• Title/Summary/Keyword: Acetate buffer

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Studies on the cellulase properties of Aspergillus clavatus from the Cellulose-Cultural Properties (섬유질 문화재로부터 분리된 Aspergillus clavatus의 섬유소분해호소에 관한 연구)

  • Chung, Hee-Jin;Han, Sung-Hee;Ahn, Hee-Kyun;Min, Kyung-Hee
    • The Korean Journal of Mycology
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    • v.15 no.1
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    • pp.29-37
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    • 1987
  • Cellulolytic mocrooganisms were isolated from the cellulose-cultural properties. Among them, Aspergillus clavatus with the highest cellulase activity was identified by the morphological characteristics and it's enzyme activities were compared on the various cultural conditions. It was found that the induction of carboxymethylcellulase(CMCase), avicelase and salicinase in CMC liquid media showed the highest enzyme acitivity on five day's cultivation at $30^{\circ}C$ and thereafter their activities were gradually decreased with time. After crude extracellular enzymes precipitated with the 70% saturated ammonium sulfate solution were dialyzed with 20 mM acetate buffer pH 6.0, each crude enzyme was examined. The optimal activities of CMCase and avicelase were both found to be at $50^{\circ}C$ and pH 6.0. Their thermal stability was appeared at the $50^{\circ}C$. CMCase and avicelase had the maxima activities with 1.5% and 2.2% substrate concentration, respectively. The concentration of 5 mM $Mg^{++}$ or $Ca^{++}$ was found to have a maximum cellulase activity and its activity was inhibited with more than 5 mM $Cu^{++}$ and $Zn^{++}$ concentration. Cellulase activity was also inhibited sensitively by the inhibitors such as 2-mercaptoethanol, iodine and sodium azide.

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The Optimum Conditions for the Simultaneous Determination of Neurotransmitters in Rat Brain Striatum by High Performance Liquid Chromatography with Electrochemical Detection (HPLC-ECD를 이용한 흰쥐 뇌의 선조체 중 신경전달물질의 동시분석시 최적 조건)

  • Kang, Jong-Seong;Mun, Min-Seon;Shin, Hyung-Seon;Lee, Soon-Chul
    • Analytical Science and Technology
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    • v.8 no.2
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    • pp.215-220
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    • 1995
  • A simple, efficient and sensitive method was described for the simultaneous determination of catecholamine, indoleamine and related metabolites from the homogenates of the rat brain striatum by HPLC-ECD. The optimum mobile phase on a reverse phase $C_{18}$ column was 35mM sodium acetate buffer(included 10mM citric acid, 0.13mM $Na_4EDTA$, 0.58mM SOS, pH3-4):MeOH=85:15. The column temperature was $30^{\circ}C$. Dopamine(DA), 3, 4-dihydroxyphenyl acetic acid(DOPAC), homovanilic acid(HVA), 5-hydroxyindole acetic acid(5-HIAA), serotonin(5-HT) and noradrenaline(NA) could be separated and analysed to very small amount. The detection limits of this method were 2~10pg per injection for all components. The effects of age and sex of rat on the contents of the catecholamines and their metabolites in rat brain striatum were studied. The levels of DA and 5-HT contents of the 7 week old female rats were higher than those of the 7 week old male rats. As the age of rat increases, the contents of DOPAC increased significantly.

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Simultaneous Determination of Quinolones in Flatfish and Egg Using liquid Chromatography with Fluorescence Detection (액체크로마토그래피를 이용한 광어 및 계란 중 퀴놀론계의 동시분석법 개발)

  • Lee, Sang-Hee;Shim, You-Shin;Kim, Hyun-Ju;Choi, Yoon-Hee;Shin, Dong-Bin
    • Journal of Food Hygiene and Safety
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    • v.23 no.4
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    • pp.324-329
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    • 2008
  • An analytical method for the simultaneous determination of nine quinolones (QNs) namely, marbofloxacin, norfloxacin(IS), ciprofloxacin, danofloxacin, enrofloxacin, sarafloxacin, difloxacin, oxolinic acid, and flumequine in flatfish and egg was developed and validated using liquid chromatography with fluorescence detection (LC-FD). The samples were extracted using a traditional liquid-liquid extraction process; deproteinization was accomplished by the addition of trichloroacetic acid and acetonitrile (ACN), and defatting was performed with hexane. Chromatographic separation was achieved on a reverse phase C8 column with gradient elution using a mobile phase of 200 mM ammonium acetate buffer (pH 4.5) and ACN. The proposed method was validated according to the CODEX guideline. Mean recoveries of QNs from flatfish and egg were 89.6-106.5% with relative standard deviations (RSDs) below 15% at three different concentrations of 50, 100 and $500{\mu}g/kg$. Linearity was obtained with a correlation coefficient ($r^2$) of 0.9989-1.0000. The LOD for the investigated QNs was $1-16{\mu}g/kg$ depending on flatfish and egg. The present method can be applied simultaneously to determine QNs in muscle of flatfish and egg.

Hyaluronidase Inhibitory Activity of Extracts from Doenjang, Chungkookjang and Miso (된장, 청국장 및 미소추출물의 Hyaluronidase 저해활성)

  • Ahn, Sun-Kyung;Hong, Kwang-Won
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.8
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    • pp.1119-1123
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    • 2005
  • The inhibitory effects of water and methanol extracts of Doenjang, Chungkookjang and Miso on bovine hyaluronidase were examined. The extracts were vacuum-dried and the resulting pellets were dissolved with 0.1 M acetate buffer with 1$\%$ DMSO to a final concentration of 50 mg/mL. The water extracts of Doenjang, Chungkookjang and Miso inhibited 62$\%$, 70$\% $, 56$\%$ of hyaluronidase activity, respectively, while the extract of crushed soybean inhibited 24$\%$ of the activity. Under the same condition, disodiumcromoglycate known as an anti-allergic drug inhibited 46$\%$ of hyaluronidase activity at the concentration of 0.35 mg/mL as a positive control. Also the methanol extracts of Doenjang, Chungkookjang and Miso inhibited 48$\%$, 52$\%$, 70$\%$ of hvaluronidase activity, respectively, while the extract of crushed soybean inhibited 46$\%$ of the activity. After heat treatment of the water extracts of Doenjang, Chungkookjang and Miso at 100"C for 10 min, hyaluronidase inhibitory effects of them were reduced approximately to half. However, hyaluronidase inhibitory effects of methanol extracts were maintained well even after heat treatment.

Amperometric Determination of Anthracycline Antibiotics with the Mercury Film Thin Layer Flow Cell (수은피막 박막흐름전지를 이용한 Anthracycline계 항생제의 전류법 정량)

  • Kim, Kyung Eun;Hahn, Younghee
    • Analytical Science and Technology
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    • v.17 no.6
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    • pp.470-475
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    • 2004
  • The mercury film thin layer flow cell (MFTLFC) which yielded the highest sensitivity for the electrochemical reduction of doxorubicin was constructed by coating the glassy carbon working electrode (GCE; $A=0.208cm^2$) with $5{\mu}L$ of HgO coating solution (0.5% HgO + 0.25% polystyrene/cyclohexanone) and subsequently followed by applying a potential of -0.40 V for 300 sec in the flow stream of an acetate buffer of pH 4.5. The voltammogram of doxorubicin reached the diffusion current plateau at -0.53 V vs. a Ag/AgCl (3 M NaCl) in the MFTLFC. The diffusion current (Id) of doxorubicin at the MFTLFC was 1.7 times greater than the Id obtained at the TLFC employing a bare glassy carbon working electrode. When the peak areas (electric charge) were plotted vs. concentrations of standard anthracyclines, the calibration factors of doxorubicin and daunorubicin were $1.12{\times}10^8{\mu}C/M$ (coefficient of determination; $R^2$: 0.969) and $0.98{\times}10^8{\mu}C/M$> ($R^2$: 0.999), respectively in the concentration range between $1.0{\times}10^{-8}M$ and $1.0{\times}10^{-6}M$.

Optimization of Submerged Culture Conditions for Protease Production and Its Enzymatic Properties (Protease 생산을 위한 최적 배양조건 및 생산된 Protease의 특성)

  • Cho, Hee-Yeon;Cho, Nam-Seok
    • Journal of the Korean Wood Science and Technology
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    • v.32 no.5
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    • pp.12-19
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    • 2004
  • This study was performed to investigate the optimum condition of protease production from submerged culture of oak mushroom (Lentinula edodes, Sanlim No. 5) and its enzymatic features. Among several combinations of media, the combination of wheat bran, corn flour, water and corn oil (WB+CF+W+ CO) yielded 84.8 U/g of maximum protease activity. This combination of ingredients, in spite of not being particularly protein-rich in comparison to the other media, allowed for good growth of the fungus and maximal protease production. Comparison of different growth medium liquids indicated that demineralized water afforded the best growth of the fungus and the highest protease activity. Acetate buffer and acidified water negatively affected The protease production peaked around 72 hr of incubation, and decreased thereafter. The molecular weights of produced protease were about 45,000 by Sephadex G-75 chromatography. The pH optimum for protease activity was 4, while maximal activity incubated at 37℃ for 1 hr was observed between pH 4~6. The optimum temperature of this protease was 55℃, and the enzyme was active over a broad temperature range (30~60℃), indicating that this protease would be suitable for a wide range of applications where. different pH and temperature are necessary, such as digestive aids, food industry, beer and tannery industries.

Purification and Characterization of Xylanase from Bacillus sp. A-6 (Bacillus sp. A-6의 Xylanase의 정제와 특성)

  • Choi, Suk-Ho
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.147-152
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    • 2009
  • A xylanase was purified from the culture supernatant of Bacillus sp. A-6 by using ultrafiltration and ion exchange chromatography on the column of SP-Sepharose using 5 mM acetate buffer, pH 5.0. The xylanase was eluted from the column at the concentration less than 0.05 M NaCl. The eluted xylanase was shown to be a single protein band in SDS-PAGE. Zymogram analysis indicated that the protein band in SDS-PAGE had the enzyme activity to hydrolyze oat spelt xylan. The molecular weights of the xylanase were 15,000 based on SDS-PAGE and 14,100 based on gel filtration chromatography. Thin layer chromatography showed that the xylanase hydrolyzed oat spelt xylan into xylobiose and high-molecular-weight xylooligosaccharides. The relative activities of the heated xylanase decreased to 80% at $40^{\circ}C$ after 7 hr and less than 40% at $60^{\circ}C$ after 1 hr.

Ion-exchange Separation and Spectrophotometric Determination of Trace Amount of Aluminium with Thorinin the Presence of Triton X-100 (Triton X-100 존재하에 Thorin에 의한 미량의 알루미늄 이온의 분광학적 정량 및 이온-교환 분리)

  • Park, Chan-Il;Cha, Ki-Won;Jung, Duck-Chae
    • Analytical Science and Technology
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    • v.12 no.6
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    • pp.515-520
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    • 1999
  • The spectrophotometric determination of Al(III) with thorin have been investigated. The optimum condition of pH, concentration of ligand and surfactant, and stability were evaluated. The thorin ligand offers selective separation of Al(III) from sample solution containing Fe(III), Ni(II), Cu(II), Pb(II) and Cu(II). Various surfactants were tested and Triton X-100 showed the best stability and the maximum absorbance in an aqueous solution of Al(III)-Thorin-Triton X-100 complex appears about 526 nm. The method was applied for the determination of Al(III) in mixed sample solution. Separation and preconcentration was performed with a short column filled with resorcinol-formaldehyde resin. Control of the pH during the column operation is essential because the adsorption capacities are very sensitive to change in pH. Their separation was carried out in 0.2 M acetic acid-sodium acetate buffer solution (pH 4.5) and 1.0 M $HNO_3$media.

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Effects of Exogenous Bovine Somatotropin on Mammary Function of Late Lactating Crossbred Holstein Cows

  • Tanwattana, P.;Chanpongsang, S.;Chaiyabutr, N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.1
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    • pp.88-95
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    • 2003
  • The objective of the present study was to determine the effect of exogenous bovine somatotropin on the mammary function in late lactating crossbred Holstein cows. Twelve 87.5% late lactating Holstein cows, approximately 30 weeks postpartum, were divided into two groups of 6 animals each. Animals in the control group were given sodium bicarbonate buffer by subcutaneous injection, while animals in the treated group were given recombinant bovine somatotropin (bST) by subcutaneous injection with 500 mg of bST (14 day prolonged-release bST). After bST injection, milk yield significantly increased from the control level on day 8 to day 20 (p<0.05) with a concomitant increase in mammary blood flow (p<0.01). An increase in mammary blood flow in response to bST treatment was greater than an increase in milk production. An increased plasma concentration of IGF-I coincided with an increase in mammary blood flow in animals treated with bST. There were no significant changes in the concentration of arterial plasma glucose concentration, the arteriovenous concentration difference (A-V difference) and mammary extraction ratio while the mammary glucose uptake increased when compared to the control group. The concentration of arterial plasma triglyceride decreased throughout the experimental period in animals give bST. The plasma concentration of acetate, and the mammary uptake for acetate significantly increased (p<0.05) after bST treatment. The action of bST did not affect the plasma concentration, A-V difference and extraction ratio across the mammary gland for $\beta$-hydroxybutyrate. The concentrations of milk fat and lactose tended to increase during bST treatment. Milk protein concentration initially increased in the first few days and decreased after bST injection when compared to the pretreated period. The present results indicated that bST could affect the mammary function in late lactating cows by increase in milk yield involving changes in both extra-mammary and intra-mammary mechanisms. The exogenous bST exerted its galactopoietic action through an increase in circulating IGF-I of the late lactating Crossbred Holstein cattle.

Solvent Fractionation of Fig Leaves and its Antimicrobial Activity (무화과 잎의 용매분획 및 항미생물 활성)

  • Kang, Seong-Kuk;Chung, Hee-Jong
    • Applied Biological Chemistry
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    • v.38 no.4
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    • pp.289-292
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    • 1995
  • Fig leaves were extracted with methanol and then fractionated with ethyl acetate and various buffers to get active fractions, and their antimicrobial activities in each fraction were determined. Acidic fraction and phenolic fraction of fig leaves showed strong antimicrobial activities, but the basic fraction and neutral fraction did not show any activities. The degree of antimicrobial activities in phenolic fraction against tested bacteria were higher than those in acidic fraction, but these against yeasts and mold were almost equivalent to those in acidic fraction. Especially, phenolic fraction showed the strongest activities against Staphylococcuns aureus and Pseudomonas aeruginosa. Although there were some differences among microorganisms, minimum inhibition concentrations(MIC) in acidic and phenolic fractions against tested microorganisms were 0.05 to 0.1 g eq./ml and 0.01 to 0.25 g eq./ml, on the basis of fresh sample weight, respectively. Minimum lethal concentrations(MLC) in acidic and phenolic fractions were 0.25 to 2.00 g eq./ml and 0.05 to 1.50 g eq./ml, respectively. These supposed the antimicrobial activities in phenolic fraction were generally higher than those in acidic fraction and also phenolic fraction had lower MIC and MLC values than acidic fraction.

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