• 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.

• 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.

• Journal of Life Science
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• v.19 no.8
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• pp.1047-1054
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• 2009

Oral squamous carcinoma (OSC) cells present resistance to chemotherapeutic agents-mediated apoptosis in the late stages of malignancy. Advances in the understanding of bacterial toxins have produced new strategies for the treatment of cancers. It was demonstrated here that Pseudomonas aeruginosa exotoxin A (PEA) significantly decreased the viability of chemoresistant YD-9 cells in the apoptosis mechanism. Apoptotic manifestations were evident through changes in nuclear morphology and generation of DNA fragmentation. PEA treatment induced caspase-3, -6 and -9 cleavage, and activation. These events preceded proteolysis of the caspase substrates poly (ADP-ribose) polymerase (PARP), DNA fragmentation factor 45 (DFF45), and lamin A in YD-9 cells. The reduction of mitochondrial membrane potential, release of cytochrome c and SmacjDlABLO from mitochondria to cytosol, andtranslocation of AlF into nucleus were shown. While p53, p21 and $14-3-3{\gamma}$ were upregulated, cyclin Band cdc2 were downregulated by PEA treatment. Taken together, PEA induces apoptosis in chemoresistant YD-9 cells via activation of caspases, mitochondrial events and regulation of cell cycle genes.

• Microbiology and Biotechnology Letters
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• v.39 no.1
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• pp.29-36
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• 2011

Pseudomonas aeruginosa is an opportunistic pathogen that causes various infections on urinary track, cornea, respiratory track, and burn wound site, and mainly relies on quorum sensing (QS) for its virulence. To control the infectivity of P. aeruginosa, we previously synthesized the structural analogues of a major QS signal, N-3-oxododecanoyl homoserine lactone (3OC12-HSL) to use as a QS inhibitor. Two of them (5b and 5f) had been confirmed to have an inhibitory effect on LasR, a major QS signal receptor of P. aeruginosa in the screening by the recombinant Escherichia coli reporter. To further evaluate these compounds, we tested their efficacy to control the QS and virulence of P. aeruginosa. Unlike the result from E. coli reporter, both 5b and 5f failed to affect the LasR activity in P. aeruginosa, but instead they selectively affected the activity of QscR, another 3OC12-HSL receptor of P. aeruginosa. Interestingly, their effect on QscR was complex and opposite to what we obtained with E. coli system. Both 5b and 5f enhanced the QscR activity at the low concentration range (< 10 ${\mu}m$), but high concentration of 5f (${\approx}$1 mM) strongly inhibited QscR. While 5b and 5f didn't affect the production of proteases, the key virulence factor, they significantly reduced the biofilm formation that is important in mediating chronic infections. Especially, 5f inhibited the initial attachment of P. aeruginosa, rather than the biofilm maturation. Based on our results, we suggest that 5f can be applied for an anti-biofilm agent without increasing virulence of P. aeruginosa.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.72.2-73
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• 2003

We found a soybean (Glycine max) cultivar 561 that was strongly resistant to a virulent bacterial strain of a Pseudomonas spp. Further identification revealed that the Pseudomonas spp. was a strain of Pseudomonas aeruginosa. Furthermore we identified specific genes involved in the resistance of soybean 561 and analyzed the pattern of gene expression against the Pseudomonas infection using differential-display reverse transcription PCR (DDRT-PCR). More than 126 cDNA fragments representing mRNAs were induced within 48 hours of bacteria inoculation. Among them, 28 cDNA fragments were cloned and sequenced. Twelve differentially displayed clones with open reading frames had unknown functions. Sixteen selected cDNA clones were homologous to known genes in the other organisms. Some of the identified cDNAs were pathogenesis-related genes (PR genes) and PR-like genes. These cDNAs included a putative calmodulin-binding protein, an endo-1,3-1,4-b-D-glucanase, a b-1,3-endoglucanase, a b-1,3-exoglucanase, a phytochelatin synthetase-like gene, a thiol pretense, a cycloartenol synthase, and a putative receptor-like sorineithreonine protein kinase. Among them, we found that four genes were putative pathogenesis-related genes (PR) induced significantly by the p. aeruginosa infection. These included a calmodulin-binding protein gene, a b-1,3-endoglucanase gene, a receptor-like sorine/threonine protein kinase gene, and pS321 (unknown function). These results suggest that the differentially expressed genes may mediate the strong resistance of soybean 561 to Pseudomonas aeruoginosa.

• Korean Journal of Microbiology
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• v.52 no.4
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• pp.455-462
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• 2016

Pseudomonas aeruginosa P-5 is an unusual organism capable of synthesizing polyhydroxyalkanoates (PHAs) consisting of 3-hydroxyvalerate (3HV) and medium-chain-length (MCL) 3-hydroxyalkanoate (3HA) monomer units when C-odd alkanoic acids are fed as the sole carbon source. Evaluation of the substrate chain-length specificity of two P. aeruginosa P-5 PHA synthases ($PhaC1_{P-5}$ and $PhaC2_{P-5}$) by heterologous expression of $PhaC1_{P-5}$ and $PhaC2_{P-5}$ genes in Pseudomonas putida GPp104 revealed that $PhaC2_{P-5}$ incorporates both 3HV and MCL 3HAs into PHA, whereas $PhaC1_{P-5}$ favors only MCL 3HAs for polymerization. In order to obtain $PhaC2_{P-5}$ mutants with altered substrate specificity, site-specific mutagenesis for $PhaC2_{P-5}$ was conducted. Amino acid substitutions of $PhaC2_{P-5}$ at two positions (Ser326Thr and Gln482Lys) were very effective for synthesizing copolymers with a higher 3HV fraction. When recombinant P. putida GPp104 harboring double mutated $phaC2_{P-5}$ gene ($phaC2_{P-5}QKST$) was grown on nonanoic acid, 2.5-fold increase of copolymer content with 3.8-fold increase of 3HV fraction was observed. The $phaC2_{P-5}QKST$-containing Ralstonia eutropha PHB-4 supplemented with valeric acid also produced copolymers consisting of 3HV and 3-hydroxyheptanoate with a high 3HV fraction. These results suggest that recombinants containing $phaC2_{P-5}QKST$ could be useful for production of new PHA copolymers with improved material properties.

• Korean Journal of Microbiology
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• v.38 no.4
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• pp.265-265
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• 2002

Iron increased the susceptibilities of clinical isolates Pseudomonas aeruginosa to quinolones. In the presence of iron, increased susceptibilities to ofloxacin were observed in twenty-six out of thirty isolates and with no change in four isolates. In the case of norfloxacin, iran increased susceptibilities of twelve isolates but did not render any change in eighteen isolates. In the case of ciprofloxacin, iron decreased the MICs (Minimal Inhibitory Concentration) of twenty isolates, increased the MIC of one isolate, and did net change the MICs of nine isolates. To find out how iron increased susceptibility to ofloxacin, bacterial cells were grown in Muller Hinton (MH) media and succinate minimal media (SMM) to induce iran acquisition systems and the intracellular ofloxacin concentrations were assayed in the presence of iron. The addition of iron to the media decreased the MICs of cells whether they were grown in MH or SMM. Siderophores, carbonyl cyanide m-chlorophenylhydrazone (an inhibiter of proton motive force), and ouabain (an inhibitor of ATPase) did not decrease the effect of iron. Results suggested that the increase in the intracellular ofloxacin concentration by iron is accomplished not by decreasing the efflux but by increasing the of ofloxacin permeability.

• Proceedings of the Microbiological Society of Korea Conference
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• 2002.10a
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• pp.40-50
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• 2002

Fruit fly, Drosophila melanogaster has developed efficient immune mechanisms to prevent microbial infection, which are consisted of cellular and humoral responses. During the systemic or local infection, two distinct pathways (Toll and Imd) play major roles in antimicrobial peptide synthesis. The Toll pathway is required to defend Gram-positive bacterial and fungal infections, whereas the Imd pathway is important in Gram-negative bacterial infection. We have shown that the infection of the opportunistic Gram-negative bacterium, Pseudomonas aeruginosa strain PA14 (PA14) into fly dorsal thorax can kill the flies within 48 h ($100\%$ mortality) in our optimized infection condition, suggesting that the PA14 strain can cause disease progress in fly model system. We found that flies carrying a constitutively activated mutant form of the Toll receptor $(Tl^{10b})$ showed increased resistance to P. aeruginosa infection and that flies carrying mutations in the Toll signaling pathway as well as in the Imd signaling pathway was more susceptible to PA14 infection. All these results imply that the Toll pathway might be important in the resistance to this pathogenic Gram-negative bacterial infection.

• Journal of Microbiology
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• v.35 no.1
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• pp.25-29
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• 1997

Microbial reduction of hexavalent (VI) to trivalent (III) chromium decreases its toxicity by two orders of magnitude. In order to investigate the nature of Cr-reduction, Cr-resistant Pseudomonas aeruginosa HP014 was isolated and tested for its reduction capability. At the concentration of 0.5 mM Cr(VI), cell growth was not inhibited by the presence of Cr(VI) in a liquid medium, and Cr(VI) reduction was accompanied by ell growth. When cell-free extract was tested, the reduction of Cr(VI) showed a saturation kinetics with the maximum specific activity of 0.33 .mu.mol min$\^$-1/ mg$\^$-1/ cell protein, and an apparent K. of 1.73 mM Cr(VI). The activity required either NADH or NADPH as an electron donor. However, NADPH gave 50% as mush activity as sequently the supernatant and pelleted membrane fractions were tested for Cr(VI) reduction activity. The supernatant of the centrifugation showed almost the same Cr(VI) reduction activity as compared with that of the cell-free extract, indicating that the Cr(VI)-reducing activity of P. aeruginosa HP-14 is due to soluble enzyme. Moreover, the activity appeared to be the highest among the known activities, suggesting that the strain might be useful for remediation of Cr(VI)-contaminated sites.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.261-264
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• 2001

Cytotoxicity and antibacterial activity of preservatives were examined. Fibroblast cell L929 was used for cytotoxicity experiment and Pseudomonas aeruginosa A TCC27853, Staphylococcus aureus ATCC25923. Trichoderma reesei ATCC6967 were used for antibacteria and antifungi. Benzalkoniumchloride(BAK) as synthetic preservative and chitosan as natural preservative were used. Minimum inhibitary concentration (MIC) of BAK was 0.1 % for P. aeruginsa and 0.001% for S. aureus and 0.1 % for T reesei MIC of chitosan was 2% for P. aeruginosa and 1 % for S. aureus.