• Title/Summary/Keyword: Pseudomonas sp. P2

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Improvement of 4-chlorobiphenyl degradation bya recombinant strain, pseudomonas sp. DJ12-C

  • Kim, Ji-Young;Kim, Young-Chang;You, Lim-Jai;Lee, Ki-Sung;Ok, Ka-Jong;Hee, Min-Kyung;Kim, Chi-Kyung
    • 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.

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Effects of Nutritional Sources on Degradation of Polychlorinated Biphenyls (PCBs) by Pseudomonas sp. P2 (Pseudomonas sp. P2에 의한 Polychlorinated Biphenyls(PCBs) 분해에 대한 영양원의 영향)

  • 최상기;금정호
    • 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.

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pKT230 벡터를 이용한 Pseudomonas sp. P20의 2,3-Dihydroxybiphenyl Dioxygenase 유전자의 클로닝

  • Kim, Ji-Young;Kim, Chi-Kyung;Ka, Jong-Ok;Min, Kyung-Hee;Park, Yong-Keun
    • Microbiology and Biotechnology Letters
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    • v.24 no.6
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    • pp.657-663
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    • 1996
  • Pseudomonas sp. P20 isolated from the polluted environment is capable of degrading biphenyl and 4-chlorobiphenyl. The pcbABCD genes responsible for degradation of biphenyl and 4-chlorobiphenyl were cloned using pBluescript SK(+) from the chromosomal DNA of Pseudomonas sp. P20 to construct pCK1 and pCK102, harbouring pcbABCD and pcbCD, respectively. The 2, 3-DHBP dioxygenase gene, pcbC, was cloned again from pCK102 by using pKT230 which is known as a shuttle vector and pKK1 hybrid plasmid was constructed. The E. coli KK1 transformant obtained by transforming the pKK1 into E. coli XL1-Blue showed 2, 3-DHBP dioxygenase activity. The specific 2, 3-DHBP dioxygenase activity of E. coli KK1 was similar to that of the E. coli CK102, but much higher than those of the natural isolates, Pseudomonas sp. DJ-12 and Pseudomonas sp. P20.

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Isolation of Marine Bacteria Killing Red Tide Microalgae II. Isolation and Algicidal Properties of Pseudomonas sp. LG-2 Possessing Killing Activity for Dinoflagellate, Prorocentrum micans (적조생물 살조세균 탐색 II. 적조생물 Prorocentrum micans 살조세균 Pseudomonas sp. LG-2의 분리와 살조특성)

  • LEE Won-Jae;PARK Young-Tae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.6
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    • pp.852-858
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    • 1998
  • We have isolated a bacterial strain that tends to kill P. micans from the mixed culture of p. minns plus seawater filtrate (poresize, 0.8 $\mu$m) collected at Masan bay in July 1996, in which the mixed culture grown in the f/2 medium. According to the experimental results of the isolated bacterium such as fatty acids analysis, morphological and biochemical characteristic tests, the strain was supposed to be a Pseudomonas and then it was named as Pseudomonas sp. LG-2. The killing effect of Pseudomonas sp. LG-2 against P. micans was proportionally increased with the concentrations of culture filtrate (pore size, 0.8 $\mu$m) is well as with the number of bacterium inoculated. In the mixed culture inoculated with $1.3\times10^6$ cells/ml of Pseudomonas sp. LG-2, the number of P. micans (2,000 cells/ml) was gradually decreased and then killed below 100 cells/ml within 7 days. In addition, the culture filtrate with $30\%$ of final concentration revealed a significant killing effect against P. micans around 3 days after culture. In the relationship between killing effects and growth stage of Pseudomonas sp. LG-2, the culture filtrate at lag phase has little effects on P. micans. In constant, the culture filtrate at mid-log phase showed the killing effect by decreasing P. micans to 112 in number within 5 days. In particular, the culture filtrate at stationary phase showed a significant killing effect against P. micans in which the majority of it was killed after 3 day culture. The species specificity of killing effects of Pseudomonas sp. LG-2 against 5 species of dinoflagellate was only found in P. micans and Scrippsiella trochoidea.

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Biodegradation of Polychlorinated Biphenyls (PCBs) within Insulating Oil by Pseudomonas sp. P2 (Pseudomonas sp. P2에 의한 절연류 내의 Polychlorinated Biphenyls (PCBs)의 분해)

  • Kim, Jung-Ho;Choi, Sang-Ki;Kim, Young-Ho
    • Journal of Environmental Health Sciences
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    • v.22 no.3
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    • pp.1-7
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    • 1996
  • Polychlorinated Biphenyls(PCBs)의 생물학적 처리가 시도되고 있으며, PCBs를 분해할 수 있는 미생물을 이용할 수 있다. 따라서 본 연구에서는 폐기된 절연유의 생물학적 처리를 위하여 PCBs를 분해하는 균을 분해하였으며, 분해된 균을 이용하여 절연유 내의 Polychlorinated Biphenyls(PCBs) 분해를 회분식 실험에서 연구하였다. 대구의 신천으로부터 유일한 탄소원으로 Biphenyl을 포함하고 있는 고체배지에서 PCBs를 분해할 수 있는 Pseudomonas sp. P2 균주를 분해하였다. PCBs의 용해도를 높이기 위해 사용된 유화제 alkyl aryl ethoxylated phosphate가 200 mg/L에서는 Pseudomonas sp. P2 균주의 성장에 영향을 미치지 않았다. 1000 mg/L의 Biphenyl과 PCBs에 Pseudomonas sp. P2를 접종하여 160시간 배양후에 Biphenyl과 PCBs의 분해가 각각 97.5%, 58.0%였다. Biphenyl 1000 mg/L에서 최대성장율($\mu_{max}$)은 0.34 $day^{-1}$, 0.26 였다. 따라서 염소가 결합되지 않은 Biphenyl는 염소가 결합된 PCBs보다 분해가 빠르게 진행되었다. 또한 Pseudomonas sp. P2는 Biphenyl과 PCBs의 분해로 부터 유도된 황색의 분해대사산물을 확인하였다. 본 연구에서는 Pseudomonas sp. P2 균주가 절연유 내의 PCBs를 분해할 수 있다는 것을 확인하였다.

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Cloning of pcb Genes in Pseudomonas sp.P20 Specifying Degradation of 4-Clorobiphenyl (4-Chlorobiphenyl을 분해하는 Pseudomonas sp. P20의 pcb 유전자군의 클로닝)

  • 남정현;김치경
    • Microbiology and Biotechnology Letters
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    • v.22 no.4
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    • pp.353-359
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    • 1994
  • Pseudomonas sp. P20 was a bacterial isolate which has the ability to degrade 4-chlorobi- phenyl(4CB) to 4-chlorobenzoic acid via the process of meta-cleavage. The recombinant plasmid pCK1 was constructed by insetting the 14-kb EcoRI fragment of the chromosomal DNA containing the 4CB-degrading genes into the vector pBluescript SK(+). Subsequently, E. coli XL1-Blue was transformed with the hybrid plasmid producing the recombinant E. coli CK1. The recombinant cells degraded 4CB and 2,3-dihydroxybiphenyl(2,3-DHBP) by the pcbAB and pcbCD gene products, respectively. The pcbC gene was expressed most abundantly at the late exponential phase in E. coli CK1 as well as in Pseudomonas sp. P20, and the level of the pcbC gene product, 2,3-DHBP dioxygenase, expressed in E. coli CK1 was about two-times higher than in Pseudomonas sp. P20. The activities of 2,3-DHBP dioxygenase on catechol and 3-methylcatechol were about 26 to 31% of its activity on 2,3-DHBP, but the enzyme did not reveal any activities on 4-methylcatechol and 4-chlorocatechol.

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Molecular Cloning and Expression of Cellulase of Gene of Pseudomonas sp. in Escherichia coli (Pseudomonas sp.의 Cellulase 유전자의 대장균에의 클로닝 및 발현)

  • 정영철;김양우;노종수;성낙계;강신권
    • Microbiology and Biotechnology Letters
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    • v.18 no.6
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    • pp.633-639
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    • 1990
  • The genes for cellulases of Pseudomonas sp. LBC505 and CYC10, potent cellulase complex-producing strains, were cloned in Escherichia coli with pUC19. Recombinant plasmids pLCl and pLC2 were isolated from transformants producing cellulase by Congo red staining, and their genes cloned were 0.7 kb and 4.6 kb HindIII fragments, respectively. The inserts of pLCl and pLC2 were hybridized to chromosomal DNAs digested with HindIII from Pseudomona~ sp. LBC505 and CYC10, respectively. Immunodiffusion assays revealed that pLC1-and pLC2-encoded cellulase showed similarity with that of host strains. About 24% of cellulase activity was observed in the extracellular fraction of E. coli carrying pLC1, and its activity was higher about 1.4 times than that of LBC505. The enzymatic properties of pLC1 and pLC2 encoded cellulase were the same as those of cellulase from host strains. HPLC analysis and substrate specificity showed that cellulases were the same as those of cellulase from host strains. HPLC analysis and substrate specificity showed that cellulases cloned were endocellulase.

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Characterization of the Quinoline-Degrading Bacterium Pseudomonas sp. NFQ-1 Isolated from Dead Coal Pit Areas (폐광지역에서 분리한 quinoline 분해 세균인 Pseudomonas sp. NFQ-1의 특성연구)

  • 윤경하;황선영;권오성;오계헌
    • KSBB Journal
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    • v.18 no.3
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    • pp.174-179
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    • 2003
  • The bacterium NFQ-1 capable of utilizing quinoline (2,3-benzopyridine) as the sole source of carbon, nitrogen and energy was enriched and isolated from soil samples of dead coal pit areas. Strain NFQ-1 was identified as Pseudomonas nitroreducens NFQ-1 by BIOLOG system, and assigned to Pseudomonas sp. NFO-1. Pseudomonas sp. NFQ-1 was used with the concentration range of 1 to 10 mM quinoline. Strain NFQ-1 could degrade 2.5 mM quinoline within 9 hours of incubation. Initial pH 8.0 in the culture was reduced to 6.8, and eventually 7.0 as the incubation was proceeding. 2-Hydroxyquinoline, the first intermediate of the degradative pathway, accumulated transiently in the growth medium. The highest concentration of quinoline (15 mM) in this work inhibited cell growth and quinoline degradation. Pseudomonas sp. NFQ-1 was able to utilize various quinoline derivatives and aromatic compounds including 2-hydroxyquinoline, p-comaric acid, benzoic acid, p-cresol, p-hydroxybenzoate, protocatechuic acid, and catechol. The specific activity of catechol oxygenases was determined to approximately 184.7 unit/㎎ for catechol 1.2-dioxygenase and 33.19 unit/㎎ for catechol 2,3-dioxygenase, respectively. As the result, it showed that strain NFQ-1 degraded quinoline via mainly orthp-cleavage pathway, and in partial meta-cleavage pathway.

Characteristics of Biosurfactant Producing Pseudomonas sp. Z1 (생물 계면활성제를 생산하는 Pseudomonas sp. Z1의 특성)

  • Chang, Dong-Ho;Ko, Eun-Jung;Park, Kyeong-Ryang
    • Journal of Life Science
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    • v.21 no.1
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    • pp.134-140
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    • 2011
  • One hundred forty five bacterial colonies which were able to degrade crude oil were isolated from soil samples that were contaminated with oil in the Daejon area. Among these colonies, one bacterial strain was selected for this study based on its low surface tension ability, and this selected bacterial strain was identified as Pseudomonas sp. Z1 through physiological-biochemical tests and analysis of its 16S rRNA sequence. Pseudomonas sp. Z1 showed a high resistance to antibiotics such as chloramphenicol and ampicillin, as well as heavy metals such as lithium, manganese, and barium. It was found that the optimal pH and temperature for biosurfactant production of Pseudomonas sp Z1 were pH 6.0-7.0 and $30^{\circ}C$, respectively. After ten hours of inoculation, the biosurfactant activity of the culture broth decreased rapidly, and had maximum surface tension (28 dyne/cm) after twenty-one hours incubation. The biosurfactant activity of the culture broth was also decreased up to 2% NaCl concentration.

Isolation of a Pseudomonas sp. Capable of Utilizing 4-Nonylphenol in the Presence of Phenol

  • Chakraborty Joydeep;Dutta Tapan K.
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1740-1746
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    • 2006
  • Enrichment techniques led to the isolation of a Pseudomonas sp. strain P2 from municipal waste-contaminated soil sample, which could utilize different isomers of a commercial mixture of 4-nonylphenol when grown in the presence of phenol. The isolate was identified as Pseudomonas sp., based on the morphological, nutritional, and biochemical characteristics and 16S rDNA sequence analysis. The ${\beta}$-ketoadipate pathway was found to be involved in the degradation of phenol by Pseudomonas sp. strain P2. Gas chromatography-mass spectrometric analysis of the culture media indicated degradation of various major isomers of 4-nonylphenol in the range of 29-50%. However, the selected ion monitoring mode of analysis of biodegraded products of 4-nonylphenol indicated the absence of any aromatic compounds other than those of the isomers of 4-nonylphenol. Moreover, Pseudomonas sp. strain P2 was incapable of utilizing various alkanes individually as sole carbon source, whereas the degradation of 4-nonylphenol was observed only when the test organism was induced with phenol, suggesting that the degradation of 4-nonylphenol was possibly initiated from the phenolic moiety of the molecule, but not from the alkyl side-chain.