• Title/Summary/Keyword: 이온 영동법

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Comparative Studies on the Enzymatic Properties of two Trypsin-like Enzymes from Menhaden, Brevoortia tyranus (혈합육어 멘헤이든의 장기조직분포Trypsin-유사효소에 관한 비교효소학적 연구)

  • PYEUN Jae-Hyeung;KIM Hyeung-Rak;GODBER J. S.
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.23 no.1
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    • pp.12-24
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    • 1990
  • Two trypsin-like enzymes, designated trypsin A and 3, purified from the intestine of menhaden by $(NH_4)_2SO_4$ fractionation, Benzamidine-Sepharose 6B affinity chromatography, DEAE-Sephacel ion exchange chromatography and Sephadex G-75 gel filtration chromatography. The two trypsins were subjected to compare the enzymatic properties of the trypsin-like enzymes from the other dark fleshed fishes. Both trypsins catalysed the hydrolysis of N$\alpha$-benzoyl-DL-arginine-p-nitroanilide and they were remarkably inhibited by several well known trypsin-inhibitors, tosyllysyl chloromethyl ketone, soybean trypsin inhibitor, be-nzamidine, leupeptin and antipain, etc. Therefore, it was ascertained that the two enzymes are serine-type trypsins. The molecular weights of these enzymes were about 25,000 and 26,200, respectively, ;Is determined by SDS-PAG electrophoresis and by Sephadex G-100 gel filtration, and the molecular weights of these two enzymes are somewhat fewer than those from the other dark fleshed fishes. Both enzymes had less basic amino acids such as arginine and Iysine, whereas they had slightly high contents of neutral amino acids, glycine, alanine and tryptophane. The enzymes showed a pH optimum of $8\~11$ at $60^{\circ}C$ against the $N\alpha$-benzoyl-DL-argi-nine-p-nitroanilide substrate and they were quite unstable above $40^{\circ}C$ and under the atidic pH region. The Km constant of the two enzymes against the $N\alpha$-benzoyl-DL-arginine-p-nitroanilide was $1.4\times10^{-4}M$ for trypsin A and $4.3\times10^{-5}M$ for trypsin B, respectively.

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Chemical Composition and Lead Isotope Ratio of Poong-Tag(Wind Bell) from Winggyeong Site, Cyeongju (경주 왕경지구 금동풍탁(金銅風鐸)의 성분조성과 납동위원소비)

  • Chung, Young-Dong;Kang, Hyung-Tae;Huh, Il-Kwon;Cho, Nam-Chul
    • Journal of Conservation Science
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    • v.19
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    • pp.67-72
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    • 2006
  • The chemical compositions and lead isotope ratio of Poong-Tag(wind bell) bell excavated from Wanggyeong site, Gyeongju have analyzed by ICP and TIMS. The analysis result of chemical composition of Poong-Tag shows that it consists of 92:4 ratio of Cu:Sn. Other 8 minor elements(Pb, Zn, Fe, Ag, Ni, As, Sb and Co) show the concentration of below 0.2% respectively. This result means that Poong-Tag have made using highly purified Cu and Sn ore. For the study of provenance of raw material with the lead isotope ratio, the origin of the raw material of Poong-Tag can not be presumed owing to beyond the scope. The observation result of microstructure of Poong-Tag show the dendrite structure made through casting process. The chemical composition, microstructure, and lead isotope ratio of Poong-Tag excavated from Wanggyeong site, Gyeongju can be used as fundamental data to compare with other Poong-Tag of different regions and periods.

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Hydrogen Isotope Exchange Reaction in Electrical Discharge through D2/H2O System (전기방전하에서 D2/H2O 반응계의 수소 동위원소 교환반응)

  • Kim, H.J.;Park, Y.D.;Lee, W.M.
    • Transactions of the Korean hydrogen and new energy society
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    • v.9 no.2
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    • pp.77-84
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    • 1998
  • Hydrogen isotope exchange in mixtures of $H_2O/D_2$, $H_2O/D_2O$, or $D_2O/H_2$ can be facilitated under electrical discharge. For example, a simple DC corona discharge through the mixture creates a plasma in which the reactants are excited energetically. The reactants in such plasma, due to increase in population of excited quantum levels or due to production of radicals or ions, undergo very rapid chemical reactions even at ambient temperature. The isotope exchange reaction of hydrogen(H) and deuterium(D) produces the third kind of heavy water(HDO) and isotopic hydrogen gas(HD), as shown in $D_2+H_2O{\rightarrow}HD$ K=11.257(at $25^{\circ}C$) The reaction products can be detected with temporal resolution using the Fourier transform infrared(FTIR) absorption spectroscopy. Since $H_2O$, $D_2O$ and HDO are all infrared active with different absorption peaks, FTIR proves to be a useful tool for monitoring the reaction. Experimental results show that the electrical method is indeed a useful means to promote the reaction, showing a better efficiency than traditional catalytic methods.

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Characterization and Action Mode of Anti-Complementary Substance Prepared from Lactobacillus plantarum (Lactobacillus plantarum 균체 중 항보체 활성물질의 특성과 작용양식)

  • Kim, Jang-Hyun;Shin, Kwang-Soon;Lee, Ho
    • Korean Journal of Food Science and Technology
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    • v.34 no.2
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    • pp.290-295
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    • 2002
  • Among 12 lactic acid bacteria examined for their abilities to activate the complement system by hemolytic complement assay $(TCH_{50})$, Lactobacillus plantarum previously isolated from Kimchi showed high anti-complementary activity. The anti-complementary activity of the cell wall fraction of L. plantarum was more potent than that of the cytosol fraction, and both activities showed dose dependency. These high activities of the cytosol and the cell wall fractions were relatively resistant to the digestion with pronase, but sharply decreased after the treatment of $NaIO_4$. These results suggested that the complement activation by the cytosol and the cell wall fractions was mainly due to their polysaccharides. By the cross-immunoelectrophoresis using anti-human C3, the C3 activation products from both fractions were identified in $Ca^{++}$-free condition. Anti-complementary activity $(ITCH_{50})$ of the cell wall fraction was retained under the same condition, whereas that of the cytosol fraction was reduced considerably. From these results, it was inferred that the mode of complement activation by the cell wall fraction was mainly via alternative pathway, and that of the cytosol fraction was via both alternative and classical pathways.

Screening of Complement-System Activating Polysaccharide from Edible Plants and Its Action Mode (식물성 식품재료로부터 보체계 활성화 다당의 검색 및 그 활성검토)

  • Shin, Kwang-Soon;Ra, Kyung-Soo;Sung, Ha-Chin;Yang, Han-Chul
    • Korean Journal of Food Science and Technology
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    • v.25 no.3
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    • pp.197-203
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    • 1993
  • Screenings were performed on edible plants to examine their complement-system activating ability (anti-complementary activity) by hemolytic complement assay $(TCH_{50})$. Among 38 kinds of plant extracts, 5 kinds showed relatively strong anti-complementary activity which decreased $TCH_{50}$ more than 60% comparison with control and the order of activity was Zingiber officinale>Colocasia antiquorum>Capsella bursapastoris>Ginkgo biloba>Alium monanthum in $1000{\mu}g/ml$. The anti-complementary activity of ZR-1 prepared from the root of Zingiber officinale which was showed the most potent activity, did not change by pronase treatment, but decreased greatly by periodate oxidation. These results indicate that not protein moiety but carbohydrate moiety in ZR-1 fraction may also contribute to the anti-complementary activity. Also, the anti-complementary activity of ZR-1 was reduced partially in the absence of the $Ca^{2+}$ ion. When crossed immunoelectrophoresis using anti-human C3 serum was carried out after incubation of normal human serum with the ZR-1 in $Ca^{2+}$ free condition, a cleavage of C3 precipitin line was observed. Furthermore this polysaccharide fraction considerably inhibited $ACH_{50}$. These results also indicate that the mode of complement activation by polysaccharide from Zingiber officinale is via not only the classical pathway but also the alternative pathway.

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Isolation and purification of a cecropin-like antimicrobial peptide from the japanese oak silkworm, Antheraea yamamai (천잠 세크로핀 항균펩타이드 분리 및 정제)

  • Kim, Seong-Ryul;Goo, Tae-Won;Choi, Kwang-Ho;Park, Seung-Won;Kim, Sung-Wan;Hwang, Jae-Sam;Kang, Seok-Woo
    • Journal of Sericultural and Entomological Science
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    • v.50 no.2
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    • pp.145-149
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    • 2012
  • Cecropin is a well-studied antimicrobial peptide that play important role as key factor in insect humoral immunity. In this study, cecropin-like antimicrobial peptide was isolated and purified from the larval haemolymph of immune-challenged japanese oak silkworm, Antheraea yamamai. To isolate antimicrobial peptide, we separated and compared acidic extracted hemolymph protein bends between control and immune-challenged larvae using SDS-PAGE analysis. In the immune hemolymph extract, but not of non-immune hemolymph, we detected differential expressed peptide band with molecular mass 4,223.01 Da. To understand this peptide better, we successfully purified this peptide using cation exchange chromatography and gel permeation chromatography. Its N-terminal amino acid sequence obtained by Edman degradation evidenced a significant degree of identity with other lepidopteran cecropins. The purified A. yamamai cecropin-like peptide showed a broad spectrum of activity against fungi, Gram-negative and Gram-positive bacteria.

In Situ Surfactant Flushing of Contaminated Site (계면 활성제를 이용한 In Situ 토양 세척)

  • 염익태;안규홍
    • Journal of Korea Soil Environment Society
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    • v.2 no.2
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    • pp.9-24
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    • 1997
  • Surfactant-aided in situ soil flushing has been proposed as an alternative for the expensive and time consuming 'pump and treat' technology in remediation of contaminated soil and groundwater Injected surfactants can effectively solubilize contaminants sorbed to the soil matrix or nonaqueous phase liquids(NAPLs) in residual saturation. The contaminants solubilized in groundwater are recovered and treated further. The theoretical background of the technology and the results of the field operations, mostly in the US. were summarized. In addition, the factors crucial to the successful application of the technology were discussed. Cost analyses and technical limitations in current applications were also discussed. In conclusion, it is likely that in situ surfactant flushing become a viable option for soil remediation in limited cases. Currently, further advances with respect to operation cost and to treatment efficiency are required for more extensive application of the technology. However, the current trends in soil remediation, specially the growing emphasis on risk based corrective action and natural attenuation, will increase the competitiveness of the technology. For example, removal of easily washable contaminants by short term soil flushing followed by long term monitoring and natural attenuation can greatly reduce the operation cost and time.

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Isolation and Structure Identification of Photosensitizer from Perilla frutescens Leaves Which Induces Apoptosis in U937 (들깻잎(Perilla frutescens)으로부터 U937 세포에 apoptosis를 유도하는 광과민성 물질의 분리 및 구조동정)

  • Ha, Jun Young;Kim, Mi Kyeong;Lee, Jun Young;Choi, Eun Bi;Hong, Chang Oh;Lee, Byong Won;Bae, Chang Hwan;Kim, Keun Ki
    • Journal of Life Science
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    • v.25 no.1
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    • pp.53-61
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    • 2015
  • In this study, we tried to separate the photosensitizer that induces apoptosis of leukemia cells (U937) from perilla leaves. Perilla leaves (Perilla frutescens Britt var. japonica Hara) are a popular vegetable in Korea, being rich in vitamins (A and E), GABA, and minerals. Dried perilla leaves were extracted with methanol to separate the photosensitizer by various chromatographic techniques. The structure of the isolated compound (PL9443) was identified by 1D-NMR, 2D-NMR, and FAB-mass spectroscopy. Absorbance of the UV-Vis spectrum was highest at 410 nm and was confirmed by the 330, 410, and 668 nm. PL9443 compound was determined to be pheophorbide, an ethyl ester having a molecular weight of 620. It was identified as a derivative compound of pheophorbide structure when magnesium comes away from a porphyrin ring. Observation of morphological changes in U937 cells following cell death induced by treated PL9443 compound revealed representative phenomena of apoptosis only in light irradiation conditions (apoptotic body, vesicle formation). Results from examining the cytotoxicity of PL9443 substance against U937 cells showed that inhibition rates of the cell growth were 99.9% with the concentration of 0.32 nM PL9443. Also, the caspase-3/7 activity was 99% against U937 cells with the concentration of 0.08 nM of PL9443 substance. The result of the electrophoresis was that a DNA ladder was formed by the PL9443. The PL9443 compound is a promising lead compound as a photosensitizer for photodynamic therapy of cancer.

The Proteinase Distributed in the Intestinal Organs of Fish 2. Characterization of the Three Alkaline Proteinases from the Pyloric Caeca of Mackerel, Scomber japonicus (어류의 장기조직에 분포하는 단백질분해효소에 관한 연구 2. 고등어 유문수조직중에 분포하는 3종 알칼리성 단백질분해효소의 특성)

  • KIM Hyeung-Rak;PYEUN Jae-Hyeung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.19 no.6
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    • pp.547-557
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    • 1986
  • The characteristics of the three alkaline proteinases, Enz. A, B and C, from the pyloric caeca of mackerel have been investigated. The optimum condition for the activity of the Enz. A, B and C was pH 9.4, 9.8 and 9.8 at $45^{\circ}C$ for $2\%$ casein solution, and was pH 9.2 10.2 and 9.8 at $45^{\circ}C$ for $5\%$ hemoglobin denatured by urea, respectively. Enz. A, B and C by heat treatment at $50^{\circ}C$ for 5 min were inactivated 90, 33 and $37\%$, respectively, over the original activity. The reaction rate of the three alkaline proteinases was constant to the reaction time to 40 min in the reaction condition of $2{\mu}g/ml$ of enzyme concentration and $2\%$ casein solution. The reaction rate equation and Km value against casein substrate determined by the method of Lineweaver and Burk were: Enz. A, Y=3.6X and $Km=5.0{\times}10^{-3}\%$; Enz. B, Y=6.0X and $Km=1.0{\times}10^{-3}\%$; Enz. C, Y=4.2X and $Km=3.6{\times}10^{-3}\%$. The three alkaline proteinases were inactivated by $Ag^+$ and $Hg^{2+}$, but activated by $Mn^{2+},\;Sn^{2+}\;and\;Pb^{2+}$, Enz. B and C were remarkably inhibited by the soybean trypsin inhibitor. Molecular weight of Enz. A, B and C determined by SDS-polyacrylamide gel electrophoresis and Sephadex G-100 gel filtration was in the range of $27,500{\pm}2,500,\;20,500{\pm}1,500\;and\;15,250{\pm}250$, respectively.

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