• Title/Summary/Keyword: Sp1-21

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Gene Cloning and Enzymatic Properties of Thermostable Laccase from Thermus thermophilus HJ6 (Thermus thermophilus HJ6 유래 내열성 laccase의 유전자 클로닝 및 효소학적 특성)

  • Lee, So-Young;Jung, Young-Hoon;Seo, Min-Ho;Jeon, Sung-Jong
    • KSBB Journal
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    • v.27 no.4
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    • pp.257-262
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    • 2012
  • The gene encoding Thermus thermophilus HJ6 laccase (Tt-laccase) was cloned, sequenced, and comprised of 1,389 nucleotides encoding a protein (462 amino acids) with a predicted molecular mass of 51,049 Da. The deduced amino acid sequence of Tt-laccase showed 99.7% and 44.3% identities to the Thermus thermophilus HB27 laccase and Synechococcus sp. RS9917 laccase, respectively. Tt-laccase gene was expressed as a fusion protein with six histidine residues in E. coli Rosetta-gami (DE3) cells, and the recombinant protein was purified to homogeneity. UV-Vis spectrum analysis revealed that the enzyme has copper atoms, a type I Cu(II) and a type III binuclear Cu(II). The optimum pH for the oxidation of guaiacol was 5.0 and the optimum temperature was $90^{\circ}C$ The half-life of heat inactivation was about 120 min at $90^{\circ}C$ The enzyme reaction was inhibited by sodium azide, L-cystein, EDTA, dithiothreitol, tropolone, and kojic acid. The enzyme oxidized various known laccase substrates, its lowest $K_m$ value being for 4-hydroxyindole, highest $k_{cat}$ value for syringaldazine, and highest $k_{cat}/K_m$ for guaiacol.

Molecular Identification of Sarcocystis grueneri in Wild Korean Water Deer (Hydropotes inermis argyropus)

  • Kim, Hye-won;Kim, Hyeon-Cheol;Ryu, Si-Yun;Choi, Kyoung-Seong;Yu, Do-Hyeon;Park, Jinho;Chae, Joon-Seok;Park, Bae-Keun
    • Parasites, Hosts and Diseases
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    • v.56 no.2
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    • pp.129-134
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    • 2018
  • The cysts of Sarcocystis grueneri were detected and characterized from the cardiac muscles of the Korean water deer (Hydropotes inermis argyropus). Of the 38 heart muscle samples examined by light microscopy, 10 were found infected with the cysts of Sarcocystis sp. The cysts appeared oval to spherical shape and measured $110-380{\mu}m$ in length and $90-170{\mu}m$ in width. A phylogenetic tree of the 18S rRNA sequences (1.5 kb) revealed a close relationship of the infected cysts to genus Sarcocystis. The 18S rRNA sequence of the infected cysts showed 100% identity to S. grueneri and 97% to S. capracanis. Here, we first report the S. grueneri infections in the Korean water deer.

Expression of Rotavirus Capsid Proteins VP6 and VP7 in Mammalian Cells Using Semliki Forest Virus-Based Expression System

  • Choi, Eun-Ah;Kim, Eun;Oh, Yoon-I;Shin, Kwang-Soon;Kim, Hyun-Soo;Kim, Chul-Joong
    • Journal of Microbiology and Biotechnology
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    • v.12 no.3
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    • pp.463-469
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    • 2002
  • Rotaviruses are the world-wide leading causative agents of severe dehydrating gastroenteritis in young children and animals. The outer capsid glycoprotein VP7 and inner capsid glycoprotein VP6 of rotaviruses are highly antigenic and immunogenic. An SFV-based expression system has recently emerged as a useful tool for heterologous protein production in mammalian cells, exhibiting a much more efficient performance compared to other gene expression systems. Accordingly, the current study adopted an SFV-based expression system to express the VP7 of a group A human rotavirus from a Korean isolate, and the VP6 of a group B bovine rotavirus from a Korean isolate, in mammalian cells. The genes of the VP6 and VP7 were inserted into the SFV expression vector pSFV-1. The RNA was transcribed in vitro from pSFV-VP6 and pSFV-VP7 using SP6 polymerase. Each RNA was then electroporated into BHK-21 cells along with pSFV-helper RNA containing the structural protein gene without the packaging signal. The expression of VP6 and VP7 in the cytoplasm was then detected by immunocytochemistry. The recombinant virus was harvested by ultracentrifugation and examined under electron microscopy. After infecting BHK-21 cells with the defective viruses, the expressed proteins were separated by SDS-PAGE and analyzed by a Western blot. The results indicate that an SFV-based expression system fur the VP6 and VP7 of rotaviruses is an efficient tool for developing a diagnostic kit and/or preventive vaccine.

Biosynthesis of Lactate-containing Polyhydroxyalkanoates in Recombinant Escherichia coli from Sucrose (재조합 대장균에서 수크로즈로부터의 젖산을 모노머로 함유한 폴리하이드록시알칸산 생산 연구)

  • Oh, Young Hoon;Kang, Kyoung-Hee;Shin, Jihoon;Song, Bong Keun;Lee, Seung Hwan;Lee, Sang Yup;Park, Si Jae
    • KSBB Journal
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    • v.29 no.6
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    • pp.443-447
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    • 2014
  • Biosynthesis of lactate-containing polyhydroxyalkanoates (PHAs) was examined in recombinant Escherichia coli W strain from sucrose. The Pseudomonas sp. MBEL 6-19 phaC1437 gene and the Clostridium propionicum pct540 gene, which encode engineered Pseudomonas sp. MBEL 6-19 PHA synthase 1 ($PhaC1_{Ps6-19}$) and engineered C. propionicum propionyl-CoA transferase ($Pct_{Cp}$), respectively, were expressed in E. coli W to construct key metabolic pathway to produce poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)]. The recombinant E. coli W expressing the phaC1437 gene and the pct540 gene could synthesize P(3HB-co-13mol%LA) up to the polymer content of 31.3 wt% when it was cultured in chemically defined MR medium containing 20 g/L of sucrose and 2 g/L of sodium 3-hydroxybutyrate. When Ralstonia eutropha phaAB genes were additionally expressed to provide 3-hydroxybutyrate-CoA (3HB-CoA) from sucrose, P(3HB-co-16mol%LA) could be synthesized from sucrose as a sole carbon source without supplement of sodium 3-hydroxybutyrate in culture medium, but the PHA content was decreased to 12.2 wt%. The molecular weight of P(3HB-co-16 mol%LA) synthesized in E. coli W using sucrose as carbon source were $1.53{\times}10^4$ ($M_n$) and $2.78{\times}10^4$ ($M_w$), respectively, which are not different from those that have previously been reported by other recombinant E. coli strains. Engineered E. coli strains developed in this study should be useful for the production of lactate-containing PHAs from sucrose, one of the most abundant and least expensive carbon sources.

Characterization of alkaline cellulase from Bacillus subtilis 4-1 isolated from Korean traditional soybean paste (전통 장류에서 분리된 알칼리성 Cellulase 생성 Bacillus subtilis 4-1 균주의 효소학적 특성)

  • Baek, Seong Yeol;Lee, You Jung;Yun, Hye Ju;Park, Hye Young;Yeo, Soo-Hwan
    • Food Science and Preservation
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    • v.21 no.3
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    • pp.442-450
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    • 2014
  • In this study, we isolated a cellulase-producing bacterium isolated from traditional Korean fermented soybean paste and investigated the effect of culture conditions on the production of cellulase. This bacterium, which was identified as Bacillus subtilis 4-1 through 16S rRNA gene sequence analysis, showed the highest cellulase activity when the cells were grown at $45^{\circ}C$ for 24 hours in the CMC medium supplemented with 1.0% of soluble starch and 0.1% yeast extract. The initial optimum pH of the medium was observed in the range of 5.0~9.0. The optimal pH and temperature for the production of cellulase from B. subtilis 4-1 were pH 9.0 and $60^{\circ}C$ respectively. In addition, the enzyme showed significant activity in the temperature range of $20{\sim}90^{\circ}C$, which indicates that B. subtilis 4-1 cellulase is an alkaline-resistance and thermo-stable enzyme. This enzyme showed higher activity with CMC as the substrate for endo-type cellulase than avicel or pNPG as the exo-type substrates for exo-type cellulase and ${\beta}$-glucosidase. These results suggest that the cellulase produced from B. subtilis 4-1 is a complex enzyme rather than a mono-enzyme.

Screening of cryoprotectants (CPAs) for cryopreservation in the Nitzschia sp. of marine microalgae (해양 규조류 Nitzschia sp.의 초저온동결보존을 위한 보존제의 영향 분석)

  • Lee, In Hye;Jeon, Ji Young;Kim, Kyeung Mi;Kang, Myung suk
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.400-408
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    • 2018
  • Biodiversity has continued to degrade in the $21^{st}$ century due to global warming occasioned by destruction of the environment around the world.. The Nagoya protocol places Korea in a unique position to effectively develop and protect its domestic genetic resources. Microalgae under study in this research contains large amount of antioxidant substances such as beta carotene and astaxanthin, that can be used as biological resource owing to the large amounts of biomass that can be secured through photosynthesis. However, it is difficult to preserve it since cryopreservation method used for long-term preservation is yet to be developed. A basic study for long term cryopreservation was carried out on Nizschia frustulum and Nitzschia amabilis which belong to marine diatoms. As cryoprotectants (CPAs), glycerol, DMSO, and methanol which penetrate into cells were prepared at 5%, 10%, and 15% concentrations each, in case of methanol, it was tested at concentrations of 5%, 10% and 12% by its nature. Two kinds of microalgae, N. frustulum and N. amabilis, were diluted with $10^2$, $10^3$ and $10^4cells\;ml^{-1}$, respectively. The highest survival rate was shown at12% concentration of methanol, and the figures were $6.94{\pm}0.31%$ in N. frustulum and $8.85{\pm}0.16%$ in N. amabilis. As a result of 3 weeks cultivation of thawed microalgae after freezing, the result is shows that N. frustulum increased about 10 times faster and N. amabilis increased about 12 times the original concentration.

The Change of Phytoplankton Community Structure and Water Quality in the Juksan Weir of the Yeongsan River Watershed (영산강수계 죽산보의 식물플랑크톤과 이화학적 변화)

  • Son, Misun;Chung, Hyeon Su;Park, Chang Hee;Park, Jong-hwan;Lim, Cheahong;Kim, Kyunghyun
    • Korean Journal of Environmental Biology
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    • v.36 no.4
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    • pp.591-600
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    • 2018
  • The objective of this study was to determine the changes in phytoplankton and long-term water quality of Juksan-Weir in Yeongsan River that took place between April 2010 and December 2015. The number of species used in this study was 288, which consisted of 6% of Cyanophyta, 26% of Bacillariophyta, 53% of Chlorophyta and the others (15%). The standing crops of phytoplankton ranged from $500cells{\cdot}mL^{-1}-29,950cells{\cdot}mL^{-1}$ with an average of $7,885cells{\cdot}mL^{-1}$. At the two site, 20 dominant genera of found. The dominant genera were 6 of Bacillariophyta, 6 of Cyanophyta, 7 of Chlorophyta and 1 of Cryptophyta. The most dominant genus among the phytoplankton was Stephanodiscus sp. (Total 59%, each 54% and 63%). The most dominant genus among the Cyanophyta was Microcystis sp., which had a cell abundance ratio of 17%. The results of two sites were 21% and 13%, and the upstream was higher than the downstream.

Fatty Acid Composition of Antarctic Toothfish Dissostichus mawsoni (남극이빨고기(Dissostichus mawsoni)의 지방산조성)

  • Lim, Chi-Won;Jo, Hyun-Su;Yeon, InJa;Seok, KyuJin;Choi, Seok-Gwan;Yoon, Na-Young;Shim, Kil-Bo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.45 no.6
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    • pp.579-585
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    • 2012
  • The proximate compositions and fatty acid profiles of Antarctic toothfish Dissostichus mawsoni that was caught in the southern Ross Sea (J, L) of the Antarctic Ocean were studied. The lipid contents of samples from J and L were 18.2 and 21.1%, respectively. The protein and ash contents were similar for samples J and L. The prominent fatty acids in the total lipids of the fish muscle were 18:1n-9, 16:1n-7, 16:0, 14:0, 18:1n-7, 20:5n-3 (eicosapentaenoic acid, EPA), and 22:6n-3 (docosahexaenoic acid, DHA). In addition, the total fatty acids of bycatch products in the toothfish stomachs (Pleuragramma antarcticum, Gerlachea australis, Pasiphaea sp., Trematomus eulepidotus, Chionodraco hamatus, Chionodraco myersi, and Neopagetopsis ionah) were determined. The prominent fatty acids in those species were 18:1n-9, 16:0, 14:0, DHA, EPA, and 18:1n-7.

The Variation of Natural Population of Pinus densiflora S. et Z. in Korea(I) -Characteristics of Needle and Wood of Chuwang-san, An-Myeon-do and Odae-san Populations- (소나무 천연집단(天然集團)의 변이(變異)에 관(關)한 연구(硏究)(I))

  • Yim, Kyong-Bin;Kim, Zin-Suh
    • Journal of Korean Society of Forest Science
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    • v.28 no.1
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    • pp.1-20
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    • 1975
  • F$\ddot{u}$r die Erforschllng der Variationen bei der Naturpopulationen der Pinus densifiora wurden drei Populationen aus Chuwangsan (Kyong-buk), Anmyondo (Chung-nam) und Odaesan (Kangwon) Ausgewiiblt. Bei je 20 Individuen jeder Population wurden die Variationen von der Eigenschaft der $\ddot{a}$usseren Gestalt, von der anatomischen und morphologischen Eigenschaft der Kiefernadel und von der Holzqualit$\ddot{a}$t an Bohrsp$\ddot{a}$nen untersucht. Das Ergebnis der Untersuchung konnte in den folgenden sechs Punkten formuliert werden. 1. Von den Eigenschaften der $\ddot{a}$usseren Gestalt ist die Geradheit des Baumstamms, der Durchmesser der Zweige und der Winkel der Verzweigung in der Odaesan Population wlinschenswerter als in anderen Populationen. 2. Es wird festgestellt, dass sich die drei Nadelmerkmale, Anzahl der Z$\ddot{a}$hne je 0.5 cm, Anzahl der Stomata-Reihen und Anzahl der Harzkan$\ddot{a}$le, sowohl zwischen den Populationen als auch zwischen den Individuen signignifikant unterscheiden. 3. Bei der Betrachtung des Harzkanal Index, der Schichtanzahl der Hypodermis und der Entfernung der Fibrovasalb$\ddot{u}$ndel wird aufgezeigt, dass sich alle drei gepr$\ddot{u}$ften Populationen geringf$\ddot{u}$gig von der normalen Kiefer unterscheiden. 4. Zwischen den Populationen ergibt sich ein Unterschied im Prozentanteil des Spatho lzes und im spezifischen Geswicht, jedoch zeigt die Breite der Jahresrings und die Tracheidenl$\ddot{a}$nge keinen Unterschied. Das spezifische Gewicht und die Tracheidenl$\ddot{a}$nge der Individuen jeder Population ist signifikant verschieden. 5. Bei dem Alter 11-20 ist der Prozentanteil des sp$\ddot{a}$tholzes am gr$\ddot{o}$ssten und wird aber danach immer geringer bis zum Alter 30; sp$\ddot{a}$ter zeigt sich dann keine sichtbare Ver$\ddot{a}$nderung mehr. Das spezifische Gewicht sich vergr$\ddot{o}$ssert bei der Zunahme des Jahresrings, nimmt aber dann in Anmyondo und Odaesan Populationen beim Alter 21-30 wieder ab. Die Tracheidenl$\ddot{a}$nge der drei Populationen besitzen die Neigung, sich nach der zunahme der Jahresringe zu vermehren. 6. Mit diesen hier behandelten Charakteren werden ganz allgemeinen die Unterschiede der drei Kiefer populationen festgestellt. Diese Ergebnisse sind sehr bemerkenswert betrachtet vom genetischen Gesichtspunkt der waldbaumpopulation.

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Biodiversity of Benthic Macroinvertebrate on Organic Rice Paddy Field (유기농업 논에서 저서성대형무척추동물의 다양성)

  • Kim, Jong-Sun;Kim, Do-Ik;Kim, Seon-Gon;Kang, Beom-Ryong;Ko, Suk-Ju;Lim, Gyeong-Ho;Kim, Hong-Jae
    • Korean Journal of Organic Agriculture
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    • v.17 no.2
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    • pp.193-209
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    • 2009
  • Total collected benthic macro invertebrates in rice paddy fields were identified 3 phylum, 5 class, 13 order, 25 family, 32 genera and 36 species, those in organic rice field were 12 order, 22 family and 28 species, and in conventional rice field were 10 order, 19 family and 25 species. The almost collected species inhabit lentic zone but Semisulcospira libertina, Corbicula fluminea, and Tipula sp. were habitat on lotic zone. The number of benthic macro invertebrates species was higher in organic paddy field than in conventional field. Eleven species were collected only on organic but eight species only on conventional field. The number of individuals were 870 on organic rice field which higher 3.6-fold on conventional field. The diversity indices of organic rice field were 1.57-2.45 and 0.71-2.61 in the first and second survey, respectively. The higher diversity indices, the lower dominance indices.

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