• Title/Summary/Keyword: Pseudomonas aeruginosa P-5

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Purification and Characterization of a Thermostable Protease from Pseudomonas aeruginosa NS-83

  • Kim, Hyung-Kwoun;Kim, Kee-Hyun;Lee, Jung-Kee;Bae, Kyung-Sook;Sung, Chang;Oh, Tae-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.4 no.2
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    • pp.113-118
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    • 1994
  • A bacterial strain NS-83 isolated from soil was able to produce an extracellular thermostable protease. The strain was identified as Pseudomonas aeruginosa based on its morphological and physiological characteristics. A thermostable protease from this strain has been purified to homogeneity as judged by SDS-PAGE and isoelectric focusing. The purification procedures included hydrophobic interaction, ion exchange, and gel filtration chromatography. The $M_r$ and the pl of the enzyme were 32,000 and 5.9, respectively. The optimal pH at 55$^{\circ}C$ and the optimal temperature at pH 7.0 were 8.0 and 60$^{\circ}C$, respectively. The D-values of the enzyme at 60, 65, and 70$^{\circ}C$ were 22, 2.1, and 0.75 hrs, respectively. The enzyme activity was significantly inhibited in the presence of 1 mM o-phenanthroline or EDTA, suggesting that the enzyme is metalloprotease. The $K_m$, and $V_{max}$ for Hammarsten casein were found to be 3.2 mg/ml and 0.918 unit/ml, respectively. These enzymatic properties were similar to those of elastase produced from P. aeruginosa IFO 3455, but the enzyme was clearly different from the reported elastase, in respect to $Ca^{++}$ effects on enzyme-thermostability. This property, together with amino acid composition analysis, confirmed that the enzyme differs from the known P. aeruginosa elastase.

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Two Cases of Invasive Pseudomonas aeruginosa Infection that Developed in the Apparently Immunocompetent Infants (면역 기능의 이상이 발견되지 않은 영아에서의 침습성 녹농균 감염증 2례)

  • Kang, Min Jae;Kim, So Hee;Kim, Nam Hee;Lee, Jin-A;Eun, Byung Wook;Choi, Eun Hwa;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
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    • v.13 no.2
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    • pp.180-185
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    • 2006
  • Invasive Pseudomonas infections most often occur in the immunocompromised patients and are associated with high mortality rate. Rarely this disease may develop in healthy infants and children. We report two cases of invasive Pseudomonas aeruginosa infections that were diagnosed in otherwise healthy infants. The first case was a previously healthy 5-month-old infant with ecthyma gangrenosum and septicemia. She presented with fever, swelling of left periorbital area and multiple erythronodular skin lesions. Each skin lesion formed a black eschar surrounded by an erythematous areola over time. Cultures of blood, urine and discharge from skin lesions grew P. aeruginosa. On the day of visit, she showed pancytopenia which was normalized after 10 days. The patient responded well to the management with ceftazidime and tobramycin. The other case was a previously healthy 9-month-old infant with community-acquired pneumonia. He was referred from an outside hospital with fever and cough. Chest x-ray revealed pneumonic infiltrations on both lower lungs with pleural effusion on the right side. Cultures of blood and pleural fluid grew P. aeruginosa. Chest CT performed on the ninth day demonstrated pneumatoceles, lung abscess and necrosis of lung parenchyma. He was managed with ceftazidime and amikacin for 50 days. No residual pulmonary complications were noted during the three month follow-up. Laboratory results to evaluate immunologic defects of phagocytic cells, complement components and T- and B-lymphocytes were all within normal range in both patients. It should be kept in mind that Pseudomonas can be, though uncommon, a cause of community-acquired invasive infections in the previously healthy infants.

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Bactericidal Efficacy of a Fumigation Disinfectant with Ortho-phenylphenol as an Active Ingredient Against Pseudomonas Aeruginosa and Enterococcus Hirae (Ortho-phenylphenol을 주성분을 하는 훈증소독제의 Pseudomonas aeruginosa와 Enterococcus hirae에 대한 살균효과)

  • Cha, Chun-Nam;Park, Eun-Kee;Kim, Yongpal;Yu, Eun-Ah;Yoo, Chang-Yeol;Hong, Il-Hwa;Kim, Suk;Lee, Hu-Jang
    • Journal of Food Hygiene and Safety
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    • v.29 no.1
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    • pp.60-66
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    • 2014
  • This test was performed to evaluate the bactericidal efficacy of a fumigation disinfectant containing 20% ortho-phenylphenol against Pseudomonas aeruginosa (P. aeruginosa) and Enterococcus hirae (E. hirae). In preliminary tests, P. aeruginosa and E. hirae working culture suspension number (N value) were $2.8{\times}10^8$ and $4.0{\times}10^8CFU/mL$, respectively. And all the colony numbers on the carriers exposed to the fumigant (n1, n2, n3) were higher than 0.5N1 (the number of bacterial test suspentions by pour plate method), 0.5N2 (the number of bacterial test suspentions by filter membrane method) and 0.5N1, respectively. In addition, the mean number of P. aeruginosa and E. hirae recovered on the control-carriers (T value) was $2.8{\times}10^8$ and $3.4{\times}10^6CFU/mL$, respectively. In the bactericidal effect of the fumigant, the reduction number of $2.8{\times}10^8$ (d value) was 6.46 and 5.19 logCFU/mL, respectively. According to the French standard for the fumigant, the d value for the effective bactericidal fumigant should be over than 5 logCFU/mL. With the results from this study, the fumigation disinfectant containing 20% ortho-phenylphenol has an effective bactericidal activity, then the fumigant can be applied to disinfect food materials and kitchen appliances contaminated with the pathogenic bacteria.

Inhibition of Verticillium Wilt in Cotton through the Application of Pseudomonas aeruginosa ZL6 Derived from Fermentation Residue of Kitchen Waste

  • Qiuhong Niu;Shengwei Lei;Guo Zhang;Guohan Wu;Zhuo Tian;Keyan Chen;Lin Zhang
    • Journal of Microbiology and Biotechnology
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    • v.34 no.5
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    • pp.1040-1050
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    • 2024
  • To isolate and analyze bacteria with Verticillium wilt-resistant properties from the fermentation residue of kitchen wastes, as well as explore their potential for new applications of the residue. A total of six bacterial strains exhibiting Verticillium wilt-resistant capabilities were isolated from the biogas residue of kitchen waste fermentation. Using a polyphasic approach, strain ZL6, which displayed the highest antagonistic activity against cotton Verticillium wilt, was identified as belonging to the Pseudomonas aeruginosa. Bioassay results demonstrated that this strain possessed robust antagonistic abilities, effectively inhibiting V. dahliae spore germination and mycelial growth. Furthermore, P. aeruginosa ZL6 exhibited high temperature resistance (42℃), nitrogen fixation, and phosphorus removal activities. Pot experiments revealed that P. aeruginosa ZL6 fermentation broth treatment achieved a 47.72% biological control effect compared to the control group. Through activity tracking and protein mass spectrometry identification, a neutral metalloproteinase (Nml) was hypothesized as the main virulence factor. The mutant strain ZL6ߡNml exhibited a significant reduction in its ability to inhibit cotton Verticillium wilt compared to the strain P. aeruginosa ZL6. While the inhibitory activities could be partially restored by a complementation of nml gene in the mutant strain ZL6CMߡNml. This research provides a theoretical foundation for the future development and application of biogas residue as biocontrol agents against Verticillium wilt and as biological preservatives for agricultural products. Additionally, this study presents a novel approach for mitigating the substantial amount of biogas residue generated from kitchen waste fermentation.

Characterization of SAL plasmid isolated from Pseudomonas putida (Pseudomonas putida에서 분리한 SAL 플라스미드의 특성)

  • 김희윤;임영복;이영록
    • Korean Journal of Microbiology
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    • v.25 no.1
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    • pp.9-16
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    • 1987
  • Three strains of bacteria utilizing salicylate, KU801(pKU5, pKU8), KU803(pKU6, pKU9), and KU806(pKU7, pKU10), were selected from the isolates and identified as Pseudomonas putida. By agarose gel electrophoresis, it was found that the strains had two plasmids each. All three strains were resistant to antibiotics such as ampicillin, tetracyclin, and chloramphenicol, and did not utilize other aromatic and aliphatic hydrocarbons examined except salicylate. The plasmids (pKU5, pKU6, and pKU7) of larger molecular weight were cured by treatment with mitomycin C and frequencies of curing were 0.4%, 1.67%, and 0.75%, respectively. Cured strains did not degrade salicylate and still had antibiotic resistances, which were identical with wild strains. The genes for salicylate degradation were proved to be enclded on thier plasmids. The molecular weights of pKU5 and pKU6 were estimated as 103.5Md, and that of pKU 7 as 101 Md. The new SAL plasmids, pKU5, pKU6, and pKU7 were transferred to P. putida and P. aeruginosa, but not to E. coli.

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Fungal-Sporulation Suppressing Substances Produced by Pseudomonas aeruginosa KMCS-1

  • Min, Bu-Yong;Shim, Jae-Young;Kim, Kun-Woo;Lee, Jong-Kyu;Choi, Hyung-Tae;Yoon, Kwon-Sang
    • Journal of Microbiology
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    • v.34 no.3
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    • pp.284-288
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    • 1996
  • Among the bacteria isolated from compost piles of cattle excretion in a pasture located at the suburbs of Chunchon city, Pseudomonas aeruginosa KMCS-1 was selected for the test of antifungal substances produced. Six fractions were separated by silica gel column chromatography, and then the antifungal activity of each fraction was assayed against Escherichia coli, Bacillus subtilis, Candida albicans, Rhizopus sp., Aspergillus nidulans, Coprinus cinereus, and Pyricularia oryzae by paper disc method. Two fractions showed significant suppressive activities against A. nidulans, C. cinereus, and P. oryzae however, their mycelial growth was not affected by neither of these fractions. Inhibitory activities of these fractions to sporulation was assayed at the concentration of 50. 25, 12. 5, and 6.25 $\mu$g/ml and the average inhibition rates against sporulation of A. nidulans, C. cinereus, and P. oryzae were 94.0, 98.3, and 77.9%, respectively. Further purification and analysis of active substances are now being conducted.

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Production, Purification, and Characterization of Antifungal Metabolite from Pseudomonas aeruginosa SD12, a New Strain Obtained from Tannery Waste Polluted Soil

  • Dharni, Seema;Alam, Mansoor;Kalani, Komal;Abdul-Khaliq, Abdul-Khaliq;Samad, Abdul;Srivastava, Santosh Kumar;Patra, Dharani Dhar
    • Journal of Microbiology and Biotechnology
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    • v.22 no.5
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    • pp.674-683
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    • 2012
  • A new strain, SD12, was isolated from tannery waste polluted soil and identified as Pseudomonas aeruginosa on the basis of phenotypic traits and by comparison of 16S rRNA sequences. This bacterium exhibited broad-spectrum antagonistic activity against phytopathogenic fungi. The strain produced phosphatases, cellulases, proteases, pectinases, and HCN and also retained its ability to produce hydroxamate-type siderophore. A bioactive metabolite was isolated from P. aeruginosa SD12 and was characterized as 1-hydroxyphenazine ((1-OH-PHZ) by nuclear magnetic resonance (NMR) spectral analysis. The strain was used as a biocontrol agent against root rot and wilt disease of pyrethrum caused by Rhizoctonia solani. The stain is also reported to increase the growth and biomass of Plantago ovata. The purified compound, 1-hydroxyphenazine, also showed broad-spectrum antagonistic activity towards a range of phytopathogenic fungi, which is the first report of its kind.

Comparison of Gamma Irradiation and Sodium Hypochlorite Treatments to Inactivate Staphylococcus aureus and Pseudomonas aeruginosa Biofilms on Stainless Steel Surfaces

  • Kim, Jang-Ho;Jo, Cheo-Run;Rho, Yong-Taek;Lee, Chun-Bok;Byun, Myung-Woo
    • Food Science and Biotechnology
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    • v.16 no.2
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    • pp.315-319
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    • 2007
  • Biofilm formation on various surfaces is a well-known phenomenon and it has caused pollution problems, health and safety hazards, and substantial economic loss in many areas including the food industry. In the present study, Gamma irradiation at a dose of 2.0 kGy reduced the bacterial counts of Staphylococcus aureus and Pseudomonas aeruginosa suspensions by 6.7 and >6.5 log CFU/mL, respectively, and 30 ppm of sodium hypochlorite effectively reduced the counts of both bacterial suspensions to below the limit of detection ($<2\;log\;CFU/cm^2$). However, in bacterial biofilms attached to stainless steel, gamma irradiation at a dose of 10.0 kGy reduced the counts of S. aureus attached fur 1 hr and overnight by ${\geq}5.1\;and\;5.0\;log\;CFU/cm^2$, respectively. Gamma irradiation at a dose of 1.0 kGy reduced the counts of P. aeruginosa counts to below the limit of detection ($<2\;log\;CFU/cm^2$). On the contrary, S. aureus and P. aeruginosa cells attached to stainless steel chips were difficult to eliminate using sodium hypochlorite. Four hundred ppm of sodium hypochlorite reduced the counts of S. aureus and P. aeruginosa attached for 1 hr by 2.5 and $3.3\;log\;CFU/cm^2$, respectively.

Evaluation of the Synergistic Effect of Mixed Cultures of White-Rot Fungus Pleurotus ostreatus and Biosurfactant-Producing Bacteria on DDT Biodegradation

  • Purnomo, Adi Setyo;Ashari, Khoirul;Hermansyah, Farizha Triyogi
    • Journal of Microbiology and Biotechnology
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    • v.27 no.7
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    • pp.1306-1315
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    • 2017
  • DDT (1,1,1-trichloro-2,2-bis(4-chlorophenyl) ethane) is one of the organic synthetic pesticides that has many negative effects for human health and the environment. The purpose of this study was to investigate the synergistic effect of mixed cutures of white-rot fungus, Pleurotus ostreatus, and biosurfactant-producing bacteria, Pseudomonas aeruginosa and Bacillus subtilis, on DDT biodegradation. Bacteria were added into the P. ostreatus culture (mycelial wet weight on average by 8.53 g) in concentrations of 1, 3, 5, and 10 ml ($1ml{\approx}1.25{\times}10^9$ bacteria cells/ml culture). DDT was degraded to approximately 19% by P. ostreatus during the 7-day incubation period. The principal result of this study was that the addition of 3 ml of P. aeruginosa into P. ostreatus culture gave the highest DDT degradation rate (approximately 86%) during the 7-day incubation period. This mixed culture combination of the fungus and bacteria also gave the best ratio of optimization of 1.91. DDD (1,1-dichloro-2,2-bis(4-chlorophenyl) ethane), DDE (1,1-dichloro-2,2-bis(4-chlorophenyl) ethylene), and DDMU (1-chloro-2,2-bis(4-chlorophenyl) ethylene) were detected as metabolic products from the DDT degradation by P. ostreatus and P. aeruginosa. The results of this study indicate that P. aeruginosa has a synergistic relationship with P. ostreatus and can be used to optimize the degradation of DDT by P. ostreatus.

Investigation of ${\beta}$-Lactamase-producing Multidrug-resistant Pseudomonas aeruginosa Isolated from Non-Tertiary Care Hospitals in Korea

  • Sohn, Eui-Suk;Yoo, Jeong-Sik;Lee, Jeom-Kyu;Lee, Kyeong-Min;Chung, Gyung-Tae;Shin, Eun-Shim;Han, Sun-Young;Lee, Sang-Hee;Kim, Joon;Lee, Yeong-Seon
    • Journal of Microbiology and Biotechnology
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    • v.17 no.10
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    • pp.1733-1737
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    • 2007
  • A total of 2,280 nonduplicate clinical isolates of Pseudomonas aeruginosa, obtained nationwide from Korean non-tertiary care hospitals from 2002 to 2005, were identified and their susceptibilities to aminoglycosides, antipseudomonal penicillins, carbapenems, cephalosporins, monobactams, and quinolones were studied, together with their production of ${\beta}$-lactamases. Using disk diffusion and minimum inhibitory concentration tests, it was found that 2.9% of isolates were multidrug-resistant (MDR) P. aeruginosa. An EDTA-disk synergy test, PCR amplification with specifically designed primers, and direct sequencing of the PCR products showed that the $bla_{OXA-10}$, $bla_{VIM-2}$, $bla_{OXA-2}$, $bla_{OXA-17}$, $bla_{PER-1}$, $bla_{SHV-12}$, and $bla_{IMP-1}$ genes were carried by 34.3%, 26.9%, 3.0%,3.0%, 1.5%, 1.5%, and 1.5% of 67 MDR P. aeruginosa isolates, respectively. The prevalence of MDR P. aeruginosa was three-fold higher, compared with that from the United States. More than two types of ${\beta}$-lactamase genes were carried by 10.4% of isolates. The most prevalent ${\beta}$-lactamase genes were $bla_{VIM-2}$ and $bla_{OXA-10}$. This study is the first description of MDR P. aeruginosa trom non-tertiary care hospitals in Korea and the coexistence of the $bla_{VIM-2}$, $bla_{IMP-1}$, or $bla_{PER-1} in these clinical isolates.