• Title/Summary/Keyword: fab E

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Isolation and Identification of Stilbene glycosides from the Bark of Pinus koraiensis (잣나무 수피의 Stilbene glycosides의 분리 및 동정)

  • Song, Hong-Keun
    • Journal of the Korean Wood Science and Technology
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    • v.29 no.4
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    • pp.97-102
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    • 2001
  • EtOAc extract from the bark of Pinus koraiensis Sieb. et Zucc was isolated by column chromatography which was packed with Sephadex LH-20 or TSK-gel HW-40F. Several stilbene glycosides were identified by $^1H{\cdot}^{13}C$-NMR, HMQC, HMBC and $FAB^+$ MS. Three stilbene glycosides, Z-pinostilbenoside, E-desoxyrhaponticin, and E-resveratroloside, were identified.

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Application of Fast Atom Bombardment Collision-induced Dissociation Tandem Mass Spectrometry for Structural identification of Glycerolipids Isolated From Marine Sponge

  • Lee, Sun-Young;Hong, Joo-Yeon;Jung, Jee-H.;Hong, Jong-Ki
    • Mass Spectrometry Letters
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    • v.2 no.1
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    • pp.8-11
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    • 2011
  • Two types of glycerolipids [monoacylglycerols (MAG) and cyclitols] were isolated by reversed phase high-performance liquid chromatography from the methanol extracts of a marine sponge, and analyzed by fast atom bombardment mass spectrometry (FAB-MS) in positive-ion mode. FAB mass spectra of these compounds yielded protonated molecules $[M + H]^+$ and abundant sodiated molecules $[M + Na]^+$ from a mixture of 3-nitrobenzyl alcohol and NaI. The structures of these compounds were elucidated by FAB-collisional-induced dissociation (CID)-tandem mass spectrometry. We carried out collision-indused dissociation (CID) of these lipids in B/E-linked scan mode. The CID B/E-linked scan of $[M + H]^+$ and $[M + Na]^+$ precursor ions resulted in the formation of numerous characteristic product ions through a series of dissociative processes. The product ions formed by charge-remote fragmentation (CRF) provided important information for the identification of the acyl chain structure substituted at the glycerol backbone. Some of the product the ions were diagnostic for the presence of a glycerol backbone or acyl chain structure.

Periplasmic Expression of a Recombinant Antibody (MabB9) in Escherichia coli

  • Chang, Hae-Choon;Kwak, Ju-Won
    • Journal of Microbiology and Biotechnology
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    • v.7 no.5
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    • pp.299-304
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    • 1997
  • Expression in the periplasm of Escherichia coli of cloned heavy and light chain cDNAs for Fab fragment of a murine monoclonal antibody MabB9 (${\gamma}2b$, K), specific for human plasma apolipoprotein B-100 of LDL, was studied. For the purpose, a vector for two-cistronic expression of the heavy chain cDNA, at the 5' terminus, and light chain cDNA, at the 3' terminus, was constructed using the signal sequences, pelB (for heavy chain) and ompA (for light chain) in a pET vector system. The constructed vector was transformed into E. coli BL21(DE3). The expressed heavy chain (25 kDa) and light chain (23 kDa) of the antibody molecule were detected in total cell extracts as well as in the periplasmic extracts of E. coli.

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Effects of Glucose and Acetic Acid on the Growth of Recombinant E.coli and the Production of Pyruvate Dehydrogenase Complex-E2 Specific Human Monoclonal Antibody (유전자 재조합 대장균의 세포성장과 Pyruvate Dehydrogenase Complex-E2 특이성 인간 모노클론 항체 생산에 대한 포도당과 초산의 영향)

  • 이미숙;전주미;차상훈;정연호
    • KSBB Journal
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    • v.15 no.5
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    • pp.482-488
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    • 2000
  • The Fab fraction of PDC-E2 specific human monoclonal antibody was produced using recombinant E. coli, and the effects of glucose and acetate were investigated to develop an optimal strategy for recombinant human antibody production. Higher glucose concentration in the culture media resulted inn higher cell growth and glucose consumption rate, which in turn resulted in an increased acetate production rate. When glucose was depleted, cells began to consume acetate as an energy source, and this consumption rate depended on the glucose concentration. When the residual glucose concentration was high, the accumulation of acetate was accelerated due to an increase in the acetate production rate and a decrease in the acetate consumption rate. Futhermore, it was found that a high accumulation of acetate, accompanied by a high glucose concentration, inhibited human antibody formation; the critical acetate concentration was $0.6g/\ell$. During production, a high glucose concentration enhanced cell growth, but inhibited antibody formation due to catabolic repression. Therefore, it is important to keep the concentration of both glucose and acetate as low as possible to increase antibody production after induction. Accordingly, it is important to accurately control the concentration of glucose and acetate in the culture media to obtain high cell densities and high productivity levels of recombinant human antibody.

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Physico-Chemical Properties and Antimicrobial Activity of Pyocyanine Produced by Pseudomonase aeruginosa KLP-2 (Pseudomonas aeruginosa KLP-2가 생산한 Pyocyanine의 항균활성 및 생리화학적 성상)

  • 박은희;이상준;차인호
    • Journal of Life Science
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    • v.11 no.5
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    • pp.483-488
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    • 2001
  • The antimicrobial substance produced by Pseudomonas aeruginosa KLP-2 strain was purified and identified. The substance was identified as a pyocyanine by the fast atom bombardment mass(FAB-MS). In physic-chemical properties, the pyocyanine was dark blue needles, and was soluble in various organic solvents such as chlorogorm, methanol, ethanol and ethyl acetae. The pyocyanine possessed a ultraviolet absorbance spectrum in methanol, 0.1 M HCl, and chlorogorm. The maximum absorption peak of the pyocyanine showed at 318 mm in methanol. The molecular formula of the pyocyanine was determined to the $C_{13}$ H$_{10}$ N$_{2}$O and protonate molecular ion species (M+H)$^{+}$ was observed at m/z 211 by FAB-MS. The pyocyanine showed antimicrobial against Bacillus cereus, Bacillus subtilis, Micrococcus luteus, Rodococcus equi, Staphylococcus aureus, Streptococcus faecalis, E. col, Legionella pneumophila, Shigella flexneri Shigella boydii, shgella sonnei, NAG Vibrio cholerae, Vibrio parahaemolyticus, Vibro vulnificus, Yersinia enterocolitica, and Saccharomyces cerevisiae. However, Salmonella spp. Shigela dysenteriae, 3 strains of Pseudomonas aeruginosa, Klebsiela pneumoniae, and Aspergillus niger were resistant to the pyocyanine. The pyocyanine showed the highest antimicrobial activity aganist Legionella pneumophila based on the size of inhibition zone by the disk contained 0.5 $\mu\textrm{g}$ of the pyocyanine.e.

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A Study on Gamma ray effects on Stress Response and Cellular Toxicity using Bacterial Cells

  • Min, Ji-Ho;Lee, Hyeon-Ju;Lee, Chang-U;Gu, Man-Bok
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.187-190
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    • 2000
  • Effects of gamma ionizing radiation on recombinant Escherichia coli cells containing stress promoters, recA, fabA, grpE, or katG, fused to luxCDABE originated from Vibrio fischeri were characterized by monitoring transcriptional responses reflected by bioluminescent output. Quantification of gamma-ray intensity may be possible using the recA and fabA promoter fusion since a linear enhancement of bioluminescence emission with increasing gamma-ray intensity was observed. Other strains sensitive to either oxidative stress (DPD2511, katG::luxCDABE) or protein-damaging stress (TV1061, grpE::luxCDABE) were also irradiated by gamma-rays, and resulted in no noticeable bioluminescent output while DPD2794 with recA promoter and DPD2540 with fabA promoter irradiated by the same intensities of gamma-rays gave a significant bioluminescent output. This indicates that the main stresses in the recombinant bacteria caused by ionizing radiation are DNA and membrane-damage, not protein- or oxidative-damage. In addition, in this study, to investigate the relationship between the radiation dose rate and bacterial responses, two recombinant Escherichia coli strains, DPD2794 and GC2, containing lac promoter fused to luxCDABE originating from Photorhapdus luminescences, were used for detecting DNA damage and cellular toxicity under various radiation dose rates. Throughout this study, it was found that these bacteria showed quantitative stress responses to DNA damage and general toxicity caused by gamma rays, depending on the radiation dose rates, indicating that the bacterial stress responses and general toxicity were seriously influenced according to radiation dose rates.

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An Unusually Stable S-Nitrosothiol from Glutathione

  • Park, Jeen-Woo;Means, Gary-E.
    • Archives of Pharmacal Research
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    • v.12 no.4
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    • pp.257-258
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    • 1989
  • Glutathione was converted by $HNO_2$ into a S-nitrosothiol which was stable in solution and atypically so even as a solid. FAB/MS and IR data have been obtained for the confirmation of structure of S-nitrosogulathione in the crystalline state.

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Fabrication of a Graphene Nanoribbon with Electron Beam Lithography Using a XR-1541/PMMA Lift-Off Process

  • Jeon, Sang-Chul;Kim, Young-Su;Lee, Dong-Kyu
    • Transactions on Electrical and Electronic Materials
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    • v.11 no.4
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    • pp.190-193
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    • 2010
  • This report covers an effective fabrication method of graphene nanoribbon for top-gated field effect transistors (FETs) utilizing electron beam lithography with a bi-layer resists (XR-1541/poly methtyl methacrylate) process. To improve the variation of the gating properties of FETs, the residues of an e beam resist on the graphene channel are successfully taken off through the combination of reactive ion etching and a lift-off process for the XR-1541 bi-layer. In order to identify the presence of graphene structures, atomic force microscopy measurement and Raman spectrum analysis are performed. We believe that the lift-off process with bi-layer resists could be a good solution to increase gate dielectric properties toward the high quality of graphene FETs.

Optimization of Etching Profile in Deep-Reactive-Ion Etching for MEMS Processes of Sensors

  • Yang, Chung Mo;Kim, Hee Yeoun;Park, Jae Hong
    • Journal of Sensor Science and Technology
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    • v.24 no.1
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    • pp.10-14
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    • 2015
  • This paper reports the results of a study on the optimization of the etching profile, which is an important factor in deep-reactive-ion etching (DRIE), i.e., dry etching. Dry etching is the key processing step necessary for the development of the Internet of Things (IoT) and various microelectromechanical sensors (MEMS). Large-area etching (open area > 20%) under a high-frequency (HF) condition with nonoptimized processing parameters results in damage to the etched sidewall. Therefore, in this study, optimization was performed under a low-frequency (LF) condition. The HF method, which is typically used for through-silicon via (TSV) technology, applies a high etch rate and cannot be easily adapted to processes sensitive to sidewall damage. The optimal etching profile was determined by controlling various parameters for the DRIE of a large Si wafer area (open area > 20%). The optimal processing condition was derived after establishing the correlations of etch rate, uniformity, and sidewall damage on a 6-in Si wafer to the parameters of coil power, run pressure, platen power for passivation etching, and $SF_6$ gas flow rate. The processing-parameter-dependent results of the experiments performed for optimization of the etching profile in terms of etch rate, uniformity, and sidewall damage in the case of large Si area etching can be summarized as follows. When LF is applied, the platen power, coil power, and $SF_6$ should be low, whereas the run pressure has little effect on the etching performance. Under the optimal LF condition of 380 Hz, the platen power, coil power, and $SF_6$ were set at 115W, 3500W, and 700 sccm, respectively. In addition, the aforementioned standard recipe was applied as follows: run pressure of 4 Pa, $C_4F_8$ content of 400 sccm, and a gas exchange interval of $SF_6/C_4F_8=2s/3s$.