• Journal of Life Science
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• v.29 no.1
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• pp.84-89
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• 2019

Previous screening of novel antibacterial agents revealed that some bacterial isolates exhibited antibiotic activity against both gram-positive and gram-negative bacteria and that they showed antibacterial activity, even against methicillin-resistant Staphylococcus aureus (MRSA). Among these isolates, one bacterial strain, BCNU 1204, was identified as Pseudomonas aeruginosa using phenetic and phylogenetic analysis, based on 16S ribosomal RNA gene sequences. The maximum productivities of antimicrobial substances of BCNU 1204 were obtained after being cultured at $35^{\circ}C$ and pH 7.0 for 4 d in King's medium B (KMB). Dichloromethane (DCM) and ethylacetate (EA) extracts of P. aeruginosa BCNU 1204 exhibited strong antimicrobial activity, particularly against gram-positive bacteria. The EA extracts exhibited broad-spectrum activity against antibiotic resistant strains. Fraction 5-2, was obtained by recycling preparative liquid chromatography (LC) and preparative thin-layer chromatography (TLC) and was identified as phenazine-1-carboxylic acid belonging to phenazines using gas chromatography and mass spectrometry (GC/MS). Its minimum inhibitory concentration (MIC) values were $25{\mu}g/ml$, $50{\mu}g/ml$, ${\geq}25{\mu}g/ml$, and ${\geq}50{\mu}g/ml$ for MRSA CCARM 3089, 3090, 3091, and 3095 strains, respectively. P. aeruginosa BCNU 1204 may be a potential resource for the development of anti-MRSA antibiotics. Additional research is required to identify the active substance from P. aeruginosa BCNU 1204.

• 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.

• Journal of Microbiology and Biotechnology
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• v.29 no.8
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• pp.1299-1309
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• 2019

As an opportunistic bacterial pathogen, Pseudomonas aeruginosa PAO1 contains two phenazine-producing gene operons, phzA1B1C1D1E1F1G1 (phz1) and phzA2B2C2D2E2F2G2 (phz2), each of which is independently capable of encoding all enzymes for biosynthesizing phenazines, including phenazine-1-carboxylic acid and its derivatives. Other previous study reported that the RpoS-deficient mutant SS24 overproduced pyocyanin, a derivative of phenazine-1-carboxylic acid. However, it is not known how RpoS mediates the expression of two phz operons and regulates pyocyanin biosynthesis in detail. In this study, with deletion of the rpoS gene in the $PA{\Delta}phz1$ mutant and the $PA{\Delta}phz2$ mutant respectively, we demonstrated that RpoS exerted opposite regulatory roles on the expression of the phz1and phz2 operons. We also confirmed that the phz1 operon played a critical role and especially biosynthesized much more phenazines than the phz2 operon when the rpoS gene was knocked out in P. aeruginosa. By constructing the translational reporter fusion vector lasR'-'lacZ and the chromosomal fusion mutant $PA{\Delta}lasR::lacZ$, we verified that RpoS deficiency caused increased expression of lasR, a transcription regulator gene in a first quorum sensing system (las) that activates overexpression of the phz1 operon, suggesting that in the absence of RpoS, LasR might act as an intermediate in overproduction of phenazine biosynthesis mediated by the phz1 operon in P. aeruginosa.

• Journal of the Korean Applied Science and Technology
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• v.33 no.2
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• pp.225-231
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• 2016

This study was conducted to investigate the antimicrobial activities of ethanol extracts (70%, v/v) from Whole, Root and Leaf stem part of dried Zostera marina. In order to use Zostera marina extract as a basic material of cosmetic component. The extracts of Zostera marina conducted an antimicrobial activity against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Propionibacterium acnes by disc diffusion method and measure clear zone. As a result, clear zone(mm) of Staphylococcus epidermidis was confirmed at $13.00{\pm}0.50mm$ and Staphylococcus aureus, Propionibacterium acnes, Pseudomonas aeruginosa have measured $11.75{\pm}0.25mm$, $12.00{\pm}0.50mm$, $12.25{\pm}0.25mm$ from Root extract part of Zostera marina. A Zostera marina extract is expectied to have antimicrobial effects.

• Microbiology and Biotechnology Letters
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• v.47 no.1
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• pp.54-63
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• 2019

Rubus coreanus Miquel (RCM), also known as Korean blackberry or bokbunja, is used as a South Korean traditional medicine to treat acne and inflammatory skin conditions. The antimicrobial activity of RCM root and its active compounds remain unclear. In this study, we prepared a 50% ethanol fraction, ethyl acetate fraction, and acid-treated ethyl acetate fraction (aglycone fraction) of RCM root, and evaluated antibacterial activities against the skin pathogens Staphylococcus aureus, Pseudomonas acnes, Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa. In a paper disc assay, all fractions of RCM root showed antimicrobial activities against the five skin pathogens. The ethyl acetate fraction displayed 6-, 12-, and 2-fold higher minimal inhibitory concentration (MIC) than the 50% ethanol fraction against S. aureus, E. coli, and P. acnes, respectively. The aglycone fraction displayed 2-fold higher MIC than methyl paraben against P. acnes, S. aureus, E. coli, and P. aeruginosa. The ethyl acetate fraction displayed a minimal bactericidal concentration (MBC) similar to that of methyl paraben, and the aglycone fraction showed 2- to 4-fold higher MBCs than those of methyl paraben. In particular, the ethyl acetate fraction was not cytotoxic and showed thermal stability after incubation at high temperatures ($60-121^{\circ}C$). Finally, the ethyl acetate fraction was separated and four components were identified: procyanidin C, propelagonidin dimer, ellagic acid, and methyl ellagic acid acetyl pentose. The compounds showed high antibacterial activities. These results suggest that RCM root is potentially applicable as a natural preservative in cosmetics.

• 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.

• Natural Product Sciences
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• v.24 no.1
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• pp.47-53
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• 2018

Callophylum symingtonianum (Guttiferae), an evergreen broad-leaved tree that usually grows in hill forests, can be found distributed in the Malay Peninsula. The barks, leaves, flowers and seeds is often used medicinally to treat diarrhea and rheumatism. In the present study, we isolated two inophyllum type coumarins, 12-O-ethylinophyllum D (1) and iso-soulattrolide (2) from the stembarks of C. symingtonianum together with their antibacterial activity. The compounds were isolated by chromatographic methods on a silica gel. The structures were established by spectroscopic methods including UV, IR, (1D and 2D) NMR and mass spectrometry as well as by comparison with several literature sources. The antibacterial activity of those compounds was tested using a disc-diffusion assay against Staphylococcus aureus, Bacillus cereus, Escherichia coli and Pseudomonas aeruginosa. Both compound exhibited mild inhibition against P. aeruginosa with both $111{\mu}g/ml$ MIC value. Compound 2 also inhibits S. aureus with $25{\mu}g/ml$ MIC value.

• Journal of the Korean Chemical Society
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• v.58 no.4
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• pp.366-376
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• 2014

Synthesis, structural characterization, and biological activity studies of novel pyrido[2,3-d]pyrimidines (10a-h, 11a-h) are described. Cyclization of cynoacetamides (4, 5) with malonitrile (7) and aldehyde (6a-h) via Hantzsch pyridine synthesis afforded cyanopyridones (8a-h, 9a-h), which on cyclization with formic acid under microwave conditions led to the final product. All the reactions are significantly faster and the isolated yields are remarkably higher in microwave conditions compared to the conventionally heated reactions. The compounds were tested in vitro for their antibacterial activity against Escherichia coli, Pseudomonas aeruginosa, Bacillus subtillus, Staphylococcus aureus, and Micrococcus luteus and antifungal activity against Trichphyton longifusus, Candida albicans, Microsporum canis, Fusarium solani. Compounds 10b, 10e, 11b and 11e exhibited good antibacterial and antifungal activities compared with standards.

• Microbiology and Biotechnology Letters
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• v.45 no.3
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• pp.209-217
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• 2017

In the enduring investigation of the bioactive microbes, Pseudomonas aeruginosa strain (referred to as CGK-KS-1 (ICTB-315)), isolated from Chumathang hot spring, Ladakh, and India, was identified to possess a major bioactive fraction with antimicrobial and anti-biofilm properties. This bioactive metabolite was purified through bioactivity-guided fractionation. The chemical structure of this major compound was elucidated as phenazine-1-carboxamide (PCN) based on $^1H$ and $^{13}C$ NMR, FT-IR, EI-HR-MS and 2D NMR spectroscopic techniques. In the current study, PCN exhibited antimicrobial activity with MIC values ranging between $1.9-3.9{\mu}g/ml$ against various test human pathogens and Xanthomonas spp. PCN showed the anti-biofilm property with the $IC_{50}$ values ranging from 17.04 to $60.7{\mu}M$ against different test pathogens. The in silico docking studies showed PCN strongly interacted with various proteins of different Xanthomonas spp. with high binding energies. We report herein for the first time the anti-biofilm property and the docking studies of PCN. The extrolite from P. aeruginosa strain CGK-KS-1 showed promising bioactivities and may be considered as a potential candidate for application in various biocontrol strategies.

• 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.