• Title/Summary/Keyword: acid-diffusion technique

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Purification and Characterization Sucrose phosohorylase in Leuconostoc mesenteroides NRRL B-1149 (Leuconostoc mesenteroides NRRL B-1149의 Sucrose phosohorylase의 분리와 특성 연구)

  • Lee Jin Ha;Park Jun Seong;Park Hyen Joung;Cho Jae Young;Choi Jeong Sik;Kim Do Man
    • KSBB Journal
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    • v.19 no.5
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    • pp.363-367
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    • 2004
  • Leuconostoc mesenteroides NRRL B-1149 produces various glucoseyltransferases for the synthesis of dextran, levan and glucose-1-phosphate using sucrose as a substrate. A sucrose phosphorylase (1149SPase) was purified from L. mesenteroides NRRL B-1149 culture by using hollow fiber filtration (30 kDa cut off), Toyopearl DEAE 650 M column chromatography and following two times of DEAE-Sepharose column chromatographies. The specific activity of the purified 1149SPase was 25.7 (U/mg) with $16\%$ yield. The 1149SPase showed a molecular size of 56 kDa on denatured $10\%$ SDS-PAGE. The N-terminal amino acid sequence of the enzyme was MEIQNKAM. The optimum pH and temperature of this enzyme were 6.2~6.5 and 37^{circ}C, respectively. It had an apparent K_{m} of 6.0 mM and K_{cat} of 1.62/s for sucrose. 1149SPase crystal was formed by hanging drop diffusion technique using 20 mM calcium chloride dihydrate, 100 mM sodium acetate trihydrate pH 4.6 and $30\%$ 2-methyl-2,4-pentanediol as vaporizing and reservation solution. The 1149SPase catalyzes transferring of glucose from isomaltose or sucrose to salicin and salicyl alcohol by disproportionation reaction or acceptor reaction and synthesized two acceptor products, respectively.

Bactericidal Application and Cytotoxic Activity of Biosynthesized Silver Nanoparticles with an Extract of the Red Seaweed Pterocladiella capillacea on the HepG2 Cell Line

  • El Kassas, Hala Yassin;Attia, Azza Ahmed
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.3
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    • pp.1299-1306
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    • 2014
  • Background: Nano-biotechnology is recognized as offering revolutionary changes in various fields of medicine. Biologically synthesized silver nanoparticles have a wide range of applications. Materials and Methods: Silver nanoparticles (AgNPs) were biosynthesized with an aqueous extract of Pterocladiella (Pterocladia) capillacea, used as a reducing and stabilizing agent, and characterized using UV-VIS spectroscopy, Fourier Transform Infra red (FT-IR) spectroscopy, transmission electron microscopy (TEM) and energy dispersive analysis (EDX). The biosynthesized AgNPs were tested for cytotoxic activity in a human hepatocellular carcinoma ($HepG_2$) cell line cultured in Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum, 1% antibiotic and antimycotic solution and 2 mM glutamine. Bacterial susceptibility to AgNPs was assessed with Staphylococcus aureus, Bacillus subtilis [Gram+ve] and Pseudomonas aeruginosa and Escherichia coli [Gram-ve]. The agar well diffusion technique was adopted to evaluate the bactericidal activity of the biosynthesized AgNPs using Ampicillin and Gentamicin as gram+ve and gram-ve antibacterial standard drugs, respectively. Results: The biosynthesized AgNPs were $11.4{\pm}3.52$ nm in diameter. FT-IR analysis showed that carbonyl groups from the amino acid residues and proteins could assist in formation and stabilization of AgNPs. The AgNPs showed potent cytotoxic activity against the human hepatocellular carcinoma ($HepG_2$) cell line at higher concentrations. The results also showed that the biosynthesized AgNPs inhibited the entire panel of tested bacteria with a marked specificity towards Bacillus subtillus. Conclusions: Cytotoxic activity of the biosynthesized AgNPs may be due to the presence of alkaloids present in the algal extract. Our AgNPs appear more bactericidal against gram-positive bacteria (B. subtillus).

Dispersion of Particle Pulse in Human Lung Airway (인체기관지내의 입자펄스 확산 실험)

  • 이진원;이동엽;추경호
    • Journal of Biomedical Engineering Research
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    • v.19 no.5
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    • pp.511-518
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    • 1998
  • In order to develop the aerosol bolus technique which is thought to be a potential tool for probing geometries or abnormalities of small airways, an experimental system of measuring fast time variations of particle concentration in the inhaled and exhaled breathing air was developed. The system generates monodisperse sebacic acrid particles of 1 micron size and 1.2 of geometric standard deviation in high concentration of $10^8$ particles/cc, delivers a short pulse of particles at the controlled instant during inhalation using a solenoid valve, and measures the fast change of particle concentration in using the laser light scattering. Successful operation of the generator and the measuring system was confirmed by smooth concentration profiles in inhalation. It was also confirmed that maintaining a constant breathing rate is essential to stable outputs and any disturbance in flow rate near the mode (maximum concentration) induces a large number of spurious peaks in the exhalation. Experimental data without strict control of breathing flow rate showed a substantial amount of scatter. The measured results showed an improvement in scatter over the existing results. When compared with theoretical predictions from 1-D convective diffusion equation and other experiments, general characteristics of dispersion for several penetration depths showed a good agreement, but there exists some difference in absolute values, which is attributed to the difference in body conditions. Improvements are needed in the theory, especially in relation to correcting for the effect of breathing flow rate.

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