• Title/Summary/Keyword: strain characterization and identification

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Characterization of Extracellular \alpha-galactosidase Produced by Streptomyces sp. YB-4. (균체외 \alpha-galactosidase를 생산하는 Streptomyces sp. YB-4의 분리 및 효소 특성)

  • 김소영;조기행;김창진;박동진;윤기홍
    • Microbiology and Biotechnology Letters
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    • v.30 no.4
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    • pp.332-338
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    • 2002
  • A strain YB-4 producing the extracellular $\alpha$-galactosidase was isolated from soil, and has been identified as Streptomyces sp. on the basis of its cultural, morphological and physiological properties. The partially purified $\alpha$-galactosidase was most active on paranitrophenyl-$\alpha$-D-galactopyranoside at pH 6.0 and 6$0^{\circ}C$. The enzyme retained 90% of its maximum activity between pH 4.0 and pH 10.0 after pre-incubation for 1 h. The enzyme was able to hydrolyze oligomeric substrates such as melibiose, raffinose and stachyose to liberate galactose residue, indicating that the $\alpha$-galactosidase of Steptomyces sp. YB-4 hydrolyzed $\alpha$-1,6 linkage.

Identification and Characterization of Aeromonas hydrophila Producing Nitrification Capability (질산화 작용이 있는 Aeromonas hydrophila의 동정 및 특성)

  • 엄미나;장재철;유영희;지의상
    • The Korean Journal of Food And Nutrition
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    • v.13 no.6
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    • pp.611-618
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    • 2000
  • For the purpose of the isolation of microorganisms which have the capability of nitrification, we isolated the microorganisms in 6 samples collected from the stream of Kyonggi area. 60 strains were isolated. The selected strain were identified as a Aeromonas hydrophila based on the data obtained from the morphological, biochemical and cultural characteristics defined experiments. Among them Aeromonas hydrophila (AH-1), (AH-3) , (AH-4), (AH-6) showed the highest nitrification capability. All isolates were resistant to amoxillin, ampicillin, cephalothin and ticarcillin. Optimum culture conditions of isolates were 37$^{\circ}C$ and 1${\times}$10$\^$8/ cells/ml for 4 hours in the nitrate medium.

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Isolation and identification of mammalian orthoreovirus type 3 from a Korean roe deer (Capreolus pygargus)

  • Yang, Dong-Kun;An, Sungjun;Park, Yeseul;Yoo, Jae Young;Park, Yu-Ri;Park, Jungwon;Kim, Jong-Taek;Ahn, Sangjin;Hyun, Bang-Hun
    • Korean Journal of Veterinary Research
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    • v.61 no.2
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    • pp.13.1-13.8
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    • 2021
  • Mammalian reovirus (MRV) causes respiratory and intestinal disease in mammals. Although MRV isolates have been reported to circulate in several animals, there are no reports on Korean MRV isolates from wildlife. We investigated the biological and molecular characteristics of Korean MRV isolates based on the nucleotide sequence of the segment 1 gene. In total, 144 swabs from wild animals were prepared for virus isolation. Based on virus isolation with specific cytopathic effects, indirect fluorescence assays, electron microscopy, and reverse transcription-polymerase chain reaction, only one isolate was confirmed to be MRV from a Korean roe deer (Capreolus pygargus). The isolate exhibited a hemagglutination activity level of 16 units with pig erythrocytes and had a maximum viral titer of 105.7 50% tissue culture infectious dose (TCID50)/mL in Vero cells at 5 days after inoculation. The nucleotide and amino-acid sequences of the partial segment S1 of the MReo2045 isolate were determined and compared with those of other MRV strains. The MReo2045 isolate had nucleotide sequences similar to MRV-3 and was most similar (96.1%) to the T3/Bat/Germany/342/08 strain, which was isolated in Germany in 2008. The MReo2045 isolate will be useful as an antigen for sero-epidemiological studies and developing diagnostic tools.

Identification of Glycine max Genes Expressed in Response to Soybean mosaic virus Infection

  • Jeong, Rae-Dong;Lim, Won-Seok;Kwon, Sang-Wook;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.21 no.1
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    • pp.47-54
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    • 2005
  • Identification of host genes involved in disease progresses and/or defense responses is one of the most critical steps leading to the elucidation of disease resistance mechanisms in plants. Soybean mosaic virus (SMV) is one of the most prevalent pathogen of soybean (Glycine max). Although the soybeans are placed one of many important crops, relatively little is known about defense mechanism. In order to obtain host genes involved in SMV disease progress and host defense especially for virus resistance, two different cloning strategies (DD RT-PCR and Subtractive hybridization) were employed to identify pathogenesis- and defenserelated genes (PRs and DRs) from susceptible (Geumjeong 1) and resistant (Geumjeong 2) cultivars against SMV strain G7H. Using these approaches, we obtained 570 genes that expressed differentially during SMV infection processes. Based upon sequence analyses, differentially expressed host genes were classified into five groups, i.e. metabolism, genetic information processing, environmental information processing, cellular processes and unclassified group. A total of 11 differentially expressed genes including protein kinase, transcription factor, other potential signaling components and resistant-like gene involved in host defense response were selected to further characterize and determine expression profiles of each selected gene. Functional characterization of these genes will likely facilitate the elucidation of defense signal transduction and biological function in SMV-infected soybean plants.

Molecular Characterization of Protease Producing Idiomarina Species Isolated from Peruvian Saline Environments

  • Flores-Fernandez, Carol N.;Chavez-Hidalgo, Elizabeth;Santos, Marco;Zavaleta, Amparo I.;Arahal, David R.
    • Microbiology and Biotechnology Letters
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    • v.47 no.3
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    • pp.401-411
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    • 2019
  • All Idiomarina species are isolated from saline environments; microorganisms in such extreme habitats develop metabolic adaptations and can produce compounds such as proteases with an industrial potential. ARDRA and 16S rRNA gene sequencing are established methods for performing phylogenetic analysis and taxonomic identification. However, 16S-23S ITS is more variable than the 16S rRNA gene within a genus, and is therefore, used as a marker to achieve a more precise identification. In this study, ten protease producing Idiomarina strains isolated from the Peruvian salterns were characterized using biochemical and molecular methods to determine their bacterial diversity and industrial potential. In addition, comparison between the length and nucleotide sequences of a 16S-23S ITS region allowed us to assess the inter and intraspecies variability. Based on the 16S rRNA gene, two species of Idiomarina were identified (I. zobellii and I. fontislapidosi). However, biochemical tests revealed that there were differences between the strains of the same species. Moreover, it was found that the ITS contains two tRNA genes, $tRNA^{Ile(GAT)}$ and $tRNA^{Ala(TGC)}$, which are separated by an ISR of a variable size between strains of I. zobellii. In one strain of I. zobellii (PM21), we found nonconserved nucleotides that were previously not reported in the $tRNA^{Ala}$ gene sequences of Idiomarina spp. Thus, based on the biochemical and molecular characteristics, we can conclude that protease producing Idiomarina strains have industrial potential; only two I. zobellii strains (PM48 and PM72) exhibited the same properties. The differences between the other strains could be explained by the presence of subspecies.

Characterization of a Phenazine and Hexanoyl Homoserine Lactone Producing Pseudomonas aurantiaca Strain PB-St2, Isolated from Sugarcane Stem

  • Mehnaz, Samina;Baig, Deeba Noreen;Jamil, Farrukh;Weselowski, Brian;Lazarovits, George
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1688-1694
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    • 2009
  • A novel strain of fluorescent pseudomonad (PB-St2) was isolated from surface-sterilized stems of sugarcane grown in Pakistan. The bacterium was identified as Pseudomonas aurantiaca on the basis of 16S rRNA gene sequence analysis and results from physiological and biochemical characteristics carried out with API50 CH and QTS 24 bacterial identification kits. Assays using substrate-specific media for enzymes revealed lipase and protease activities but cellulase, chitinase, or pectinase were not detected. The bacterium was unable to solubilize phosphate or produce indole acetic acid. However, it did produce HCN, siderophores, and homoserine lactones. In dual culture assays on agar, the bacterium showed antifungal activity against an important pathogen of sugarcane in Pakistan, namely Colletotrichum falcatum, as well as for pathogenic isolates of Fusarium oxysporium and F. lateritium but not against F. solani. The antifungal metabolites were identified using thin-layer chromatography, UV spectra, and MALDI-TOFF spectra and shown to be phenazine-1-carboxylic acid (PCA), 2-hydroxyphenazine (2-OH-PHZ), and N-hexanoyl homoserine lactone (HHL) (assessed using only TLC data). The capacity of this bacterium to produce HCN and 2-OH-PHZ, as well as to inhibit the growth of C. falcatum, has not been previously reported.

Enhanced Production of Astaxanthin in Paracoccus haeundaensis Strain by Physical and Chemical Mutagenesis (물리·화학적 돌연변이 유도를 통한 Paracoccus haeundaensis의 astaxanthin 생산량 증대)

  • Seo, Yong Bae;Jeong, Tae Hyug;Choi, Seong Seok;Lim, Han Kyu;Kim, Gun-Do
    • Journal of Life Science
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    • v.27 no.3
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    • pp.339-345
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    • 2017
  • Carotenoids are natural lipid-soluble pigments, which are produced primarily by bacteria, algae, and plants. Many studies have focused on the identification, production, and utilization of natural sources of astaxanthin from algae, yeast, and crustacean byproducts as an alternative to the synthetic pigment, which is mostly used today. The aim of the present study was to identify a mutant of Paracoccus haeundaensis by exposure to UV and ethyl methanesulfonate (EMS). The mutant was then exposed to nutrient stress conditions to isolate an astaxanthin-hyperproducing strain, followed by characterization of the mutant. The survival rate decreased in accordance with an increase in the UV exposure time and an increase in the EMS concentration. A mutant of the original P. haeundaensis strain was identified that showed hyperproduction of astaxanthin following exposure to UV irradiation (20 min) and EMS treatment (0.4 M concentration). The optimal culture conditions for the PUE mutant were $25^{\circ}C$, pH 7-8, and 3% NaCl. The effects of various carbon and nitrogen sources on the growth and astaxanthin production of PUE were examined. The addition of 1% raffinose and 3% potassium nitrate influenced cell growth and astaxanthin production. The selected mutant exhibited an increase of 1.58 folds in astaxanthin content compared to initial wild type strain. A genetically stable mutant strain obtained using mutagen (UV irradiation and EMS treatment) may be a suitable candidate for further industrial scale production of astaxanthin.

Keratitis by Acanthamoeba triangularis: Report of Cases and Characterization of Isolates

  • Xuan, Ying-Hua;Chung, Byung-Suk;Hong, Yeon-Chul;Kong, Hyun-Hee;Hahn, Tae-Won;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.46 no.3
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    • pp.157-164
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    • 2008
  • Three Acanthamoeba isolates (KA/E9, KA/E17, and KA/E23) from patients with keratitis were identified as Acanthamoeba triangularis by analysis of their molecular characteristics, a species not previously recognized to be a corneal pathogen. Epidemiologic significance of A. triangularis as a keratopathogen in Korea has been discussed. Morphologic features of Acanthamoeba cysts were examined under a microscope with differential interference contrast (DIC) optics. Mitochondrial DNA (mtDNA) of the ocular isolates KA/E9, KA/E17, and KA/E23 were digested with restriction enzymes, and the restriction patterns were compared with those of reference strains. Complete nuclear 188 and mitochondrial (mt) 16S rDNA sequences were subjected to phylogenetic analysis and species identification. mtDNA RFLP of 3 isolates showed very similar patterns to those of SH621, the type strain of A. triangularis. 16S and 18S rDNA sequence analysis confirmed 3 isolates to be A. triangularis. 18S rDNA sequence differences of the isolates were 1.3% to 1.6% and those of 16S rDNA, 0.4% to 0.9% from A. triangularis SH621. To the best of our knowledge, this is the first report, confirmed by 18S and 16S rDNA sequence analysis, of keratitis caused by A. triangularis of which the type strain was isolated from human feces. Six isolates of A. triangularis had been reported from contaminated contact lens cases in southeastern Korea.

Characterization and Xylanase Productivity of Streptomyces sp. YB914 (Xylanase를 생산하는 Streptomyces sp. YB914의 특성과 효소 생산성)

  • Yoon, Ki-Hong
    • Microbiology and Biotechnology Letters
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    • v.37 no.4
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    • pp.383-388
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    • 2009
  • A strain YB914 was isolated from soil as a producer of the extracellular xylanase, which catalyzes the hydrolysis of oat spelt xylan. The strain YB914 was identified as Streptomyces sp. on the basis of its morphological, cultural and biochemical properties. The xylanase of culture filtrate was the most active at $55^{\circ}C$ and pH 5.5, and retained 80% of its maximum activity at the range of pH 4.5~7.0. In order to optimize the culture medium for xylanase production, ingredients of G.S.S medium were replaced by several carbohydrates. The carbohydrates such as oat spelt xylan, corn cob xylan, wheat bran and lactose increased the xylanase productivity of Streptomyces sp. YB914. However, xylanase production was greatly repressed by galactose or arabinose. The maximum xylanase productivity was reached to 48 U/mL in the modified medium containing 1% oat spelt xylan and 1.5% lactose.

Characterization and Xylanse Productivity of Streptomyces sp. WL-2 (Xylanase 생산균 Streptomyces sp. WL-2의 특성과 효소 생산성)

  • Lee Eun-Hee;Kim Chang-Jin;Yoon Ki-Hong
    • Microbiology and Biotechnology Letters
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    • v.33 no.3
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    • pp.178-183
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    • 2005
  • A strain WL-2 was isolated from soil as a producer of the extracellular xylanase, which catalyzes the hydrolysis of oat spelt xylan. The strain WL-2 was identified as Streptomyces sp. on the basis of its 16S rRNA sequence, morphology, cultural and physiological properties. The xylanase of culture filtrate was the most active at $60^{\circ}C$ and pH 6.0, and retained $90{\%}$ of its maximum activity at range of pH $4.5{\~}6.5$. In order to optimize the culture medium for xylanase production, ingredients of G.S.S medium were replaced by several carbohydrates. The carbohydrates such as ${\alpha}-cellulose$, oat spelt xylan and maltose increased dramatically the xylanase productivity of Streptomyces sp. WL-2. The maximum xylanase productivity was reached to 120 U/ml in the modified medium containing $1{\%}\;\alpha-cellulose$ and $1\%}$ maltose.