• Title/Summary/Keyword: Pseudomonas aeruginosa D2D2

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Cloning of hadA-like Sigma Factor Gene from Streptomyces coelicolor A3(2) (Streptomyces coelicolor A3(2)에서 hrdA유사 Sigma 인자 유전자의 클로닝)

  • Hahn, Ji-Sook;Cho, Eun-Jung;Roe, Jung-Hye
    • Korean Journal of Microbiology
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    • v.32 no.4
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    • pp.264-270
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    • 1994
  • A gene coding for a novel putative $\sigma$ factor of RNA polymerase has been identified from Streptomyces coelicolor A3(2) using Escherichia coli rpoS gene fragment as a probe. The 486 bp rpoS gene fragment was amplified from E. coli genomic DNA by PCR with two synthetic oligonucleotides, the sequences of which were deduced from the amino acid sequences in the regions 2.3 and 4.2 conserved among various bacterial factors. When E. coli genomic DNA fragments were hybridized with cloned rpoS probe, only one band corresponding to rpoS gene (3.2 kb PvuII fragment or 2.3 kb KpnI fragment) was detected. In S. coelicolor, however, two bands were detected both in PvuII digested DNA and SalI digested DNA. 3.5 kb PvuII fragment which binds the rpoS gene probe was cloned (pMS1) from the sublibrary, and the nucleotide sequences of 1.0 kb BamH'/HincII subclone (pBH2) was partially determined. The nucleotide sequences revealed extensive similarity to other $\sigma$ factor genes of S. coelicolor (hrdA, hrdB, hrdC, hrdD), S. aureofaciens (hrdA, hrdB, hrdC, hrdD), Synechococcus species, Pseudomonas aeruginosa, Stigmatella aurantiaca, and Anabaena species. The nucleotide sequences in regions 1.2 and 4 were compared with the corresponding regions of 5 known ${\sigma}$ factor genes of S. coelicolor by multiple alignment. It turned out that the cloned gene is most closely related to hrdA showing 88% amino acid similarity in region 1.2 and 75% in region 4.

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Evaluation of Antioxidant, Cytoprotective and Antimicrobial Activities of the Extract and Fractions Obtained from Young Shoots of Nypa Fruticans Wurmb (니파야자(Nypa fruticans Wurmb) 싹 추출물 및 분획물의 항산화, 세포 보호 및 항균 효과에 관한 평가)

  • Shin, Hyuk Soo;Lee, Yoon Joo;Kim, Ji Woong;Song, Ba Reum;Lee, Sang Lae;Park, Soo Nam
    • Korean Journal of Pharmacognosy
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    • v.49 no.2
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    • pp.155-164
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    • 2018
  • Nypa fruticans Wurmb is a species of palm, which is widely distributed in the mangrove forest of Southeast Asia. Various parts of N. fruticans has been used as a traditional medicinal plant. However, the physiological activities of N. fruticans has not yet been clarified well. Therefore, in this study, the 50% ethanol extract and its aqueous and ethyl acetate fractions of young shoots of N. fruticans were investigated for their antioxidant, cytoprotective effect, and antimicrobial activities. Every sample possessed very high free radical and various ROS scavenging capacities assessed by employing different in vitro assays such as $DPPH^{\cdot}$, $O_2^{{\cdot}-}$, ${\cdot}OH$, and $^1O_2$ scavenging activities. Based on these results, the cytoprotective effect was investigated using the oxidative hemolysis of erythrocyte. We found that the extract and fractions provide a greater protective effect compared with (+)-${\alpha}$-tocopherol. Furthermore, antimicrobial activities were confirmed against skin pathogens by broth microdilution assay. The ethyl acetate fraction had much higher antimicrobial activities than methyl paraben against Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, and Candida albicans. Taken together, our results indicated that the young shoots of N. fruticans may have the potential role as a natural active ingredient through their antioxidant, cytoprotective effect, and antimicrobial activities.

The Antimicrobial Activity of Fermented Extracts from Korean Dendropanax morbifera (국내 황칠나무 발효 추출물의 항균력 평가)

  • Lee, Jae-Yeul;Park, Tae-Hee;Park, Se-Ho;Yang, Seun-Ah;Jhee, Kwang-Hwan
    • Journal of Life Science
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    • v.29 no.1
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    • pp.29-36
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    • 2019
  • We investigated the fermentation conditions for extracts of leave/branches and sap from Korean Dendropanax morbifera (D. morbifera) using Lactobacillus plantarum (L. plantarum) ilchiwhangchil 1785 and L. plantarum ilchiwhangchil 2020. Log growth phase cultured L. plantarum ilchiwhangchil 1785 and L. plantarum ilchiwhangchil 2020 were used for fermentation. The pH and growth of the microorganisms in broth were monitored during the fermentation period. The results revealed that the optimum fermentation conditions for 20 wt% of leave/branches extracts and 1 wt% of sap extract was 2 days incubation at $37^{\circ}C$. The minimum inhibitory concentration (MIC) method and a disk diffusion assay were used to evaluate the antimicrobial activity of the fermented extracts of the leave/branches and sap against Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. The antimicrobial activity increased in all three strains grown on the medium containing fermented extracts of the leave/branches and sap as compared with that of the strains grown on medium containing non fermented extracts. Furthermore, the antimicrobial activity increased in proportion to the contents of the fermented extracts. Our data suggest that fermented extracts of leave/branches and sap of D. morbifera have applications as natural bio functional materials, such as preservatives, cosmetic materials, and natural packaging materials.

Elution of amikacin and vancomycin from a calcium sulfate/chitosan bone scaffold

  • Doty, Heather A.;Courtney, Harry S.;Jennings, Jessica A.;Haggard, Warren O.;Bumgardner, Joel D.
    • Biomaterials and Biomechanics in Bioengineering
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    • v.2 no.3
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    • pp.159-172
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    • 2015
  • Treatment of polymicrobial infected musculoskeletal defects continues to be a challenge in orthopaedics. This research investigated single and dual-delivery of two antibiotics, vancomycin and amikacin, targeting different classes of microorganism from a biodegradable calcium sulfate-chitosan-nHA microsphere composite scaffold. The addition of chitosan-nHA was included to provide additional structure for cellular attachment and as a secondary drug-loading device. All scaffolds exhibited an initial burst of antibiotics, but groups containing chitosan reduced the burst for amikacin at 1hr by 50%, and vancomycin by 14-25% over the first 2 days. Extended elution was present in groups containing chitosan; amikacin was above MIC ($2-4{\mu}g/mL$, Pseudomonas aeruginosa) for 7-42 days and vancomycin was above MIC ($0.5-1{\mu}g/mL$ Staphylococcus aureus) for 42 days. The antibiotic activity of the eluates was tested against S. aureus and P. aeruginosa. The elution from the dual-loaded scaffold was most effective against S. aureus (bacteriostatic 34 days and bactericidal 27 days), compared to vancomycin-loaded scaffolds (bacteriostatic and bactericidal 14 days). The dual- and amikacin-loaded scaffolds were effective against P. aeruginosa, but eluates exhibited very short antibacterial properties; only 24 hours bacteriostatic and 1-5 hours bactericidal activity. For all groups, vancomycin recovery was near 100% whereas the amikacin recovery was 41%. In conclusion, in the presence of chitosan-nHA microspheres, the dual-antibiotic loaded scaffold was able to sustain an extended vancomycin elution longer than individually loaded scaffolds. The composite scaffold shows promise as a dual-drug delivery system for infected orthopaedic wounds and overcomes some deficits of other dual-delivery systems by extending the antibiotic release.

Comparative Study on the Antimicrobial Activity of Glycyrrhiza uralensis and Glycyrrhiza glabra Extracts with Various Countries of Origin as Natural Antiseptics (원산지별 감초 추출물의 항균 활성 비교 및 천연방부제로써의 효능 연구)

  • Kim, Hye Jin;Bae, Jeong Yun;Jang, Ha Na;Park, Soo Nam
    • Microbiology and Biotechnology Letters
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    • v.41 no.3
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    • pp.358-366
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    • 2013
  • The aim of this study was to evaluate the antimicrobial activities of Glycyrrhiza uralensis and Glycyrrhiza glabra extracts with various countries of origin. Three samples of licorice with various origins (Korea, China, and Uzbekistan) were evaluated for their antimicrobial activities against six skin microflora. The bioassay applied for determining the antimicrobial effects included the disc diffusion assay, minimum inhibitory concentration, and challenge test. The ethyl acetate fractions of G. uralensis and G. glabra extracts showed significant antimicrobial activities against two gram-positive (Bacillus subtilis, Propionibacterium acnes) and two gram-negative (Escherichia coli, Pseudomonas aeruginosa) bacteria. These samples had much more intensive antimicrobial activities than synthetic preservatives on B. subtilis, P. acnes, and P. aeruginosa, especially. Korean licorice showed the highest antimicrobial activity amongst the samples tested. In view of the observed inhibitory features of these G. uralensis and G. glabra extracts, it is suggested that they could be used as natural antiseptics against bacterial contamination in cosmetics and foods, instead of the common synthetic preservatives currently employed.

Antimicrobial Activity of Niaouli (Melaleuca quinquenervia) Leaf Extracts against Skin Flora (피부 상재균에 대한 니아울리 잎 추출물의 항균활성)

  • Jang, Ha Na;Park, Soo Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.40 no.3
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    • pp.313-320
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    • 2014
  • In this study, the antimicrobial activity of niaouli leaf extracts was evaluated against skin flora. The skin flora used for experiments were three gram-positive bacteria such as Bacillus subtilis (B. subtilis), Staphylococcus aureus (S. aureus), Propionibacterium acnes (P. acnes), and two gram-negative, Escherichia coli (E. coli), Pseudomonas aeruginosa( P. aeruginosa), and the yeast, Plasmodium ovale (P. ovale). The bioassay applied for determining the antimicrobial effects of niouli leaf extracts or fraction included the disc diffusion assay and broth dilution assay. Minimum inhibitory concentration (MIC) values of 50% ethanol extract on B. subtilis, S. aureus, P. acnes, E. coli and P. aeruginosa were 0.25%, 0.50%, 1.00%, 0.13% and 0.25% respectively and the MIC values of water fraction were 0.25%, 0.25%, 4,00%, 0.25% and 0.25%. P. ovale did not show antimicrobial activities. The MIC values of methyl paraben used as positive control indicated 0.25%, 0.25%, 0.25%, 0.13% and 0.50%. Also, Minimum bactericidal concentration (MBC) values of 50% ethanol extract were 2.00%, 2.00%, 1.00%, 0.50% and 2.00% individually and the MBC values of water fraction were 0.50%, 0.25%, 4.00%, 0.50% and 1.00%. The MBC values of methyl paraben indicated 1.00%, 0.500%, 0.50%, 0.50% and 1.00%. These results showed that water fraction was as good as methyl paraben except for P. acnes. The 50% ethanol extract also showed activity similar with it. Thus, it is concluded that the 50% ethanol extract/fraction of niaouli could be applicable to cosmetics as a natural preservatives effective in antimicrobial activity against skin flora.

Antimicrobial Activities of Eight Compounds Purified from the Roots of Polygala tenuifolia Willdenow and the Aerial Bulbils of Dioscorea batatas Decene (원지와 영여자로부터 분리한 8종 화합물의 항균효과)

  • Chin, Hwi-Seung;Son, Rak-Ho;Lee, Yong-Hwa;Ham, Ah-Rom;Mar, Woong-Chon;Kim, Won-Ki;Nam, Kung-Woo
    • YAKHAK HOEJI
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    • v.54 no.2
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    • pp.106-111
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    • 2010
  • The chemical structures of eight compounds purified from two plants (Polygala tenuifolia Willdenow and Dioscorea batatas Decene) were determined and their anti-microbial activity against three microbial strains (Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans) was tested. The three micro organisms were cultured in 96-well plates or Petri dishes without (control) or with the eight compounds added at concentrations of 100 to 0.01 ${\mu}M$ (wt/vol). The growth of the microorganisms in the medium was examined after a 24-h incubation. The inhibitory effect of each compound on the growth of the microorganisms was calculated from the optical density measured at 595 nm, turbidity, and size of the inhibition zone around the treated paper disc. The minimum inhiitory concentration (MIC) of compounds 4 to 7 against S. aureus was 0.08, 0.05, 1.3 and 0.02 ${\mu}M$, respectively, and 0.09, 0.1, 0.2 and 100 ${\mu}M$ against C. albicans. The $IC_{50}$ (50% inhibition) values of compounds 5 and 6 were 3.1 and 6.4 ${\mu}M$ against S. aureus, respectively, and 10 and 2.4 ${\mu}M$ against C. albicans. Therefore, compounds 4 to 6 were the most potent anti-microbial agents among the eight compounds tested.

Isolation and Characterization of Lactic Acid Bacteria with Angiotensin-Converting Enzyme Inhibitory and Antioxidative Activities (안지오텐신 전환효소 저해 활성 및 항산화 활성을 가진 젖산균의 분리 및 특성)

  • Park, Sung-Bo;Kim, Jeong-Do;Lee, Na-Ri;Jeong, Jin-Ha;Jeong, Seong-Yun;Lee, Hee-Seob;Hwang, Dae-Youn;Lee, Jong-Sup;Son, Hong-Joo
    • Journal of Life Science
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    • v.21 no.10
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    • pp.1428-1433
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    • 2011
  • In this study, we isolated and characterized plant-associated lactic acid bacteria which are able to produce angiotensin-converting enzyme (ACE) inhibitory and antioxidative activities. Five lactic acid bacteria were isolated from plants (grape and leek), a plant-associated fermentative product (Kimchi) and Korean traditional alcohol (Dongdongju). Strains K-1 and K-21 from Kimchi, strain L-5 from leek, strain G-3 from grape, and strain D-3 from Dongdongju were identified as Pediococcus pentosaceus, Lactobacillus plantarum, Weissella cibaria, L. plantarum, and L. brevis, respectively, by 16S rRNA gene analysis. ACE inhibitory activities of isolated strains ranged from 44.3 to 71.9% in the MRS broth. G-3, L-5 and K-1 strains especially showed high ACE inhibitory activities (59.8-98.69%) in the MRS broth containing skim milk. DPPH radical scavenging activities of the strains were in the range of 42.5-82.7%. All strains showed varying levels of resistance in artificial gastric fluid (pH 2.5), retaining viability ranging from 42.2 to 88.1% after 3 hr of incubation. All strains showed high resistance to 0.3% oxgall after 24 hr of incubation; survival rates were in the range of 55.4-112.8%. Isolated strains were found to be antagonistic to some pathogens including Pseudomonas aeruginosa.

In vitro and in vivo Antibacterial Activities of the New Quinolone, DWQ-013 (새로운 퀴놀론계 항균제 DWQ-013의 항균작용)

  • Yu, Young-Hyo;Park, Nam-Jun;Kim, Byung-O;Choi, Moon-Jung;Shim, Jeom-Soon;Kang, Tae-Chung;Lee, Jae-Wook;Kim, Dae-Young
    • YAKHAK HOEJI
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    • v.38 no.3
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    • pp.265-273
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    • 1994
  • ln vitro and in vivo antibacterial activities of DWQ-013(1-cyclopropyl-6,8-difluoro-7-(3-methylthiomethylpyrrolidinyl)-1,4-dihydro-4-oxo-3-quinolinecarboxylic acid), a new fluoroquinolone antibacterial agent, were compared with those of ciprofloxacin, sparfloxacin and ofloxacin against aerobic and anaerobic standard strains and clinical isolates. DWQ-013 had a broad spectrum and potent antibacterial activity against Gram-positive and Gram-negative bacteria. The antibacterial activity of DWQ-013 against Staphylococcus aureus was equal to that of sparfloxacin(SPFX) and superior to those of ciprofloxacin(CPFX). The antibacterial activity against Gram-negative bacteria was slightly lower than those of ciprofloxacin and sparfloxacin. MIC of DWQ-013 against Pseudomonas aeruginosa$(0.781{\sim}1.563\;{\mu}g/ml)$ was usually equal to that of sparfloxacin$(0.781\;{\mu}g/ml)$ and was inferior to that of ciprofloxacin$(0.098\;{\mu}g/ml)$. The number of viable cells was decreased rapidly after addition of DWQ-013 at concentration of $1{\sim}2$ folds of MIC.

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유기용매 내에서 중쇄지방질의 합성

  • Gwon, Dae-Yeong
    • Bulletin of Food Technology
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    • v.7 no.2
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    • pp.64-73
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    • 1994
  • Using 20 lipases from various microbial origins medium chain glycerides, namely, mono-, di-, and tri-carproyl glycerols from glycerol and acid were synthesized in isooctane. Enzyme reaction was performed at 0.35 M of capric acid, 0.025M of glycerol and the same mass of silica gel to remove water in 5ml of isooctane with 30mg of lyophilized lipase. Among 20 lipases, eleven lipases showed good synthetic activities, especially lipase from Pseudomonas aeruginosa (Lipase PS), Rhizomucor miehei origined lipase and Chromobacterium viscosum lipase (Lipase CV) showed good activities for production of tricaproylglycerol, while Lipase OF-360 (origined from Candida rugosa) and Lipase D (Rhizopus delemar) were good for production of dicaprolyglycerol. The lipases, especially Lipase PS, have high thermal stability at $ 60^{circ}C$, and optimum pH of lyophilization for dehydrating the lipase was pH 6.5.

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