• Title/Summary/Keyword: Disc assay

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Purification and Some Properties of the polyphenol Oxidase form Ascidian, Halocynthia roretzi

  • Jeon, Byeong-Jun;Lee, Kang-Ho;Ryu, Hong-Soo;You, Byeong-Jin
    • Preventive Nutrition and Food Science
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    • v.1 no.1
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    • pp.111-116
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    • 1996
  • Polyphenol oxidase(PPO) isolated from the crude extract of ascidian, Halocynthia roretzi, showed higher affinity for catechol than tyrosine or DL-DOPA. Successful enzyme assay could be performed at $25^{\circ}C$, 10min. by mixing 0.2ml of crude enzyme extract with 2.8ml of 0.13M catechol in 0.1M sodium phosphate buffer(pH 6.4). The specific activity of PPO which had been purified with a combination of ammonium sulfate treatment, ion exchange chromatography on DEAE-cellulose, and gel filtration on Sepharose 6B was 13-fold disc gel electrophoresis. The activity of PPO was stable from pH 5.0 to 8.0 and showed the peak activity at pH 6.4 .The optimum reaction temperature for PPO oxidation on catechol was 35$^{\circ}C$ and those enzyme were heat stable up to 4$0^{\circ}C$. Molecular weigth of the enzyme was estimated about 170kDa. One molecule was found to be composed of gour subunits. Two of them had molecular weigh of 55kDa and the others 30kDa. The {TEX}$K_{m}${/TEX} values, {TEX}$V_{max}${/TEX} and catalytic efficiency({TEX}$V_{max}${/TEX}/{TEX}$K_{m}${/TEX}) for catechol were 0.12mM, 2.5mM/liter/min. and {TEX}$0.18min^{-1}${/TEX} respectively. The substrate affinity and electrophorectic pattern suggested that the enzyme of ascidian was considered to be not tyosine but catechol oxidase.

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Antimicrobial efficacy and safety analysis of zinc oxide nanoparticles against water borne pathogens

  • Supraja, Nookala;Avinash, B.;Prasad, T.N.V.K.V.
    • Advances in nano research
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    • v.5 no.2
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    • pp.127-140
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    • 2017
  • Metal nanoparticles have been intensively studied within the past decade. Nano-sized materials have been an important subject in basic and applied sciences. Zinc oxide nanoparticles have received considerable attention due to their unique antibacterial, antifungal, and UV filtering properties, high catalytic and photochemical activity. In this study, microbiological aspects of scale formation in PVC pipelines bacteria and fungi were isolated. In the emerging issue of increased multi-resistant properties in water borne pathogens, zinc oxide (ZnO) nanoparticle are being used increasingly as antimicrobial agents. Thus, the minimum bactericidal concentration (MBC) and minimum fungal concentration of ZnO nanoparticles towards pathogens microbe were examined in this study. The results obtained suggested that ZnO nanoparticles exhibit a good anti fungal activity than bactericidal effect towards all pathogens tested in in-vitro disc diffusion method (170 ppm, 100 ppm and 30 ppm). ZnO nanoparticles can be a potential antimicrobial agent due to its low cost of production and high effectiveness in antimicrobial properties, which may find wide applications in various industries to address safety issues. Stable ZnO nanoparticles were prepared and their shape and size distribution characterized by Dynamic light scattering (35.7 nm) and transmission electron microscopic TEM study for morphology identification (20 nm), UV-visible spectroscopy (230 nm), X-ray diffraction (FWHM of more intense peak corresponding to 101 planes located at $36.33^{\circ}$ using Scherrer's formula), FT-IR (Amines, Alcohols, Carbonyl and Nitrate ions), Zeta potential (-28.8). The antimicrobial activity of ZnO nanoparticles was investigated against Bacteria and Fungi present in drinking water PVC pipelines biofilm. In these tests, Muller Hinton agar plates were used and ZnO nanoparticles of various concentrations were supplemented in solid medium.

Chemical Composition and Antimicrobial Efficiency of Swietenia macrophylla Seed Extract on Clinical Wound Pathogens

  • Gopalan, Hanan Kumar;Md Hanafiah, Nor Faizzah;Ring, Leong Chean;Tan, Wen-Nee;Wahidin, Suzana;Hway, Teo Siew;Yenn, Tong Woei
    • Natural Product Sciences
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    • v.25 no.1
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    • pp.38-43
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    • 2019
  • Microbial wound infection prolonged the hospitalization and increase the cost for wound management. Silver is commonly used as antimicrobial wound dressing. However, it causes several adverse side effects. Hence, this study was aimed to evaluate the antimicrobial efficiency of Swietenia macrophylla seed extract on clinical wound pathogens. Besides, the bioactive constituents of the seed extract were also determined. S. macrophylla seeds were extracted with methanol by maceration method. The seed extract inhibited 5 test bacteria and 1 yeast on disc diffusion assay. The antibacterial activity was broad spectrum, as the extract inhibited both Gram positive and Gram negative bacteria. On kill curve analysis, the antibacterial activity of the seed extract was concentration-dependent, the increase of extract concentration resulted in more reduction of bacterial growth. The extract also caused 99.9% growth reduction of Bacillus subtilis relative to control. A total of 21 compounds were detected in gas chromatography- mass spectrometry analysis. The predominant compounds present in the extract were oleic acid (18.56%) and linoleic acid (17.72%). In conclusion, the methanolic extract of S. macrophylla seeds exhibited significant antimicrobial activity on clinical wound pathogens. Further investigations should be conducted to purify other bioactive compounds from the seeds of S. macrophylla.

Isolation and Characterization of Bioactive Compounds from Root of Rubus coreanus Miquel and their Antimicrobial Activity

  • Jang, Ha Na;Ha, Ji Hoon;Lee, Yoon Ju;Fu, Min Min;Park, Soo Nam
    • 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.

Anticaries Effect of Ethanol Extract of Terminalia chebula

  • Lee, Moonkyung;Hwang, Young Sun
    • Journal of dental hygiene science
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    • v.21 no.2
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    • pp.119-126
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    • 2021
  • Background: Dental caries is mainly composed of various cellular components and is deposited around the tooth surface and gums, causing a number of periodontal diseases. Streptococcus mutans is commonly found in the human oral cavity and is a significant contributor to tooth decay. The use of antibacterial ingredients in oral hygiene products has demonstrated usefulness in the management of dental caries. This study investigated the anticaries effect of the ethanol extract of Terminalia chebula (EETC) against S. mutans and their cytotoxicity to gingival epithelial cells. Methods: The EETC was prepared from T. chebula fruit using ethanol extraction. Disk diffusion, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and colony forming unit (CFU) were analyzed to investigate the antimicrobial activity of the EETC. Glucan formation was measured using the filtrate of the bacterial culture medium and sucrose. Gene expression was analyzed using reverse transcription-polymerase chain reaction (RT-PCR). Cytotoxicity was analyzed via the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Results: The antibacterial activity of the EETC was explored using disc diffusion and CFU measurements. The MIC and MBC of the EETC were 10 and 20 ㎍/ml, respectively. EETC treatment decreased insoluble glucan formation by S. mutans enzymes and also resulted in reduced glycosyltransferase B (gtf B), gtf C, gtf D, and fructosyltransferase (ftf), expressions on RT-PCR. In addition, at effective antibacterial concentrations, EETC treatment was not cytotoxic to gingival epithelial cells. Conclusion: These results demonstrate that the EETC is an effective anticaries ingredient with low cytotoxicity to gingival epithelial cells. The EETC may be useful in antibacterial oral hygiene products for the management of dental caries.

Antibacterial and Antioxidative Activities of Epimedium koreanum Nakai Extracts (삼지구엽초 추출물의 항균 및 항산화 활성에 대한 연구)

  • Won, Doo Hyun;Gu, Hyun A;Kim, Hye Jin;Han, Saet Byeol;Park, Jino;Park, Soo Nam
    • Microbiology and Biotechnology Letters
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    • v.41 no.3
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    • pp.284-291
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    • 2013
  • In this study, the antibacterial and antioxidative activities of Epimedium koreanum Nakai were investigated for applications as cosmetic ingredients. Minimum inhibitory concentrations (MICs) of fraction-bacterium, that showed high antibacterial activity from disc diffusion assay on human skin pathogens, were tested. The ethyl acetate fraction on Staphylococcus aureus, Bacillus subtilis, Propionibacterium acnes and 50% ethanol extract on S. aureus exhibited higher antibacterial activities than methyl paraben, well known as a preservative. The DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging activities of 3 fractions of E. koreanum Nakai were lower than (+)-${\alpha}$-tocopherol, known as a typical antioxidant. From the results of the scavenging activities of various ROS generated in $Fe^{3+}-EDTA/H_2O_2$ systems ($OSC_{50}$), 50% ethanol extract ($OSC_{50}=2.46{\pm}0.06{\mu}g/ml$) and aglycone fraction ($OSC_{50}=1.45{\pm}0.02{\mu}g/ml$) showed high activities similar to L-ascorbic acid ($OSC_{50}=1.50{\pm}0.85{\mu}g/ml$), used as reference. The cellular protective effects (${\tau}_{50}$) on photohemolysis by $^1O_2$ generated by photosensitization reaction were tested. The cellular protective effect of 50% ethanol extract (${\tau}_{50}=37.0{\pm}0.3$ min) was similar to (+)-${\alpha}$-tocopherol (${\tau}_{50}=38.0{\pm}1.8$ min), used as reference. In particular, the ${\tau}_{50}$ of aglycone fraction results were $165.9{\pm}7.2$ min. This is a high cellular protective effect, more than 4 times that of (+)-${\alpha}$-tocopherol. These results indicate that E. koreanum Nakai extract, and its fractions, could be utilized as a cosmetic ingredient possessing antibacterial and antioxidative activities.

Evaluation of Anti-oxidant, Anti-microbial and Anti-thrombosis Activities of Fruit, Seed and Pomace of Schizandra chinensis Baillon (오미자 열매, 씨, 착즙 후 박의 항산화, 항균 및 항혈전 활성 평가)

  • Kim, Mi-Sun;Sung, Hwa-Jung;Park, Jong-Yi;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.27 no.2
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    • pp.131-138
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    • 2017
  • In this study, for the efficient use of the byproduct of the omija (Schizandra chinensis Baillon: SC) processing industry, the ethanol extracts of the fruit (F), seed (S), and pomace (P) of SC were prepared, and their useful bioactivities were evaluated. For F-SC, S-SC, and P-SC, the extraction yields were 28.3%, 22.1%, and 7.2%, respectively, and the polyphenol contents were 8.81, 37.22, and 9.20 mg/g, respectively. The total flavonoid content in P-SC (4.31 mg/g) was 3.5-fold higher than that in F-SC (0.76 mg/g). In an antioxidation activity assay, P-SC showed stronger radical scavenging activities against DPPH anion, ABTS cation, and nitrite and stronger reducing power activities than the other extracts. The calculated concentration required for 50% radical scavenging activity, $RC_{50}s$, of P-SC for DPPH anion, ABTS cation, and nitrite was 226.2, 192.5, and $92.5{\mu}g/ml$, respectively. In an antimicrobial activity assay, F-SC, S-SC, and P-SC showed similarly strong growth inhibitions against Bacillus subtilis and P. vulgaris at a concentration of 0.5 mg/disc. F-SC and P-SC showed 15-fold extended time in thrombin, prothrombin, and activated partial thromboplastin time assays at a concentration of 5 mg/ml. The anticoagulation activity of P-SC (2.5 mg/ml) was comparable to that of aspirin (1.5 mg/ml). Furthermore, F-SC and S-SC showed very good platelet aggregation inhibitory activities. F-SC, S-SC, and P-SC did not show significant hemolysis against human red blood cell up to a concentration of 0.5 mg/ml. These results suggest that S-SC and P-SC, both of which are byproducts of the omija processing industry, show strong potential as novel antioxidant, antimicrobial, and antithrombosis agents.

Antimicrobial and Hemolytic Activity of Oriental Medicinal Herbs (한약재의 항균 활성 및 인간 적혈구 용혈 활성)

  • Ryu, Hee-Young;Ahn, Seon-Mi;Shin, Yong-Kyu;Sohn, Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.38 no.2
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    • pp.190-197
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    • 2010
  • To develop the safe and natural antimicrobial agents, the 68 ethanol extracts from the 61 different kinds of oriental herbal medicine were prepared and their antimicrobial activities were evaluated. The herbal medicine used were from China (46 kinds), South Korea (14 kinds), North Korea (5 kinds) and Vietnam (3 kinds), respectively, and the root (27 species) was popular part in this study. The average water content and extraction ratio for ethanol were 7.10% and 6.75%, respectively. Determination of antimicrobial activity by disc-diffusion assay at 0.5 mg/disc concentration showed that the extract of Angelica tenuissima Nakai (china), Illicium verum, Junci medulla, Rhus javanica L., Salvia miltiorrhiza Bunge and Syzygium aromaticum has strong antimicrobial activities against different food spoilage and pathogenic bacteria and fungi. Determination of MIC and MBC/MFC further showed that the extract of Syzygium aromaticum has MIC of 1.25 mg/mL and MBC/MFC of 1.25~5.00 mg/mL against Listeria monocytogenes, Bacillus subtilis, Staphylococcus epidermidis, Staphylococcus aureus, Salmonella typhimurium, Proteus vulgaris, Escherichia coli, Pseudomonas aeruginosa, Candida albicans and Saccharomyces cerevisiae. And, the extract of Junci medulla, Rhus javanica L. and Salvia miltiorrhiza Bunge showed strong antibacterial activities with MIC of 0.08~0.63 mg/mL and MBC/MFC of 0.08~2.50 mg/mL against the tested bacteria except E. coli and P. aeruginosa. In a while, the results of hemolytic activity of 68 different herbal extracts against human red blood cells showed that the extract of Angelica tenuissima Nakai has hemolytic activity at 0.5 mg/mL concentration. Therefore, Illicium verum, Junci medulla, Rhus javanica L., Salvia miltiorrhiza Bunge and Syzygium aromaticum were finally selected for natural antimicrobial resources. Further research on active substances and the mode of action of the selected herbal medicine is necessary.

The Limnological Survey of Major Lakes in Korea (4): Lake Juam (국내 주요 호수의 육수학적 조사(4) : 주암호)

  • Kim, Bom-Chul;Heo, Woo-Myung;Lim, Byung-Jin;Hwang, Gil-Son;Choi, Kwang-Soon;Choi, Jong-Soo;Park, Ju-Hyun
    • Korean Journal of Ecology and Environment
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    • v.34 no.1 s.93
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    • pp.30-44
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    • 2001
  • In this study limnological characteristics of Lake Juam was surveyed from June 1993 to May 1994 in order to provides important information regarding water resources. Secchi disc transparency, epilimnetic chlorophyll a (chi-a), total nitrogen (TN), total phosphorus (TP) concentration and primary productivity were in the range of $2.0{\sim}4.5\;m$, $0.9{\sim}13.6\;mgChl/m^3$, 0.78$\{sim}$2.32 mgN/l, $11{\sim}56\;mgP/m^3$, $270{\sim}2.160\;mgCm^{-2}\;day^{-1}$, respectively. On the basis of TP, Chl-a and Secchi disc depth, the trophic state of Lake Juam can be classied as mesotrophic lake. The phosphorus inputs from non-point sources are concentrated in heavy rain episodes during the monsoon season. As a result, phosphorus concentration are higher in summer than in winter. TP loading from the watershed were estimated to be $0.9\;gPm^{-2}yr^{-1}$, which correspond to a boundary of the critical loading ($1.0\;gPm^{-2}yr^{-1}$) for eutrophication. From the results of the algal assay, both phosphous and nitrogen act as limiting nutrients in algal growth. The seasonal succession of phytoplankton community structure in Lake Juam was similar to that observed in other temperate lakes. Diatoms (Asterionella formosa and Aulacoseira granulate var. angustissima)fujacofeira BraHuJafa uar. aHgusHrsiaia) weredominant in spring and winter, cyanobacteria) were dominant in warm season. The organic carbon, nitrogen and phosphorus content of lake sediment were $9.5{\sim}14.0\;mgC/g$, $1.01{\sim}1.82\;mgN/g$ and $0.51{\sim}0.65\;mgP/g$, respectively. The allochthonous organic carbon loading from the watershed and autochthonous organic carbon loading by primary production of phytoplankton were determined to be 1,122 tC/yr and 6,718 tC/yr, respectively. To prevent eutrophication of Lake Juam, nutrient management of watershed should be focus on reduction of fertilizer application, proper treatment of manure, and conservation of topsoil as well as point source.

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Antimicrobial Activity of Extracts from Different Parts and Essential Oil from Pinus densiflora on Skin Pathogens (소나무 부위별 추출물 및 essential oil의 피부상재균에 대한 항균 활성)

  • Park, Sun-Hee;Kim, Koth-Bong-Woo-Ri;Kim, Min-Ji;Choi, Jung-Su;Cho, Young-Je;Ahn, Dong-Hyun
    • Journal of Life Science
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    • v.27 no.6
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    • pp.646-651
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    • 2017
  • This study was carried out to examine the antimicrobial activity of 70% ethanol and hot water extracts from different parts and essential oil in Pinus densiflora on skin pathogens such as Staphylococcus epiermidis, S. aureus, Propionibacterium acnes, Candida albicans, C. tropicalis, and Trichophyton rubrum. The antimicrobial activities of extracts and essential oil were tested by paper disc assay and minimum inhibitory concentration test. The ethanol extract of pine pollen showed antimicrobial activity against C. tropicalis and T. rubrum. Ethanol extract of pine needle and pine gnarl showed antimicrobial activity against S. aureus, C. tropicalis, and P. acnes. Essential oil from pine needle exhibited antimicrobial activity against C. albicans, C. tropicalis, and P. acnes. The minimum inhibitory concentration of ethanol extracts of pine needle and pine gnarl against P. acnes and C. tropicalis ranged from 0.002% to 0.0063%. The lowest minimum inhibitory concentration (0.025%) against C. albicans and C. tropicalis was obtained from essential oil. These results indicate that ethanol extracts of pine pollen, pine needle, pine gnarl, and essential oil of pine needle could be applicable to control the skin infection pathogens. Especially, ethanol extract of pine gnarl had a broad spectrum of antimicrobial activity and pine extracts and essential oil exhibited higher antimicrobial activity with Candida sp. and P acnes.