• Title/Summary/Keyword: Molecular Characteristics

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Spectrofluorometric Characteristics of the N-Terminal Domain of Riboflavin Synthase (아미노-말단 리보플라빈 생성효소 단백질의 형광 특성)

  • Kim, Ryu-Ryun;Yi, Jeong-Hwan;Nam, Ki-Seok;Ko, Kyung-Won;Lee, Chan-Yong
    • Korean Journal of Microbiology
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    • v.47 no.1
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    • pp.14-21
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    • 2011
  • Riboflavin synthase catalyzes the formation of one molecule of each riboflavin and 5-amino-6-ribitylamino-2,4-pyrimidinedione by the transfer of a 4-carbon moiety between two molecules of the substrates, 6,7-dimetyl-8-ribityllumazine. The most remarkable feature is the sequence similarity between the N-terminal half (1-97) and the C-terminal half domain (99-213). To investigate the structure and fluorescent characteristics of the N-terminal half of riboflavin synthase (N-RS) in Escherichia coli, more than 10 mutant genes coding for the mutated N-terminal domain of riboflavin synthase were generated by polymerase chain reaction. The genes coding for the proteins were inserted into pQE vector designed for easy purification of protein by 6X-His tagging system, expressed, and the proteins were purified. Almost all mutated N-terminal domain of riboflavin synthases bind to 6,7-dimethyl-8-ribityllumazine and riboflavin as fluorescent ligands. However, N-RS C47D and N-RS ET66,67DQ mutant proteins show colorless, indicating that fluorescent ligands were dissociated during purification. In addition, most mutated proteins show low fluorescent intensity comparing to N-RS wild type, whereas N-RS C48S posses stronger fluorescent intensity than that of wild type protein. Based on this result, N-RS C48S can be used as the tool for high throughput screening system for searching for the compound with inhibitory effect for the riboflavin synthase.

A study of morphological characteristics and hybridization on Lepista nuda (민자주방망이버섯(Lepista nuda )의 형태적 특성 및 교배에 관한 연구)

  • Jung, Won Soon;Kim, Jong Bong
    • Journal of Mushroom
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    • v.11 no.1
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    • pp.1-8
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    • 2013
  • In this study, twelve of Lepista nuda were collected from various localities in Korea. Also thirteen exotic L. nuda species were collected from Japan, France, Switzerland and Portugal. Spores were isolated under optical microscope. These spores were placed on the surface of YM medium for inducing to germination. Eleven mating-groups were selected by morphological characters of fruit body such as size, color and stipe patterns. Intra-isolate crosses were made between two single-spore isolates derived from mating-groups. Also, dikaryotic crossing using the isolates from L. nuda were carried out to evaluated tetrakaryon formation. Cross-mating compatibility tests also verified its dikaryotic state by microscopic or molecular genetic observation of clamp connection and Random Amplified Polymorphic DNA (RAPD) band pattern. To analyze the growth rate of hybrids and parents mycelium in dikaryons obtained from compatible mating groups were placed on PDA medium. Intra-isolate crosses determined eleven mating-groups within L. nuda. The typical clamp connection were mostly observed in mating-groups of Korean L. nuda in $K1{\times}K2$, $K1{\times}K3$, $K1{\times}K4$, $K1{\times}K6$, $K1{\times}K5$, $K2{\times}K4$, $K2{\times}K3$, $K2{\times}K6$, $K3{\times}K4$, $K4{\times}K5$, and $K4{\times}K6$. Korean L. nuda type of dikaryon, shown to cross-incompatibility with L. sordida, it seemed that mating induce more rapidly than wild types in a view of growth rate. In conclusion, it would be useful to improve mass production with better morphological characteristics through a special mating of L. nuda.

Expression of Clostridium thermocellum Endoglucanase Gene in Lactobacillus bulgaricus and Lactobacillus plantarum and in vitro Survival Characteristics of the Transformed Lactobacilli (Lactobacillus bulgaricus와 Lactobacillus plantarum 균주에서 Clostridium thermocellum 유래 endoglucanase의 발현과 발현 유산균의 in vitro 생존 특성)

  • Cho, J.S.;Kang, S.H.;Lee, H.G.;Lee, H.J.;Woo, J.H.;Moon, Y.S.;Yang, C.J.;Choi, Y.J.
    • Journal of Animal Science and Technology
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    • v.45 no.4
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    • pp.659-666
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    • 2003
  • Endoglucanase A from Clostridium thermocellum which is resistant to pancreatic proteinase was selected out of numbers cellulases then were expressed in lactobacilli. Recombinant lactobacilli expression vector, pSD1, harboring the endoglucanase gene from C. thermocellum under the control of its own promoter, was constructed. Both L. bulgaricus and L. plantarum were electrotransformed with pSD1. The endoglucanase activities of 0.120 and 0.144 U/ml were found in culture media of L. bulgaricus and L. plantarum containing pSD1, respectively. In vitro survival characteristics of the transformed lactobacilli were tested. Both L. bulgaricus and L. plantarum showed a similar resistance to low pH 3. Moreover, L. plantarum was bile-salt resistant in the presence of 0.3 and 1% oxgall. L. bulgaricus and L. plantarum showed a rather homogenous resistant pattern against the tested antibiotics. Both of the strains were resistant to amikacin, gentamicin, streptomycin, kanamycin, and colistin.

The Nutritive Values and Manufacture of Total Mixed Fermentation Feeds using Green Forage Crops and RiCE-straw (청예 사료작물과 볏짚을 이용한 완전배합발효사료의 제조와 영양적가치)

  • Lee, H.J.;Cho, K.K.;Kim, W.H.;Kim, Hyeon-Seop;Kim, J.S.;Hang, S.H.;Woo, J.H.;Lee, H.G.;Choi, Y.J.
    • Journal of Animal Science and Technology
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    • v.44 no.1
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    • pp.75-86
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    • 2002
  • Adequate forage amounts in ruminant animal are necessary for proper ruminal function in dairy cow. This study was carried out to determine the effect of total mixed fermentation feeds made by different green forage crops and grain processings on chemical compositions, RFV (relative feed value) and ruminal characteristics in sheep. The experiment was arranged in a split plot design with 4 replications. The main plot consisted of 6 kinds of green forages (corn, grass, rye, rape, alfalfa and oat) and the sub plots three different grain processings such as non-milling, half milling (7mm mesh over), and regular milling (7mm mesh below). And the different TMFFs (total mixed fermentation feeds) were analyzed for chemical composition and fed to 8 ruminally fistulated sheep for ruminal charactics and palatability. RFV, daily feed intake, acetate/propionate ratio of the rape-TMFFs were higher compared with the other treatment. Ruminal content of VFA (volatile fatty acid) of corn-TMFF was highest as 90.19 mmol% and pH of the feed was lowest as 3.82. But, acetic acid, propionic acid and butyric acid were no difference among treatments. In conclusion, the effect of grain proceeding was not appeared but if consider of only RFV, palatability and dry matter disappearance, grade of TMFF was improved in order of rape-, corn-, alfalfa-, grass-, oat- and rye-TMFF.

Establishment and Characterization of Multipotent Germ Line Stem Cells (MGSCs) from Neonatal Mouse Testis (신생 생쥐 고환에서 기인한 다분화능 생식줄기세포주의 확립 및 특성 분석)

  • Han, Sang-Chul;Song, Haeng-Seok;Jun, Jin-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.35 no.1
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    • pp.39-48
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    • 2008
  • Objective: The aim of this study was to investigate whether multipotent germline stem cells (MGSCs) can be established from neonatal mouse testis. Methods: Various cells containing MGSCs were collected from neonatal testis of ICR mice and allocated to plates for in vitro culture. After 7 days in culture, the cells were passed to a fresh culture plate and continuously cultured. From the third or fourth passage, the presumed MGSCs were cultured and maintained on mitomycin C-inactivated STO feeder cells. The MGSCs were cultured in a condition where mouse embryonic stem cells (ESCs) are cultured. Characteristics of the MGSCs were evaluated by RT-PCR, immunocytochemistry, alkaline phosphatase activity, karyotyping, and transmission electron microscopy. Results: Two MGSCs lines were established from 9 pooled sets of neonatal testicular cells. MGSCs colonies were morphologically undistinguishable from ESCs colonies and both MGSC lines as well as ESCs expressed undifferentiated stem cell markers, such as Thy-1, Oct-4, Nanog, Sox2 and alkaline phosphatase. Fine structure of undifferentiated MGSCs were similar to those of ESCs and 60% of MGSCs (12/20) had normal karyotype at passage 10. They were able to form embryoid bodies (EBs) and MGSC-derived EBs expressed marker genes of three germ layers. Conclusion: We could establish the MGSCs from neonatal mouse testis and they were differentiated to multipotent lineages of three germ layers. Molecular characteristics of MGSCs were similar to those of ESCs. Our results suggest a possibility that multipotent stem cells derived from testis, the MGSCs, could replace the ESCs in biotechnology and regenerative medicine.

Characteristics of the yellowfin sole and dover sole skins as processing material of gelatin (젤라틴의 원료로서 가자미류 껍질의 성상)

  • Kim, Jin-Soo;Kim, Jeong-Gyun;Cho, Soon-Yeong;Ha, Jin-Hwan;Lee, Eung-Ho
    • Applied Biological Chemistry
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    • v.36 no.4
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    • pp.290-295
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    • 1993
  • In order to utilize effectively fish skin from fish processing, characteristics of the yellowfin sole and dover sole skins were investigated. In the yellowfin sole, the crude protein content and yield of fish skin used for the preparation of gelatin were 22.3% and 11.3%, respectively and in the dover sole, 17.2% and 8.9%, respectively. In the yellowfin sole skin, the soluble and insoluble collagen occupied 66.1% and 33.9%, respectively and in the dover sole skin, 78.8% and 21.1%, respectively. No difference in the amino acid composition between soluble and insoluble collagen was detected. The sum of proline and hydroxyproline content in the collagen extracted from fish skin was lower than that of those from pork skin or bone. The molecular weight of the two major subunits from the soluble collagen in the yellowfin sole skin were found to be 143 KDa and 202 KDa. Those in the dover sole skin were 142 KDa and 207 KDa. The physico-chemical properties such as the melting point and gelling point of yellowfin sole skin gelatin were superior to those of dover sole skin gelatin.

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Occurrence and characterization of oseltamivir-resistant influenza virus in children between 2007-2008 and 2008-2009 seasons

  • Kim, Seoung Geun;Hwang, Yoon Ha;Shin, Yung Hae;Kim, Sung Won;Jung, Woo Sik;Kim, Sung Mi;Oh, Jae Min;Lee, Na Young;Kim, Mun Ju;Cho, Kyung Soon;Park, Yeon Gyeong;Min, Sang Kee;Lee, Chang Kyu;Kim, Jun Sub;Kang, Chun;Lee, Joo Yeon;Huh, Man Kyu;Kim, Chang Hoon
    • Clinical and Experimental Pediatrics
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    • v.56 no.4
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    • pp.165-175
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    • 2013
  • Purpose: There was a global increase in the prevalence of oseltamivir-resistant influenza viruses during the 2007-2008 influenza season. This study was conducted to investigate the occurrence and characteristics of oseltamivir-resistant influenza viruses during the 2007-2008 and 2008-2009 influenza seasons among patients who were treated with oseltamivir (group A) and those that did not receive oseltamivir (group B). Methods: A prospective study was conducted on 321 pediatric patients who were hospitalized because of influenza during the 2007-2008 and 2008-2009 influenza seasons. Drug resistance tests were conducted on influenza viruses isolated from 91 patients. Results: There was no significant difference between the clinical characteristics of groups A and B during both seasons. Influenza A/H1N1, isolated from both groups A and B during the 2007-2008 and 2008-2009 periods, was not resistant to zanamivir. However, phenotypic analysis of the virus revealed a high oseltamivir $IC_{50}$ range and that H275Y substitution of the neuraminidase (NA) gene and partial variation of the hemagglutinin (HA) gene did not affect its antigenicity to the HA vaccine even though group A had a shorter hospitalization duration and fewer lower respiratory tract complications than group B. In addition, there was no significant difference in the clinical manifestations between oseltamivir-susceptible and oseltamivir-resistant strains of influenza A/H1N1. Conclusion: Establishment of guidelines to efficiently treat influenza with oseltamivir, a commonly used drug for treating influenza in Korean pediatric patients, and a treatment strategy with a new therapeutic agent is required.

Photocatalytic Oxidation of Han River Humic Substances and Change of Their Characteristics by $TiO_2/UV$ in a Rotating Photoreactor ($TiO_2/UV$ 회전반응기를 이용한 한강 휴믹물질의 광촉매산화 처리 및 특성 변화)

  • Shin, Jee-Won;Kim, Hyun-Chul;Han, Ihn-Sup
    • Journal of Korean Society of Environmental Engineers
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    • v.27 no.10
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    • pp.1129-1135
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    • 2005
  • In this study. the applicability of a rotating reactor for the oxidative removal of aqueous humic substances extracted from the Han River in Seoul, Korea was investigated. As air blowing for proper mixing of $TiO_2$ photocatalyst could inhibit UV-irradiation between a UV lamp and photocatalyst by air bubbles, a rotating reactor with some baffles was used for better UV-irradiation effect in this study. Han River humic substances are different from the other commercial humic substances(e.g., from Aldrich and International Humic Substance Society). Their characteristics were investigated with structural and spectroscopic analyses using FT-IR(Fourier transform-infrared), and $^{13}C$-NMR (nuclear magnetic resonance). The humic substances were extracted by XAD-7HP and treated with $TiO_2$-coated hollow beads under UV-A and UV-C irradiation in order to solve problems of separation and recovery of photocatalyst after reaction. At approximately 5 mg/L of initial TOC concentration, pH 3 and $2.0\;g-TiO_2/L$ dose, photocatalytic oxidation of Han River humic substances showed the optimum removal efficiency. Also, UV-C and UV-A lamps showed similar TOC removal efficiency. However, under UV-C irradiation, Han River humic substances were degraded to smaller compounds and increased the proportion of low molecular weight fractions compared to UV-A.

Effect of H2O2 Injection and Temperature Changing on the Organic Carbon Fraction in Chromatogram Dissolved Organic Carbon (CDOC) from Thermal Pretreatment (H2O2 주입과 온도변화가 열적 전처리 후 발생 슬러리의 CDOC 유기탄소분율에 미치는 영향평가)

  • Kim, Hee-Joong;Kim, Tae-Kyoung;Kim, Youn-Kwon
    • Journal of Korean Society of Environmental Engineers
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    • v.38 no.3
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    • pp.110-116
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    • 2016
  • Biogas yields point of view, the possibility of reusing excess sludge treated by thermal pretreatment for the purpose of improving the efficiency of the anaerobic digestion process has been investigated in recent year. Thermal pretreatment technology was considered as a pretreatment technique to improve excess sludge properties because of the solubilization of particulate organics. As a view point of sludge reduction and recycle, however, many researchers focused on the ability of particulate hydrolysis and COD solubilization under a high temperature, and few reports have addressed on the physical/chemical characteristics changing. This research was performed to evaluate the effects of a various temperature and chemical additives on carbon formation and fractionation in treated slurry from thermal pretreatment. Based on the results, it was revealed that oxidants injection and temperature changing have significantly caused the change of carbon fractions in slurry from thermal pretreament. Especially, it was considered that the efficiencies of particle hydrolysis increased with the increase of the reaction temperature. Low molecular weight(Mw < 350 g/mol) organic carbon formation increased with the increase of oxidants injection. It was expected that results of this research will provide an overview of the characteristics of thermal pretreatement for excess sludge reduction and recycle.

Investigation on Reaction Products From Oxidative Coupling Reactions of 1-Naphthol By Manganese Oxide (망간산화물에 의한 1-Naphthol의 산화-결합 반응에 따른 반응산물 연구)

  • Lim, Dong-Min;Lee, Doo-Hee;Kang, Ki-Hoon;Shin, Hyun-Snag
    • Journal of Korean Society of Environmental Engineers
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    • v.29 no.9
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    • pp.989-996
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    • 2007
  • In this study, abiotic transformation of 1-naphthol(1-NP) via oxidative-coupling reaction and its reaction products were investigated in the presence of Mn oxides. The reaction products were characterized for their relative polarity using solvent extraction experiment and reverse-phase HPLC, and for structure using CCMS and LC/MS, and for absorption characteristics using UV-Vis spectrometry. The reaction products present in aqueous phase were more polar than parent naphthol and comprised of 1,4-naphthoquinon(1,4-NPQ) and oligomers such as dimers and trimers. Hydrophilic component present in water phase after solvent$(CH_2Cl_2)$ extractions was identified as naphthol polymerized products having molecular weight(m/z) ranging from 400 to 2,000, and showed similar UV-Vis. absorption characteristics to that of foil fulvic acid. Transformation of 1,4-NPQ, which is non-reactive to Mn oxide, to the polymerized products via cross-coupling reaction in the presence of 1-NP was also verified. In this experimental conditions(20.5 mg/L, 1-NP, 2.5 g/L $MnO_2$, pH 5), the transformation of 1-NP into the oligomers and polymerized products were about 83% of initial 1-NP concentrations, and more than 30% of the reaction products was estimated to be water insoluble fractions, not extracted by $H_2O$ methanol. Results from this study suggest that Mn oxide-mediated treatment of naphthol contaminated soils can achieve risk reduction through the formation of oligomers md polymer precipitation.