• Title/Summary/Keyword: 16S-rRNA

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Identification and Antioxidant Activity of Marine Actinomycetes Streptomyces sp. ACT-1 (해양방선균 Streptomyces sp. ACT-1의 동정 및 항산화 활성)

  • Kim, Man-Chul;Kim, Ju-Sang;Kim, Yun-Beom;Harikrishnan, Ramasamy;Han, Yong-Jae;Heo, Moon-Soo
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.397-403
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    • 2009
  • For the research of the natural antioxidant from marine sources, an antioxidant-producing marine actinomycetes was isolated from sea water in Jeju coastal area. The strain was identified based on 16S rDNA sequencing, the morphology by a method of scanning electron microscopy, physiological and biochemical characteristics and cellular fatty acid analysis. The isolated strain ACT-1 cell size was $0.5\sim1.0{\mu}m$ and gram positive, aerobic, nonmotile, substrate mycelium are red and gray aerial mycelium. 16S rRNA sequence analysis showed that were Gram-positive bacteria grouped on Streptomyces genus. Results of cellular fatty acid analysis showed that major cellular fatty acids were $C_{15:0}$ anteiso (39.33%), $C_{16:1}$ cis 9 (11.96%), $C_{16:0}$ (13.08%) and $C_{17:0}$ anteiso (10.99%). Finally, strain was identified Streptomyces sp. ACT-1. The antioxidant activity of methanol extract from Streptomyces sp. ACT-1 was evaluated by measuring DPPH, hydroxyl, and alkyl radical scavenging activity using an electron spin resonance (ESR) spectrometer. DPPH radical scavenging activity of SBME-1 (Streptomyces broth methanol extract) was 67% at $1,000{\mu}g$/ml. Hydroxyl radical scavenging activity of SBME-1 was 84% at $500{\mu}g$/ml. Alkyl radical scavenging activity of SBME-1 was 71% at $1,000{\mu}g$/ml.

Isolation and Enzyme Production of a Neutral Protease-Producing Strain, Bacillus sp. DS-1. (Neutral Pretense를 생산하는 Bacillus sp. DS-1 균주의 분리와 효소 생산성)

  • 전대식;강대경;김하근
    • Microbiology and Biotechnology Letters
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    • v.30 no.4
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    • pp.346-351
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    • 2002
  • A bacterium producing the neutral pretense was isolated from soil, and was identified as Bacillus sp. DS-1 by 16S rRNA sequence comparison and biochemical determinations. The production of protease from Bacillus sp. DS-1 was increased 20% and 30% by the additions of 1% glucose and 1% yeast extract, respectively. The optimum pH and temperature for the protease activity were pH 7.0 and 55$^{\circ}C$. Bacillus sp. DS-1 produced a metalloprotease as a major protease in culture medium, since the pretense activity in culture supernatant was inhibited by the presence of 1 mM EDTA significantly.

Purification and Identification of Paenibacillus sp., Isolated from Diseased Larvae of Allomyrina dichotoma (Linnaeus, 1771) (Coleoptera: Scarabaeidae) in Insect Farms

  • Kang, Tae Hwa;Han, Sang Hoon;Weon, Hang Yeon;Lee, Young Bo;Kim, Namjung;Nam, Sung Hee;Park, Hae Chul
    • International Journal of Industrial Entomology and Biomaterials
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    • v.25 no.2
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    • pp.195-203
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    • 2012
  • In reared populations of Allomyrina dichotoma, commercial insects, the skin of last instar larvae was changed softer with opaque white, and infested grubs eventually died. To clarify the cause of the symptom, we collected the larvae of A. dichotoma from five farms and examined their intestinal bacterial florae using pyrosequencing technique. From those results, a member of Paenibacillus was found only in the larvae showing the symptom of disease. Through PCR analysis using a Paenibacillus specific primer set, we obtained the partial 16S rRNA gene sequence and confirmed the microbe as Paenibacillus sp. For clear identification, a whole guts was extracted from each larva showing the sign of the disease and incubated at $70^{\circ}C$ for 15 min to isolate spore forming bacteria. After then, each content of guts was cultured on $MYPGP_{NAL}$ agar medium($12.5{\mu}g/ml$ of nalidixic acid) at $30^{\circ}C$. The 16S rRNA gene sequence analysis for the isolated bacteria showed that they were closely related to P. rigui(97.9% similarity), to P. chinjuensis(96.1% similarity), and to P. soli(95.3% similarity). Additional tests including API test and cellular fatty acid composition analysis were performed, but the strain couldn't be identified at species level, suggesting it may represent novel species of the genus Paenibacillus.

Metagenomic Approach to Identifying Foodborne Pathogens on Chinese Cabbage

  • Kim, Daeho;Hong, Sanghyun;Kim, You-Tae;Ryu, Sangryeol;Kim, Hyeun Bum;Lee, Ju-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.28 no.2
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    • pp.227-235
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    • 2018
  • Foodborne illness represents a major threat to public health and is frequently attributed to pathogenic microorganisms on fresh produce. Recurrent outbreaks often come from vegetables that are grown close to or within the ground. Therefore, the first step to understanding the public health risk of microorganisms on fresh vegetables is to identify and describe microbial communities. We investigated the phyllospheres on Chinese cabbage (Brassica rapa subsp. pekinensis, N = 54). 16S rRNA gene amplicon sequencing targeting the V5-V6 region of 16S rRNA genes was conducted by employing the Illumina MiSeq system. Sequence quality was assessed, and phylogenetic assessments were performed using the RDP classifier implemented in QIIME with a bootstrap cutoff of 80%. Principal coordinate analysis was performed using a weighted Fast UniFrac matrix. The average number of sequence reads generated per sample was 34,584. At the phylum level, bacterial communities were composed primarily of Proteobacteria and Bacteroidetes. The most abundant genera on Chinese cabbages were Chryseobacterium, Aurantimonadaceae_g, Sphingomonas, and Pseudomonas. Diverse potential pathogens, such as Pantoea, Erwinia, Klebsiella, Yersinia, Bacillus, Staphylococcus, Salmonella, and Clostridium were also detected from the samples. Although further epidemiological studies will be required to determine whether the detected potential pathogens are associated with foodborne illness, our results imply that a metagenomic approach can be used to detect pathogenic bacteria on fresh vegetables.

Design, Optimization and Verification of 16S rRNA Oligonucleotide Probes of Fluorescence in-situ Hybridization for Targeting Clostridium spp. and Clostridium kluyveri

  • Hu, Lintao;Huang, Jun;Li, Hui;Jin, Yao;Wu, Chongde;Zhou, Rongqing
    • Journal of Microbiology and Biotechnology
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    • v.28 no.11
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    • pp.1823-1833
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    • 2018
  • Fluorescence in-situ hybridization (FISH) is a common and popular method used to investigate microbial communities in natural and engineered environments. In this study, two specific 16S rRNA-targeted oligonucleotide probes, CLZ and KCLZ, were designed and verified to quantify the genus Clostridium and the species Clostridium kluyveri. The optimal concentration of hybridization buffer solution for both probes was 30% (w/v). The specificity of the designed probes was high due to the use of pellets from pure reference strains. Feasibility was tested using samples of Chinese liquor from the famed Luzhou manufacturing cellar. The effectiveness of detecting target cells appears to vary widely in different environments. In pit mud, the detection effectiveness of the target cell by probes CLZ and KCLZ was 49.11% and 32.14%, respectively. Quantitative analysis by FISH technique of microbes in pit mud and fermented grains showed consistency with the results detected by qPCR and PCR-DGGE techniques, which showed that the probes CLZ and KCLZ were suitable to analyze the biomass of Clostridium spp. and C. kluyveri during liquor fermentation. Therefore, this study provides a method for quantitative analysis of Clostridium spp. and C. kluyveri and monitoring their community dynamics in microecosystems.

The microbial diversity analysis of the Korea traditional post-fermented tea (Chungtaejeon) (한국 전통 미생물발효차(청태전)의 미생물 군집분석)

  • Kim, Byung-Hyuk;Jang, Jong-Ok;Kang, Zion;Joa, Jae Ho;Moon, Doo-Gyung
    • Korean Journal of Microbiology
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    • v.53 no.3
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    • pp.170-179
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    • 2017
  • Tea is the most popular beverage in the world. In fact, there are mainly three different kinds of tea (Green tea, black tea, and post-fermented tea). Post-fermented tea is produced by the microbial fermentation process using sun-dried green tea leaves (Camellia sinensis) as the raw material. Chungtaejeon was a traditional tea introduced in the age of the ancient three states and is the only "Ddeok-cha or Don-cha" culture in the world that survived on the southwestern shore of Republic of Korea. In this study, the structures of the bacterial community involved in the production of oriental traditional post-fermented tea (Chungtaejeon) were investigated using 16S rRNA gene analysis. The 16S rRNA gene analysis of dominant microbial bacteria in post-fermented tea confirmed the presence of Pantoea sp., and Klebsiella oxytoca. Phylogenetic analysis suggested that the taxonomic affiliation of the dominant species in the post-fermented tea was ${\gamma}$-proteobacteria. As a result of the microbial community size analysis, it was confirmed that the size of the microbial communities of Chungtaejeon was the largest compared to other teas

Characterization of Vibrio ponticus KJS1 Isolated from Larvae of Maroon clownfish, Premnas biaculeatus (Maroon clownfish, Premnas biaculeatus 자어로부터 분리한 Vibrio ponticus KJS1의 특성)

  • Kim, Jong-Su;Rho, Sum;Kang, Bong-Jo
    • Journal of fish pathology
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    • v.20 no.1
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    • pp.25-31
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    • 2007
  • Vibriosis is one of the most important diseases affecting marin fish. In this study, a bacterium belonging to the Vibrio were isolated from maroon clownfish (Premnas biaculeatus) larvae (3~7 days after hatching). The bacterium was identified as Vibrio ponticus KJS1 by 16S rRNA gene sequence analysis. The result of biochemical test using the GN2 microplate (BioLog Inc, USA) showed that it has the ability to use 35 substrates including dextrin and sucrose was detected, could not degrade 52 substrates including α-cyclodextrin, and showed the weak ability to use 7 substrates including glycogen. It was sensitive to oxolinic acid, flumequine, ciprofloxacine, ofloxacin, norfloxacin, nalidixic acid, pefloxacin, and doxycycline hydrochloride.

Web-based Research Assistant Tools for Analysis of Microbial Diversity (미생물 다양성 분석을 위한 웹기반의 생물정보도구 개발)

  • Kang, Byeong-Chul;Kim, Hyun-Jin;Park, Jun-Hyung;Park, Hee-Kyung;Kim, Cheol-Min
    • Journal of the Korean Institute of Intelligent Systems
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    • v.14 no.5
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    • pp.545-550
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    • 2004
  • The study of available genotypes (biodiversity analysis) in bacterial communities is of growing importance in several fields such as ecology, environmental technology, clinical diagnostics, etc. These culture-independent genotyping techniques, especially amplifying 16S rRNA genes, attempt to overcome some shortcomings of conventional cultivation method. Biodiversity analysis based on molecular technique were laborious for base-calling chromatogram, trimming primer sites, correcting strand directions, electing representative operation taxonomic units (OTU), etc. Also, biologists wanted intuitively to confirm results of the above processes. For making up these demands, we developed the web application based on Folder-Process-Filter (FPF) modeling with correspondence to classical Model-View-Controller model. The model of web application leads to keep virtues of simplicity and directness for development and management of the stepwise web interfaces. The web application was developed in Perl and CGI on Linux workstation. It can be freely accessed from http://home.pusan.ac.kr/~genome/tools/rat.htm.

Characteristics and Antibiotic Susceptibility of Imipenem-Resistant Clinical Isolates Producing Carbapenemase (Carbapenemase를 생산하는 imipenem 내성 세균의 특성 및 항생제 감수성)

  • Choe, Han-Na;Park, Chul;Kim, Hyung-Rak;Baik, Keun-Sik;Kim, Se-Na;Seong, Chi-Nam
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1214-1220
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    • 2010
  • Imipenem-resistant bacteria were isolated from clinical specimens taken from hospitalized patients in Suncheon, Korea. Fifty-four isolates were phylogenetically analyzed based on 16S rRNA gene and gyrB gene sequence comparisons. Isolates were affiliated with Pseudomonas aeruginosa (30 strains; 55.6%), Acinetobacter baumannii (21; 38.9%), Enterobacter hormaechei (2) and Pseudomonas putida (2). Twenty-two isolates produced metallo-$\beta$-lactamase (MBL); 12 Acinetobacter baumannii strains, 7 Pseudomonas aeruginosa strains, 2 P. putida strains and 1 Enterobacter hormaechei strain. Antibiotic susceptibility of the isolates was determined using the disc diffusion method and Vitek system. Strains producing metallo-$\beta$-lactamase (type IMP & VIM) were more resistant to antibiotics ceftazidime, aztreonam, amikacin and gentamicin than to strains producing OXA and SHV type of $\beta$-lactamase.

Occurrence of Natural Hybrid between Oplegnathus fasciatus and Oplegnathus punctatus from the South Sea of Korea (한국 남해에서 출현한 돌돔 (Oplegnathus fasciatus)과 강담돔 (Oplegnathus punctatus) 사이의 자연교잡종)

  • Kwun, Hyuck-Joon;Kim, Jin-Koo
    • Korean Journal of Ichthyology
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    • v.22 no.3
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    • pp.201-205
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    • 2010
  • One specimen of a natural hybrid of an Oplegnathus (Oplegnathus fasciatus $\times$ Oplegnathus punctatus) was found in Tongyeong, Korea in August 2008. We, herein, describe its morphological and genetic characteristics and compare them with those of O. fasciatus and O. punctatus. In morphology, the hybrid showed many distinctive black rounded blotches on body sides and four faint vertical bars, being in those features similar to O. punctatus. Although the counts and measurements of the hybrid mostly overlapped between O. fasciatus and O. punctatus, the Oplegnathus hybrid resembled O. punctatus in the ratio of pelvic-fin length in standard length: Oplegnathus hybrid (26.7%) was closer to O. punctatus (26.4%) than to O. fasciatus (17.2~23.6%). In genetics, as a result of analysis of 510 base pair sequences of mitochondrial DNA 16S rRNA, the hybrid was closer to O. fasciatus (d=0.000~0.010) than to O. punctatus (d=0.020). Our results suggest that the natural hybridization represented by the subject specimen occurred between an O. fasciatus female and an O. punctatus male.