• Title/Summary/Keyword: solid medium

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Studies on the ${\beta}-Tyrosinase$ -Part 1. On the Enzymological Characteristics of ${\beta}-Tyrosinase$- (${\beta}-Tyrosinase$에 관한 연구 -제1보, ${\beta}-Tyrosinase$의 효소학적(酵素學的) 성질(性質)에 대하여-)

  • Kim, Chan-Jo;Nagasawa, Toru;Tani, Yoshiki;Yamada, Hideaki
    • Applied Biological Chemistry
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    • v.22 no.4
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    • pp.191-197
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    • 1979
  • ${\beta}-Tyrosinase$ was purified and crystallized from cells of Escherichia intermedia A-21 grown in a medium supplemented with 0.2% L-tyrosine. Molecular weight of its subunit, Km value and absorption spectra were determined. Crystallization methods were also studied to eliminate any unnecessary procedures. The results obtained were as follows: 1. The purification procedure included ammonium sulfate fractionation, dialysis against potassium phosphate buffer, pH 6.0 and pH 7.0, and DEAE-Sephadex column chromatography. In the column chromatography, 11 mg of protein was applied per ml of DEAE-Sephadex for efficiency. 2. Steps of protamine sulfate treatment and Sephadex G-150 gel filtration could be eliminated for this enzyme from the known procedures. 3. The purified enzyme was dissolved in 0.01M potassium phosphate buffer containing 2-mercaptoethanol, with a concentration of 20mg/ml. Crystalline enzyme, which appears as hexagonal rods, was obtained by adding solid fine powdered ammonium sulfate to the enzyme solution. 4. Absorption maxima of the enzyme appeared at 340 and 430nm when associated with pyridoxal phosphate. 5. Km value of the enzyme for L-tyrosine was $2.31{\times}10^{-4}M$ and the molecular weight of its subunit was determined by SDS-polyacrylamide electrophoresis to be approximately 50,000.

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Morphological and Physiological Characteristics of Acclimated Liriodendron tulipifera Tree Produced by Several Types of In Vitro Germination Culture (백합나무 체세포배 기내발아 방식에 따른 순화묘의 형태적 및 생리적 특성)

  • An, Chan Hoon;Yi, Jae Seon;Moon, Heung Kyu;Kim, Yong Wook
    • Journal of Korean Society of Forest Science
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    • v.101 no.1
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    • pp.158-162
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    • 2012
  • The acclimatization of in vitro propagated plants is an important step to produce vigorous plants for clonal forestry and in vitro micro-environment may affect the growth in ex vitro condition. To monitor in vitro environmental effects on the growth in ex vitro condition, several culture systems such semi-solid medium(SS), temporary immersion bioreactor(TIB) and continuous immersion bioreactor(CIB) culture types were tested to compare for the growth of acclimated plants of Liriodendron tulipifera. Results suggested that morphological characters, stomatal conductance, evapotranspiration and chlorophyll contents of acclimated plants were affected by the different of in vitro culture conditions. CIB type of culture was resulted to the lowest value in the biomass of acclimated plants. Net photosynthsis rate of CIB was the same level as those of SS and TIB. However, stomatal conductance, evapotranspiration and $CO_2$ partial pressure in the intercellular air space were lower than those of SS and TIB. The amounts of chlorophyll a, b and carotenoids were also lower than those of the other two culture systems. TIB, showing a little lower or higher value than SS in many growth character, is recommended rather than CIB to produce healthy yellow poplar plants in ex situ condition.

Gene Structure and Function of fkhE, a Forkhead Gene in a Filamentous Fungus Aspergillus nidulans (Aspergillus nidulans forkhead 유전자 fkhE의 구조와 기능 분석)

  • Park, Mi-Hye;Kim, Hyoun-Young;Kim, Jong-Hwa;Han, Kap-Hoon
    • The Korean Journal of Mycology
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    • v.38 no.2
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    • pp.160-166
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    • 2010
  • A homothallic filamentous fungus Aspergillus nidulans has been used as the a model organism for studying growth and development for eukaryotic system. Various studies about specific transcription factors have been performed for elucidating the molecular mechanisms of growth, asexual and sexual developmental processes. Among them, the fkhE gene (AN2025.3) is located in chromosome VII and contains an ORF encoding 718 amino acid polypeptide intervening with two short introns. The cDNA sequencing revealed that at least four types of alternative splicing events were occurred when the fkhE gene was transcribed. The putative FkhE polypeptide contains a conserved forkhead domain and a bipartite nuclear localization signal at it's N-terminus and C-terminus, respectively. Deletion of fkhE resulted in impaired conidiophore formation in a solid medium. However, the sexual developmental process or cleistothecia formation was normal. Furthermore, fkhE deletion mutant produced conidiophores and conidia under the submerged culture, indicating that the fkhE gene is involved in asexual developmental process similar to the fkhF gene.

3D Finite Element Analysis of Lateral Loaded Pile using Beam and Rigid Link (빔요소와 Rigid 링크를 이용한 수평하중에 대한 말뚝 거동 3차원 유한요소해석)

  • Park, Du-Hee;Park, Jong-Bae;Kim, Sang-Yeon;Park, Yong-Boo
    • Land and Housing Review
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    • v.4 no.3
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    • pp.271-277
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    • 2013
  • The BNWF (Beam on Nonlinear Winkler Foundation) model is one of the simplest idealizations for a pile embedded in soil as it ignores the continuity of the soil. This method is difficult to model the behavior of pile group foundation subjected to lateral loading. The limitation can be overcome with the utilization of the finite element method (FEM) or finite different method (FDM) to represent a pile element embedded in a soil medium. Both the ground and piles are modeled with soild elements. The solid elements, which do not have rotational degree of freedom, is not appropriate for modeling piles. It can be overcome by substantially increasing the number of elements, which can be prohibitive for 3D modeling. This paper used the beam element and rigid link incorporated in the OpenSees to model the pile. The accuracy of the model is validated through comparison with lateral load test and BNWF analysis. It is shown that the method can capture the measured behavior accurately. It is therefore recommended to be used in group pile analyses.

Micro-Spot Atmospheric Pressure Plasma Production for the Biomedical Applications

  • Hirata, T.;Tsutsui, C.;Yokoi, Y.;Sakatani, Y.;Mori, A.;Horii, A.;Yamamoto, T.;Taguchi, A.
    • Proceedings of the Korean Vacuum Society Conference
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    • 2010.02a
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    • pp.44-45
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    • 2010
  • We are currently conducting studies on culturing and biocompatibility assessment of various cells such as neural stem cells and induced pluripotent stem cells(IPS cells) on carbon nanotube (CNT), on nerve regeneration electrodes, and on silicon wafers with a focus on developing nerve integrated CNT based bio devices for interfacing with living organisms, in order to develop brain-machine interfaces (BMI). In addition, we are carried out the chemical modification of carbon nanotube (mainly SWCNTs)-based bio-nanosensors by the plasma ion irradiation (plasma activation) method, and provide a characteristic evaluation of a bio-nanosensor using bovine serum albumin (BSA)/anti-BSA binding and oligonucleotide hybridization. On the other hand, the researches in the case of "novel plasma" have been widely conducted in the fields of chemistry, solid physics, and nanomaterial science. From the above-mentioned background, we are conducting basic experiments on direct irradiation of body tissues and cells using a micro-spot atmospheric pressure plasma source. The device is a coaxial structure having a tungsten wire installed inside a glass capillary, and a grounded ring electrode wrapped on the outside. The conditions of plasma generation are as follows: applied voltage: 5-9 kV, frequency: 1-3 kHz, helium (He) gas flow: 1-1.5 L/min, and plasma irradiation time: 1-300 sec. The experiment was conducted by preparing a culture medium containing mouse fibroblasts (NIH3T3) on a culture dish. A culture dish irradiated with plasma was introduced into a $CO_2$-incubator. The small animals used in the experiment involving plasma irradiation into living tissue were rat, rabbit, and pick and are deeply anesthetized with the gas anesthesia. According to the dependency of cell numbers against the plasma irradiation time, when only He gas was flowed, the growth of cells was inhibited as the floatation of cells caused by gas agitation inside the culture was promoted. On the other hand, there was no floatation of cells and healthy growth was observed when plasma was irradiated. Furthermore, in an experiment testing the effects of plasma irradiation on rats that were artificially given burn wounds, no evidence of electric shock injuries was found in the irradiated areas. In fact, the observed evidence of healing and improvements of the burn wounds suggested the presence of healing effects due to the growth factors in the tissues. Therefore, it appears that the interaction due to ion/radicalcollisions causes a substantial effect on the proliferation of growth factors such as epidermal growth factor (EGF), nerve growth factor (NGF), and transforming growth factor (TGF) that are present in the cells.

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The Effects of Different Feeding Conditions on Growth, Cast Production and Conversion Efficiency of Organic Matter to the Earthworm(Eisenia foetida L.) (먹이조건의 차이가 지렁이 생육, 분립생산량 및 체조직으로의 유기물 전환효율에 미치는 영향)

  • Lee, Ji-Young;Lee, Ju-Sam
    • Korean Journal of Organic Agriculture
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    • v.16 no.3
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    • pp.287-298
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    • 2008
  • This experiment was carried out to investigate the effect of different feed conditions on growth, cast production and conversion efficiency of organic matter to the earthworm. The experiment was tested on cow manure(CM) mixed with rice hulls (RH) or rice straw(RS). The mixture ratios were designed as one time, two times and three times of cow manure volume, respectively. The CM and RH mixtures resulted better on growth rate, reproductive efficiency and wormcast production than those for CM and RS mixtures. Especially on three times of RH mixture showed the highest growth characteristics compared to the other mixtures. Although both were between 20 and 34 on carbon and nitrogen ratio, the result inferred that the difference on the growth might have been caused by feeding conditions. On the mixtures of cow manure with rice hulls(CM+RH) was significantly higher on values on the conversion rate and conversion efficiency of organic matter to earthworm tissues than the mixtures of cow manure with rice straw(CM+RS). The most concerned point of the mixture of earthworm feed is that high contents of volatile solid and total carbon that increases in conversion efficiency of organic matter to earthworm tissues, and also causes the increase in growth rate and reproductive efficiency. The wormcast could be used as a valuable plant growth medium or soil conditioner for sustainable agriculture and it may be due to their high qualities of physico-chemical properties.

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Isolation and Characterization of Sulfur-oxidizing Denitrifying Bacteria Utilizing Thiosulfate as an Electron Donor (황(thiosulfate)을 이용하는 탈질 미생물의 분리 및 특성 파악)

  • Oh, Sang-Eun;Joo, Jin-Ho;Yang, Jae E
    • Korean Journal of Soil Science and Fertilizer
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    • v.39 no.6
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    • pp.409-414
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    • 2006
  • Sulfur-oxidizing bacteria were enumerated and isolated from a steady-state anaerobic master culture reactor (MCR) operated for over six months under a semi-continuous mode and nitrate-limiting conditions using thiosulfate as an electron donor. Most are Gram-negative bacteria, which have sizes up to 12 m. Strains AD1 and AD2 were isolated from the plate count agar (PCA), and strains BD1 and BD2 from the solid thiosulfate/nitrate medium. Based on the morphological, physiological, FAME and 16S rDNA sequence analyses, the two dominant strains, AD1 and AD2, were identified as Paracoccus denitrificans and Paracoccus versutus (formerly Thiobacillus versutus), respectively. From the physiological results, glucose was assimilated by both strains AD1 and AD2. Heterotrophic growth of strains AD1 and AD2 could be a more efficient way of obtaining a greater amount of biomass for use as an inoculum. Even though facultative autotrophic bacteria grow under heterotrophic conditions, autotrophic denitrification would not be reduced.

Effect of Shade Levels on Growth and Fruit Blight of 'Campbell Early' Grapes (포도 '캠벨얼리' 차광수준이 수체생장과 열매마름 증상에 미치는 영향)

  • Kim, Byeong-Sam;Cho, Kyung-Chul;Hwang, In-Taek;Choi, Hyun-Sug;Jung, Seok-Kyu
    • Korean Journal of Organic Agriculture
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    • v.27 no.1
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    • pp.45-56
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    • 2019
  • The study I was initiated in six private 'Campbell Early' vineyards in parts of Jeollanam-do province to relieve symptom of fruit blight mostly due to a high temperature occurred in Summer. High percentage of fruit blight was observed for medium growth of grape trees non-irrigated. In the study II, grape trees treated with 0%, 30% and 60% shade films were investigated for the tree responses, including fruit blight symptom in the research plots. Harvesting time was advanced approximately two weeks by the 30% shade treatment. High percentage of shades increased total shoot length per tree and decreased shoot diameter, with the greatest number of shoots observed for the 30% shade-treated grapes. Light intensity in the tree canopy was approximately 26% decreased by 30% shade treatment. Marketable harvested fruits per tree were 50.6 of 30% shade treatment, 33.6 of 0% shade, and 42.8 of 60% shade. The 30% shade treatment decreased to 11.8% of cluster blight per tree and to 11.2% of berry blight per cluster. The 30% shade treatment increased cluster weight, berry weight, soluble solid contents, and anthocyanin contents.

Determination of Siderophore from Bacillus Mojavensis Using Liquid Chromatography quadrupole Time-of-flight Tandem Mass Spectrometry (액체크로마토그래피-사중극 비행시간형 탠덤질량분석기를 이용한 Bacillus mojavensis 균주 속 사이드로포어 규명)

  • Cheon, Hae In;Yeo, Mi Seon;Kim, Kang Min;Kang, Jae Seon;Pyo, Jaesung
    • Journal of Life Science
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    • v.29 no.2
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    • pp.198-201
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    • 2019
  • Recently, it has been reported that Bacillus mojavensis possesses antifungal properties and plant growth-promoting activities, which are similar to the characteristics of siderophore. In this study, the siderophore produced by B. mojavensis was assessed using a solid phase extraction (SPE) cartridge and liquid chromatography quadrupole time-of-flight tandem mass spectrometry (Q-TOF MS/MS). After B. mojavensis was incubated in phenol medium for 16 hr and lyophilized, the sample was dissolved in water and loaded to an SPE cartridge to remove interferences. The cartridge was washed with 5% methanol in water and eluted with 2% formic acid in methanol sequentially. The eluted solution was evaporated under a stream of nitrogen gas and reconstituted in methanol. The reconstituted sample was filtered, and $1{\mu}l$ of the sample was assessed using Q-TOF MS/MS. The mass spectrometer was operated using the positive electrospray ionization mode. Based on the mass spectrum and tandem mass spectrum, the siderophore produced by B. mojavensis was bacillibactin, one of the catechol types of siderophore with a molecular weight of 882.2556. This siderophore analysis could provide a justification for the study of B. mojavensis as a functional food and for pharmaceutical applications.

Purification and Biochemical Characterization of β-agarase Produced by Marine Microorganism Cellulophga sp. J9-3 (해양미생물 Cellulophga sp. J9-3이 생산하는 베타-아가레이즈의 분리 및 생화학적 특성)

  • Kim, Da Som;Kim, Jong-Hee;Chi, Won-Jae
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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    • pp.329-336
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    • 2021
  • Cellulophga sp. J9-3, is a gram-negative, aerobic marine bacterium belonging to the family Flavobacteriaceae. In addition to cellulose degradability, the J9-3 strain is also capable of hydrolyzing agar in the solid and liquid medium, and the production of agarase in the presence of agarose can be remarkably induced by the bacterium. From the cell culture broth of Cellulophga sp. J9-3, ammonium sulfate precipitation and three kinds of column chromatography were successively performed to purify a specific agarase protein, the AgaJ93. Purified AgaJ93 showed the strongest hydrolyzing activity towards agarose (approximately 22%), and even displayed activity towards starch. AgaJ93 hydrolyzed agarose into neoagarotetraose and neoagarohexaose via various oligosaccharide intermediates, indicating that AgaJ93 is an endo-type β-agarase. AgaJ93 showed maximum activity at a pH of 7.0 and temperature of 35 ℃. Its activity increased by more than six times in the presence of Co2+ ions. The N-terminal sequence of AgaJ93 showed 82% homology with the heat-resistant endo-type β-agarase Aga2 of Cellulophaga sp. W5C. However, the biochemical properties of the two enzymes were different. Therefore, AgaJ93 is expected to be a novel agarose, different from the previously reported β-agarases.