• Journal of Microbiology
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• v.35 no.1
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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.

• Microbiology and Biotechnology Letters
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• v.34 no.3
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• pp.185-195
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• 2006

Phytoremediation is an economical and environmentally friendly bioremediation technique using plants which can increase the microbial population in soil. Unlike other pollutants such as heavy metals, poly-chlorinated biphenyl, trichloroethylene, perchloroethylene and so on, petroleum hydrocarbons are relatively easily degradable by soil microbes. For successful phytoremediation of soil contaminated with petroleum hydrocarbons, it is important to select plants with high removal efficiency through microbial degradation. In this study, we clarified the roles of plants and rhizobacteria and identified their species effective on phytore-mediation by reviewing the papers previously reported. Plants and rhizobacteria can degrade and remove the petroleum hydrocarbons directly and indirectly by stimulating each other's degradation activity. The preferred plant species are alfalfa, ryegrass, tall fescue, poplar, corn, etc. The microorganisms with a potential to degrade hydrocarbons mostly belong to Pseudomonas spp., Bacillus spp., and Alcaligenes spp. It has been reported that the elimination efficiency of hydrocarbons by soil microorganisms can be improved when plants were simultaneously applied. For more efficient restoration, it's necessary to understand the plant-rhizobacteria interaction and to select the suitable plant and microorganism species.

• Journal of Environmental Science International
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• v.5 no.5
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• pp.611-617
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• 1996

The effects of nutritional sources on growth of Pseudomonas sp. P2 were investigated in medium containing biphentyl as a carbon source. To determine characterization of Pseudomonas sp. P2, the incubation time was determined to 100 h of the log phase in the growth curve. The optimal compositions for the growth of Pseudomonas sp. P2 degrading polychlorinated biphenyls (PCBs) were 1000 mg/L $NH_4NO_3$, 1000mg/L KH_2PO_4$, 100mg/L MgSO_4$.$7H_2O$, 30mg/L $CaCl_2$.$2H_2O$, 200mg/L NaCl, and 10mg/L $FeSO_4$.$7H_2O$. Pseudomonas sp. P2 showed the degradability of 59.3%, 57.6%, 51.4%, and 48.7% at 500mg/L, 1000mg/L, 1500mg/L, and 2000mg/L of the PCBs within insulating oil after 100 h incubation under the optimum conditions, respectively.

• Journal of Microbiology
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• v.34 no.3
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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.

• Journal of Korea Foresty Energy
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• v.17 no.1
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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.

• Journal of Life Science
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• v.19 no.5
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• pp.659-663
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• 2009

Bacterial stain 3Y was isolated from a site that was contaminated with diesel for more than 15 years. The strain could grow on various petroleum using hydrocarbons as the sole carbon source. The strain grew not only on aliphatic hydrocarbons but also on aromatic hydrocarbons. 3Y grew on aliphatic petroleum hydrocarbons hexane or hexadecane, and aromatic petroleum hydrocarbons BTEX, phenol, biphenyl, or phenanthrene. The strain showed aromatic ring dioxygenase and meta-cleavage dioxygenase activities as determined by tests using indole and catechol. Aromatic ring dioxygenase is involved in the initial step of biodegradation of aromatic hydrocarbons while meta-cleavage dioxygenase catalyzes the cleavage of the benzene ring. Based on a nucleotide sequence analysis of its 16S rRNA gene, 3Y belongs to the genus Sphingomonas. A phylogenetic tress was constructed based on the nucleotide sequences of closest relatives of 3Y and petroleum hydrocarbon degrading sphingomonads. 3Y was in a cluster that was different from the cluster that contained well-known sphingomonads. The results of this study suggest that 3Y has the potential to cleanup oil-contaminated sites. Further investigation is warranted to optimize conditions to degrade petroleum hydrocarbons by the strain to develop a better bioremediation strategy.

• Microbiology and Biotechnology Letters
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• v.33 no.3
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• pp.215-221
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• 2005

It was characterized the hexane biodegradation and mineralization using a hexane-degrading consortium, and analyzed its bacterial community structure by 16S rDNA PCR-DGGE (denaturing gradient gel electrophoresis). The specific growth rate (${\mu}_{max}$) of the hexane-degrading consortium was 0.2 $h^{-1}$ in mineral salt medium supplemented with hexane as a sole carbon source. The maximum degradation rate ($V_{max}$) and saturation constant ($K_{s}$) of hexane of the consortium are 460 ${\mu}mol{\cdot}g-DCW^{-1}{\cdot}h^{-1}$ and 25.87 mM, respectively. In addition, this consortium could mineralize $49.1{\%}$ of $^{14}C$-hexane to $^{14}CO_2$, and $43.6{\%}$ of $^{14}C$-hexane) was used for the growth of biomass. The clones isolated from the DGGE bands were closely related to the bacteria which were capable of degrading pollutants such as oil, biphenyl, PCE, and waste gases. The hexane-degrading consortium obtained in this study can be applied for the biological treatment of hexane.

• Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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• 1998.11a
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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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• v.18 no.1
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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.

• Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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• 1998.06a
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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.