• Title/Summary/Keyword: Pseudomonas sp. P20

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Isolation and Characterization of Yam-Putrefactive Psychrotrophic Bacteria from Rotted Yam (생마 저온부패 원인세균의 분리 및 부패균의 특성)

  • Ryu Hee-Young;Kim Young-Sook;Park Sang-Jo;Lee Bong-Ho;Kwon Soon-Tae;Sohn Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.34 no.2
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    • pp.109-114
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    • 2006
  • Yam has been recognized as healthy food due to its various biological activities, such as anti-obesity, antimicrobial, anticancer and immuno-stimulation activities, and its consumption has been increased during last decades. In this study, to investigate low-temperature, long-term storage of yam and to develop processed yam products, yam-putrefactive psychrotrophic bacteria were isolated from rotted yam and identified based on BBL identification system, fatty acid analysis in cell membrane and 16S rDNA sequence analysis. The putrefaction activity of isolated thirteen bacteria was evaluated using yam-slices (NaOCl-treated, autoclaved yam and without treatment), and YAM-10 and YAM-12 were identified as major psychrotrophic putrefactive bacteria. Both YAM-10 (Pseudomonas cepacia) and YAM-12 (Pseudomonas rhodesiae) bacteria grew well at 4$\sim$12$^{\circ}C$ and showed strong activity of polymer degrading enzymes, especially amylase, carboxy methyl cellulase and xylanase, at 20$^{\circ}C$. But they failed to grow at acidic pH (<5) or alkaline pH (>10). Our results suggested that the control of psychrotrophic Pseudomonas sp. by pH change and inhibition of polymer degrading enzymes, such as amy-lase, are necessary to long-term storage of yam.

Isolation and Identification of Lipolytic Enzyme Producing Pseudomonas sp. OME and Optimization of Cultural Conditions (지방분해효소 생산균 Pseudomonas sp. OME 의 분리 동정 및 배양조건 최적화)

  • Kumar, G.Satheesh;Reddy, T. Kiran;Madhavi, B.;Teja, P.Charan;Chandra, M.Subhosh;Choi, Yong-Lark
    • Journal of Life Science
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    • v.20 no.5
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    • pp.662-669
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    • 2010
  • Lipolytic enzyme-producing bacteria were isolated from edible oil mill effluents on tributyrin agar medium. The shake-flask-scale studies yielded a promising isolate and it was identified as Pseudomonas sp. An OME using various microbiological observations such as cultural, microscopic, and biochemical tests was undertaken and confirmed using PIBWIN bacterial identification software. Lipolytic enzyme production was screened with oils such as sunflower, caster, coconut, tributyrin, and olive. Amongst these, olive oil showed an increased lipase production 6.1 U/ml. In view of the highest lipolytic enzyme production with olive oil, further optimizations were carried out using olive oil as a carbon source. Lipolytic enzyme production was optimized by a conventional 'one variable at a time' approach and the significant factors were further analyzed statistically using response surface methodology (RSM). The effect of physical factors such as incubation time, temperature, initial medium pH, and nutritional factors such as concentration of olive oil and yeast extract were examined for lipase production. Lipolytic enzyme secretion was strongly affected by three variables (incubation time, concentration of yeast extract and olive oil). Therefore, the interaction of these three factors was further optimized using response surface methodology. The optimized conditions of lipase production using response surface methodology yielded a maximum of 9.62 U/ml with optimum conditions for incubation, yeast extract and olive oil concentrations were found to be 48 hr, 0.3 g. and 0.9 ml. respectively.

Production Conditions and Characterization of ${\beta}$-Lactamase Inhibitor from Pseudomonas sp. X-8 (슈도모나스 sp. X-8의 베타락타마제 억제제의 생산 조건과 특성)

  • Kim, Kyoung-Ja;Kim, Tae-Sung
    • YAKHAK HOEJI
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    • v.41 no.5
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    • pp.658-665
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    • 1997
  • Identification of a soil microorganism strain X-8, producer of ${\beta}$-lactamase inhibitor, based on its morphological, physiological, biochemical and chemotaxonomical characteristics was performed. The strain X-8 was identified as Pseudomonas sp. The beta-lactamase inhibitor produced by this strain was highly achieved in fermentation medium contained glucose 0.5%, urea 0.25%, $K_2HPO_4{\cdot}3H_2O\;0.5%,\;MgSO_4{\cdot}7H_2O\;0.5%,\;FeSO_4{\cdot}7H_2O\;0.01%,\;CuSO_4,\;ZnSO_4,\;MnSO_4\;0.02%$. The beta-lactamase inhibitor was not extracted by organic solvent such as n-butanol and ethyl acetate but remained in aqueous layer. The n-butanol extract showed antimicrobial activity against M. smegmatis. The ${\beta}$-lactamase inhibitor was stable at pH 7.0~8.0 and 4$^{\circ}C$ for 24h. The ${\beta}$-lactamase inhibitor was bound on ion exchanger Diaion WA-30 and HP-20 and eluted with 2N-$NH_4OH$ and acetone, respectively.

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In-situ Bioremediation of Total Petroleum Hydrocarbons-Contaminated Soil by Pseudomonas Species (토양 내 TPH(Total Petroleum Hydrocarbons)의 생물학적 분해 연구)

  • Kim, Jee-Young;Lee, Sang-Seob
    • Microbiology and Biotechnology Letters
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    • v.39 no.2
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    • pp.161-167
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    • 2011
  • We previously showed that five strains belonging to Pseudomonas could remove TPH (Total Petroleum Hydrocarbons) efficiently when they are applied to TPH-contaminated soil. We optimized the bioremediation condition using different hydrocarbons and nutrients conditions to improve the efficiency. We setup lab-scale column bioreactor to monitor TPH and diesel removal efficiency. When we applied five Pseudomonas sp. mixtures to 25,000 $mg{\cdot}kg^{-1}$ TPH-contaminated soil (diesel 10,000 $mg{\cdot}kg^{-1}$, kerosene 10,000 $mg{\cdot}kg^{-1}$, gasoline 5,000 $mg{\cdot}kg^{-1}$) with the optimum condition, 76.3% of TPH removal efficiency was shown for 25 days. Meanwhile, in the application of five Pseudomonas sp. mixtures to 20,000 $mg{\cdot}kg^{-1}$ diesel-contaminated soil with the optimum condition, 99.2% of diesel removal efficiency was shown for 40 days. In the application to lab-scale bioreactor with five high efficiency bacteria, 88.5% of TPH removal efficiency was shown for 45 days. Based on the results from this study, we confirmed that this mixed Pseudomonas sp. consortium might improve the bioremediation of TPH in contaminated soil, the efficacy can be controlled by improving the nutrients. We also confirmed that the nutrients and oxygen for biodegradation of TPH could contribute on the management and control of applications of these strains for the study of bioremediation of TPH-contaminated soil.

Induction of Defense Related Enzymes and Pathogenesis Related Proteins in Pseudomonas fluorescens-Treated Chickpea in Response to Infection by Fusarium oxysporum f. sp. ciceri

  • Saikia, Ratul;Kumar, Rakesh;Singh, Tanuja;Srivastava, Alok K.;Arora, Dilip K.;Lee, Min-Woong
    • Mycobiology
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    • v.32 no.1
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    • pp.47-53
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    • 2004
  • Pseudomonas fluorescens 1-94 induced systemic resistance in chickpea against Fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceri by the synthesis and accumulation of phenolic compounds, phenylalanine ammonia lyase(PAL) and pathogenesis related(PR) proteins(chitinase, $\beta$-1,3-glucanase and peroxidase). Time-course accumulation of these enzymes in chickpea plants inoculated with P. fluorescens was significantly(LSD, P=0.05) higher than control. Maximum activities of PR-proteins were recorded at 3 days after inoculation in all induced plants; thereafter, the activity decreased progressively. Five PR peroxidases detected in induced chickpea plants. Molecular mass of these purified peroxidases was 20, 29, 43, 66 and 97 kDa. Purified peroxidases showed antifungal activity against plant pathogenic fungi.

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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Characterization of Nitroreductase Purified from TNT-degrading Bacterium, Pseudomonas sp. HK-6. (폭약 TNT를 분해하는 세균인 Pseudomonas SP. HK-6에서 분리정제된 Nitroreductase의 특성연구)

  • 호은미;강형일;오계헌
    • Microbiology and Biotechnology Letters
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    • v.32 no.3
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    • pp.230-237
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    • 2004
  • In this study nitroreductase from Pseudomonas sp. HK-6 capable of degrading 2,4,6-trinitrotoluene (TNT) was characterized. Through a series of purification process including ammonium sulfate precipitation, DEAE-sepharose, and Q-sepharose, three different fractions I, II, and III having the enzyme activity of NTRs whose molecular weights were approximately 27 kDa were detected in fractions from HK-6 cells. Specific activity of the three fractions were approximately 4.85 unit/mg, 5.47 unit/mg, and 5.01 unit/mg, and concentrated to 9.0-, 10.1-, and 9.3-fold compared to crude extract, respectively. The optimal pH and temperature for the three NTR fractions were approximately 7.5 and $30^{\circ}C$, respectively. Metal ions, $Ag^{+}$ , $Cu^{ 2+}$, $Hg^{2+}$ inhibited approximately 70% of enzymes activities of all NTR, while $Fe^{2+}$ did not stimulate or inhibit the activities. Monitoring the effect of chemicals on the enzyme activity revealed that those NTR fractions lost enzyme activity in presence of $\beta$-mercaptoethanol, but were a little influenced by dithiothreitol, EDTA and NaCl. The three NTR fractions demonstrated enzyme activities for nitrobenzene and RDX as well as TNT.

Purification and Comparison of Properties of the C-Terminus Truncated Agarase of Pseudomonas sp. W7

  • Yoon, Soo-Cheol;Lee, Jong-Hee;Ahn, Sun-Hee;Lee, Eun-Mi;Park, Eun-Mi;Kong, In-Soo
    • Journal of Microbiology and Biotechnology
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    • v.13 no.5
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    • pp.767-772
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    • 2003
  • Three plasmids derived from the ${\beta}-agarase$ gene (PjaA) of Pseudomonas sp. W7 were expressed in Escherichia coli AD494(DE3) pLysS with lactose as an inducer. These products corresponded to the complete (PjaA) and the two C-terminal truncated (PjaAI and PjaAII) forms of ${\beta}-agarase$. The PjaAI and the PjaAII were originated from exonuclease L treatment from PjaA by deleting 127 and 182 amino acid residues-encoded nucleic acids at 3' region, respectively. The molecular weights of the purified proteins were 71 kDa, 58 kDa, and 50 kDa on SDS-PAGE, respectively. The $K_m$ value of PjaAI was lower than that of the PjaA, and the catalytic efficiency ($k_{cat}/K_m$) of PjaAI was increased to 5 times. The enzyme of PjaAI retained more than 90% activity at $50^{\circ}C$. In contrast to the PjaAI, the remaining activity of the PjaA was only 20% at the same temperature.

A Study on Bioremediation of Fish Farm Sediment Using CaO2 by Enhancement of Indigenous Microbial Activity (어류양식장 저질개선을 위한 과산화칼슘 투입에 의한 생태 환경변화 관찰)

  • Cho, Daechul;Bae, Hwan-Jin;Kwon, Sung-Hyun
    • Journal of Environmental Science International
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    • v.21 no.10
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    • pp.1187-1193
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    • 2012
  • The aim of this research is to enhance the bottom environment of Geoje fish farm that has been severely contaminated. Treatment of microbial agent and/or calcium oxide significantly changed that environment: in ignition loss, either treatment (25% or 21%) showed better than mixed treatment (13.2%). In COD, the oxygen releasing agent or mixed treatment reduced the index by more than 20%. In T-P and T-N, the effects of $CaO_2$ on them were overwhelming (50% or more) meanwhile that of the microbial agent on them was less than 20%. Also, $CaO_2$ influenced on the microbial flora: Desulfobvibrio thermophilus, a sulfate reducing bacterium decreased in number, considering the increase of pH and rise of redox potential. In contrast, Pseudomonas sp., Pseudoalteromonas sp., Pseudomonas aeruginosa were remarkably dominant over other species with mixed treatment as a PCA analysis confirmed it.

Isolation and Characterization of Aerobic Trichloroethylene Cometabolizing Bacterium (호기적 Trichloroethylene 공동대사 세균의 분리 및 특성)

  • 김호성;박근태;손홍주;박성훈;이상준
    • Journal of Environmental Science International
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    • v.10 no.2
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    • pp.99-103
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    • 2001
  • Several microorganisms which degrade phenol and trichloroethylene(TCE) were isolated from the activated sludge of a wastewater treatment plant. Among them, one isolate EL-04J showed the highest degradability and was identified as a Pseudomonas species according to morphological, cultural and biochemical properties. The phenol-induced cells of Pseudomonas EL-04J, which were preincubated in the mineral salts medium containing phenol as a sole carbon source, degraded 90% of 25$\mu$M TCE within 20h. This strain could also utilize some of methylated phenol derivatives (o-cresol, m-cresol and p-cresol) as the sole source of carbon and energy. Cresol-induced cells of Pseudomonas EL-04J also cometabolized TCE.

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