• Title/Summary/Keyword: Pseudomonas isolate

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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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Isolation and Utilization of Antagonistic Pseudomonas fluorescens from Soils for the Protection of Soybean Sprouts Rot (콩나물 부패병 방제를 위해 토양으로부터 분리한 길항균 Pseudomonas fluorescens의 이용)

  • Kim, Jin-Ho;Joo, Gil-Jae;Choi, Yong-Hwa
    • Korean Journal of Environmental Agriculture
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    • v.20 no.1
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    • pp.50-56
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    • 2001
  • Thirty-three bacterial and fungal strains were isolated from the rotten soybeans and soybean sprouts to isolate pathogenic microorganisms which cause soybean sprouts rot during soybean sprouts cultivation. In pathogenicity tests of the isolates on soybean sprouts, two isolates(K-17 and K-28) caused soybean sprouts rot and were identified as Erwinia carotovora and Fusarium sp., respectively. To isolate antagonists aganist K-17 and K-28 pathogens, bacteria were isolated from various soybean-cultivated soils and screened by the inhibition zone method. A bacterial isolate(J-232) which inhibited growth of both pathogens was identified as Pseudomonas fluorescens and further examined. The culture filtrate of P. fluorescens J-232 (dilution rate of 500 times) inhibited the growth of Erwinia carotovora K-17 and Fusarium sp. K-28 both on potato dextrose agar medium and on soybean sprouts cultivated in vessel. The development of soybean sprouts rots was observed during cultivation by inoculation of soybean seeds with culture filtrate of both pathogens. The combined inoculation of soybean seeds with culture filtrate of antagonistic bacterium and that of pathogens prevented soybean sprouts rot, and the growth of soybean sprouts was similar to that of control. The soybean sprouts inoculated with antagonists culture filtrate alone did not develop soybean sprouts rot, and the growth of the seedlings was shown to be slightly promoted as compared with that of control.

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Production of Antibacterial Substance, and Changes in Peroxidase nd Polyphenoloxidase Activities in Tobacco Plants Inoculated with Avirulent Isolate of Pseudomonas solanacearum (비병원성 Pseudomonas solanacearum 접종에 의한 담배내 항균물질생성과 Peroxidase 및 Polyphenoloxidase의 변화)

  • Yi Young Keun;Min Tae Gi;Park Won Mok
    • Korean Journal Plant Pathology
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    • v.3 no.3
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    • pp.203-209
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    • 1987
  • The substances obtained from the leaf, stem and root of tobacco plants inoculated with avirulent and virulent isolates of Pseudomonas solanacearum were at R_f\;0.6$ and R_f\;0.9$ on TLC plate, respectively. Both substances showed antibacterial activities not only on P. solanacearum but also on Erwinia carotovora subsp. carotovora and Escherichia coli in vitro. However, the antibacterial substances were not detectable from the filtrate of the autoclaves tobacco sap medium, in which the avirulent or virulent bacterium was cultured for 3 days. Peroxidase and poly phenoloxidase activities and their isozyme patterns did not differ significantly between plants treated with the virulent and avirulent isolates, or between the susceptible cultivar BY 4 and the resistant cultivar NC 82. However, activities of the two enzymes were increased in leaves of the susceptible cultivar BY 4 treated with either the virulent or the avirulent isolate.

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Isolation and Characterization of a Marine Bacterium, Pseudomonas sp. YJ-1 with Anti-Methicillin Resistant Staphylococcus aureus Activity (항 Methicillin Resistant Staphylococcus aureus 활성을 가지는 해양미생물 Pseudomonas sp. YJ-1의 분리와 특성)

  • Woo, Ye-Ju;Jeong, Seong-Yun
    • Korean Journal of Environmental Biology
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    • v.35 no.4
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    • pp.694-705
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    • 2017
  • The aim of this study was to isolate and identify marine bacterium with anti-methicillin-resistant Staphylococcus aureus (MRSA) activity, and to purify the anti-MRSA compound, as well as to determine its activity and synergistic effects. Among the marine bacteria isolated in this study, the YJ-1 isolate had the strongest anti-MRSA activity. The YJ-1 isolate was identified on the basis of its biochemical characteristics and an analysis of 16S rRNA gene sequences. The YJ-1 isolate showed over 99.2% homology with Pseudomonas stutzeri, and was designated as a Pseudomonas sp. YJ-1. The optimal culture conditions were $25^{\circ}C$ and initial pH 7.0. For the purification of the anti-MRSA compounds, the YJ-1 was cultured in Pa PES-II medium, and the culture filtrates were extracted by ethyl acetate, hexane, and 80% MeOH. The 80% MeOH fraction was separated by a $C_{18}$ ODS column, silica gel chromatography and a reverse phase HPLC, to yield three anti-MRSA agents, the MR1, MR2, and MR3 compounds. When the MR1 compound of $250{\mu}g\;mL^{-1}$ concentration was applied to the MRSA cells, over 95% of bacterial cells was killed within 48 hr. Compared with vancomycin and ampicillin, the MR1 compound showed significant anti-MRSA activity. In addition, the anti-MRSA activity was increased by dose and time dependent manners. Furthermore, the combination of an MR1 compound with vancomycin produced a more rapid decrease in the MRSA cells than did the MR1 compound alone. Taken together, our results suggest that the Pseudomonas sp. YJ-1 and its anti-MRSA compounds could be employed as a natural antibacterial agent in MRSA infections.

Isolation and Characterization of Pseudomonas sp. P2 Degrading Polychlorinated Biphenyls (PCBs)

  • Kim, Jung Ho;Sang Ki Choi;Moon Ki Park;Young Ho Kim;Seung Kyo Suh;Cheol Joo Woo;Heui Dong park
    • Journal of Microbiology and Biotechnology
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    • v.6 no.3
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    • pp.167-172
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    • 1996
  • The bacterial strain P2 degrading polychlorinated biphenyls (PCBs) was isolated from the soil around the Shinchun stream in Taegu after enrichment culture in a media containing biphenyl as the sole carbon source. The isolate was identified as a strain of Pseudomonas sp. based on its morphological and physiological characteristics. The optimal conditions of initial pH of media and temperature for growth were 7.0 and $30^{\circ}C$, respectively. Degradation of biphenyl and PCBs was confirmed by GC during the culture of Pseudomonas sp. P2 in a media containing them at a concentration of 500 mg/I. It was observed that Pseudomonas sp. P2 could degrade 97.0$%$ of biphenyl and 60.0$%$ of PCBs after 160 h culture.

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Genome Analysis of Naphthalene-Degrading Pseudomonas sp. AS1 Harboring the Megaplasmid pAS1

  • Kim, Jisun;Park, Woojun
    • Journal of Microbiology and Biotechnology
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    • v.28 no.2
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    • pp.330-337
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    • 2018
  • Polycyclic aromatic hydrocarbons (PAHs), including naphthalene, are widely distributed in nature. Naphthalene has been regarded as a model PAH compound for investigating the mechanisms of bacterial PAH biodegradation. Pseudomonas sp. AS1 isolated from an arseniccontaminated site is capable of growing on various aromatic compounds such as naphthalene, salicylate, and catechol, but not on gentisate. The genome of strain AS1 consists of a 6,126,864 bp circular chromosome and the 81,841 bp circular plasmid pAS1. Pseudomonas sp. AS1 has multiple dioxygenases and related enzymes involved in the degradation of aromatic compounds, which might contribute to the metabolic versatility of this isolate. The pAS1 plasmid exhibits extremely high similarity in size and sequences to the well-known naphthalene-degrading plasmid pDTG1 in Pseudomonas putida strain NCIB 9816-4. Two gene clusters involved in the naphthalene degradation pathway were identified on pAS1. The expression of several nah genes on the plasmid was upregulated by more than 2-fold when naphthalene was used as a sole carbon source. Strains have been isolated at different times and places with different characteristics, but similar genes involved in the degradation of aromatic compounds have been identified on their plasmids, which suggests that the transmissibility of the plasmids might play an important role in the adaptation of the microorganisms to mineralize the compounds.

Isolation and Characterization of Pseudomonas putida N3 Degrading Naphthalene (Naphthalene을 분해하는 Pseudomonas putida N3의 분리 및 특성)

  • 고영희;하일호;배경숙
    • Microbiology and Biotechnology Letters
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    • v.16 no.3
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    • pp.199-204
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    • 1988
  • A strain capable of growth on naphthalene minimal medium was isolated from soil by selective enrichment culture and identified as Pseudomonas putida N3 according to its morphological and physiological characteristics. The optimum pH and temperature for growth of the isolate were 7.0 and 3$0^{\circ}C$, respectively. This strain was resistant to ampicillin, chloramphenicol, kanamycin and streptomycin but. sensitive to tetracycline and rifampicin. Of the naphthalene related compounds, 1, 5-dihydroxynaphthalene was more easily utilized than naphthalene due to its solubility. And catechol was degraded through meta-cleavage pathway. A 110 Kb plasmid which encodes for a single set of enzymes responsible for the degradation of naphthalene was obtained.

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Isolation and Characterization of Pseudomonas sp. KM10, a Cadmium- and Mercury-resistant, and Phenol-degrading Bacterium

  • Yoon, Kyung-Pyo
    • Journal of Microbiology and Biotechnology
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    • v.8 no.4
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    • pp.388-398
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    • 1998
  • A bacterium which is resistant to both mercury and cadmium, and also capable of utilizing phenol as a carbon and energy source, was isolated from the Kumho River sediments near Kangchang Bridge, Taegu, Korea. The isolate was labeled Pseudomonas sp. KM10 and characterized. The bacteria grew in 4 mM $CdCl_2$and in $70{\mu}M$ $HgCl_2$. The bacteria efficiently removed over 90% of 1 g/l phenol within 30 h. In the presence of 1.250 g/l phenol, the growth of the microorganism was slightly retarded and the microorganism could not tolerate 1.5 g/l phenol. Curing of plasmid from the bacteria was carried out to generate a plasmidless strain. Subsequent experiments localized the genes for phenol degradation in plasmid and the genes for mercury resistance and cadmium resistance on the chromosome. Dot hybridization and Southern hybridization under low stringent conditions were performed to identify the DNA homology. These results showed significant homologies between the some sequence of the chromosome of Pseudomonas sp. KM10 and merR of Shigella flexneri R 100, and between the some sequence of the chromosome of Pseudomonas sp. KM10 and cadA of Staphylococcus aureus pI258. The mechanism of cadmium resistance was efflux, similar to that of S. aureus pI258 cadA, and the mechanism of mercury resistance was volatilization, similar to that of S. flexneri R100 mer.

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