• Title/Summary/Keyword: C4 column

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Studies on the Production of Thermostable Amylase. Part 1. Optimal Culture Conditions and Purification of Enzyme. (내열성 Amylase의 생산에 관한 연구 (제1보) 최적배양조건과 효소의 정제)

  • 오두환;이강표;변유량;유주현
    • Microbiology and Biotechnology Letters
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    • v.9 no.2
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    • pp.91-97
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    • 1981
  • A thermophilic soil isolate Bacillus sp. Y-127 was selected for the production of thermostable amylase. The strain was used for the enzyme production and the thermostable amylase was characterized. The optimum cultural conditions for the enzyme production were 6$0^{\circ}C$ at pH 7.0 for 32 hours using a mineral medium containing 2% soluble starch and 0.2% yeast extract. The extra-cellular enzyme was purified about 123-folds with about 6% recovery. The purified enzyme was stable at pH between 4.0 and 7.0, and temperature up to 6$0^{\circ}C$.

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Purification and Properties of Extracellular Adenine Deaminase from Nocardioides sp. J-257L (Nocardioides sp. J-275L이 생산하는 세포외 Adenine Deaminase의 정제 및 성질)

  • 전홍기;박정혜;김태숙
    • Korean Journal of Microbiology
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    • v.25 no.3
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    • pp.221-228
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    • 1987
  • The extracellular adenine deaminase from Nocardioides sp. J-275L was purified by the following techniques: ammonium sulfate fractionation, DEAE-Cellulose, DEAE-Sephadex A-50 column chromatography, and Sephacryl S-200 superfine gel filtration. The enzyme was partially purified about 3889.5-fold with about 5.2% yield by these procedures. The molecular weight of the enzyme was 39,000 by a calibrated Sephacryl S-200 superfine column chromatography. The enzyme was stable at pH 7.5 and up to $40^{\circ}C$. Glycerol was effective on the stabilization of the enzyme during storage. The optimum pH and temperature of the enzyme were around pH 7.5 and $40^{\circ}C$, respectively. The apparent Michaelis constant Km of the enzyme for adenine was $7.4\times 10^{-5}$M. The purine analogues, 6-chloropurine, 2,6-diaminopurine, 6-bromopurine, 4-aminopyrazolo [3.4-d]pyrimidine, and 8-azaadenine were substrates for the enzyme. 6-Dimethylaminopurine was a competitive inhibitor of the enzyme. The enzyme was inhibited by 1mM of $Cu^{2+}, Fe^{3+}, Pb^{2+}, Hg^{2+}$, and $Ag^{+}$, and 1mM of $\alpha$,$\alpha$'-dipyridyl, pentachlorophenol, and pCMB.

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A New Gas-Chromatograghic Method of Organic Elemental Analysis (가스크로마토그래피에 依한 微量元素分析)

  • Kim, You-Sun;Son, Youn-Soo;Choi, Q.Won
    • Journal of the Korean Chemical Society
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    • v.8 no.4
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    • pp.188-191
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    • 1964
  • A new gas-chromatographic method for determining carbon and hydrogen in organic compounds has been developed. After sample combustion was performed in a regular analytical combustion tube with an internal oxidant (a mixture of silver oxide and manganese dioxide) under a helium flow, the water produced was converted to acetylene by passing through a calcium carbide tube. The carbon dioxide and acetylene were trapped by a molecular sieve 5A column at room temperature. The trapped gases were released under programmed temperature raise up to $340^{\circ}C$ and the released gases were passed through a silica gel column. The adsorption of $CO_2$ and $C_2H_2$ in the molecular sieve 5A trapping column were found to be quantitative and the silica gel column showed an excellent resolution of $CO_2$ and $C_2H_2$ for analytical purpose. The analytical results for various known compounds based on the out-put of the thermal conductivity cell calibrated for the amounts of carbon and hydrogen contents in benzoic acid, showed average errors ${\pm}0.5%$ and ${\pm}0.33%$ for carbon and hydrogen, respectively.

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The Development of Analysis System for Genes Related Disease Using Chemical Properties of DHPLC (DHPLC의 화학적 특성을 이용한 질병 유전자의 분석 시스템 개발)

  • Kim, Jong-Gyu;Nam, Yun-Hyeong;Park, Sang-Beom;Lee, Jae-Sik;Gang, Won
    • Journal of the Korean Chemical Society
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    • v.50 no.2
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    • pp.116-122
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    • 2006
  • In this study we extracted DNA from 100 tissues of breast cancer patients and 103 normals. Then we confirmed single-nucleotide polymorphism(SNP) using PCR-DHPLC(polymerase chain reaction-denaturing high performance liquid chromatogrphy).Also, we studied SNP of samples using several columns to identify relation between packing materials of column and resolution.As a result, we identified 4 C/A, C/G genotypes(4%) in exon 5 and 37 T del genotypes(37%) in exon 8 among 100 breast cancer tissues and 2 in exon 5, 9 in exon 8 among 103 normal samples.In resolution test, we confirmed that PS-DVB(poly styrene-divinylbenzen) column is more efficient than C18 column.

Purification and Characteristics of Adenylate Kinase from Extreme Thermophile Thermus caldophilus GK-24 (고도 호열성균 Thermus caldophilus Adenylate Kinase의 정제와 성질)

  • ;Ohta Takahisa
    • Microbiology and Biotechnology Letters
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    • v.16 no.5
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    • pp.393-397
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    • 1988
  • The adenylate kinase was purified from an extreme thermophile by adenosine-pentaphospho-adenosine elution from phosphocellulose column. The molecular weight was estimated to be 22,000 by SDS-PAGE and gel filtration. The optimum temperature of the enzyme activity was 8$0^{\circ}C$ and the activation energy was given as 22.4 kcal/mole. The enzyme even showed full activity after incubation at 9$0^{\circ}C$ or in 6M guanidine-HCI at 3$0^{\circ}C$ and retained 75% of its original activity even after 1 hour at 10$0^{\circ}C$. The Michaelis constants of the enzymes for AMP, ADP, and ATP were 0.01mM, 0.017mM and 0.067mM, respectively.

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Characteristics of Liquid-Liquid Direct Contact Heat Exchanger for a Solar System (태양열 이용을 위한 직접접촉식 액-액 열교환기 특성)

  • 강인석;김종보;강용혁;곽희열
    • Transactions of the Korean Society of Mechanical Engineers
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    • v.18 no.12
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    • pp.3276-3286
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    • 1994
  • In most direct contact liquid-liquid heat exchangers, oil or hydrocarbon with a density less than water is normally used as dispersed working fluid. The main difficulty that arises with this arrangement lies in the control of the interface at the top of the column. When it is connected with a solar collector which uses water as its working fluid, the main difficulties arise from the fact that the water can be frozen during winter time. In order to solve these problems and to demonstrate the technical feasibility of a direct contact liquid-liquid heat exchanger, liquids heavier than water with low freezing temperature has been utilized as dispersed phase liquids in a small laboratory scale model made of pyrex glass. In the present investigation, dimethyl phthalate(C/sub 6/H/sub 4/)COOCH/sub 3/)/sub 2/) and diethyl phthalate (C/sub 6/H/sub 4/(CO/sub 2/C/sub 2/H/sub 5/)/sub 2/) are utilized as heavy dispersed phase working fluids. The results of the present investigation the technical in the utilization of heavier dispersed working liquid in the spray-column liquid-liquid heat exchanger for a solar system. The overall average temperature difference along the column is found to be almost half of the initial temperature difference between the dispersed and the continuous phase. Despite the fact that the two phthalates tested in the experiment differ significantly in some of their physical properties, the volumetric heat transfer coefficients in terms of dispersed fluid superficial velocities were found to be similar for both phthalates tested.

A Study on the Hysteretic Behavior of High Strength Steel Fiber Reinforced Concrete Beam-Column Joint (강섬유 보강 고강도 철근 콘크리트 보-기둥 접합부의 이력 거동에 관한 연구)

  • 오경남;이정한;유영찬;이원호;정헌수
    • Proceedings of the Korea Concrete Institute Conference
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    • 1999.10a
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    • pp.533-536
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    • 1999
  • The primary purpose of this study is to investigate experimentally the effect of steel fiber reinforcement on the total energy dissipation capacity of R/C flexural members and to make a contribution to the construction of 40~60 story R/C high rise building by developing the new materials and reinforcing details which can improve the seismic performance of high-strength R/C beam-column joints. Experimental research was carried out on 4 type specimen under cyclic loading. Main variables are steel fiber reinforcement, intermediate reinforcements and yield strength of rebars. From the test results, steel fiber reinforcement can improve the ductility of R/C flexural members.

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Quantitative Analysis of Lignan Compounds in Shred and Smoke of 'Balloon-Flower' Cigarettes (도라지담배 각초 및 연기중의 Lignan화합물 정량)

  • 나효환;손현주;백순옥;복진영
    • Journal of the Korean Society of Tobacco Science
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    • v.11 no.2
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    • pp.225-232
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    • 1989
  • Deoxyschizandrin, gomisin N, schizandrin, wuweizisu C, gomisin A and angeloylgomisin H were isolated from fruits of Schisandra chinensis BAILLON, and the lignan contents in shred and smoke of 'Balloon-Flower' cigarettes were quantitatively analyzed by capillary-GC(FID). The GC column was SPB-1 fused silica capillay (0.25mm id$\times$30m, Supelco) and the column oven temperature was programmed from 20$0^{\circ}C$ to 30$0^{\circ}C$ at the rate of 4$^{\circ}C$/min. In the shred of 'Balloon-Flower' cigarettes deoxyschizandrin. gomisin N, schizandrin, gomisin A and angeloylgomisin H were detected and schizandrein contets were the highest among them, 22.77$\mu\textrm{g}$/cig., gomisin N, schizandrin and gomisin A were 0.023, 0.054, 0.0849 and 0.167$\mu\textrm{g}$/cig. respectively, and angeloylgomisin H was not detected.

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Electrospray-Mass Spectrometric Analysis of Plasma Pyrophosphates Separated on a Multi-Modal Liquid Chromatographic Column

  • Lee, Su-Hyeon;Lee, Jeong-Ae;Lee, Won-Yong;Chung, Bong-Chul;Choi, Man-Ho
    • Mass Spectrometry Letters
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    • v.2 no.4
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    • pp.92-95
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    • 2011
  • Pyrophosphates are the key intermediates in the biosynthesis of isoprenoids, and their concentrations could reveal the benefits of statins in cardiovascular diseases. Quantitative analysis of five pyrophosphates, including isopentenyl pyrophosphate (IPP), dimethylallyl pyrophosphate (DMAPP), geranyl pyrophosphate (GPP), farnesyl pyrophosphate (FPP), and geranylgeranyl pyrophosphate (GGPP), was performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in negative ionization mode. After dilution with methanol, samples were separated on a 3 ${\mu}m$ particle multi-modal $C_{18}$ column ($50{\times}2$ mm) and quantified within 10 min. The gradient elution consists of 10 mM ammonium bicarbonate and 0.5% triethylamine (TEA) in water and 0.1% TEA in 80% acetonitrile was used at the flow rate of 0.4 mL/min. Overall recoveries were 51.4-106.6%, while the limit of quantification was 0.05 ${\mu}g$/mL for GPP and FPP and 0.1 ${\mu}g$/mL for IPP, DMAPP, and GGPP. The precision (% CV) and accuracy (% bias) of the assay were 1.9-12.3% and 89.6-111.8%, respectively, in 0.05-10 ${\mu}g$/mL calibration ranges ($R^2$ > 0.993). The devised LC-MS/MS technique with the multi-modal $C_{18}$ column can be used to estimate the biological activity of pyrophosphates in plasma and may be applicable to cardiovascular events with cholesterol metabolism as well as the drug efficacy of statins.

A Role and Properties of $C_{1}$ Enriched Cellulase Fraction from Anaerobic Clostridium thermocellum in Cellulose Degradation (섬유소 분해시 혐기성 Clostridium thermocellum이 생산하는 Cellulase의 $C_{1}$ 성분의 역할과 성질)

  • 이용현;심욱한;신현동
    • Korean Journal of Microbiology
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    • v.25 no.4
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    • pp.293-303
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    • 1987
  • A $C_{1}$ enriched cellulase fraction was separated from culture filtrate of anaerobic Clostridium thermocellum by hydroxyapatite column chromatography. The separated fraction showed strong synergistic action with $C_{x}$ component (endo-$\beta$-1, 4-glucanase) in digestion of crystalline cellulose, similar to the other aerobic cellulolytic microorganisms. Unlike the $C_{x}$ component the $C_{1}$ enriched fraction was rapidly inactivated by oxidation at the atmospheric condition. The enzyme activity was significantly enhanced by the addition of reducing agents, especially $\beta$-mercaptoethanol, which indicates that a $C_{1}$ component has a lot of sulfhydryl groups essential for the enzyme activity. The effect of metal ions on $C_{1}$ activity was also investigated. The $C_{1}$ fraction was found to be thermally stable compare to endo-$\beta$-1,4-glucanase. Optimal temperature and pH were found to be $60^{\circ}C$ and 6.0, respectively.

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