• Title/Summary/Keyword: 16S rDNA sequences

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Statistical Optimization for Production of Carboxymethylcellulase from Rice Hulls by a Newly Isolated Marine Microorganism Bacillus licheniformis LBH-52 Using Response Surface Method (통계학적인 방법과 왕겨를 기질로 사용하여 해양에서 분리한 Bacillus licheniformis LBH-52 를 사용한 carboxymethylcellualse의 생산조건 최적화)

  • Kim, Hye-Jin;Gao, Wa;Chung, Chung-Han;Lee, Jin-Woo
    • Journal of Life Science
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    • v.21 no.8
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    • pp.1083-1093
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    • 2011
  • A microorganism utilizing rice hulls as a substrate for the production of carboxymethylcellulase (CMCase) was isolated from seawater and identified as Bacillus lincheniformis by analyses of its 16S rDNA sequences. The optimal carbon and nitrogen sources for production of CMCase were found to be rice hulls and ammonium nitrate. The optimal conditions for cell growth and the production of CMCase by B. lincheniformis LBH-52 were investigated using the response surface method (RSM). The analysis of variance (ANOVA) of results from central composite design (CCD) indicated that a highly significant factor ("probe>F" less than 0.0001) for cell growth was rice hulls, whereas those for production of CMCase were rice hulls and initial pH of the medium. The optimal conditions of rice hulls, ammonium nitrate, initial pH, and temperature for cell growth extracted by Design Expert Software were 48.7 g/l, 1.8 g/l, 6.6, and 35.7$^{\circ}C$, respectively, whereas those for the production of CMCase were 43.2 g/l, 1.1 g/l, 6.8, and 35.7$^{\circ}C$. The maximal production of CMCase by B. lincheniformis LBH-52 from rice hulls under optimized conditions was 79.6 U/ml in a 7 l bioreactor. In this study, rice hulls and ammonium nitrate were developed to be substrates for the production of CMCase by a newly isolated marine microorganism, and the time for production of CMCase was reduced to 3 days using a bacterial strain with submerged fermentation.

Bacterial Leaf Spot of English Ivy Caused by Xanthomonas hortorum pv. hederae (Xanthomonas hortorum pv. hederae에 의한 헤데라 세균성점무의병)

  • Lee, Seung-Don;Lee, Jung-Hee;Han, Kyoung-Suk;Seo, Sang-Tae;Kim, Yong-Ki;Heu, Sung-Gi;Ra, Dong-Soo
    • Research in Plant Disease
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    • v.13 no.1
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    • pp.61-65
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    • 2007
  • English ivy (Hedera helix) in Araliaceae family is an evergreen climbing vine. A severe bacterial disease of English ivy was observed and investigated in January 2005. Initial symptoms on the leaves appeared as small water-soaked lesions on the lower surface. As the spots enlarged, the center became brown to brownish black and greenish-brown water-soaked, irregular margins surrounded the center, The spots developed into large irregular blotches, sometimes 5$\sim$10 mm in diameter, then coalesced. Finally, the water-soaked margins raised, dried out, became corky and broke in the center. A bacterial organism, isolated from the advancing margins of the lesions, was tested for its pathogenicity according to the Koch's postulates and biochemical and physiological tests identified the isolated bacterium as a Xanthomonas. The representative Xanthomonas strains (SL4821 and SL4822) isolated from English ivy were compared with a reference strain X. hortorum pv. hederae for fatty acid profiles, metabolic fingerprints and 16s rDNA sequences, showing that all outcomes were indistinguishable between the representative and reference strains. This is the first report of bacterial leaf spot of English ivy in Korea.

Identification and Physiological Characters of Intestinal Bacteria of the Black Soldier Fly, Hermetia illucens (아메리카동애등에 장내세균 동정과 생리적 특징)

  • Kim, Eunsung;Park, Jiyeong;Lee, Sanghoon;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.53 no.1
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    • pp.15-26
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    • 2014
  • The black soldier fly, Hermetia illucens, larvae may depend on indigenous bacteria in the intestine to feed and digest diverse food sources. To prove this hypothesis, we isolated and identified the intestinal bacteria of the black soldier fly for their digestive and antimicrobial abilities. The last instar larvae had long digestive tracts, which were about seven times longer than its body length. An individual of H. illucens larvae possessed a total of $5.0{\pm}10^6$ bacteria in the whole intestine, of which more than 98% bacteria were located in the hindgut. Three different bacterial isolates cultured on nutrient agar (NA) medium were detected in the intestine and identified as Morganella morganii, Providencia rettgeri and Bacillus halodurans by Biolog microbial identification system. Analysis of 16S rDNA sequences of the intestinal bacteria detected the additional bacteria of Proteus mirabilis, Providencia alcalifaciens, and Providencia sp. These intestinal bacteria cultured on NA medium exhibited high resistance to 4 antibiotics and inhibited growth of other microbes which are mainly plant pathogens. Also, these bacteria exhibited catalytic activities to degrade cellulose, lipid, proteins, and carbohydrates. These results suggest that H. illucens larvae possess intestinal bacteria that may play crucial roles in their digestive physiology.

Characterization and Selection of Lactic Acid Bacteria Producing ${\beta}-Galactosidase$ (${\beta}-Galactosidase$ 생산 유산균 선별 및 특성 조사)

  • Lee, Young-Ki;Choi, Susanna;Park, Young-Il;Park, Chan-Sun;Yoon, Byung-Dae;Hwang, Yun-Sik;Kim, Hee-Sik
    • Korean Journal of Microbiology
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    • v.42 no.3
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    • pp.216-222
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    • 2006
  • This study was carried out to select the lactic acid bacteria producing ${\beta}-galactosidase$ (lactase) and investigate the properties of the ${\beta}-galactosidase$. About 100 strains of lactic acid bacteria showing blue colony on the MRS agar medium containing X-gal were isolated from several kinds of Kimchi. Among them, 2 strains were selected as potential ${\beta}-galactosidase$ producers. The selected strains, ET-1 and LA-12, were identified as Lactobacillus fermentum and L. acidophilus, respectively by the analysis of 16S rDNA sequences. They showed relatively high ${\beta}-galactosidase$ activity and cellular viability. Their ${\beta}-galactosidase$ showed the highest activity at $55^{\circ}C$. And the optimum pHs of the enzymes produced by ET-1 and LA-12 were pH 5.5 and pH 7.0, respectively. They were also highly resistant to artificial gastric juice and bile. Two selected strains showed little change of viable cell number for 3 hr incubation in artificial gastric juice, and maintained the viable cell number at $10^8CFU/ml$ for 24 hr in 0.3% oxgall after incubation for 2 hours in artificial gastric juice. Based on these results, ET-1 and LA-12 are expected to be applied in dairy industry.

Diversity and Polymorphism in AHL-Lactonase Gene (aiiA) of Bacillus

  • Huma, Nusrat;Shankar, Pratap;Kushwah, Jyoti;Bhushan, Ashish;Joshi, Jayadev;Mukherjee, Tanmoy;Raju, Sajan C.;Purohit, Hemant J.;Kalia, Vipin Chandra
    • Journal of Microbiology and Biotechnology
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    • v.21 no.10
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    • pp.1001-1011
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    • 2011
  • To explore bacterial diversity for elucidating genetic variability in acylhomoserine lactone (AHL) lactonase structure, we screened 800 bacterial strains. It revealed the presence of a quorum quenching (QQ) AHL-lactonase gene (aiiA) in 42 strains. These 42 strains were identified using rrs (16S rDNA) sequencing as Bacillus strains, predominantly B. cereus. An in silico restriction endonuclease (RE) digestion of 22 AHL lactonase gene (aiiA) sequences (from NCBI database) belonging to 9 different genera, along with 42 aiiA gene sequences from different Bacillus spp. (isolated here) with 14 type II REs, revealed distinct patterns of fragments (nucleotide length and order) with four REs; AluI, DpnII, RsaI, and Tru9I. Our study reflects on the biodiversity of aiiA among Bacillus species. Bacillus sp. strain MBG11 with polymorphism (115Alanine > Valine) may confer increased stability to AHL lactonase, and can be a potential candidate for heterologous expression and mass production. Microbes with ability to produce AHL-lactonases degrade quorum sensing signals such as AHL by opening of the lactone ring. The naturally occurring diversity of QQ molecules provides opportunities to use them for preventing bacterial infections, spoilage of food, and bioremediation.

Xanthomonas euvesicatoria Causes Bacterial Spot Disease on Pepper Plant in Korea

  • Kyeon, Min-Seong;Son, Soo-Hyeong;Noh, Young-Hee;Kim, Yong-Eon;Lee, Hyok-In;Cha, Jae-Soon
    • The Plant Pathology Journal
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    • v.32 no.5
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    • pp.431-440
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    • 2016
  • In 2004, bacterial spot-causing xanthomonads (BSX) were reclassified into 4 species-Xanthomonas euvesicatoria, X. vesicatoria, X. perforans, and X. gardneri. Bacterial spot disease on pepper plant in Korea is known to be caused by both X. axonopodis pv. vesicatoria and X. vesicatoria. Here, we reidentified the pathogen causing bacterial spots on pepper plant based on the new classification. Accordingly, 72 pathogenic isolates were obtained from the lesions on pepper plants at 42 different locations. All isolates were negative for pectolytic activity. Five isolates were positive for amylolytic activity. All of the Korean pepper isolates had a 32 kDa-protein unique to X. euvesicatoria and had the same band pattern of the rpoB gene as that of X. euvesicatoria and X. perforans as indicated by PCR-restriction fragment length polymorphism analysis. A phylogenetic tree of 16S rDNA sequences showed that all of the Korean pepper plant isolates fit into the same group as did all the reference strains of X. euvesicatoria and X. perforans. A phylogenetic tree of the nucleotide sequences of 3 housekeeping genes-gapA, gyrB, and lepA showed that all of the Korean pepper plant isolates fit into the same group as did all of the references strains of X. euvesicatoria. Based on the phenotypic and genotypic characteristics, we identified the pathogen as X. euvesicatoria. Neither X. vesicatoria, the known pathogen of pepper bacterial spot, nor X. perforans, the known pathogen of tomato plant, was isolated. Thus, we suggest that the pathogen causing bacterial spot disease of pepper plants in Korea is X. euvesicatoria.

Molecular Identification and Morphological Descriptions of the Eggs, Larvae and Juvenile of the Previously Unrecorded Species Acanthaphritis unoorum (Perciformes, Percophidae) in Korean Waters (한국산 1미기록종, Acanthaphritis unoorum (농어목, 꼬리점눈퉁이과)의 어란 및 자치어의 분자동정 및 형태기재)

  • Heo, Sung-Hyun;Ban, Tae Woo;Kim, Jin-Koo;Ji, Hwan-Sung;Moon, Seong Yong
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.52 no.1
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    • pp.67-73
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    • 2019
  • We presented detailed morphological descriptions of the eggs, larvae and juvenile of Acanthaphritis unoorum based on specimens collected with bongo nets from Korean waters during the period May 2017-July 2018. We collected 18 individuals including eggs (n= 4, 0.77-0.85 mm in egg diameter), preflexion larvae (n= 6, 4.11-6.31 mm in standard length, SL), flexion larvae (n= 4, 6.60-7.82 mm SL), postflexion larvae (n= 3, 8.94-13.46 mm SL), and one juvenile (n= 1, 14.67 mm SL). The mitochondrial (mt) DNA 16S rRNA sequences of the eggs, and the cytochrome c oxidase subunit I (COI) sequences of the larvae were identical to those of A. unoorum adults (genetic distances <0.01). The A. unoorum larvae and the juvenile that we collected were morphologically similar to those of Dactylopsaron dimorphicum, but the A. unoorum specimens were readily distinguishable by the presence of lateral melanophores. This is the first record of A. unoorum in Korean waters. We propose a new Korean name for A. unoorum: "O-ri-bu-ri-nuntung-i".

Isolation and Characterization of Six Microorganisms from the Digestive Tract of the Cricket Gryllus bimaculatus (쌍별귀뚜라미(Gryllus bimaculatus) 소화기관에서 분리한 6종류의 특성규명)

  • Kwon, Kisang;Lee, Eun Ryeong;Yoo, Bo-Kyung;Ko, Young Hwa;Shin, Hyojung;Choi, Ji-Young;Kwon, O-Yu
    • Journal of Life Science
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    • v.27 no.9
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    • pp.1040-1046
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    • 2017
  • We describe the isolation and characterization of six different intestinal microorganisms from the digestive tract of the cricket Gryllus bimaculatus. Based on 16S rRNA gene sequences, we obtained six isolates belonging to four different genera: Staphylococcus, Bacillus, Citrobacter, and Proteus. All the isolates were resistant to ampicillin. Ampicillin is an irreversible inhibitor of the enzymeetranspeptidase, which is needed to make bacterial cell walls. None of the isolates were resistant to kanamycin, which binds to the 30S subunit of the bacterial ribosome and then inhibits total protein synthesis. Gram staining was conducted, in addition to morphological classification under a microscope. Four grampositive isolates and two gram-negative isolates were detected. The gram-positive isolates were GL1 (round shaped, 2 am in diameter), GL2 (rod shaped, $2.5{\mu}m$ in length), GL3 (rod shaped, $2{\mu}m$ in length), and GL4 (round shaped, $1.5{\mu}m$ in diameter). The gram-negative isolates were GL5 (rod shaped, $2{\mu}m$ in length) and GL6 (rod-shaped, $2.5{\mu}m$ in length). Notably, two of the isolates, GL2 and GL4, secreted specific extracellular proteins. These were determined by MALDI-TOF-MS spectral analysis to be a 87 kDa collagenase, 56 kDa hypothetical protein, and 200 kDa hypothetical protein. The six isolates in this study could be used for various biotechnological applications and pest management, both in the field and in greenhouse systems. In addition, it would be interesting to determine the relationship between these isolates and their host.

Effects of Diverse Water Pipe Materials on Bacterial Communities and Water Quality in the Annular Reactor

  • Jang, Hyun-Jung;Choi, Young-June;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.21 no.2
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    • pp.115-123
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    • 2011
  • To investigate the effects of pipe materials on biofilm accumulation and water quality, an annular reactor with the sample coupons of four pipe materials (steel, copper, stainless steel, and polyvinyl chloride) was operated under hydraulic conditions similar to a real plumbing system for 15 months. The bacterial concentrations were substantially increased in the steel and copper reactors with progression of corrosion, whereas those in stainless steel (STS) and polyvinyl chloride (PVC) reactors were affected mainly by water temperature. The heterotrophic plate count (HPC) of biofilms was about 100 times higher on steel pipe than other pipes throughout the experiment, with the STS pipe showing the lowest bacterial number at the end of the operation. Analysis of the 16S rDNA sequences of 176 cultivated isolates revealed that 66.5% was Proteobacteria and the others included unclassified bacteria, Actinobacteria, and Bacilli. Regardless of the pipe materials, Sphingomonas was the predominant species in all biofilms. PCR-DGGE analysis showed that steel pipe exhibited the highest bacterial diversity among the metallic pipes, and the DGGE profile of biofilm on PVC showed three additional bands not detected from the profiles of the metallic materials. Environmental scanning electron microscopy showed that corrosion level and biofilm accumulation were the least in the STS coupon. These results suggest that the STS pipe is the best material for plumbing systems in terms of the microbiological aspects of water quality.

Detection of Denitrifying Bacteria in Groundwater by PCR (PCR을 이용한 지하수 내의 탈질화 세균의 검출)

  • Shin, Kyu-Chul;Suh, Mi-Yeon;Han, Myung-Soo;Choi, Yong-Keel
    • Korean Journal of Environmental Biology
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    • v.19 no.4
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    • pp.321-324
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    • 2001
  • Groundwater samples were collected at 6 sites in Seoul area. DNA extraction from the sample was performed by the boiling method. Samples were boiled with guanidinium thyocyanate and phenol-chloroform. One set of primer was designed for amplification of 16S rDNA. For detection of denitrifying bacteria in groundwater sample, the author used primer sets consensus regions in gene sequences encoding the two forms of nitrite reductase (NIR), a key enzyme in the denitrification pathway. Two sets of PCR primer were designed to amplify $cd_1$-and Cu-nir. We confirmed the existence of denitrifying bacteria in 3 sites using $cd_1$-nir primer and in 4 sites using Cu-nir primer.

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