• Title/Summary/Keyword: Sequence of 16S rDNA.

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Isolation and Identification of Streptomyces sp. Producing Anti-vancomycin Resistant Staphylococcus aureus Substance (반코마이신 내성 Staphylococcus aureus 억제 물질 생산 Streptomyces sp.의 분리 및 동정)

  • Oh Se-Teak;Lee Jun-Jae;Lee Ji-Youn;Kim Jin-Kyu;Yang Si-Yong;Kim Yang-Soo;Song Min-Dong
    • Microbiology and Biotechnology Letters
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    • v.33 no.2
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    • pp.90-95
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    • 2005
  • An Actinomycetes producing an anti-VRSA (vancomycin-resistant Staphylococcus aureus) substance was isolated from soil. The cultural, morphological, physiological and phylogenetic analyses of an isolated strain were investigated for identification. Cultural characteristics based on ISP (International Streptomyces Project) were as follows: white aerial mycelium, yellow reverse side, and good growth on various medium. Also, the isolate did not produce the soluble pigment. Morphological characteristics were showed cylindrical spore chain and smooth spore surface by SEM (Scanning Electron Microscope). Physiological characteristics were showed LL-type by DAP isomer analysis and detected glycine, glutamic acid and alanine. A phylogenetic analysis of the 16S rDNA provided a clue that the isolated strain was actually a member of the genus Streptomyces, because the determined sequence exhibited a higher homology with Streptomyces echinatus. The isolate was identified to be a genus of Streptomyces sp.. The optimal culture conditions for the maximum production of anti-VRSA substance by Streptomyces sp. were attained in a culture medium composed of $2.0\%$ (w/v) glucose, and $0.4\%$ (w/v) yeast extract. The anti-VRSA substance was highly produced after 5 days of culture. Optimal pH and temperature conditions for the production of anti-VRSA substance were pH 7.0 and $28^{\circ}C$, respectively.

Physiological Characteristics and Production of Folic Acid of Lactobacillus plantarum JA71 Isolated from Jeotgal, a Traditional Korean Fermented Seafood

  • Park, Sun-Young;Do, Jeong-Ryong;Kim, Young-Jin;Kim, Kee-Sung;Lim, Sang-Dong
    • Food Science of Animal Resources
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    • v.34 no.1
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    • pp.106-114
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    • 2014
  • Folic acid, one of the B group of vitamins, is an essential substance for maintaining the functions of the nervous system, and is also known to decrease the level of homocysteine in plasma. Homocysteine influences the lowering of the cognitive function in humans, and especially in elderly people. In order to determine the strains with a strong capacity to produce folic acid, 190 bacteria were isolated from various kinds of jeotgal and chungkuk-jang. In our test experiment, JA71 was found to contain $9.03{\mu}g/mL$ of folic acid after 24 h of incubation in an MRS broth. This showed that JA71 has the highest folic acid production ability compared to the other lactic acid bacteria that were isolated. JA71 was identified as Lactobacillus plantarum by the result of API carbohydrate fermentation pattern and 16s rDNA sequence. JA71 was investigated for its physiological characteristics. The optimum growth temperature of JA71 was $37^{\circ}C$, and the cultures took 12 h to reach pH 4.4. JA71 proved more sensitive to bacitracin when compared with fifteen different antibiotics, and showed most resistance to neomycin and vancomycin. Moreover, it was comparatively tolerant of bile juice and acid, and displayed resistance to Escherichia coli, Salmonella Typhimurium, and Staphylococcus aureus with restraint rates of 60.4%, 96.7%, and 76.2%, respectively. These results demonstrate that JA71 could be an excellent strain for application to functional products.

Bacterial Communities of Biofilms Sampled from Seepage Groundwater Contaminated with Petroleum Oil

  • CHO WONSIL;LEE EUN-HEE;SHIM EUN-HWA;KIM JAISOO;RYU HEE WOOK;CHO KYUNG-SUK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.952-964
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    • 2005
  • The diesel-degrading activities of biofilms sampled from petroleum-contaminated groundwaters in urban subway drainage systems were examined in liquid cultures, and the microbial populations of the biofilms were characterized by denaturing gel gradient electrophoresis (DGGE) and 16S rDNA sequence analysis. Biofilm samples derived from two sites (19 K and 20 K) at subway Station N and Station I could degrade around $80\%$ of applied diesel within 20 and 40 days, respectively, at $15^{\circ}C$, and these results were strongly correlated with the growth patterns of the biofilms. The closest phylogenetic neighbor of a dominant component in the 19 K biofilm was Thiothrix fructosivorans strain Q ($100\%$ similarity). Four dominant strains in the 20 K biofilm were closely related to Thiothrix fructosivorans strain Q ($100\%$ similarity), Thiothrix sp. CC-5 ($100\%$ similarity), Sphaerotilus sp. IF14 ($99\%$ similarity), and Cytophaga-Flexibacter-Bacterioides (CFB) group bacterium RW262 ($98\%$ similarity). Three dominant members in the Station I biofilms were very similar to uncultured Cytophagales clone CRE-PA82 ($91\%$ similarity), Pseudomonas sp. WDL5 ($97\%$ similarity), and uncultured CFB group bacterium LCK-64 ($94\%$ similarity). The microbial components of the biofilms differed depending on the sampling site. This is the first report on the isolation of clones highly similar to Thiothrix fructosivorans and Thiothrix sp. from biofilms in petroleum-polluted groundwaters, and the first evidence that these organisms may play major roles in petroleum degradation and/or biofilm-development.

Diagnosis on sudden death cases during summer season and isolation of Clostridium novyi (하절기 급사 돼지의 Clostridium novyi 진단 및 분리)

  • Jeong, Chang-Gi;Seo, Byoung-Joo;Kim, Won-Il
    • Korean Journal of Veterinary Service
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    • v.39 no.2
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    • pp.131-136
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    • 2016
  • Clostridium novyi (C. novyi) is a gram positive, non-capsulated, motile, obligatory anaerobe that produces endospores. Both C. novyi type A and B produce a bacteriophage encoded lethal alpha toxin which belongs to a family of large clostridial cytotoxins. These large clostridial cytotoxins of C. novyi bind to the uncharacterized receptors on host vascular endothelial cells, which leads to the loss of integrity of the vascular endothelium with subsequent edema, refractory hypotension, organ failure, and sudden death. A total of 13 sudden death cases were submitted to Chonbuk National University-Veterinary Diagnostic Center between June and October, 2015. The samples, mainly liver, were collected in sterile vials after necropsy and processed within 12~24 hours for diagnosis, isolation and identification of C. novyi. All of the 4 gram positive samples showed amplification by PCR. Out of 4 positive samples, 3 were detected to be C. novyi type B and 1 was detected as C. novyi type A. Based on the 16S rDNA sequence analysis, 1 case (150564) showed 99% similarity with C. novyi type A while other 3 cases (150388, 150557 and 150775) presented 99% similarity with C. novyi type B. Based on the results, C. novyi was found to be prevalent in Korean pig farms and causes sudden death to finishing pigs or sows during summer season. Thus, C. novyi should be considered for differential diagnosis on sudden death cases during the summer season.

Probiotic Potential of Enterococcus faecium Isolated from Chicken Cecum with Immunomodulating Activity and Promoting Longevity in Caenorhabditis elegans

  • Sim, Insuk;Park, Keun-Tae;Kwon, Gayeung;Koh, Jong-Ho;Lim, Young-Hee
    • Journal of Microbiology and Biotechnology
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    • v.28 no.6
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    • pp.883-892
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    • 2018
  • Probiotics, including Enterococcus faecium, confer a health benefit on the host. An Enterococcus strain was isolated from healthy chicken cecum, identified as E. faecium by 16S rDNA gene sequence analysis, and designated as E. faecium L11. To evaluate the potential of E. faecium L11 as a probiotic, the gastrointestinal tolerance, immunomodulatory activity, and lifespan extension properties of the strain were assayed. E. faecium L11 showed >66% and >62% survival in artificial gastric juice (0.3% pepsin, pH 2.5) and simulated small intestinal juice (0.5% bile salt and 0.1% pancreatin), respectively. Heat-killed E. faecium L11 significantly (p < 0.05) increased immune cell proliferation compared with controls, and stimulated the production of cytokines (IL-6 and $TNF-{\alpha}$) by activated macrophages obtained from ICR mice. In addition, E. faecium L11 showed a protective effect against Salmonella Typhimurium infection in Caenorhabditis elegans. In addition, feeding E. faecium L11 significantly (p < 0.05) extended the lifespan of C. elegans compared with the control. Furthermore, genes related to aging and host defense were upregulated in E. faecium L11-fed worms. In conclusion, E. faecium L11, which prolongs the lifespan of C. elegans, may be a potent probiotic supplement for livestock.

Characterization and Enhanced Production of Enterocin HJ35 by Enterococcus faecium HJ35 Isolated from Human Skin

  • Yoon Yoh Chang;Park Hye Jung;Lee Na-Kyoung;Paik Hyun-Dong
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.4
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    • pp.296-303
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    • 2005
  • A strain named as HJ35 was isolated from the skin of sixty-five men and fourteen women for acne therapy, in order to find an effective antimicrobial agent against Propionibacterium acnes. Isolate HJ35 was identified as Enterococcus faecium based on 16 rDNA sequence and produced enterocin HJ35 having antimicrobial activities against most lactic acid bacteria, En­terococcus spp., Staphylococcus aureus, S. epidermidis, Clostridium perfringens, some bacilli, Mi­crococcus flavus, Listeria monocytogenes, L. ivanovii, Escherichia coli, Pseudomonas fluorescens and Propionibacterium acnes, in the modified well diffusion method. Especially, enterocin HJ35 showed a bactericidal activity against Propionibacterium acnes P1. The antimicrobial activity of enterocin HJ35 was disappeared completely with the use of protease XIV. But enterocin HJ35 activity is very stable at high temperature (up to $100^{\circ}C$ for 30 min), in wide range of pH (3.0${\~}$9.0), and by treatment with organic solvents. The apparent molecular mass of enterocin HJ35 was estimated to be approximately 4${\~}$4.5 kDa on detection of its bactericidal activity after SDS-PAGE. In batch fermentation of E. faecium HJ35, enterocin HJ35 was produced at the mid­log growth phase, and its maximum production was obtained up to 2,300 AU/mL at the late stationary phase. By employing fed-batch fermentation, the enhanced production of enterocin HJ35 was achieved up to 12,800 AU/mL by feeding with 10 g/L glucose or 6 g/L lactate.

Characterization of Three Antifungal Calcite-Forming Bacteria, Arthrobacter nicotianae KNUC2100, Bacillus thuringiensis KNUC2103, and Stenotrophomonas maltophilia KNUC2106, Derived from the Korean Islands, Dokdo and Their Application on Mortar

  • Park, Jong-Myong;Park, Sung-Jin;Ghim, Sa-Youl
    • Journal of Microbiology and Biotechnology
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    • v.23 no.9
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    • pp.1269-1278
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    • 2013
  • Crack remediation on the surface of cement mortar using microbiological calcium carbonate ($CaCO_3$) precipitation (MICP) has been investigated as a microbial sealing agent on construction materials. However, MICP research has never acknowledged the antifungal properties of calcite-forming bacteria (CFB). Since fungal colonization on concrete surfaces can trigger biodeterioration processes, fungi on concrete buildings have to be prevented. Therefore, to develop a microbial sealing agent that has antifungal properties to remediate cement cracks without deteriorative fungal colonization, we introduced an antifungal CFB isolated from oceanic islands (Dokdo islands, territory of South Korea, located at the edge of the East Sea in Korea.). The isolation of CFB was done using B4 or urea-$CaCl_2$ media. Furthermore, antifungal assays were done using the pairing culture and disk diffusion methods. Five isolated CFB showed $CaCO_3$ precipitation and antifungal activities against deteriorative fungal strains. Subsequently, five candidate bacteria were identified using 16S rDNA sequence analysis. Crack remediation, fungi growth inhibition, and water permeability reduction of antifungal CFB-treated cement surfaces were tested. All antifungal CFB showed crack remediation abilities, but only three strains (KNUC2100, 2103, and 2106) reduced the water permeability. Furthermore, these three strains showed fungi growth inhibition. This paper is the first application research of CFB that have antifungal activity, for an eco-friendly improvement of construction materials.

Cytotoxic, Antioxidative, and ACE Inhibiting Activities of Dolsan Leaf Mustard Juice (DLMJ) Treated with Lactic Acid Bacteria

  • Yoo Eun-Jeong;Lim Hyun-Soo;Park Kyung-Ok;Choi Myeong-Rak
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.1
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    • pp.60-66
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    • 2005
  • This study was performed to know whether there is any change of physiological activity in DLMJ which is inoculated by lactic acid bacteria. Lactic acid bacteria were isolated from Dolsan leaf mustard Kimchi (DLMK) at $20^{\circ}C$. In the optimum ripening period, the population of Leuconostoc and Lactobacilli in the DLMK were found to be high. The Leuconostoc, Lactobacilli and Lactococci strains were identified as Leuconostoc mesenteroides, Leuconostoc gelidum, Weissella confusa, Lactobacillus plantarum, Lactobacillus raffinolactis, Lactococcus lactis and Weissella confusa using the Biolog system. The most predominant strain which was isolated from DLMK was Weissella confusa. As the results of the phylogenetic analysis using 16s rDNA sequence, the Weissella confusa turned out to be Weissella kimchii, with 99.0% similarity. To investigated the change of physiological activity in DLMJ by lactic acid bacteria, 7 predominant strains inoculated to DLMJ (Dolsan Leaf Mustard Juice). The cytotoxicity was found to be under $19.55\%$ all cases. Also, the antioxidative activity of the DLMJ treated with lactic acid bacteria was very low, which might have been due to the reduced antioxidative phytochemicals during the preparation of the sterile sample. The ACE inhibiting activity of DLMJ by inoculation with Weissella kimchii was shown to be the highest ($94.0\%$). This could be that the degradation of sulfur containing materials in DLMJ by Weissella kimchii gave rise to ACE inhibiting activity.

Application of Gamma Ray Irradiation to the Microbiological Safety of Dried Seafood Products (건조 수산물의 미생물학적 안전성 확보를 위한 감마선 조사 기술의 이용)

  • Choi, Jong-Il;Kim, Hyun-Joo;Kim, Jae-Hun;Ahn, Dong-Hyun;Chun, Byung-Soo;Lee, Ju-Woon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.43 no.2
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    • pp.169-173
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    • 2010
  • This study evaluated the effects of gamma ray irradiation on the safety of dried seafood products. Dried salted squid, Engraulis japonica, Hizikia fusiformis, Mytilus coruscus, and Porphyra tenera were gamma-irradiated at doses of 0, 1, 3, and 5 kGy. The total bacterial populations were then enumerated on total plate count agar, and bacteria isolated from the samples were identified by 16S rDNA sequencing. In addition, the isolated strains were inoculated in the products to determine $D_{10}$ values. The total bacterial populations in the dried seafood products ranged from 3.40 to 6.59 log CFU/g, and those of yeasts and molds ranged from 2.21 to 4.56 log CFU/g. The sequence analysis identified Staphylococcus sp. as the most common species in the dried seafood products, except for dried P. tenera. The $D_{10}$ values of the contaminating bacteria were lower than 0.7 kGy, except for Deinococcus sp., which had a value of 1.39 kGy. This study demonstrated that gamma irradiation could be used to improve the safety of dried seafood products.

Physiological Characteristics and GABA Production of Lactobacillus plantarum K255 Isolated from Kimchi

  • Park, Sun-Young;Kim, Kee-Sung;Lee, Myung-Ki;Lim, Sang-Dong
    • Food Science of Animal Resources
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    • v.33 no.5
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    • pp.595-602
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    • 2013
  • As a major inhibitory neurotransmitter of the central nervous system in animals, ${\gamma}$-aminobutyric acid (GABA) has several physiological functions, such as anti-hypertensive, diuretic, tranquilizer and anti-stress effects in human. In order to determine strains with high GABA producing ability and glutamate decarboxylase (GAD) activity, 273 bacteria were isolated from various types of Kimchi. Strain K255 contained $386.37{\mu}g/mL$ of GABA in MRS broth containing 1% MSG, $600.63{\mu}g/mL$ of GABA in MRS broth containing 2% MSG and $821.24{\mu}g/mL$ of GABA in MRS broth containing 3% MSG. It showed that K255 had the highest GABA production ability compared to other commercial lactic acid bacteria. K255 was identified as Lactobacillus plantarum based on its API carbohydrate fermentation pattern and 16S rDNA sequence. K255 was investigated for its physiological characteristics. The optimum growth temperature of K255 was $37^{\circ}C$and cultures took 13 h to reach the pH 4.4. K255 showed more sensitive to bacitracin in a comparison of fifteen different antibiotics, and showed most resistance to kanamycin and vancomycin. Moreover, it was comparatively tolerant to bile juice and acid and displayed resistance to Escherichia coli, Salmonella Typhimurium, Staphylococcus aureus, with rates of 30.8%, 29.7%, and 23.4% respectively. These results demonstrate that K255 could be an excellent strain for the production of functional products.