• Title/Summary/Keyword: 16s rRNA sequence

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Characterization of a Novel Fibrinolytic Enzyme, BsfA, from Bacillus subtilis ZA400 in Kimchi Reveals Its Pertinence to Thrombosis Treatment

  • Ahn, Min-Ju;Ku, Hye-Jin;Lee, Se-Hui;Lee, Ju-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.25 no.12
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    • pp.2090-2099
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    • 2015
  • Recently, the cardiovascular disease has been widely problematic in humans probably due to fibrin formation via the unbalanced Western style diet. Although direct (human plasmin) and indirect methods (plasminogen activators) have been available, bacterial enzyme methods have been studied because of their cheap and mass production. To detect a novel bacterial fibrinolytic enzyme, 111 bacterial strains with fibrinolytic activity were selected from kimchi. Among them, 14 strains were selected because of their stronger activity than 0.02 U of plasmin. Their 16S rRNA sequence analysis revealed that they belong to Bacillus, Leuconostoc, Propionibacterium, Weissella, Staphylococcus, and Bifidobacterium. The strain B. subtilis ZA400, with the highest fibrinolytic activity, was selected and the gene encoding fibrinolytic enzyme (bsfA) was cloned and expressed in the E. coli overexpression system. The purified enzyme was analyzed with SDS-PAGE, western blot, and MALDI-TOF analyses, showing to be 28.4 kDa. Subsequently, the BsfA was characterized to be stable under various stress conditions such as temperature (4-40oC), metal ions (Mn2+, Ca2+, K2+, and Mg2+), and inhibitors (EDTA and SDS), suggesting that BsfA could be a good candidate for development of a novel fibrinolytic enzyme for thrombosis treatment and may even be useful as a new bacterial starter for manufacturing functional fermented foods.

Characterization of Acetobacter pomorum KJY8 Isolated from Korean Traditional Vinegar

  • Baek, Chang Ho;Park, Eun-Hee;Baek, Seong Yeol;Jeong, Seok-Tae;Kim, Myoung-Dong;Kwon, Joong-Ho;Jeong, Yong-Jin;Yeo, Soo-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.24 no.12
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    • pp.1679-1684
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    • 2014
  • Acetobacter sp. strains were isolated from traditional vinegar collected in Daegu city and Gyeongbuk province. The strain KJY8 showing a high acetic acid productivity was isolated and characterized by phenotypic, chemotaxonomic, and phylogenetic inference based on 16S rRNA sequence analysis. The chemotaxonomic and phylogenetic analyses revealed the isolate to be a strain of Acetobacter pomorum. The isolate showed a G+C content of 60.8 mol%. It contained $\small{LL}$-diaminopimelic acid ($\small{LL}$-$A_2pm$) as the cell wall amino acid and ubiquinone $Q_9$ (H6) as the major quinone. The predominant cellular fatty acids were $C_{18:1}w9c$, w12t, and w7c. Strain KJY8 grew rapidly on glucose-yeast extract (GYC) agar and formed pale white colonies with smooth to rough surfaces. The optimum cultivation conditions for acetic acid production by the KJY8 strain were $20^{\circ}C$ and pH 3.0, with an initial ethanol concentration of 9% (w/v) to produce an acetic acid concentration of 8% (w/v).

Isolation of an Indigenous Imidacloprid-Degrading Bacterium and Imidacloprid Bioremediation Under Simulated In Situ and Ex Situ Conditions

  • Hu, Guiping;Zhao, Yan;Liu, Bo;Song, Fengqing;You, Minsheng
    • Journal of Microbiology and Biotechnology
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    • v.23 no.11
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    • pp.1617-1626
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    • 2013
  • The Bacterial community structure and its complexity of the enrichment culture during the isolation and screening of imidacloprid-degrading strain were studied using denaturating gradient gel electrophoresis analysis. The dominant bacteria in the original tea rhizosphere soil were uncultured bacteria, Rhizobium sp., Sinorhizobium, Ochrobactrum sp., Alcaligenes, Bacillus sp., Bacterium, Klebsiella sp., and Ensifer adhaerens. The bacterial community structure was altered extensively and its complexity reduced during the enrichment process, and four culturable bacteria, Ochrobactrum sp., Rhizobium sp., Geobacillus stearothermophilus, and Alcaligenes faecalis, remained in the final enrichment. Only one indigenous strain, BCL-1, with imidacloprid-degrading potential, was isolated from the sixth enrichment culture. This isolate was a gram-negative rod-shaped bacterium and identified as the genus Ochrobactrum based on its morphological, physiological, and biochemical properties and its 16S rRNA gene sequence. The degradation test showed that approximately 67.67% of the imidacloprid (50 mg/l) was degraded within 48 h by strain BCL-1. The optimum conditions for degradation were a pH of 8 and $30^{\circ}C$. The simulation of imidacloprid bioremediation by strain BCL-1 in soil demonstrated that the best performance in situ (tea soil) resulted in the degradation of 92.44% of the imidacloprid (100 mg/g) within 20 days, which was better than those observed in the ex situ simulations that were 64.66% (cabbage soil), 41.15% (potato soil), and 54.15% (tomato soil).

Screening of Antifungal Bacillus spp. against Alternaria Blight Pathogen (Alternaria panax) and Anthracnose Pathogen (Colletotrichum gloeosporioides) of Ginseng (인삼 점무늬병균(Alternaria panax)과 탄저병균(Colletotrichum gloeosporioides)에 대한 길항미생물 Bacillus spp. 선발)

  • Lee, Hye-Jin;Park, Kee-Choon;Lee, Seung-Ho;Bang, Kyong-Hwan;Park, Hong-Woo;Hyun, Dong-Yun;Kang, Seung-Weon;Cha, Seon-Woo;Chung, Ill-Min
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.5
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    • pp.339-344
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    • 2012
  • This study was carried out to research microorganisms having the antifungal activity against ginseng Alternaria blight pathogen Alternaria panax and ginseng anthracnose pathogen Colletotrichum gloeosporioides. Eleven Bacillus strains. were isolated from Korean traditional soybean paste and Kimchi. Among the 11 isolates, DJ5, KC1, KC2 and KC4 showing antagonistic activity on the mycelial growth of A. panax and C. gloeosporioides in pairing culture were finally selected as the antagonistic microorganisms. Based on 16s rRNA sequence and phylogenetic tree analysis, they were identified as Bacillus spp.. The selected microorganisms were investigated antagonistic activity by measured leaf-segment colonization in pot test. When Bacillus sp. were injected after A. panax treatment, KC1, KC2 and KC4 showed similar effect to chemical pesticides treated control. To measure preventive effect of Bacillus sp, antagonistic microorganisms were injected and C. gloeosporioides was treated in pot. When measuring the effectiveness for the prevention of Anthracnose, All Bacillus spp. showed approximately 83~90 % degree of superior preventive effect. In general, The four Bacillus spp. isolated from Korean traditional fermented foods showed therapeutic effect of Alternaria blight and preventive effect of Anthracnose.

Nontuberculous Mycobacterial Lung Disease Caused by Mycobacterium lentiflavum in a Patient with Bronchiectasis

  • Jeong, Byeong-Ho;Song, Jae-Uk;Kim, Wooyoul;Han, Seo Goo;Ko, Yousang;Song, Junwhi;Chang, Boksoon;Hong, Goohyeon;Kim, Su-Young;Choi, Go-Eun;Shin, Sung Jae;Koh, Won-Jung
    • Tuberculosis and Respiratory Diseases
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    • v.74 no.4
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    • pp.187-190
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    • 2013
  • We report a rare case of lung disease caused by Mycobacterium lentiflavum in a previously healthy woman. A 54-year-old woman was referred to our hospital due to chronic cough and sputum. A computed tomography scan of the chest revealed bilateral bronchiectasis with bronchiolitis in the right middle lobe and the lingular division of the left upper lobe. Nontuberculous mycobacteria were isolated twice from three expectorated sputum specimens. All isolates were identified as M. lentiflavum by multilocus sequence analysis based on rpoB, hsp65, and 16S rRNA fragments. To the best of our knowledge, this is the first documented case of M. lentiflavum lung disease in an immunocompetent adult in Korea.

Life History and Systematic Studies of Pseudothrix borealis gen. et sp. nov. (=North Pacific Capsosiphon groenlandicus, Ulotrichaceae, Chlorophyta)

  • Hanic, Louis A.;Lindstrom, Sandra C.
    • ALGAE
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    • v.23 no.2
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    • pp.119-133
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    • 2008
  • We cultured a tubular marine green alga, originally identified as Capsosiphon groenlandicus (J. Agardh) K.L. Vinogradova, from Amaknak Island, Alaska. The alga had an alternation of heteromorphic generations in which tubular monoecious fronds produced quadriflagellate zoospores and/or biflagellate isogametes. The gametes fused to produce cysts or Codiolum-like zygotes with long, tortuous stalks. Cysts and codiola produced 8-16 aplanospores, which germinated in situ to yield upright fronds. Fronds arising from both aplanospores and zoospores displayed a distinctive development in which non-septate colorless rhizoids from the base of the initially uniseriate, Ulothrix-like filament were transformed into septate uniseriate Ulothrix-like photosynthetic filaments. These transformed filaments then developed new basal non-septate rhizoids. This pattern of rhizoids becoming filaments, which then produced new rhizoids, was repeated to yield a tuft of up to 50 fronds. Periclinal and longitudinal divisions occurred in each filament, starting basally, until the mature tubular thallus was achieved. Pyrenoid ultrastructure revealed several short inward extensions of chloroplast lamellae, each of which was surrounded by pyrenoglobuli. Analysis of ribosomal SSU and ITS sequences placed this alga in the family Ulotrichaceae, order Ulotrichales, together with but as a distinct species from North Atlantic Capsosiphon groenlandicus. Analysis of a partial ITS sequence from authentic Capsosiphon fulvescens, the current name of the type of the genus Capsosiphon, indicated that neither our material nor C. groenlandicus belongs in that genus, and we propose a new genus, Pseudothrix, to accommodate both species. We propose P. borealis for the North Pacific entity formerly called C. groenlandicus and make the new combination P. groenlandica for the Atlantic species.

Antitumor activity of Bacillus subtilis SW-1 isolated from Jeotgal (젓갈에서 분리한 Bacillus subtilis SW-1의 항암활성)

  • 박종기;조용운;최영우;정영기;갈상완
    • Journal of Life Science
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    • v.14 no.5
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    • pp.815-820
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    • 2004
  • A bacterum containing antitumor activity was isolated from traditional korean food, Jeotgal. Through the 16s rRNA sequence analysis, the bacterium was identitied as a strain of Bacillus subtilis SW-l. The best culture condition for antitumor activity of the bacterium is 3% of soluble starch and 1 % of yeast extract as corbon and nitrogen sources, respectively. Cytotoxicitic concentrations of the culture supernatant of B. subtilis SW-1 against cancer cell lines, A549 and SK-OV3 were 30 ul/ml and 40 ul/ml, respectively, as $IC_{50}$/ values. In DNA fragmentation assay, the culture supernatant showed the programmed cell death (apoptosis) to cause degrading the chromosomal DNA like ladder. Taken together, the culture supernatant of the B. subtilis SW-1 has some possibility to be used as an antitumor agent.

Isolation and Characterization of Oligotrophic Bacteria Possessing Induced Systemic Disease Resistance against Plant Pathogens

  • Han, Song-Hee;Kang, Beom-Ryong;Lee, Jang-Hoon;Kim, Hyun-Jung;Park, Ju-Yeon;Kim, Jeong-Jun;Kim, Young-Cheol
    • The Plant Pathology Journal
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    • v.28 no.1
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    • pp.68-74
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    • 2012
  • Biocontrol microbes have mainly been screened among large collections of microorganisms $via.$ nutrient-rich $in$ $vitro$ assays to identify novel and effective isolates. However, thus far, isolates from only a few genera, mainly spore-forming bacilli, have been commercially developed. In order to isolate field-effective biocontrol microbes, we screened for more than 200 oligotrophic bacterial strains, isolated from rhizospheres of various soil samples in Korea, which induced systemic resistance against the soft-rot disease caused by $Pectobacterium$ $carotovorum$ SCC1; we subsequently conducted in $planta$ bioassay screening. Two oligotrophic bacterial strains were selected for induced systemic disease resistance against the $Tobacco$ $Mosaic$ $Virus$ and the gray mold disease caused by $Botrytis$ $cinerea$. The oligotrophic bacterial strains were identified as $Pseudomonas$ $manteilii$ B001 and $Bacillus$ $cereus$ C003 by biochemical analysis and the phylogenetic analysis of the 16S rRNA sequence. These bacterial strains did not exhibit any antifungal activities against plant pathogenic fungi but evidenced several other beneficial biocontrol traits, including phosphate solubilization and gelatin utilization. Collectively, our results indicate that the isolated oligotrophic bacterial strains possessing induced systemic disease resistance could provide useful tools as effective biopesticides and might be successfully used as cost-effective and preventive biocontrol agents in the field.

Diversity and Characterization of Endophytic Bacteria Associated with Tidal Flat Plants and their Antagonistic Effects on Oomycetous Plant Pathogens

  • Bibi, Fehmida;Yasir, Muhammad;Song, Geun-Cheol;Lee, Sang-Yeol;Chung, Young-Ryun
    • The Plant Pathology Journal
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    • v.28 no.1
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    • pp.20-31
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    • 2012
  • Endophytic bacterial communities of tidal flat plants antagonistic to oomycete plant pathogens were studied by the isolation of 256 root colonizing endophytic bacteria from surface-disinfected root tissues of six plants ($Rosa$ $rugosa$, $Suaeda$ $maritima$, $Vitex$ $rotundifolia$, $Carex$ $scabrifolia$, $Glehnia$ $littoralis$ and $Elymus$ $mollis$) growing in a tidal flat area of Namhae Island, Korea. To understand the antagonistic potential, an $in$ $vitro$ antagonistic assay was performed to characterize and identify strains that were antagonistic to the oomycete plant pathogens $Phytophthora$ $capsici$ and $Pythium$ $ultimum$ from the total population. Nine percent of the total number of isolated bacteria exhibited in vitro inhibitory activity against target plant pathogenic oomycetes. Taxonomic and phylogenetic placement of the antagonistic bacteria was investigated by analysis of the 16S rRNA gene sequences. The sequence analysis classified the antagonistic strains into four major classes of the domain bacteria ($Firmicutes$, ${\alpha}-Proteobacteria$, ${\gamma}-Proteobacteria$ and $Actinomycetes$) and 10 different genera. Further production of secondary metabolites, hydrolytic enzymes and plant growth promoting traits were determined for the putative new species of antagonistic endophytic bacteria. These new strains could not be identified as known species of ${\alpha}-Proteobacteria$, and so may represent novel bacterial taxa. The unexpected high antagonistic bacterial diversity associated with the tidal flat plants may be indicative of their importance in tidal flat plants as a promising source of novel antimicrobial compounds and biocontrol agents.

TPH Removal of the Biodegradation Process Using 4 Indigenous Microorganisms for the Diesel Contaminated Soil in a Military Camp (디젤로 오염된 군부대 토양에 대하여 토착미생물 4종을 이용한 생분해법의 TPH 제거 효율 규명)

  • Park, Min-Ho;Lee, Min-Hee
    • Journal of Soil and Groundwater Environment
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    • v.17 no.3
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    • pp.49-58
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    • 2012
  • Batch experiments using indigenous and commercialized adventive microorganisms were performed to investigate the feasibility of the biodegradation process for the diesel contaminated soil, which was taken in US Military Camp 'Hialeah', Korea. TPH concentration of the soil was determined as 3,819 mg/kg. Four indigenous microorganisms having high TPH degradation activity were isolated from the soil and by 16S rRNA gene sequence analysis, they were identified as Arthrobacter sp., Burkholderia sp., Cupriavidus sp. and Bacillus sp.. Two kinds of commercialized solutions cultured with adventive microorganisms were also used for the experiments. Various biodegradation conditions such as the amount of microorganism, water content and the temperature were applied to decide the optimal bioavailability condition in the experiments. In the case of soils without additional microorganisms (on the natural attenuation condition), 35% of initial TPH was removed from the soil by inhabitant microorganisms in soil for 30 days. When the commercialized microorganism cultured solutions were added into the soil, their average TPH removal efficiencies were 64%, and 54%, respectively, which were higher than that without additional microorganisms. When indigenous microorganisms isolated from the contaminated soil were added into the soil, TPH removal efficiency increased up to 95% (for Bacillus sp.). According to the calculation of the average biodegradation rates for Bacillus sp., the remediation goal (87% of the removal efficiency: 500 mg/kg) for the soil would reach within 24 days. Results suggested that TPH removal efficiency of biodegradation by injecting indigenous microorganisms is better than those by injecting commercialized adventive microorganisms and only by using the natural attenuation.