• Journal of Microbiology and Biotechnology
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• 제20권10호
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• pp.1397-1402
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• 2010

Papyriflavonol A (PapA), a prenylated flavonoid [5,7,3',4'-tetrahydroxy-6,5'-di-(${\gamma},{\gamma}$-dimethylallyl)-flavonol], was isolated from the root barks of Broussonetia papyrifera. Our previous study showed that PapA has a broad-spectrum antimicrobial activity against pathogenic bacteria and fungi. In this study, the mode of action of PapA against Candida albicans was investigated to evaluate PapA as an antifungal agent. The minimal inhibitory concentration (MIC) values were 10~25 ${\mu}g/ml$ for C. albicans and Saccharomyces cerevisiae, Gram-negative bacteria (Escherichia coli and Salmonella typhimurium), and Gram-positive bacteria (Staphylococcus epidermidis and Staphylococcus aureus). The kinetics of cell growth inhibition, scanning electron microscopy, and measurement of plasma membrane florescence anisotrophy revealed that the antifungal activity of PapA against C. albicans and S. cerevisiae is mediated by its ability to disrupt the cell membrane integrity. Compared with amphotericin B, a cell-membrane-disrupting polyene antibiotic, the hemolytic toxicity of PapA was negligible. At 10~25 ${\mu}g/ml$ of MIC levels for the tested strains, the hemolysis ratio of human erythrocytes was less than 5%. Our results suggest that PapA could be a therapeutic fungicidal agent having potential as a broad spectrum antimicrobial agent.

• Journal of Applied Biological Chemistry
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• 제52권2호
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• pp.65-69
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• 2009

본 연구에서는 청목노상 추출물로부터 Helicobacter pylori를 억제하는 물질을 정제 및 동정을 실시하였다. 80% 에탄올 추출물의 페놀성 물질의 함량은 16.1 mg/g으로 측정되었으며, H. pylori 저해활성은 clear zone이 15 mm로 나타났다. 저해물질의 분리는 $C_{18}$ column을 이용하여 분리한 결과 9개의 fraction으로 분리되었으며, H. pylori 저해활성을 나타낸 fraction 7과 8을 MCI-gel CHP-20 column을 이용하여 normal phase type인 ethanol 100%에서 0%로 농도를 감소시키며 10 mL/min의 유속으로 용출한 결과 6개의 단일 물질 분획을 얻을 수 있었다. 그 중 세 가지의 물질이 H. pylori저해활성을 보여 FAB-Mass, $^1H$와 $^{13}C$ NMR과 IR spectrum을 사용하여 구조 동정한 결과 chlorogenic acid, caffeic acid 및 rosmarinic acid인 것으로 확인되었다.

• Journal of Microbiology and Biotechnology
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• 제28권9호
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• pp.1563-1572
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• 2018

Gold nanoparticles (AuNP) and their conjugates have been gaining a great deal of recognition in the medical field. Meanwhile, extended-spectrum ${\beta}$-lactamases (ESBL)-producing bacteria are also demonstrating a challenging problem for health care. The aim of this study was the biosynthesis of AuNP using Rosa damascenes petal extract and conjugation of ceftriaxone antibiotic (Cef-AuNP) in inhibiting ESBL-producing bacteria and study of in vitro anticancer activity. Characterization of the synthesized AuNP and Cef-AuNP was studied. ESBL-producing strains, Acinetobacter baumannii ACI1 and Pseudomonas aeruginosa PSE4 were used for testing the efficacy of Cef-AuNP. The cells of MCF-7 breast cancer were treated with previous AuNP and Cef-AuNP at different time intervals. Cytotoxicity effects of apoptosis and its molecular mechanism were evaluated. Ultraviolet-visible spectroscopy and Fourier transform infrared spectroscopy established the formation of AuNP and Cef-AuNP. Transmission electron microscope demonstrated that the formed nanoparticles were of different shapes with sizes of 15~35 nm and conjugation was established by a slight increase in size. Minimum inhibitory concentration (MIC) values of Cef-AuNP against tested strains were obtained as 3.6 and $4{\mu}g/ml$, respectively. Cef-AuNP demonstrated a decrease in the MIC of ceftriaxone down to more than 27 folds on the studied strains. The biosynthesized AuNP displayed apoptotic and time-dependent cytotoxic effects in the cells of MCF-7 at a concentration of $0.1{\mu}g/ml$ medium. The Cef-AuNP have low significant effects on MCF-7 cells. These results enhance the conjugating utility in old unresponsive ceftriaxone with AuNP to restore its efficiency against otherwise resistant bacterial pathogens. Additionally, AuNP may be used as an alternative chemotherapeutic treatment of MCF-7 cancer cells.

• 한국식물병리학회지
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• 제14권3호
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• pp.191-202
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• 1998

About 300 actinomycetes were isolated from two forest and one sea-shore soil and tested for inhibitory effects on mycelial growth of six plant pathogenic fungi Magnaporthe grisea, Alternaria mali, Colletotrichum gloeosporioides, Phytophthora capsici, Fusarium oxysporum f. sp. cucumerinum, and Rhizoctonia solani. Among 300 actinomycetes tested, only 16 actinomycetes showed the antifungal activity against the test fungi. Isolate NH 50 was selected for production and purification of antifungal antibiotic substances. Actinomycete isolate NH 50 displayed the broad antifungal spectra against 11 plant pathogenic fungi. To identify actinomycete isolate NH 50, cultural characteristics on various agar media, diaminopimelic acid type, and morphological characteristics by scanning electron microscopy were examined. As a result, actinomycete isolate NH 50 was classified as a rare actinomycete that had LL-DAP type and did not produce spores. After incubation of isolate NH 50 in yeast extract-malt extract-dextrose broth, antifungal compound NH-B1 that inhibited mycelial growth of some plant pathogenic fungi was purified from the methanol eluates of XAD-16 resins by a series of purification procedures, i.e., silica gel flash chromatography, C18 flash chromatography, Sephadex LH-20 column chromatography, silica gel medium pressure liquid chromatography (MPLC), C18 MPLC, and high pressure liquid chromatography (HPLC). UV spectrum and 1HNMR spectrum of antifungal compound NH-B1 dissolved in methanol were examined. The antibiotic NH-B1 showed the major peaks at 230 and 271.2nm. Based on the data of 1H-NMR spectrum, NH-B1 was confirmed to be an extremely complex polymer of sugars called polysaccharides. The antibiotic NH-B1 showed strong antifungal activity against Alternaria solani and Cercospora kikuchi, but weak activity against M. grisea.

• 한국동물위생학회지
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• 제34권1호
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• pp.37-43
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• 2011

This study was conducted to investigate the prevalence and genotypes of extended-spectrum ${\beta}$-lactamase (ESBL) in 377 Escherichia coli isolated from healthy and sick animals. Two isolates (0.5%), each of which were isolated from diseased swine and chicken, respectively, were confirmed as ESBL producing isolates by double disk synergy test, and showed a multidrug resistant phenotype. Minimum inhibitory concentration of cefotaxime for the two ESBL producing isolates were 3~4 times higher than those of ceftazidime, respectively. By PCR and sequencing, one isolate from swine have both $bla_{CTX-15}$ and $bla_{TEM-1}$, and one isolate from chicken have $bla_{CTX-15}$ and $bla_{TEM-116}$. Also, these genes were transferred to E. coli J53 by conjugation. These two isolates showed unrelated pulsed-field gel electrophoresis. To our knowledge, this is the first time that $bla_{TEM-116}$ gene was identified in E. coli isolated from animals in Korea. These results suggest more prudent use of third- generation cephalosporins, and surveillance and monitoring for ESBL producing E. coli in both animals and their environments should be necessary.

• 미생물학회지
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• 제40권4호
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• pp.295-301
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• 2004

최근 extended-spectrum $\beta-lactamase$ 생산 임상 균주의 급속한 증가와 확산은 매우 심각한 문제를 야기하고 있다. 이에 국내에서 extended-spectrum $\beta-lactamase$ 생산 임상 균주의 발생율을 파악하고자 국내 한 대학병원에 입원한 환자로부터 대장균을 분리하여 항생제 감수성 검사를 실시하였다. 충 233균주 중 184균주 $(78.9\%)$가 ampicillin에 대해 내성을 나타냈으며, 80균주$(34.3\%)$가 cephalothin에, 93균주$(39.9\%)$가 gentamicin에, 64균주$(27.5\%)$가 norfloxacin에 대해 내성을 나타내었다. 이중 17균주$(7.3\%)$가 double disk synergy test에 의해 양성반응을 나타낸 것으로 확인되었고, 이들에 대해서 6가지 항생제에 대한 최소 억제 농도를 추가적으로 시험한 결과, 13 균주가 4가지 이상의 다른 계열의 항생제에 대한 다중 약제 내성인 것으로 나타났다. Isoelectric focusing gel electrophoresis에 의한 pI값과 DNA 염기서열을 분석한 결과 5균주가 TEM-1,1균주가 TEM-15,1 균주가 TEM-20,4균주가 TEM-52,2균주가 TEM-1과 AmpC, 1균주가 TEM-1과 OXA-30,1 균주가 TEM-1과 OXA-33,1 균주가 TEM-1, CTX-M-3, AmpC, 그리 고 1 균주가 AmpC를 생산하는 것으로 나타났으며, SHV를 생산하는 균주는 없었다. 이들의 항생제내성 유전자가 전달되는지 확인한 결과 동물로부터 분리된 대장균 (CCARM No.1203)에 extended-spectrum $\beta-lactamase$생산 유전자가 전달되는 것을 확인하였다. Random amplified polymorphic DNA와 pulsed field gel electrophoresis분석을 사용하여 genomic DNA에 대한 유전형을 분석한 결과 균주간의 유전적 연관성은 매우 낮은 것으로 나타나 한 병원에서 발견되는 균주는 clonal spread에 의한 것이라는 일반적인 보고와 다른 결과를 얻었다.

• 한국응용과학기술학회지
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• 제15권3호
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• pp.27-31
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• 1998

Antioxodative substances in Mulberry leaves were examined. Antioxidative substances in Mulberry leaves were extracted by 80% methanol agueous solution. Antioxidative activity of extract was determined by examining hydrogen donating ability on 1,1-diphenyl-2-picrylhydrazyl (DPPH) and the inhibitory effect on the formation of the peroxide from Linoleic acid in the test tube at $50^{\circ}C$. Antioxidative substance were, then, separated and indentified by thin layer chromatography(TLC), UV-Vis spectrum and High performance liquid chromatography(HPLC) methods. Hydrogen donating ability on DPPH and antioxidative ability on linoleic acid of the extracted antioxidative substance were higher than those of 100ppm butylated hydroxy toluene(BHT). The extracted antioxidative substances were separated by TLC using ethylacetate : chloroform : formic acid : water(8 : 1 : 1 : 1 v/v) as a solvent, and a spot at Rf=0.35 was detected. The spot was scraped from the plate, and extrated by methanol. The extract was analyzed by UV-Vis spetra and HPLC, and chlorogenic acid was identified as a antioxidative substance.

• 한국미생물·생명공학회지
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• 제24권3호
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• pp.380-383
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• 1996

Adenosine deaminase inhibitor was extracted from culture broth of Streptomyces sp. strain V-8 with ethylacetate. The ethylacetate extract showed the characteristic UV absorption spectrum of actinomycins at 440-450 nm. The ethylacetate extract was compared with respect to inhibitory behavior against adenosine deaminase from calf intestinal mucosa with actinomycin D, -C complex and actinomycin V. The Ki values for actnomycin D, -C complex, and actinomycin V against adenosine deaminase were determined to be 9.9 $\times$ 10$^{-6}$ M, 9.6 $\times$ 10$^{-6}$ M and 9.3 $\times$ 10$^{-6}$ M, respectively. The Ki value for the ethylacetate extract of culture broth against adenosine deaminase was determined to be 5.7 $\times$ 10$^{-6}$ M. The kinetic parameters of actinomycin D, -C complex, -V and ethylacetate extract of culture broth for adenosine deaminase were as follows:I$_{50}$ = 1.5 $\times$ 10$^{-5}$ M (actinomycin D), 2.7 $\times$ 10$^{-5}$ M (actinomycin C complex), 3.5 $\times$ 10$^{-5}$ M (actinomycin V), 8.9 $\times$ 10$^{-6}$ M (ethylacetate extract of culture broth). The adenosine deaminase was inhibited noncompetitively by ethylacetate extract of culture broth as well as by actinomycin D, -C complex and actinomycin V.

• 한국식품위생안전성학회지
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• 제10권1호
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• pp.33-39
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• 1995

The antibacterial and antifungal effect of grapefruit seed extract(GFSE) was investigated for its purpose of application to a diverse spectrum of field as sanitizers, disinfectants and preservatives. GFSE showed coparatively high content of such flavoniods as naringin and hesperidin and ascorbic acid. GFSE containing a low level of organic acids is a heavy viscous and water-soluble liquid. As a result of the antimicrobial test of GFSE, Bacillus subtilis and Aspergillus oryzae did not survive at detectable levels when treated with more than 100 ppm of GFSE. The minimum inhibitory concentrations of GFSE for a wide variety of pathogenic and putrefactive bacteria, fungi and yeasts were 100 ppm and 250 ppm, respectively. In the comparable electron micrograph of microbial cells treated with GFSE or not, we could conclude that GFSE destroy microorganisms by disrupting the functions of the cell wall membrane and microbial spores.

• Preventive Nutrition and Food Science
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• 제2권4호
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• pp.310-315
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• 1997

Wild propionibacteria isolated from different commercial swiss cheese samples were tested for antimicrobial activities. In initial screening, six of these Propionibacterium isolates showed antagonistic activity against 10 selected indicator organisms by the deferred method. In next, only two Propionibacterium strains JW6 and JW14 showed antibacterial activity in the cell-free supernatants by the modified well diffusion method. Propionibacterium strains JW6 and JW14 were finally identified as bacteriocin producers which exhibited a bactericidal effect against closely related species. The antimicrobial substances were proteins, since their activities were completely destroyed following several degradative enzyme treatments. The bacteriocins showed a narrow inhibitory spectrum of activity against two propionibacteria and two bacilli of strains tested in this study.