• Title/Summary/Keyword: Mono Q

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Purification and Characterization of NADH-Dependent Cr(VI) Reductase from Escherichia coli ATCD33456

  • Bae, Woo-Chul;Kang, Tae-Gu;Jung, Jae-Han;Park, Chul-Jae;Choi, Sung-Chan;Jeong, Byeong-Chul
    • Journal of Microbiology and Biotechnology
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    • v.10 no.5
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    • pp.580-586
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    • 2000
  • A soluble Cr(VI) reductase was purified from the Cr(VI) reducing strain Escherichia coli ATCC33456 by ammonium sulfate fractionation, and chromatographies on Q-Sepharose FF, Cibacron blue 3GA dye affinity, Mono-Q 5/5, and Superdex 200 HR 10/30 columns. The estimated molecular mass of the purified enzyme was 27 kDa on SDS-polyacrylamide gel electrophoresis and 54 kDa on gel filtration, thus indicating a dimeric structure. The isoelectric point of the enzyme was pH 4.85. The optimum reaction pH and storage pH were both 7.0, the optimum reaction temperature was $37^{\circ}C$, and the storage temperature was $4^{\circ}C$. NADH and NADPH both served as electron donors for the reductase, with $V_{max}$ of 68.3 ${\mu}M$ Cr(VI)/min/mg protein and Km of 7.6 $\mu$M using HADH, and Vmax of 42.3 ${\mu}M$ Cr(VI)/min/mg protein and Km of 14.6 $\muM$ using NADPH. When 1 mM EDTA was added, the Cr(VI) reducing activity increased 4-fold.

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Purification and Characterization of a Protease from Korean Pear (Pyrus serotina L.) as Meat Tenderizer

  • Guan, Hao-Li;Mandal, P.K.;Lim, Hee-Kyong;Baatartsogt, Oyungerel;Lee, Chi-Ho;Jeon, Gwang-Joo;Choe, Il-Shin;Choi, Kang-Duk
    • Food Science of Animal Resources
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    • v.29 no.2
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    • pp.157-163
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    • 2009
  • This study was conducted for the isolation, purification, and characterization of a protease from Korean pear, to see its proteolytic activity on chicken actomyosin and to find the optimum pH and temperature of activity on chicken actomyosin. The protease was isolated from crude extract of Korean pear by ammonium sulfate precipitation. Further purification was done by DEAE-Sepharose ion-exchange chromatography, Mono-Q and Mini-Q column chromatography. The purified enzyme gave a single protein band on SDS polyacrylamide gel electrophoresis and the molecular weight was found to be 38 kDa. The specific activity of purified enzyme was 34,907 unit/mg with 25 fold purification and the yield was 2%. The purified enzyme incubated with chicken actomyosin showed high activity. The optimum pH and temperature for enzyme activity on chicken actomyosin were 6.5 and $70^{\circ}C$, respectively. A protease was purified from Korean pear for the first time and characterized. It was found to be promising for meat tenderization.

Use Impacts on Environmental Deteriorations of Trail in Sobaeksan National Park (소백산국립공원 등산로의 환경훼손에 대한 이용영향)

  • 권태호;오구균;이준우
    • Korean Journal of Environment and Ecology
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    • v.6 no.2
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    • pp.168-179
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    • 1993
  • Use impacts on environmental deteriorations of trail were studied on the three major trails of Sobaeksan National Park in 1992. The entire width and bare width of trail as the trail condition were significantly greater on the more heavily used trail. Maximum depth of trail was not so great in spite of steeper grade of trail in comparison with the other National Parks. Percentages of deepening. rock-exposed. diverged points as the deterioration types of trail which were surveyed at the total of 105 points were high and trail conditions were significantly different from those of non-deteriorated points. On the Ridge trail. the damaged area more severe than Class 4 reaches about 10,335$m^2$ and the deterioration is accelerated. The dominant trees of the the upper layer in trail edge vegetation are changed from Q. mongolica. Aar mono to Q. mongolica for Huibang trail. and from Pinus densiflora. Q. mongolica to P. densiflora and to Q. mongolica for Biro trail as altitude increases. Rhododendron schlippenbachii. Weigelu subsessilis. Salix hulteni. Rubus crataeglfolius were classified for tolerant species and R. coreanus. Vaccinium koreanum for intolerant species to use impacts. Highly competetive species on the Ridge trail were grouped R. schlippenbachii. W. subsessilis. Rubus crataegifolius and Symplocos chinensis for. pilosa.

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Phytosociological Community Classification of Mountain Ridge from Guryongryeong to Mt. Yaksu in the Baekdudaegan, Korea (백두대간의 구룡령에서 약수산 마루금의 식생구조 특성에 관한 연구)

  • An, Hyun-Chul;Choo, Gab-Chul;Park, Sam-Bong;Cho, Hyun-Seo;An, Jong-Bin;Park, Jeong-Geun;Ha, Hyoun Woo;Kim, Jin Joong;Kim, Bong-Gyu
    • Korean Journal of Environment and Ecology
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    • v.28 no.6
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    • pp.741-750
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    • 2014
  • To investigate the vegetation structure of mountain ridge from Guryongryeong to Mt. Yaksu, 22 plots ($100m^2$) installed with random sampling method were surveyed. Three groups of Quercus mongolica-Acer pseudosieboldianum community, Q. mongolica community, Cornus controversa-Q. mongolica community were classified by cluster analysis. Q. mongolica was a major woody plant species in the ridge area from Guryongryeong to Yaksusan and Carpinus cordata and C. controversa was partly occupied in some area. High positive correlations showed between Q. mongolica and Symplocos chinensis for. pilosa, Rhododendron schlippenbachii; Tilia amurensis and Tilia mandshurica, Symplocos chinensis for. pilosa; Tilia mandshurica and S. chinensis for. pilosa, R. schlippenbachii; Betula costata and Acer mono; Symplocos chinensis for. pilosa and Rhododendron schlippenbachii, and relatively high negative correlations showed between A. pseudosieboldianum and S. chinensis for. pilosa, R. schlippenbachii. Species diversity(H') of investigated groups were ranged 0.8170~1.1446 and it was lower compared to those of the ridge area of the national parks in Baekdudaegan.

A Phytosociological Comparison of Forest Vegetation between Igneous and Sedimentary Rock Areas in Kyungpook Province, South Korea (경북 일원의 화성암 산지와 퇴적암 산지의 삼림식생에 대한 식생학적 비교)

  • 제갈재철;김종원
    • The Korean Journal of Ecology
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    • v.26 no.1
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    • pp.23-28
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    • 2003
  • In order to identify plant species and communities characteristic of lithospheric condition, forest vegetations of igneous(mainly granite) and sedimentary(mainly sandstone) areas in Kyungpook province were compared. We collected 108 phytosociological releves from the older forests over 20 years old. 444 vascular plant species and 25 plant communities were identified. Results of Principal Coordinates Analysis (PCA) indicated that lithospheric conditions and human impacts were the most important factors related to the plant community diversity and species richness. In the igneous rock areas net contribution degree (rNCD) of 139 species including Quercus serrata, Q. mongolica, Lindera obtusiloba and Fraxinus sieboldiana increased and those of Carpinus laxiflora and Styrax japonica decreased. In the sedimentary rock areas, to a lesser extent, 96 species including Q. aliena, Zanthoxylum schinifolium, Potentilla fragarioides var. major, Rhus javanica, and Plectranthus japonicus increased, whereas character species of Lindero-Quercion mongolicae such as Q. mongolica, Q. serrata, Rhododendron schlippenbachii, Acer mono and Styrax obassia decreased, and particularily the species Sasa borealis, the Alangium-Zelkova serrata community and the Philadelphus-Fraxinus mandshurica community were absent. The vegetation of sedimentary rock area is characterized as a regional type shaping dwarf pine forests and sparse and light broad-leaved forests.

The Characteristics of Longitudinal Permeability and Hydraulic Resistance in Stem of Acer mono (고로쇠나무 줄기의 통수성(通水性)과 통수저항(通水抵抗)의 특성(特性))

  • Kim, Sun-Hee;Han, Sang-Sup
    • Journal of Korean Society of Forest Science
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    • v.89 no.4
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    • pp.488-496
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    • 2000
  • This study was carried out to investigate the characteristics of longitudinal permeability and hydraulic resistance, and to compare the longitudinal permeability ($K_E$) calculated by the Hagen-Poiseuille's law (Siau, 1971) and the longitudinal permeability (K) measured in sapwood of Acer mono stem. The volume flow rate (Q) in a vessel was $0.80{\times}10^{-4}cm^3/sec$ and the hydraulic resistance ($R_S$) to viscous flow through a vessel was, on average, $1.37{\times}10^{10}dyn{\cdot}sec{\cdot}cm^{-3}{\cdot}cm^{-2}$. The average value of volume flow rate ($Q_N$) through the cross section of sapwood was $0.32cm^3{\cdot}sec^{-1}{\cdot}cm^{-2}$, and the average resistance ($R_{SN}$) was $3.59{\times}10^6dyn{\cdot}sec{\cdot}cm^{-3}{\cdot}cm^{-2}$. The values of K decreased as the diameter of stem increases, which was attributable to variations in the number of vessel per unit area rather than in vessel diameter, and to different resistances in the conducting tissues of each part of the stem. The average value of K measured at breast height was 31% of average value of $K_E$. The $K/K_E$ ratios were 100% in 4 to 6 year-old stems and more than 90% in 7 to 27 year-old stems. The $K/K_E$ ratio decreased as the age of stems increases, and was not more than 20% in near-ground parts of stem.

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Quantitative Analysis of Oligosaccharide Structure of Glycoproteins

  • Chang, Kern-Hee;Tamao Endo;Kim, Jung-How
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.2
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    • pp.136-140
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    • 2000
  • A sensitive and quantiative method for the structural analysis of oligosaccharide was established for the glycoform analysis of glconproproteins. Inthis study, n-linked oligosaccharides of human IgG and bovine transferin were analyzed for the evaluation of the methydrate moiety ofthe method. Chrbohydrate moiety of glycoprotein was relased by hydrazinolysis and purified by paper chromatography. The oligosaccharides were labeled with a fluorescent bye, 2-aminobenzamide, for the enhancement of detection sensitivity. sialylated (acidic) oligosaccharides were separated from neutral oligosaccharide by employing a strong anion-exchange column(MonoQ) followed by the treatment with sialidase. Enzymatically desiayated fractions and neutral fractions of oligosaccharides were applied to normal-phase HPLC to resolve the peaks according to glucose unit (GU). The structure of separated molecules was further determined by sequential digestion with exoglycosidases. As a result, disialylated biantennary complextype oligosaccharide was found to be a major sugar chain in bovine transferrin (63%). In human IgG, core fucosylated asialobiantennary complex oligosaccharides were dominant. These results coincided well with reported results.

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Purification and characterization of biochemical properties of hemolysin from Vibrio fluvialis (Vibrio fluvialis 유래의 hemolysin 정제와 생화학적 특성)

  • 이종희;한정현;안선희;김선회;이은미;공인수
    • Journal of Life Science
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    • v.12 no.4
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    • pp.490-495
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    • 2002
  • Hemolysin (VFH) of V. fluvialis, which is a pathogenic bacteria, causing watery diarrhea with vomiting, abdominal croup, was purified. V. fluvialis was cultivated in BHI medium and the culture supematant was precipitated by ammonium sulfate. The protein was purified by chromatographies on columns of DEAE-cellulose and Mono-Q. Molecular weight of the purified VFH was estimated as 79kDa by SDS-PAGE. The optimal temperature for a maximum hemolytic activity was at around 35$^{\circ}C$ and the activity was decreased at 4$0^{\circ}C$ Cytotoxicity of VFH was also investigated using RTG-2 cell line. LDH assay study showed that 50$\mu\textrm{g}$/m1 of VFH release 80% of total cellular LDH (lactate dehydrogenase) from RTG-2 cell and microscopic observation also showed the morphological change of cell.

Production and Characterization of Manganese Peroxidase from the White Rot Fungus Pleurotus ostreatus in Liquid Culture (액체배양한 느타리 버섯균(Pleurotus ostreatus)으로부터 망간퍼옥시데이즈의 생산 및 특성)

  • Lee, Jae-Sung;Ha, Hyo-Cheol
    • Applied Biological Chemistry
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    • v.47 no.1
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    • pp.22-26
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    • 2004
  • The ligninolytic basidiomycete, Pleurotus ostreatus K-2946, was produced a manganese peroxidase (MnP) activity when grown in liquid culture with glucose-yeast-peptone (G-Y-P) medium. However, lignin peroxidase (LiP) was not detected in this culture medium. The purification progress of MnP was purified that included chromatography on Sepharose CL-6B, Superdex 75 prep grade and Mono-Q. MnP purified by column chromatography, was 36400 dalton and a pI of 3.95. The optimal pH and temperature of the purified MnP activity were 5.0 and $55^{\circ}C$. The characteristics of MnP produced was quite similar to those of MnP 3 isoenzyme produced by other strains of P. ostreatus.

Purification and Properties of Laccase of the White-rot Basidiomycete Coriolus hirsutus

  • Lee, Yeo-Jin;Shin, Kwang-Soo
    • Journal of Microbiology
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    • v.37 no.3
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    • pp.148-153
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    • 1999
  • Laccase produced by Coriolus hirsutus was purified to electrophoretic homogeneity by acetone precipitation, Sephacryl S-2000 HR chromatography, DEAE Sepharose CL-6B chromatography, and Mono Q HR 5/5 chromatography. The purification of laccase was 46.6-fold with an overall yield of 23.7%. Laccase from this fungus was a monomeric glycoprotein with 16% carbohydrate content, and has an isoelectric point of 4.2, and molecular mass of 78 kDa, respectively. The N-terminal amino acid sequence of the enzyme showed significant homology to hoste of laccases from Coriolus versicolor, Pycnoporus cinnabarius, and an unidentified basidiomycete, PM1. The highest rate of 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) oxidation by laccase was reached at 45$^{\circ}C$, and te pH optima of the enzyme varied depending on the substrate in the range of 2.0 to 4.5. The enzyme was stable at 60$^{\circ}C$ for 5 h and lost 80% activity at 80$^{\circ}C$ in 30 min. The enzyme oxidized a variety of usual laccase substrates including lignin-related phenol, and had the highest affinity toward ABTS. Under standard assay conditions, the apparent Km value of the enzyme toward ABTS was 8.1 ${\mu}$M. The enzyme was completely inhibited by L-cysteine and sodium azide, but not by potassium cyanide, SDS, ad thiourea.

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