• Title/Summary/Keyword: Pseudomonas sp

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Characterization of Naphthalene-Degrading Pseudomonas Species Isolated from Pollutant-Contaminated Sites: Oxidative Stress During their Growth on Naphthalene

  • Kang, Yoon-Suk;Kim, Young-Jun;Jeon, Che-Ok;Park, Woo-Jun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1819-1825
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    • 2006
  • Four naphthalene-degrading bacteria (Pseudomonas sp. strains O1, W1, As1, and G1) were isolated feom pollutant-contaminated sites. Examination of their substrate utilization and analyses of key naphthalene-catabolic regulatory genes revealed that the pathway and regulation of naphthalene-degradation in all four strains resemble those of NAH7 from P. putida G7. Superoxide anion production, superoxide dismutase activity, and catalase activity during their growth on naphthalene-amended medium increased significantly, compared with those with glucose-amended medium. Addition of ascorbate, an antioxidant, or ferrous iron ($Fe^{2+}$) increased the growth rates of all tested microorganisms on naphthalene. Northern blot and HPLC analyses showed that both nahA gene expression and naphthalene degradation increased under those conditions. Our data suggest that naphthalene degradation can impose severe oxidative stress, and defenses against oxidative stress would play an important role in the metabolism of naphthalene.

A Study on the Isolation of the Oil-degradation Microbes and Treatment Efficiency in the Oil Contaminated Soil with Peat Moss (유류오염토양에서 유류분해 미생물의 분리 및 peat moss를 이용한 오염토양 처리에 관한 연구)

  • Chun, Mi-Hee;Son, Hee-Jeong;Kim, Chul
    • Journal of Environmental Health Sciences
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    • v.33 no.5
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    • pp.462-469
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    • 2007
  • Isolation and application of oil-degradation microbes from the oil-contaminated soil and the determination of optimal operation conditions about the peat moss, the addition for the oil-biodegradation. After all experiments, we have acquired three important conclusions: First, we found out the 4 microbes, Pseudomonas fluorescens, Pseudomonas aeruinosa, Kurtia sp., Bacillus ceres, with excellent capability for the oil-degradation; Second, the optimal operating conditions of the peat moss for TPH treatment were pH $7{\sim}8$, temperature $25{\sim}30^{\circ}C$, water content 20%, mixing 2 times/ day, addition volume 2%; Third, in case of the application to the oil-contaminated soil with 4 mixed microbes, the removal efficiency of TPH was increased from 54% to 83% in oil-contaminated soil and from 65% to 85% in oil-contaminated soil with the peat moss.

Culture-Based and Denaturing Gradient Gel Electrophoresis Analysis of the Bacterial Community Structure from the Intestinal Tracts of Earthworms (Eisenia fetida)

  • Hong, Sung-Wook;Kim, In-Su;Lee, Ju-Sam;Chung, Kun-Sub
    • Journal of Microbiology and Biotechnology
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    • v.21 no.9
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    • pp.885-892
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    • 2011
  • The bacterial communities in the intestinal tracts of earthworm were investigated by culture-dependent and -independent approaches. In total, 72 and 55 pure cultures were isolated from the intestinal tracts of earthworms under aerobic and anaerobic conditions, respectively. Aerobic bacteria were classified as Aeromonas (40%), Bacillus (37%), Photobacterium (10%), Pseudomonas (7%), and Shewanella (6%). Anaerobic bacteria were classified as Aeromonas (52%), Bacillus (27%), Shewanella (12%), Paenibacillus (5%), Clostridium (2%), and Cellulosimicrobium (2%). The dominant microorganisms were Aeromonas and Bacillus species under both aerobic and anaerobic conditions. In all, 39 DNA fragments were identified by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis. Aeromonas sp. was the dominant microorganism in feeds, intestinal tracts, and casts of earthworms. The DGGE band intensity of Aeromonas from feeds, intestinal tracts, and casts of earthworms was 12.8%, 14.7%, and 15.1%, respectively. The other strains identified were Bacillus, Clostridium, Enterobacter, Photobacterium, Pseudomonas, Shewanella, Streptomyces, uncultured Chloroflexi bacterium, and uncultured bacterium. These results suggest that PCR-DGGE analysis was more efficient than the culturedependent approach for the investigation of bacterial diversity and the identification of unculturable microorganisms.

Purification and Characterization of Chitinase from a New Species Strain, Pseudomonas sp. TKU008

  • Wang, San-Lang;Lin, Bo-Shyun;Liang, Tzu-Wen;Wang, Chuan-Lu;Wu, Pei-Chen;Liu, Je-Ruei
    • Journal of Microbiology and Biotechnology
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    • v.20 no.6
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    • pp.1001-1005
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    • 2010
  • The chitinase-producing strain TKU008 was isolated from soil in Taiwan, and it was identified as a new species of Pseudomonas. The culture condition suitable for production of chitinase was found to be shaking at $30^{\circ}C$ for 4 days in 100 ml of medium containing 1% shrimp and crab shell powder, 0.1% $K_2HPO_4$, and 0.05% $MgSO_4{\cdot}7H_2O$ (pH 7). The TKU008 chitinase was suppressed by the simultaneously existing protease, which also showed the maximum activity at the fourth day of incubation. The molecular mass of the chitinase was estimated to be 40 kDa by SDS-PAGE. The optimum pH, optimum temperature, pH stability, and thermal stability of the chitinase were pH 7, $50^{\circ}C$, pH 6-7, and <$50^{\circ}C$, respectively. The chitinase was completely inhibited by $Mn^{2+}$ and $Cu^{2+}$. The results of peptide mass mapping showed that 11 tryptic peptides of the chitinase were identical to the chitinase CW from Bacillus cereus (GenBank Accession No. gi 45827175) with a 32% sequence coverage.

Cloning and Characterization of the Genes Responsible for Degradation of 4-Chlorobenzoic Acid (4-Chlorobenzoic Acid 분해유전자의 클로닝과 유전학적 특성)

  • 이익근;김종우;김치경
    • Korean Journal of Microbiology
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    • v.28 no.1
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    • pp.41-46
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    • 1990
  • A bacterial isolate of DJ-12 capable of degrading 4-chlorobenzoic acid (4CBA) as well as 4-chlorobiphenyl (4CB) was used in this study. Its biodegradability of 4CBA was tested and the location of the genes coding for degradation of 4CBA was investigated by the nethod of in vivo cloning. The genes were found to be existed in the plasmid of pDJ121 which is about 65kb in size and which has 9, 11, 10, and 19 restriction sites for EcoRI, HindIII, SalI, and PstI, respectively. The hybrid plasmid of pDK450 was constructed by ligation of the EcoRI fragments of pDJ121 with pKT230 as a vector. In the recombinant cells selected through transformation of the hybrid vector into Pseudomonas putida KT2440, the 4CBA-degrading genes of DJ-12 were proved to be cloned and expressed in the Pseudomonas sp.

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Isolation, Characterization and Numerical Taxonomy of Novel Oxalate-oxidizing Bacteria

  • Sahin, Nurettin;Gokler, Isa;Tamer, Abdurrahman
    • Journal of Microbiology
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    • v.40 no.2
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    • pp.109-118
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    • 2002
  • The present work is aimed at providing additional new pure cultures of oxalate utilizing bacteria and its preliminary characterization for further work in the field of oxalate-metabolism and taxonomic studies. The taxonomy of 14 mesophilic, aerobic oxalotrophic bacteria isolated by an enrichment culture technique from soils rhizosphers, and the juice of the petiole/stem tissue of plants was investigated. Isolates were characterized with 95 morphological, biochemical and physiological tests. Cellular lipid components and carotenoids of isolates were also studied as an aid to taxonomic characterization. All isolates were Gram-negative, oxidase and catalase positive and no growth factors were required. In addition to oxalates, some of the strains grow on methanol and/or formate. The taxonomic similarities among isolates, reference strains or previously reported oxalotrophic bacteria were analysed by using the Simple Matching (S/ sub SM/) and Jaccard (S$\_$J/) Coefficients. Clustering was performed by using the unweighted pair group method with arithmetic averages (UPGMA) algorithm. The oxalotrophic strains formed five major and two single-member clusters at the 70-86% similarity level. Based on the numerical taxonomy, isolates were separated into three phenotypic groups. Pink-pigmented strains belonged to Methylobacterium extorquens, yellow-pigmented strains were most similar to Pseudomonas sp. YOx and Xanthobacter autorophicus, and heterogeneous non-pigmented strains were closely related to genera Azospirillum, Ancylobacter, Burkholderia and Pseudomonas. New strains belonged to the genera Pseudomonas, Azospirillum and Ancylobacter that differ taxonomically from other known oxalate oxidizers were obtained. Numerical analysis indicated that some strains of the yellow-pigmented and nonpigmented clusters might represent new species.

Characterization and Expression of Antibacterial Protein Gene, Nuecin (곤충세포주에서 누에신 단백질의 발현 및 성상구명)

  • 윤은영;구태원;황재삼;김상현;강석우;김근영;진병래
    • Journal of Sericultural and Entomological Science
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    • v.44 no.2
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    • pp.64-68
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    • 2002
  • The antibacterial protein gene, nuecin was expressed in Sf9 cells using baculovirus expression vector system (BEVS). The antibactetial activity of mature nuecin against Pectobacterium carotovorum subsp. carotovorum, Ralstonia solanacearum and Pseudomonas tolaasii was significantly high, demonstrating that nuecin had a wider antibacterial spectrum on gram negative and positive bacteria. The result appears to be superior to other antibacterial peptide, attacin. The nuecin was purified by SP-sepharose and Mono Q HR ion-exchange chromatography, and then by Superdex 200 HR 10/30 column. The purified nuecin is quite stable at 80$\^{C}$ and 100$\^{C}$ for several hours of incubation and in a wide pH range (pH 2-12).

Isolation and Identification of a Histamine-degrading Barteria from Salted Mackerel (자반고등어에서 histamine 분해능을 가진 세균의 분리 동정)

  • Hwang Su-Jung;Kim Young-Man
    • Journal of Life Science
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    • v.15 no.5 s.72
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    • pp.743-748
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    • 2005
  • Histamine can be produced at early spoilage stage through decarboxylation of histidine in red-flesh fish by Proteus morganii, Hafnia alvei or Klebsiella pneumoniae. Allergic food poisoning is resulted from the histamine produced when the freshness of Mackerel degrades. Conversely it has been reported that there are bacteria which decompose histamine at the later stage. We isolated histamine decomposers from salted mackerel and studied the characteristics to help establish hygienic measure to prevent outbreak of salted mackerel food poisoning. All the samples were purchased through local supermarket. Histamine decomposers were isolated using restriction medium using histamine 10 species were selected. Identification of these isolates were carried out by the comparison of 16S rDNA partial sequence; as a result, we identified Pseudomonas putida strain RA2 and Halomonas marina, Uncultured Arctic sea ice bacterium clone ARKXV1/2-136, Halomonas venusta, Psychrobacter sp. HS5323, Pseudomonas putida KT2440, Rhodococcus erythropolis, Klebsiella terrigena (Raoultella terrigena), Alteromonadaceae bacterium T1, Shewanella massilia with homology of $100\%,{\;}100\%,{\;}99\%,{\;}99\%,{\;}99\%,{\;}99\%,{\;}100\%,{\;}95\%,{\;}99\%,{\;}and{\;}100\%$respectively. Turbidometry determination method and enzymic method were employed to determine the ability of histamine decomposition. Among those species Shewanella massilia showed the highest in ability of histamine decomposition. From these results we confirmed various histamine decomposer were present in salted mackerel product in the market.

Associated Nitrogen Fixation in the Rhizosphere of Rice in Saline and Reclaimed Saline Paddy Soil -IV. Chemotaxis of Associative N2-fixing Bacteria to Single or Combined Sugars and Root Exudates (간척지토양(干拓地土壤)의 수도근권(水稻根圈)에서 협생질소고정(協生窒素固定)에 관(關)한 연구(硏究) -제사보(第四報). 협생질소고정균(協生窒素固定菌)의 단당(單糖) 및 복합당류(複合糖類)와 근분비물질(根分泌物質)에 대(對)한 화학주화성(化學走化性))

  • Lee, Sang-Kyu;Suh, Jang-Sun;Ko, Jae-Young
    • Korean Journal of Soil Science and Fertilizer
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    • v.20 no.4
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    • pp.359-367
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    • 1987
  • A method for evaluating bacterial chemotactic responses toward several single of combined sugars and sterile mucilage from the different rice cultivars had been tested. Bacterial genus of Azospirillum, Pseudomonas and Agrobacterium were specially identified from the histosphere of different rice cultivars and graminea grasses in saline and reclaimed saline paddy soil. To evaluate chemotaxis of these strains a modification of Fendrik channel method was used. Under this condition Azospirillum lipoferum Ecc 3-1 reacted stereoisomerically fomulating the single migration ring while Agrobacterium radiobacter Ecc 1-1 and Pseudomonas sp Ecc 4-1 did not. Strains specificities of chemotaxis to the single sugar such as D(+)-glucose and D(+)-fructose were less prominent than malic and citric acid. Chemotactic responses to the combined sugar such as D-galacturonic acid and the L-aspartate were found high attracting reaction than other combined sugars. Chemotaxis of associative $N_2$-fixing bacteria to the root exudates of different rice cultivars were differed among bacterial strains and rice cultivars.

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Removal Characteristics of Geosmin in a Slow Sand Filteration Process (완속 모래여과 공정에서의 Geosmin 제거 특성)

  • Son, Hee-Jong;Yoom, Hoon-Sik;Jang, Seong-Ho
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.8
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    • pp.754-760
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    • 2010
  • Geosmin removal by biodegradation was investigated in lab-scale slow sand filtration column with different empty bed contact times (EBCTs) and water temperature. Schmutzdecke layer was built up after 30 days operation and biomass and activity were $4.5{\times}10^6\;CFU/g$ and $3.42\;mg{\cdot}C/m^3{\cdot}hr$, respectively. The attached bio-film microorganisms in schmutzdecke layer were isolated and identified. The dominant species was Pseudomonas sp. that had occupied 56%. Removal efficiencies of dissolved organic carbon (DOC) and geosmin were 27% and 95% after 30 days operation. In lab-scale slow sand filtration column, geosmin and DOC removal efficiencies were 62% and 10% at $5^{\circ}C$, respectively. And increasing water temperature ($15^{\circ}C$ and $25^{\circ}C$) increased the geosmin and DOC removal efficiencies (88~100% and 25~42%) in lab-scale slow sand filtration column. Geosmin and DOC biodegradation rates (k) in the schmutzdecke layer (in the upper 5 cm filter bed) were $1.842{\sim}15.965\;hr^{-1}$1 and $0.253{\sim}1.123\;hr^{-1}$, respectively. It were about 18~32 times and 20~51 times of the rates in the deeper filter bed (5~60 cm).