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• 1998

Electroluminescent(EL) devices based on organic thin films have attracted lots of interests in large-area light-emitting display. One of the problems of such device is a lifetime, where a degradation of the cell is possibly due to an organic layers thickness, morphology and interface with electrode. In this study, light-omitting organic electroluminescent devices were fabricated using Alq$_3$(8-hydroxyquinolinate aluminum) and TPD(N,N'-diphenyl-N,N'-bis(3-methylphenyl(1-1\`-biphenyl]-4,4'-diamine). Where Alq$_3$ is an electron-transport and emissive layer, TPD is a hole-transport layer. The cell structure is ITO/TPD/Alq$_3$/Al and the cell is fabricated by vacuum evaporation method. In a measurement of current-voltage characteristics, we obtained a turn-on voltage at about 9 V. We also investigated stability of the devices using buffer layer with blend of PEI (Poly ether imide) and TPD by varying mot ratios between ITO and Alq$_3$. In current-voltage characteristics measurement, we obtained the turn-on voltage at about 6 V and observed an anomalous behavior at 3∼4 V. And we used other buffer layer of PEDT(3,4-pyrazino-3',4'-ethylenedithio-2,2',5,5'-tetrathiafulvalenium) with ITO/PEDT/TPD/Alq$_3$Al structure. We observed a surface morphology by AFM(Atomic Force Microscopy), UV/visible absorption spectrum, and PL(Photoluminescence) spectrum. We obtained the UV/visible absorption peak at 358nm in TPD and at 359nm in Alq$_3$, and the PL peaks at 410nm in TPD and at 510nm in Alq$_3$. We also studied EL spectrum in the cell structure of ITO/(TPD+PEI)/Alq$_3$/Al.