• Title/Summary/Keyword: polyamine

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Selective Trace Analysis of Mercury (II) Ions in Aqueous Media Using SERS-Based Aptamer Sensor

  • Lee, Chank-Il;Choo, Jae-Bum
    • Bulletin of the Korean Chemical Society
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    • v.32 no.6
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    • pp.2003-2007
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    • 2011
  • We report a highly sensitive surface-enhanced Raman scattering (SERS) platform for the selective trace analysis of mercury (II) ions in drinkable water using aptamer-conjugated silver nanoparticles. Here, an aptamer designed to specifically bind to $Hg^{2+}$ ions in aqueous solution was labelled with a TAMRA moiety at the 5' end and used as a Raman reporter. Polyamine spermine tetrahydrochloride (spermine) was used to promote surface adsorption of the aptamer probes onto the silver nanoparticles. When $Hg^{2+}$ ions are added to the system, binding of $Hg^{2+}$ with T-T pairs results in a conformational rearrangement of the aptamer to form a hairpin structure. As a result of the reduced of electrostatic repulsion between silver nanoparticles, aggregation of silver nanoparticles occurs, and the SERS signal is significantly increased upon the addition of $Hg^{2+}$ ions. Under optimized assay conditions, the concentration limit of detection was estimated to be 5 nM, and this satisfies a limit of detection below the EPA defined limit of 10 nM in drinkable water.

Purification and Characterization of S-adenosylmethionine Synthetase from Soybean (Glycine max) Axes

  • Kim, Dae-Gun;Park, Tae-Jin;Kim, Jong-Yeol;Cho, Young-Dong
    • BMB Reports
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    • v.28 no.2
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    • pp.100-106
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    • 1995
  • S-adenosylmethionine (SAM) synthetase was purified to homogeneity from soybean (Glycine max) axes. The enzyme was purified 216-fold with a 1.5% yield by ammonium sulfate fractionation, acetone fractionation, ion exchange chromatography with DEAE-sephacel, gel filtration with Sephacryl S-300, and afffinity chromatography with ATP-agarose. The enzyme activity reached a maximum 3 days after germination. SAM synthetase had a subunit molecular weight of 57,000 daltons from a silver stained single band on SDS-PAGE. The molecular weight of the enzyme was 110,000 daltons from Sephacryl S-300 gel filtration. The enzyme was composed of two identical subunits. The $K_m$ values of the enzyme for L-methionine and ATP were 1.81 and 1.53 mM, respectively. The enzymatic activity was not affected by polyamines, agmatine, or SAM analogues, but was inhibited by SAM. The inhibition pattern was showed non-competitive for L-methionine and uncompetitive for ATP. The activity of SAM synthetase was inhibited by thiol-blocking reagents. The enzyme was induced by treatment with $10^{-3}$ M putrescine at germination. Experimental data revealed a possible novel regulation mechanism of polyamine biosynthesis through several endogenous intermediates.

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Purification and Characterization of Soybean Cotyledonary Spermidine Dehydrogenase

  • Park, Sung-Joon;Cho, Young-Dong
    • BMB Reports
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    • v.28 no.5
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    • pp.408-413
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    • 1995
  • Decrease in the amount of cotyledonary spermidine in Glycine max under anaerobic conditions related to an increase in spermidine dehydrogenase. Under the same conditions, no enzymatic activity of diamine oxidase was observed. Exposure of Glycine max both to spermidine and 1,3-diaminopropane under anaerobic conditions resulted in a decrease in spermidine contents. Correlated with the decrease in spermidine contents, there was a drastic increase in spermidine dehydrogenase. The molecular weight of the purified enzyme estimated by Sephacryl S-300 gel column and SDS gel electrophoresis were 130,000 dalton and 65,000 dalton, respectively, indicating that the enzyme is a dimer. The optimal pH for activity was 9.3. The $K_m$ value for spermidine was 0.61 mM. Neither metal ions nor polyamine and derivatives affected enzymatic activity, but the enzyme was inhibited by DTNB, NEM and PCMB, suggesting that a cysteine residue of the enzyme is associated with or involved in enzyme activity. To our knowledge, this is the first report describing properties of the enzyme from plants. Considered together, the data in this paper indicate that both spermidine and 1,3-diaminopropane, novel activators, enhance the spermidine dehydrogenase activity and control the intracellular spermidine contents.

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An Investigation for the Adsorption of Heavy Metal Ions by Polyamine Organic Adsorbent from the Aqueous Solution - The Influence of Molecular Weight and Degree of Deacetylation of Chitosan - (수용액 중에서 Polyamine계 유기응집제를 이용한 중금속 이온의 흡착 - 키토산의 분자량과 탈아세틸화도 -)

  • Park, Young-Mi;Jeon, Dong-Won
    • Fashion & Textile Research Journal
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    • v.8 no.4
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    • pp.458-464
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    • 2006
  • The adsorption ability of heavy metal ions from the aqueous solution by chitosan, which it is well known natural biopolymer, has been investigated. The fundamental study in this research is focusing on the physicochemical adsorption utilizing the chitosan as a organic chelating adsorbent, adsorb especially heavy metal ions from the waste liquid solution. The adsorption ability of the chitosan between metal ions, having different characteristics with Mw of 188,600, 297,200, and 504,200 g/mol and degree of deacetylation (DD) of 86.92% and 100% were investigated targeting on the $Ni^{2+}$, $Co^{2+}$, $Zn^{2+}$, and $Pb^{2+}$ ions, respectively. The uptake of heavy metal ions with chitosan was performed by atomic absorption flame emission spectrophotometer (AAS) as conducted residual metal ions. It was found that chitosan has an strong adsorption capacity for some metals under certain conditions. Chitosan, which have 100% degree of deacetylation showed high adsorption recovery ratio and have an affinity for all kinds of heavy metals. In contrast, the molecular weight of chitosan was not completely affected on metal ion adsorption.

Induction of Anticarcinogenic Enzymes of Waxy Brown Rice Cultured with Phellinus igniarius 26005

  • Park, Ki-Bum;Ha, Hyo-Cheol;Kim, So-Yeun;Kim, Hyo-Jeong;Lee, Jae-Sung
    • Mycobiology
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    • v.30 no.4
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    • pp.213-218
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    • 2002
  • The induction of NAD(P)H: quinone oxidoreductase(QR), glutathione S-transferase(GST), and glutathione(GSH) levels in hepa1c1c7 cells(murine hepatoma) by waxy brown rice cultured with Phellinus igniarius to induce anticarcinogenic enzymes were measured. In addition, the inhibition of polyamines metabolism was tested with the growth of Acanthamoeba castellanii. The result shows that QR, GST activities, and GSH levels of experimental animals were increased much more by feeding the methanol extract of waxy brown rice cultured with Phellinus igniarius than those of the rats received the ethanol of uncultured brown rice. The growth of A. castellanii was inhibited mostly at 40 mg/3 ml concentration of methanol extract of waxy brown rice cultured with P. gniarius. The results suggested that waxy brown rice cultured with P. igniarius possess chemopreventive activity by inducing anticarcinogenic enzymes and inhibiting polyamine metabolism.

Comparison of Biochemical and Immunological Properties Between Rat and Nicotiana glutinosa Ornithine Decarboxylase

  • Lee, Yong-Sun;Cho, Young-Dong
    • BMB Reports
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    • v.34 no.5
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    • pp.408-414
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    • 2001
  • Ornithine decarboxylase (EC 4.1.1.17) is an essential enzyme for polyamine synthesis and growth in mammalian cells and plants. We compared the biochemical and immunological properties of rat and Nicotiana glutinosa ODC by cloning and expressing the recombinant proteins. The primary amino acid sequence between rat and N. glutinosa ODC had a 40% homology The molecular weight of the overexpressed rat ODC was 53 kDa, and that of N. glutinosa was 46.5 kDa. Adding 1 mM of putrescine to the enzyme reaction mixture inhibited both rat and N. glutinosa ODC activity to 30%. Agmatine had an inhibitory effect only on N. glutinosa ODC. Cysteine and lysine modifying reagents reduced both ODC activities, verifying the key roles of cysteine and lysine residues in the catalytic mechanism of ODC. ELISA was performed to characterize the immunological difference between the rat and plant ODC. Both the rat and N. glutinosa ODC were recognized by the polyclonal antibody that was raised against purified N. glutinosa ODC, but the rat ODC was 50-fold less sensitive to the antibody binding. These results indicate that even though both ODCs have the same evolutionary origin, there seems to be a structural distinction between the species.

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Cure Reactions of Epoxy/Anhydride/(Polyamide Copolymer) Blends

  • Youngson Choe;Kim, Wonho
    • Macromolecular Research
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    • v.10 no.5
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    • pp.259-265
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    • 2002
  • The cure kinetics of blends of epoxy (DGEBA, diglycidyl ether of bisphenol A)/anhydride resin with polyamide copolymer, poly(dimmer acid-co-alkyl polyamine), were studied using differential scanning calorimetry (DSC) under isothermal condition. On increasing the amount of polyamide copolymer in the blends, the reaction rate was increased and the final cure conversion was decreased. Lower values of final cure conversions in the epoxy/(polyamide copolymer) blends indicate that polyamide hinders the cure reaction between the epoxy and the curing agent. The value of the reaction order, m, for the initial autocatalytic reaction was not affected by blending polyamide copolymer with epoxy resin, and the value was approximately 1.3, whereas the reaction order, n, for the general n-th order of reaction was increased by increasing the amount of polyamide copolymer in the blends, and the value increased from 1.6 to 4.0. A diffusion-controlled reaction was observed as the cure conversion increased and the rate equation was successfully analyzed by incorporating the diffusion control term for the epoxy/anhydride/(polyamide copolymer) blends. Complete miscibility was observed in the uncured blends of epoxy/(polyamide copolymer) up to 120 $^{\circ}C$, but phase separations occurred in the early stages of the curing process at higher temperatures than 120 "C. During the curing process, the cure reaction involving the functional group in polyamide copolymer was detected on a DSC thermogram.gram.

Effect of Thesium Chinense Turczaninow on Breast Cancer Chemopreventive enzyme activity in In vitro (In vitro에서의 댑싸리하고초의 유방암예방효소 활성에 미치는 영향)

  • Shon, Yun-Hee;Kim, Mee-Kyung;Park, Sun-Dong;Nam, Kyung-Soo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.3
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    • pp.675-679
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    • 2006
  • The effect of water extract from Thesium chinese Turczaninow (TCTW) on proliferation of human breast cancer cells, nitric oxide production, nitric oxide synthase expression, and ornithine decarboxylase activity was tested. TCTW inhibited the growth of both estrogen-independent MDA-MB-231 and estrogen-dependent MCF-7 human breast cancer cells. Lipopolysaccharide-induced nitric oxide (NO) production was significantly reduced by TCTW at the concentrations of 1.0 (p<0.05) and 5.0 mg/ml (p<0.005). Expression of inducible nitric oxide synthase (iNOS) was also suppressed with the treatment of TCTW in Western blot analysis. TCTW inhibited induction of ornithine decarboxylase by 12-O-tetradecanoylphorbol-13-acetate (TPA), a key enzyme of polyamine biosynthesis, which is enhanced in tumor promotion. Therefore, TCTW is worth further investigation with respect to breast cancer chernoprevention or therapy.

Effect of Prunella vulgaris L. on Chemopreventive Enzymes of Colorectal Cancer (꿀풀하고초가 직장암 예방효소 활성에 미치는 영향)

  • Shon, Yun-Hee;Seo, Jae-Beom;Nam, Kyung-Soo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.1
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    • pp.126-130
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    • 2008
  • Water extract from Prunella vulgaris L. (PVW) was tested for colon cancer chemopreventive activity by measuring the activities of cytochrome P450 1A1, phase Ⅱ detoxification enzyme [quinone reductase (QR) and glutathione S-transferase (GST)] and ornithine decarboxylase (ODC) and glutathione (GSH) levels in cultured human colorectal adenocarcinoma HT-29 cells. PVW significantly inhibited 7,12-dimethylbenz[a]anthracene (DMBA)-induced cytochrome P450 1A1 activity at 10 and 50 ${\mu}g/ml$. PVW induced QR activity in a dose-dependent manner over a concentration range of $1{\sim}50\;{\mu}g/ml$. GST activity was also induced with the treatment of PVW in HT-29 cells. In addition GSH levels were increased with PVW. PVW inhibited ODC activity, a key enzyme of polyamine biosynthesis, which is enhanced in tumor promotion. These results suggest that Prunella vulgaris L. has colon cancer chemopreventive activity by inhibiting cytochrome P450 1A1 and ODC activities and by increasing phase Ⅱ enzyme activity and GSH levels.

In Situ Detection of the Onset of Phase Separation and Gelation in Epoxy/Anhydride/Thermoplastic Blends

  • Choe, Young-Son;Kim, Min-Young;Kim, Won-Ho
    • Macromolecular Research
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    • v.11 no.4
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    • pp.267-272
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    • 2003
  • The isothermal cure reactions of blends of epoxy (DGEBA, diglycidyl ether of bisphenol A)/anhydride resin with polyamide copolymer (poly(dimmer acid-co-alkyl polyamine)) or PEI were studied using differential scanning calorimetry (DSC). Rheological measurements have been made to investigate the viscosity and mechanical relaxation behavior of the blends. The reaction rate and the final cure conversion were decreased with increasing the amount of thermoplastics in the blends. Lower values of final cure conversions in the epoxy/thermoplastic blends indicate that thermoplastics hinder the cure reaction between the epoxy and the curing agent. Complete miscibility was observed in the uncured blends of epoxy/thermoplastics up to $120^{\circ}C$ but phase separations occurred in the early stages of the curing process at higher temperatures than $120^{\circ}C$. According to the rheological measurement results, a rise of G' and G" at the onset of phase separation is seen. A rise of G' and G" is not observed for neat epoxy system since no phase separation is seen during cure reaction. At the onset of phase separation the rheological behavior was influenced by the amount of thermoplastics in the epoxy/thermoplastic blends, and the onset of phase separation can be detected by rheological measurements.