• Title/Summary/Keyword: rRNA sequence

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Examination of Antimicrobial Activity by Phaeobacter inhibens KJ-2 Isolated from a Marine Organism (해양 생물에서 분리된 Phaeobacter inhibens KJ-2의 항균 활성)

  • Kim, Yun-Beom;Kim, Dong-Hwi;Heo, Moon-Soo
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1161-1167
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    • 2017
  • In order to find a new antimicrobial bacterium, we performed screening for antimicrobial activity of bacteria isolated from the eggs of a sea hare. The newly identified strain was designated as Phaeobacter inhibens KJ-2, based on the biochemical characterization and 16S rRNA gene sequence analysis. A colony of P. inhibens KJ-2 showed a circular and ruler-like smooth form at the edge, and a brown color. However, when maintained with a longer incubation time, its coloring was transformed into dark brown. From the result of SEM, P. inhibens KJ-2 is a bacillus which has a length of $0.8{\sim}1.0{\mu}m$ and a width of $0.4{\sim}0.6{\mu}m$. The optimal growth and antimicrobial activity were observed by shaking the culture for 24 hr at $20^{\circ}C$, which showed potent activity against pathogenic bacteria including Vibrio logei, Vibrio campbellii, Vibrio mimicus, Vibrio vulnificus, and Vibrio salmonicida. The antimicrobial activity was proportional to the amount of produced acylated homoserine lactones (AHLs). Therefore, we suggest that production of antimicrobial materials from P. inhibens KJ-2 is regulated by Quorum sensing (QS).

Isolation of Aeromonas sobria Containing Hemolysin Gene from Arowana (Scleropages formosus) (Arowana(Scleropages formosus)에서 Hemolysin Gene을 지닌 Aeromonas sobria 분리 및 특성)

  • Jun, Jin-Woo;Kim, Ji-Hyung;Casiano, Choresca Jr.;Dennis, K. Gomez;Shin, Sang-Phil;Han, Jee-Eun;Park, Se-Chang
    • Journal of Veterinary Clinics
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    • v.27 no.1
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    • pp.62-65
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    • 2010
  • Arowana (Scleropages formosus) is the most valuable group of ornamental fishes and very much in demand in the ornamental fish trade and commands high price ranging from hundreds to thousands of dollars per fish. In this paper, we described a case of mortality of arowana from a private aquarium in Korea. A bacterial pathogen from fish organs (brain, kidney, liver) was cultured, identified and confirmed using Vitek System 2, API 20E test, multiplex PCR and 16S rRNA gene sequencing. The morphological and biochemical properties of the bacterium isolated from the brain, kidney and liver of the fish were similar to Aeromonas sobria. Positive amplification products using the multiplex PCR assay for detection of A. sobria were obtained from these organs. The 16S rRNA gene of the isolates from fish was identical and exhibited 100% sequence similarity with A. sobria (AY987762.1) strain available from GenBank. This bacterium contained hemolysin gene, a virulence factor that plays an important role in outbreaks of disease and is pathogenic to humans as well as in fish. Although this opportunistic bacterium was isolated from a fish without any external symptoms, this pathogen may act as a reservoir and enhance chances of zoonosis to human such as during handling.

Purification and Characterization of the Bacteriocin Produced by Lactococcus sp. KD 28 Isolated from Kimchi (김치에서 분리한 Lactococcus lactis가 생산하는 박테리오신의 정제 및 특성)

  • Lee, Ji-Young;Choi, Nack-Shick;Chun, Sung-Sik;Moon, Ja-Young;Kang, Dae-Ook
    • Journal of Life Science
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    • v.25 no.2
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    • pp.180-188
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    • 2015
  • The bacterial strain isolated from Kimchi showed antibacterial activity against Micrococcus luteus IAM 1056. The selected strain was identified as Lactococcus lactis by 16S rRNA nucleotide sequence analysis and named as Lactococcus sp. KD 28. The treatment of culture supernatant with proteinase K removed antibacterial activity, indicating its proteinaceous nature, a bacteriocin. This bacteriocin was sensitive to hydrolytic enzymes such as ${\alpha}$-chymotrypsion, trypsin, proteinase K, lipase, ${\alpha}$-amylase and subtilisin A. The bacteriocin was highly thermostable and resistant to heating at $80^{\circ}C$ for up to an hour but 50 % of the total activity was remained at $100^{\circ}C$ for 30 min. The pH range from 2.0 to 8.0 had no effect on bacteriocin activity and it was not affected by solvents such as acetonitrile, isopropanol, methanol, chloroform and acetone up to 50% concentration. The bacteriocin showed antibacterial activity against M. luteus IAM 1056, Lactobacillus delbrueckii subsp. lactis KCTC 1058, Enterococcus faecium KCTC 3095, Bacillus cereus KCTC 1013, B. subtilis KCTC 1023, Listeria ivanovii subsp. ivanovii KCTC 3444, Staphylococcus aureus subsp. aureus KCTC 1916, B. megaterium KCTC 1098 and B. sphaericus KCTC 1184. The bacteriocin was purified through ammonium sulfate concentration, SP-Sepharose chromatography and RP-HPLC. The molecular weight was estimated to be about 3.4 kDa by tricine-SDS-PAGE analysis.

Analysis of Rhizosphere Soil Bacterial Communities on Seonginbong, Ulleungdo Island (울릉도 성인봉의 근권 토양 세균군집 분석)

  • Nam, Yoon-Jong;Yoon, Hyeokjun;Kim, Hyun;Kim, Jong-Guk
    • Journal of Life Science
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    • v.25 no.3
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    • pp.323-328
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    • 2015
  • The study of microbial diversity and richness in soil samples from a volcanic island named Ulleungdo, located east of South Korea. The soil bacterial communities on the Ulleungdo were analyzed using pyrosequencing method based on 16S rRNA gene. There were 1,613 operational taxonomic units (OUT) form soil sample. From results of a BLASTN search against the EzTaxon-e database, the validated reads (obtained after sequence preprocessing) were almost all classified at the phylum level. Proteobacteria was the most dominant phylum with 48.28%, followed by acidobacteria (26.30%), actionbacteria (6.89%), Chloroflexi (4.58), Planctomycetes (4.56%), Nitrospirae (1.83%), Bacteroidetes (1.51%), Verrucomicrobia (1.48%), and Gemmatimonadetes (1.11%). α-proteobacteria was the most dominant class with 36.07% followed by Acidobacteria_c (10.65%), Solibacteres (10.64%), δ-proteobacteria (4.42%), γ-proteobacteria (4.29%), Planctomycetacia (4.16%), Actinobacteria_c (4.00%), Betaproteobacteria (3.50%), EU686603_c (2.97%), Ktedonobacteria (2.91%), Acidimicrobiia (1.32%), Verrucomicrobiae (1.27%), Gemmatimonadetes_c (1.11%), Sphingobacteria (1.09%), and GU444092_c (1.06%). Bradyrhizobiaceae was the most dominant family with 22.83% followed by Acidobacteriaceae (10.62%), EU445199_f (5.72%), Planctomycetaceae (4.03%), Solibacteraceae (3.63%), FM209092_f (3.58%), Steroidobacter_f (2.81%), EU686603_f (2.73%), Hyphomicrobiaceae (2.33%), Ktedonobacteraceae (1.75%), AF498716_f (1.46%), Rhizomicrobium_f (1.03%), and Mycobacteriaceae (1.01%). Differences in the diversity of bacterial communities have more to do with geography than the impact on environmental factors and also the type of vegetation seems to affect the diversity of bacterial communities.

Microbiological and chemical properties of sourdough fermented with probiotic lactic acid bacteria (프로바이오틱 유산균으로 제조한 사워도우의 미생물학적 및 이화학적 특성)

  • Lim, Eun-Seo
    • Korean Journal of Microbiology
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    • v.52 no.1
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    • pp.84-97
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    • 2016
  • Isolates from Korean fermented soybean paste were identified as Enterococcus faecium SBP12, Pediococcus halophilus SBP20, Lactobacillus fermentum SBP33, Leuconostoc mesenteroides SBP37, Pediococcus pentosaceus SBP41, Lactobacillus brevis SBP49, Lactobacillus acidophilus SBP55, and Enterococcus faecalis SBP58 according to conventional morphological and biochemical characteristics, carbohydrate fermentation profiling, and 16S rRNA sequence comparison. Strain SBP20, SBP33, SBP49, and SBP55 showed very resistance to simulated gastric and intestinal juices with final populations exceeding 6 log CFU/ml, whereas cells of SBP12 and SBP58 after exposure to low pH were dramatically decreased within 2 h. Among 4 strains having good tolerance to gastrointestinal conditions, the high adhesive ability to HT-29 cells, antibiotic resistance, and antimicrobial activity against food-borne pathogens Bacillus cereus ATCC 11778 and Staphylococcus aureus ATCC 6538 were observed with SBP49 and SBP55, therefore, these two strains were confirmed as putative probiotic candidates. There was no significant difference between the sourdoughs fermented with SBP49 and SBP55 with respect to the values of pH, total titratable acidity, and viable cell count. During sourdough fermentation, SBP49 strain produced significantly greater amounts of lactic acid than SBP55 strain, which secreted large quantities of hydrogen peroxide. SBP49 and SBP55 strains producing the antimicrobial substances such as lactic acid, hydrogen peroxide, and bacteriocin effectively inhibited B. cereus and S. aureus inoculated in the sourdough.

Biodiversity and Phylogenetic Analysis of Streptomyces Collected from Bamboo Forest Soil (대나무 산림토양으로부터 수집한 Streptomyces 속 방선균의 계통학적 다양성)

  • Lee, Hyo-Jin;Whang, Kyung-Sook
    • Korean Journal of Microbiology
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    • v.46 no.3
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    • pp.262-269
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    • 2010
  • To investigate a quantitative evaluation of the actinobacteria, we have collected samples from various kinds of bamboo forest soil. Each different layers contained $2.7{\times}10^6-2.7{\times}10^8$ CFU/g of actinobacteria which was the highest in litter layers of Sasa boreali forest soil. We obtained 330 actinobacteria from different layers of bamboo forest soil; litter (100 strains), humus (70 strains), and rhizosphere soil (160 strains). Based on the colony morphology (aerial mycelium, substrate mycelium, and soluble pigment), isolates were divided into thirty-six groups and we selected 50 representative isolates. 16S rRNA gene sequence analysis showed Streptomyces was major actinobacteria (94%) and they were categorized as cluster I (2 strains), II (35 strains), III (6 strains), and IV (7 strains), respectively. The diversity index of 50 Streptomyces collected from the bamboo forest soil was calculated with the Shannon-Wiener method. Bamboo litter showed higher diversity index level of 3.33 than that of humus and rhizosphere soil. Also, antibiotic activities of our isolates were investigated against Botrytis cinerea, Xanthomonas campestris, Xanthomonas axonopodis pv. vesicatoria, and Bacillus cereus and found in 74, 16, 25, and 24 strains, respectively.

Isolation of bacteriophage-resistant Pseudomonas tolaasii strains and their pathogenic characters (박테리오파지 저항성을 갖는 Pseudomonas tolaasii 변이주 분리 및 이들의 병원특성)

  • Park, Soo-Jin;Han, Ji-Hye;Kim, Young-Kee
    • Journal of Applied Biological Chemistry
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    • v.59 no.4
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    • pp.351-356
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    • 2016
  • Bacterial blotch caused by Pseudomonas tolaasii is one of the major diseases of oyster mushroom, Pleurotus ostreatus. Application of bacteriophages is a very useful tool to decrease the density of pathogens and it has been successful to making disease-free cultivation area, known as phage therapy. Effect of phages on pathogen sterilization is very limited to the specific host strains. Minor variations of the host strains may cause changes in phage sensitivity. The phage-resistant strains of P. tolaasii were isolated and their pathogenic characters were investigated to improve the effectiveness of phage therapy. In the phylogenetic analysis, both phage-resistant strains and the corresponding host strains were identical based on the sequence comparison of 16S rRNA genes. The pathogenic characters, such as hemolytic activity and brown blotch formation, were measured on the phage-resistant strains and no correlation between phage-resistance and pathogenic characters was observed. Nevertheless, pathogenic characters were sometimes changed in the phage-resistant strains depending on the host strains. In order to make the phage therapy successful, the bacteriophages having a wide host range should be isolated.

Isolation and Characterization of Marine Bacterial Strain SH-1 Producing Agar-Degrading Enzymes (한천 분해효소를 생산하는 해양 미생물 SH-1의 분리 및 특성 분석)

  • Lee, Jae-Hag;Lee, Soon-Youl
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.324-330
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    • 2014
  • A marine bacterial strain producing agar-degrading enzymes was isolated from a mud flat in Jeboo-do (Korea) using a selective artificial sea water (ASW) agar plate containing agar as the sole carbon source. The isolate, designated as SH-1, was gram-negative, aerobic, and motile with single polar flagellum. 16S rRNA gene sequence similarity analysis showed the isolate SH-1 had the highest homology (96.5%) to marine bacterium Neiella marina J221. Cells could grow at $28-37^{\circ}C$ but not at $42^{\circ}C$, and the agarase activity of the cell culture supernatant was higher when grown at $28^{\circ}C$ than when grown at $37^{\circ}C$. Cells could grow when concentrations of 1-5% (w/v) NaCl were added to the growth media with the best growth observed at 3% NaCl, and the agardegrading enzyme activity of the cell culture supernatant was best when grown at 3% NaCl-containing growth media under the conditions we examined. The crude enzyme prepared from 48-h culture broth of strain SH-1 exhibited an optimum pH and temperature for agar-degrading activity at 7.0 and $40^{\circ}C$, respectively. Zymogram analysis of the crude supernatant and cell extract showed that strain SH-1 produced at least 3 agar-degrading enzymes with molecular weights of 15, 35, and 52 KD. Thinlayer chromatography (TLC) analysis also suggested that HS-1 produces ${\beta}$-agarase to degrade agarose to neoagarooligosaccharides.

Isolation and Identification of a Marine Bacterium, Pseudomonas sp. BK1 Producing Extracellular Enzymes Capable of Decomposing Multiple Complex Polysaccharides (복합 다당류 분해 효소들을 생산하는 해양미생물 Pseudomonas sp. BK1의 분리 및 특성)

  • Kim, Beom-Kyu;Jeon, Beong-Sam;Cha, Jae-Young;Park, Jeong-Won;Kim, Sam-Woong;Kim, Ji-Yoon;Park, Yong-Lark;Cho, Young-Su;Song, Jae-Young
    • Journal of Life Science
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    • v.13 no.6
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    • pp.871-878
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    • 2003
  • A marine bacterium (strain BKl) that produces extracellular enzymes capable of decomposing complex polysac-charides, such as agar, chitin, carboxymethylcellulose, xylan and mannan, was isolated from the marine red alga Porphyra dentata. Strain BKl was gram-negative, aerobic, catalase- and oxidase-positive, polarly flagellated bacilli that produce gelatinase and urease, but not decarboxylases. The G+C content of the DNA was 51.6 mol%. The major isoprenoid quinone component was identified as an ubiquinone-8, and the major cellular fatty acids were C16:0, C16:1 w6c and C18:1 w7c. Comparative 16S rRNA sequence analysis placed strain BK1 with members of the genus Pseudomonas. On the basis of phenotypic and genotypic data, the strain BK1 was shown to be a member of the subgroup of Pseudomonas, and named as Pseudomonas sp. BK1.

Characterization of Biogenic Amine-reducing Pediococcus pentosaceus Isolated from Traditionally Fermented Soybean Products (전통 장류에서 분리한 Biogenic Amines 저감 유산균 Pediococcus pentosaceus의 분리 및 특성)

  • Oh, HyeonHwa;Ryu, MyeongSeon;Heo, Jun;Jeon, SaeBom;Kim, Young Sang;Jeong, DoYoun;Uhm, Tai-Boong
    • Korean Journal of Microbiology
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    • v.50 no.4
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    • pp.319-326
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    • 2014
  • Two bacterial strains, named as LE17 and LE22, were isolated from traditionally fermented soybean products in order to select lactic acid bacteria for the reduction of biogenic amines and harmful bacteria. Both strains were identified as Pediococcus pentosaceus by 16S rRNA sequence analysis and additional biochemical tests. The strain LE17 reduced the amines by 13.7% for histamine and by 25.9% for tyramine, when it grew in minimal synthetic media containing 0.1% (w/v) histamine and 0.1% tyramine at $30^{\circ}C$ for 48 h, while the strain LE22 reduced the amines by 23.7% for histamine and by 15.7% for tyramine. Both strains also had broad inhibition spectra against pathogens. Considering their properties, they could be used as starters for industrial soybean fermentation.