• Title/Summary/Keyword: isolated set

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Crystallization and Preliminary X-Ray Crystallographic Analysis of PAS Factor from Vibrio vulnificus

  • Lee, Jun Hyuck;Kim, Soo Young;Rho, Seong-Hwan;Im, Young Jun;Kim, Young Ran;Kim, Mun-Kyoung;Kang, Gil Bu;Rhee, Joon Haeng;Eom, Soo Hyun
    • Molecules and Cells
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    • v.20 no.3
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    • pp.361-363
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    • 2005
  • Plasmid Achromobacter secretion (PAS) factor is a putative secretion factor that induces the secretion of periplasmic proteins. PAS factor from Vibrio vulnificus was crystallized at 294 K by the hanging drop vapor-diffusion method. It was isolated as a monomer during the purification procedures. The native crystal belongs to the F222 space group with unit cell parameters a = 56.1, b = 74.4, $c=80.0{\AA}$, ${\alpha}={\beta}={\gamma}=90^{\circ}$. The crystal was soaked in cryoprotectant containing 1 M NaBr for 1 h for MAD phasing. The diffraction limit of the Br-MAD data set was $1.9{\AA}$ using synchrotron X-ray irradiation at beam line BL-18B at the Photon Factory, Japan.

Characterization of a metalloprotease from an isolate Bacillus thuringiensis 29-126 in animal feces collected from a zoological garden in Japan

  • Lee, Eun Seok;Lee, Hyun Woo;Lee, Dong-Hyun;Kim, Hoon
    • Journal of Applied Biological Chemistry
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    • v.59 no.4
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    • pp.373-377
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    • 2016
  • An extracellular metalloprotease, Btmp, was partially purified from the culture supernatant of Bacillus thuringiensis 29-126, isolated from animal feces collected in a zoological garden in Japan, by ultrafiltration, ammonium sulfate precipitation, and a set of chromatography on Sephadex G-75 and High-Q. The molecular mass of the protease was estimated to be 60 kDa by SDS-PAGE. The enzyme showed optimum activity at $50^{\circ}C$ and pH 6.0, and had a half-life of 14 min at $50^{\circ}C$. The enzyme activity was not influenced by $Na^+$, $K^+$, $As^+$, $Mg^{+2}$, $Ca^{2+}$, $Ba^{2+}$, and phenylmethylsulfonyl fluoride, but it was moderately inhibited by $Zn^{+2}$ at a concentration of 1.0 mM, while the activity was significantly inhibited to less than 50 % by $Cu^{2+}$, $Co^{2+}$, $Cd^{2+}$, and ethylenediaminetetraacetic acid. Interestingly, the enzyme was activated to 178 % by 1.0 mM of $Mn^{2+}$. From these results, it may be suggested that the protease is a novel extracellular manganeseactivated metalloprotease.

Effects of Vanadate on Activity of $Ca^{++}$Activated ATPase and $Ca^{++}$ Uptake of Sarcoplasmic Reticulum in Rat Skeletal Muscle (Vanadate가 골격근 sarcoplasmic reticulum의 $Ca^{++}-ATPase$$Ca^{++}\;uptake$에 미치는 영향)

  • Joo, Soon-Jae;Han, Kyung-Hee;Leem, Joong-Woo;Kang, Doo-Hee
    • The Korean Journal of Physiology
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    • v.20 no.2
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    • pp.157-164
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    • 1986
  • Since it has been reported that vanadate inhibits $Ca^{++}-ATPase$ activity without affecting $Ca^{++}$ uptake, this study was undertaken to investigate the effects of vanadate on $Ca^{++}-ATPase$ activity and $Ca^{++}$ uptake in the sarcoplasmic reticulum of rat skeletal muscle. The following results were obtained. 1) $Ca^{++}$ activated ATPase activity of the intact sarcoplasmic reticulum was significantly inhibited when vanadate was added to the incubation medium at concentration greater than $10^{-6}\;M$. However $Mg^{++}$-ATPase activity of the intact SR was not affected by vanadate at concentrations ranging from $10^{-7}\;to\;10^{-4}\;M.$ Similarly, $Ca^{++}-ATPase$ activity in sonicated sarcoplasmic reticulum was significantly reduced by vanadate at a concentration $10^{-7}$ M or higher. 2) The uptake of $Ca^{++}$ by isolated sarcoplasmic reticulum was also inhibited by vanadate under the conditions where the turnover rate of $Ca^{++}-ATPase$ was made to increase. These results suggest that the inhibition of $Ca^{++}$ uptake by vanadate may be correlated with that of $Ca^{++}-ATPase$ if experimental conditions are properly set.

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Enhancement of Herboxidiene Production in Streptomyces chromofuscus ATCC 49982

  • Jha, Amit Kumar;Lamichhane, Janardan;Sohng, Jae Kyung
    • Journal of Microbiology and Biotechnology
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    • v.24 no.1
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    • pp.52-58
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    • 2014
  • Structurally, herboxidiene contains the tetrahydropyran acetic acid moiety and a side chain including a conjugated diene, and has been isolated from Streptomyces chromofuscus ATCC 49982. Its production was significantly elevated nearly 13.5-fold (0.74 g/l) in a medium supplemented with glycerol (medium No. 6A6), and was more efficacious (1.08 g/l; 19.8-fold) in fed-batch fermentation at 36 h in medium No. 6A6, from Streptomyces chromofuscus. For further enhancement, regulatory genes metK1-sp and afsR-sp from Streptomyces peucetius were overexpressed using an expression vector, pIBR25, and similarly ACCase from Streptomyces coelicolor and two genes, metK1-sp and afsR-sp, were also overexpressed using an integration vector, pSET152, under the control of the strong $ermE^*$ promoter in Streptomyces chromofuscus. Only the recombinant strains S. chromofuscus SIBR, S. chromofuscus GIBR, and S. chromofuscus AFS produced more herboxidiene than the parental strain in optimized medium No. 6A6 with an increment of 1.32-fold (0.976 g/l), 3.85-fold (2.849 g/l), and 1.7-fold(1.258 g/l) respectively.

Correlation Between Sorangium cellulosum Subgroups and Their Potential for Secondary Metabolite Production

  • Lee, Chayul;An, Dongju;Lee, Hanbit;Cho, Kyungyun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.3
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    • pp.297-303
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    • 2013
  • Phylogenetic analysis of the groEL1 and xynB1 gene sequences from Sorangium cellulosum strains isolated in Korea previously revealed the existence of at least 5 subgroups (A-E). In the present study, we used sequence analysis of polymerase chain reaction-amplified biosynthetic genes of strains from the 5 subgroups to indicate correlations between S. cellulosum subgroups and their secondary metabolic gene categories. We detected putative biosynthetic genes for disorazol, epothilone, ambruticin, and soraphen in group A, group C, group D, and group E strains, respectively. With the exception of KYC3204, culture extracts from group A, group B, and group C strains exhibited no noticeable antimicrobial inhibitory activities. By contrast, culture extracts from group D strains inhibited the growth of Candida albicans, whereas culture extracts from group E strains inhibited the growth of C. albicans and Staphylococcus aureus. High performance liquid chromatography analysis of the culture extracts from the strains of each subgroup revealed unique peak patterns. Our findings indicate the existence of at least 5 subgroups of S. cellulosum strains, each of which has the potential to produce a unique set of secondary metabolites.

Local Similarity based Document Layout Analysis using Improved ARLSA

  • Kim, Gwangbok;Kim, SooHyung;Na, InSeop
    • International Journal of Contents
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    • v.11 no.2
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    • pp.15-19
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    • 2015
  • In this paper, we propose an efficient document layout analysis algorithm that includes table detection. Typical methods of document layout analysis use the height and gap between words or columns. To correspond to the various styles and sizes of documents, we propose an algorithm that uses the mean value of the distance transform representing thickness and compare with components in the local area. With this algorithm, we combine a table detection algorithm using the same feature as that of the text classifier. Table candidates, separators, and big components are isolated from the image using Connected Component Analysis (CCA) and distance transform. The key idea of text classification is that the characteristics of the text parallel components that have a similar thickness and height. In order to estimate local similarity, we detect a text region using an adaptive searching window size. An improved adaptive run-length smoothing algorithm (ARLSA) was proposed to create the proper boundary of a text zone and non-text zone. Results from experiments on the ICDAR2009 page segmentation competition test set and our dataset demonstrate the superiority of our dataset through f-measure comparison with other algorithms.

Development and characterization of 21 microsatellite markers in Daphne kiusiana, an evergreen broad-leaved shrub endemic to Korea and Japan

  • Lee, Jung-Hyun;Cho, Won-Bum;Yang, Sungyu;Han, Eun-Kyeong;Lyu, Eun-Seo;Kim, Wook Jin;Moon, Byeong Cheol;Choi, Goya
    • Korean Journal of Plant Taxonomy
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    • v.47 no.1
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    • pp.6-10
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    • 2017
  • Microsatellite markers were isolated for Daphne kiusiana var. kiusiana (Thymelaeaceae), an evergreen broad-leaved shrub endemic to Korea and Japan. Because its populations in Jeju Island are morphologically controversial, and consistently threatened by anthropogenic pressures, taxonomic delimitation and conservation effort are required at the genetic level. We developed 21 polymorphic microsatellite loci from Next Generation Sequencing data. The primer set included di-, tri-, and tetra-nucleotide repeats. Variability in the markers was tested for 80 individuals of D. kiusiana from three natural populations in Jeju Island and Japan. Among the 21 loci, three were unavailable for population JKJU of Japan. The Neighbor-Joining tree based on microsatellite markers described here classified the three populations into two groups according to geographical or morphological traits. These will be a powerful genetics tool for determining the taxonomic boundary and establishing suitable conservation strategies for D. kiusiana in Jeju Island.

Optimization of Ascorbic Acid-2-Phosphate Production from Ascorbic Acid Using Resting Cell of Brevundimonas diminuta

  • Shin, Woo-Jung;Kim, Byung-Yong;Bang, Won-Gi
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.769-773
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    • 2007
  • With the aim to produce ascorbic acid-2-phosphate(AsA-2-P) from L-ascorbic acid(AsA, Vitamin C), nine bacteria conferring the ability to transform AsA to AsA-2-P were isolated from soil samples alongside known strains from culture collections. Most isolates were classified to the genus Brevundimonas by 16S phylogenetic analysis. Among them, Brevundimonas diminuta KACC 10306 was selected as the experimental strain because of its the highest productivity of AsA-2-P. The optimum set of conditions for the AsA-2-P production from AsA using resting cells as the source of the enzyme was also investigated. The optimum cultivation time was 16 h and the cell concentration was 120g/l(wet weight). The optimum concentrations of AsA and pyrophosphate were 550mM and 450mM, respectively. The most effective buffer was 50mM sodium formate. The optimum pH was 4.5 and temperature was $40^{\circ}C$. Under the above conditions, 27.5g/l of AsA-2-P was produced from AsA after 36 h of incubation, which corresponded to a 19.7% conversion efficiency based on the initial concentration of AsA.

Seed Transmission of Tomato yellow leaf curl virus in White Soybean (Glycine max)

  • Kil, Eui-Joon;Park, Jungho;Choi, Hong-Soo;Kim, Chang-Seok;Lee, Sukchan
    • The Plant Pathology Journal
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    • v.33 no.4
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    • pp.424-428
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    • 2017
  • Tomato yellow leaf curl virus (TYLCV) infection of the common bean (Phaseolus vulgaris) has been reported, but soybean (Glycine max) has not previously been identified as a TYLCV host. Five cultivars of white soybean were agro-inoculated using an infectious TYLCV clone. At 30 days post-inoculation, they showed infection rates of 25% to 100%. Typical TYLCV symptoms were not observed in any inoculated plants. To examine whether TYLCV was transmitted in soybean seeds, DNA was isolated from bundles of five randomly selected seeds from TYLCV-inoculated soybean plants and amplified with a TYLCV-specific primer set. With the exception of one bundle, all bundles of seeds were verified to be TYLCV-infected. Virus dissemination was also confirmed in three of the 14 bunches. Viral replication was also identified in seeds and seedlings. This is the first report demonstrating that soybean is a TYLCV host, and that TYLCV is a seed-transmissible virus in white soybean.

Effect of the Long-term Application of Organic Matters on Microbial Diversity in Upland Soils (유기물 장기 연용이 밭토양 미생물의 다양성에 미치는 영향)

  • Suh, Jang-Sun;Kwon, Jang-Sik;Noh, Hyung-Jun
    • Korean Journal of Soil Science and Fertilizer
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    • v.43 no.6
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    • pp.987-994
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    • 2010
  • To investigate the effect of long term application of organic matter in upland soils, plots for treatments of NPK, NPK+pig manure compost, rape seed cake, rice straw compost, and green manure were set up. Populations of Bacillus and Gram negative bacteria were high in the plot treated with green manure application, but microbial biomass was increased with chemical fertilizer or pig manure compost in upland soils. Activities of phosphomonoesterase and dehydrogenase were high with organic matter application comparing to control. Cluster patterns analysed using phospholipid fatty acid of plots treated with rice straw and or pig manure compost were clearly different comparing with other treatments. Dominant bacteria in upland soils were Bacillus flexus, B. subtilis and B. megaterium. And the strains isolated from upland soils had amylase, protease and lipase activities.