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

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Identification of Mycobacterium avium complex (MAC) Clinical Strains to a Species Level by Sequencing and PCR-SSCP Analysis of rpoB DNA (비결핵항산성균의 rpoB DNA 염기서열과 SSCP pattern 분석에 따른 Mycobacterium avium complex (MAC) 임상분리균주의 동정)

  • Kim, Bum-Joon;Lee, Seung-Hyun;Lee, Kuen-Hwa;Park, Chung-Kyu;Choi, Myung-Sik;Kim, Ik-Sang;Choi, Sung-Bai;Hwang, Eung-Su;Cha, Chang-Yong;Kim, Sang-Jae;Bai, Gill-Han;Kook, Yoon-Hoh
    • The Journal of the Korean Society for Microbiology
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    • v.34 no.5
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    • pp.491-500
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    • 1999
  • A recent study showed that comparative sequence analysis of rpoB DNAs could reveal natural relationships in genus Mycobacterium [J Clin Microbial. 37 (6). 1999]. rpoB DNAs showed interspecies variation and intraspecies conservation. Based on these data, we developed polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) protocols which enable species differentiation in genus Mycobacterium. When this assay was applied to 24 clinical isolates identified as M. avium complex (MAC) by biochemical test, these were successfully differentiated into M. avium and M. intracellulare. These results were concordant with those obtained by 16s rDNA analysis. It is the first report that PCR-SSCP analysis of rpoB DNA could be used for species differentiation of MAC strains.

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Isolation and Characterization of a Feather Degrading Alkalophilic Streptomyces sp. TBG-S13A5 and its Keratinolytic Properties

  • Indhuja, Selvaraj;Shiburaj, Sugathan;Pradeep, Nediyaparambu Sukumaran;Thankamani, Vaidyanathan;Abraham, Teruvath Koshy
    • Microbiology and Biotechnology Letters
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    • v.40 no.4
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    • pp.303-309
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    • 2012
  • Keratinases are of particular interest because of their action on insoluble keratins and generally on a broad range of protein substrates. Alkalophilic and neutrophilic actinomycete strains isolated from different soil samples, rich in keratinaceous substances were screened for keratinolytic activity. An alkalophilic isolate, TBG-S13A5, was found to possess good keratinolytic activity and was able to utilize feather as the sole carbon and nitrogen source. TBG-S13A5 exhibited an off-white aerial mass color with a rectus-flexibilis type of spore chain. The morphological, microscopical and biochemical characters were comparable with that of Streptomyces albidoflavus. Fatty acid methyl ester profiling (FAME) and 16S rDNA sequence analysis confirmed its identity as a strain of S. albidoflavus. Under submerged fermentation conditions, maximum protease production was recorded on the $5^{th}$ day of incubation at $30^{\circ}C$, using basal broth of pH 9.0 with 0.25% (w/v) white chicken feather. This strain could affect feather degradation when the initial pH was 8 and above and maximum protease production was recorded when the initial pH was around 10.5. The effectiveness of the crude enzyme in destaining and leather dehairing were also demonstrated.

Probiotic Potential of Indigenous Bacillus sp. BCNU 9028 Isolated from Meju (메주로부터 분리한 토착 Bacillus sp. BCNU 9028의 프로바이오틱스로서 이용 가능성)

  • Shin, Hwa-Jin;Bang, Ji-Hun;Choi, Hye-Jung;Kim, Dong-Wan;Ahn, Cheol-Soo;Jeong, Young-Kee;Joo, Woo-Hong
    • Journal of Life Science
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    • v.22 no.5
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    • pp.605-612
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    • 2012
  • Spore-forming bacteria are being used as probiotic supplements for human and animal use, due to their low pH stability and ability to survive the gastric barrier. In this study, the BCNU 9028 strain was screened from meju, a Korean fermented soybean food starter. Biochemical and physiological characteristics, as well as 16S rDNA sequence analyses, indicate that this strain belongs to the genus $Bacillus$. $Bacillus$ sp. BCNU 9028 showed a 92% survivability at pH 2.5 and could also withstand 0.3% ox bile. Furthermore, it was postulated that $Bacillus$ sp. BCNU 9028 could prevent biofilm formation and adherence of food-borne pathogens such as $Listeria$ $monocytogenes$, $S.$ $aureus$ and $E.$ $coli$ on the basis of its autoaggregation and coaggregation capacity with food-borne pathogens. It was shown that BCNU 9028 has good abilities to adhere to the intestinal tract from its hydrophobic character (63.3%). The $Bacillus$ sp. BCNU 9028 strain especially elicited antibacterial activity against both Gram-positive and -negative pathogens. These findings suggested that the $Bacillus$ sp. BCNU 9028 strain could be used as a potential probiotic.

Characterization and Organic Hydrocarbons Degradation Potential of Euryhaline Marine Microorganism, Bacillus sp. EBW4 Isolated from Polychaete (Perinereis aibuhitensis) (갯지렁이(Perinereis aibuhitensis)에서 분리한 광염성 해양 미생물 Bacillus sp. EBW4의 특성 및 유기물 분해능 분석)

  • Shin, Seyeon;Yundendorj, Khorloo;Lee, Sang-Suk;Lee, Dong-Heon;Kang, Kyoung-Ho;Kahng, Hyung-Yeel
    • Korean Journal of Microbiology
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    • v.49 no.1
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    • pp.38-45
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    • 2013
  • In this study, euryhaline marine microorganism, Bacillus sp. strain EBW4 isolated from polychaete (Perinereis aibuhitensis) of Suncheon Bay was physiologically, biochemically and genetically characterized. Based on 16S rRNA sequence, EBW14 was found to share 98.25% similarity with Bacillus hemicentroti $JSM076093^T$, 97.96% similarity with Bacillus hwajinponensis SW-$72^T$ and 96.28% similarity with B. algicoa $KMM3737^T$, respectively. The temperature range for the growth of strain EBW4 was $4-40^{\circ}C$, NaCl concentration range 0-17% and pH range pH 5-9, revealing that EBW4 was euryhaline bacterium. Major fatty acids in strain EBW4 were composed of anteiso $C_{15:0}$ (48.2%), iso $C_{16:0}$ (12.1%), anteiso $C_{17:0}$ (11.6%) and iso $C_{14:0}$ (9.4%). EBW4 was found to have DNase, amylase, protease and lipase for the degradation of macromolecules such as DNA, carbohydrates, proteins, lipids, etc. The enzyme activities of alkaline phosphatase, esterase (C4), leucine arylamidase and ${\alpha}$-chymotrypsin were also found in strain EBW4. Analysis of the biodegradation ability of EBW4 for organic hydrocarbons under different salinity conditions using synthetic water waste revealed that EBW4 exhibited the ability to degrade organic hydrocarbons very quickly, suggesting strain EBW4 may be a good candidate for the application to various industries.

Isolation, Screening, and Identification of Actinomycetes with Antifungal and Enzyme Activity Assays against Colletotrichum dematium of Sarcandra glabra

  • Song, Lisha;Jiang, Ni;Wei, Shugen;Lan, Zuzai;Pan, Limei
    • Mycobiology
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    • v.48 no.1
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    • pp.37-43
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    • 2020
  • A serious leaf disease caused by Colletotrichum dematium was found during the cultivation of Sarcandra glabra in Jingxi, Rong'an, and Donglan Counties in Guangxi Province, which inflicted huge losses to plant productivity. Biological control gradually became an effective control method for plant pathogens. Many studies showed that the application of actinomycetes in biological control has been effective. Therefore, it may be of great significance to study the application of actinomycetes on controlling the diseases caused by S. glabra. Strains of antifungal actinomycetes capable of inhibiting C. dematium were identified, isolated and screened from healthy plants tissues and the rhizospheres in soils containing S. glabra. In this study, 15 actinomycetes strains were isolated and among these, strains JT-2F, DT-3F, and JJ-3F, appeared to show antagonistic effects against anthracnose of S. glabra. The strains JT-2F and DT-3F were isolated from soil, while JJ-3F was isolated from plant stems. The antagonism rate of strain JT-2F was 86.75%, which was the highest value among the three strains. Additionally, the JT-2F strain also had the strongest antagonistic activity when the antagonistic activities were tested against seven plant pathogens. Strain JT-2F is able to produce proteases and cellulase to degrade the protein and cellulose components of cell walls of C. dematium, respectively. This results in mycelia damage which leads to inhibition of the growth of C. dematium. Strain JT-2F was identified as Streptomyces tsukiyonensis based on morphological traits and 16S rDNA sequence analysis.

Isolation and Characterization of a Protease-Producing Bacterium, Bacillus amyloliquefaciens P27 from Meju as a Probiotic Starter for Fermented Meat Products

  • Lee, Mi-Sun;Lee, Na-Kyoung;Chang, Kyung-Hoon;Choi, Shin-Yang;Song, Chi-Kwang;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.30 no.5
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    • pp.804-810
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    • 2010
  • This study was performed to select protease-producing Bacillus sp. as a potential probiotic starter for fermented meat products. In order to isolate protease-producing bacterium from meju, measured the diameter of the clear zone on agar plate (TSA, 1% (w/v) skim milk) and analyzed for intracellular protease activity, then 10 Bacillus-like strains were isolated. Three Bacillus-like strains (P19, P27, and P33) among 10 strains were able to tolerate in acidic condition (TSB, pH 2.5, 2 h incubation). These 3 strains were showed antimicrobial activity against food-borne pathogenic bacteria. These vegetative cells of 3 strains were showed a survival rate of 0.04% to 0.08% under the artificial gastric acidic condition (TSB, pH 2.5 with 1% (w/v) pepsin), but spore-forming cells were 56.29% to 84.77%. Vegetative cells of 3 strains were the least bile-resistant, while spore-forming cells of 3 strains showed higher survival rate more than 76% under artificial bile condition (TSB, 0.1% (w/v) oxgall bile). In these strains, P27 strain was finally selected as a good probiotic strain. P27 strain was tentatively identified as Bacillus amyloliquefaciens by API CHB kit and 16S rDNA sequence analysis. The results of this study suggest that B. amyloliquefaciens P27 can be used as a potential probiotic starter for fermented meat product.

Selection of the Auxin and ACC Deaminase Producing Plant Growth Promoting Rhizobacteria from the Coastal Sand Dune Plants (Auxin과 ACC Deaminase를 생산하는 사구식물 복원용 근권세균의 선발)

  • Lim, Jong-Hui;Kim, Jong-Guk;Kim, Sang-Dal
    • Microbiology and Biotechnology Letters
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    • v.36 no.4
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    • pp.268-275
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    • 2008
  • In order to develop the multi-functional rhizobacteria that can exert positive effect on the growth of plants growing in the coastal sand dune located along East Coast of Korea, rhizospheral bacteria of 11 different plants from this area were isolated 1,330 rhizobacteria. Among these, 23 strains were able to produce auxin and had spectrum of antagonism toward various phytopathogenic microbes. To know the mechanism of this antifungal activity, these 23 strains were subjected to further analyses; 19 strains of these produced siderophore as determined by color reaction on CAS-blue plate, 4 strains produced antifungal cellulase as judged by color change on CMC-Congo red plate, 17 strains were able to utilized insoluble phosphate salts, also determined by clear zone formation on PVK medium. Identification of the strain was assigned to all 23 strains by l6s rDNA sequence analysed, and all were identified to be in the genus of Bacillus and Pseudomonas. One strain of these, denoted Pseudomonas fluorescens IB4-14, showed ACC deaminase activity which is known to be involved in the resistance of environmental stress such as salt and drought. Also, P. fluorescens IB4-l4 showed the germination stimulation and roots growth promoting activity on the in vivo assay of Lysimachia mauritiana Lam. (spoonleaf yellow loosestrife).

Screening and Identification of Alkaline Protease Inhibitor-Producing Marine-derived Actinomyces. (Alkaline protease inhibitor를 생산하는 해양유래 방선균의 탐색 및 동정)

  • Kang, Sung-Il;Kong, Jai-Yul;Choi, Yeung-Joon;Kim, Min-Yong;Son, Hong-Joo
    • Journal of Life Science
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    • v.18 no.4
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    • pp.482-487
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    • 2008
  • In this study, we screened and identified the bacterial strain showing high alkaline pretense inhibitor activity from marine environment. Nine bacterial strains with alkaline pretense inhibitor activity were isolated from marine sediments. Among them, strain C12 had the highest alkaline pretense inhibitor activity and was selected for further taxonomical study. On the basis of morphological and physiological characteristics, strain C12 was identified as the genus Streptomyces. A phylogenetic analysis of the 165 rDNA showed that the isolated strain was actually a member of the genus Streptomyces, because the determined sequence exhibited a higher homology with Streptomyces thermocarboxydus. Morphological characteristics showed cylindrical spore chain and smooth spore surface by scanning electron microscope. Strain C12 was grown on all media except for ISP 9 agar. This strain could be grown in the medium containing up to 9% NaCl.

Antifungal Activity of Lactic Acid Bacteria Strains Isolated from Natural Honey against Pathogenic Candida Species

  • Bulgasem, Bulgasem Y.;Lani, Mohd Nizam;Hassan, Zaiton;Yusoff, Wan Mohtar Wan;Fnaish, Sumaya G.
    • Mycobiology
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    • v.44 no.4
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    • pp.302-309
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    • 2016
  • The role of lactic acid bacteria (LAB) in honey as antifungal activity has received little attention and their mechanism of inhibitory of fungi is not fully understood. In this study, LAB were isolated from honey samples from Malaysia, Libya, Saudi Arabia, and Yemen. Twenty-five isolates were confirmed LAB by catalase test and Gram staining, and were screened for antifungal activity. Four LAB showed inhibitory activity against Candida spp. using the dual agar overlay method. And they were identified as Lactobacillus plantarum HS isolated from Al-Seder honey, Lactobacillus curvatus HH isolated from Al-Hanon honey, Pediococcus acidilactici HC isolated from Tualang honey and Pediococcus pentosaceus HM isolated from Al-Maray honey by the 16S rDNA sequence. The growth of Candida glabrata ATCC 2001 was strongly inhibited (>15.0 mm) and (10~15 mm) by the isolates of L. curvatus HH and P. pentosaceus HM, respectively. The antifungal activity of the crude supernatant (cell free supernatant, CFS) was evaluated using well diffusion method. The CFS showed high antifungal activity against Candida spp. especially The CFS of L. curvatus HH was significantly (p < 0.05) inhibited growth of C. glabrata ATCC 2001, C. parapsilosis ATCC 2201, and C. tropicalis ATCC 750 with inhibitory zone 22.0, 15.6, and 14.7 mm, respectively. While CFS of P. pentosaceus HM was significantly (p < 0.05) effective against C. krusei, C. glabrata, and C. albicans with inhibition zone 17.2, 16.0, and 13.3 mm, respectively. The results indicated that LAB isolated from honey produced compounds which can be used to inhibit the growth of the pathogenic Candida species.

Community Structure of Arbuscular Mycorrhizal Fungi in the Islands of Chungnam, Korea (충남 섬 지역 근권 토양의 수지상균근균 군집 구조)

  • Lee, Jeong-Youn;Lee, Eun-Hwa;Ka, Kang-Hyeon;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.44 no.1
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    • pp.16-22
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    • 2016
  • Five islands (Sinjindo, Mado, Daenanjido, Wonsando, and Sapsido) and the coastal area (Muchangpo) in Chungnam, Korea, were selected to determine the diversity of arbuscular mycorrhizal (AM) fungi. Soil-inhabiting AM fungi were isolated and identified on the basis of morphological characteristics and sequence analyses of 18s rDNA. The differences in the fungal community structures were compared among sites. As a result, 24 species of AM fungi were identified, of which two species of AM fungi, Acaulospora brasiliensis and Redeckera fulvum, were isolated for the first time in Korea. This study revealed that AM fungal spore abundance was low and the genus Acaulospora was dominant in most of the islands. AM fungal community structures in five Islands were highly similar. However, the coastal area, Muchangpo, had different AM fungal community structure from the islands.