• Title/Summary/Keyword: H-NMR spectroscopy

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Purification and Characterization of a Thermostable Laccase from Trametes trogii and Its Ability in Modification of Kraft Lignin

  • Ai, Ming-Qiang;Wang, Fang-Fang;Huang, Feng
    • Journal of Microbiology and Biotechnology
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    • v.25 no.8
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    • pp.1361-1370
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    • 2015
  • A blue laccase was purified from a white rot fungus of Trametes trogii, which was a monomeric protein of 64 kDa as determined by SDS-PAGE. The enzyme acted optimally at a pH of 2.2 to 4.5 and a temperature of 70℃ and showed high thermal stability, with a half-life of 1.6 h at 60℃. A broad range of substrates, including the non-phenolic azo dye methyl red, was oxidized by the laccase, and the laccase exhibited high affinity towards ABTS and syringaldazine. Moreover, the laccase was fairly metal-tolerant. A high-molecular-weight kraft lignin was effectively polymerized by the laccase, with a maximum of 6.4-fold increase in weight-average molecular weight, as demonstrated by gel permeation chromatography. Notable structural changes in the polymerized lignin were detected by Fourier transform infrared spectroscopy and 1H NMR spectroscopy. This revealed an increase in condensed structures as well as carbonyl and aliphatic hydroxyl groups. Simultaneously, phenolic hydroxyl and methoxy groups decreased. These results suggested the potential use of the laccase in lignin modification.

Study on preparation and characterization of PPV LB films. (Poly(p-phenylenevinylene) (PPV) LB 막의 제작 및 특성 연구)

  • Kim, J.H.;Kim, K.S.;Kang, W.H.;Sohn, M.H.;Kim, Y.C.;Kim, Y.K.;Sohn, B.C.
    • Proceedings of the KIEE Conference
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    • 1994.11a
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    • pp.219-221
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    • 1994
  • Oil-soluble poly(p-phenylene(1-methoxyethylene)), as a precursor of poly(p-phenylenevinylene) (PPV), was synthesized add identified with NMR and FT-IR spectroscopy. The PPV films were prepared by PPV precursor films with a thermal treatment at $250^{\circ}C$ under vacuum, where the PPV precursor films wets formed on various substrates by using Langmuir-Blodgett(LB) method. The characterization of these films was carried out by FT-IR spectroscopy, UV-VIS absorption spectroscopym, and photoluminescence (PL). Atomic Force Microscopy (AFM) has been used to investigate He surface morphology of PPV films.

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Combining Information of Common Metabolites Reveals Global Differences between Colorectal Cancerous and Normal Tissues

  • Chae, Young-Kee;Kang, Woo-Young;Kim, Seong-Hwan;Joo, Jong-Eun;Han, Joon-Kil;Hong, Boo-Whan
    • Bulletin of the Korean Chemical Society
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    • v.31 no.2
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    • pp.379-383
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    • 2010
  • Metabolites of colorectal cancer tissues from 12 patients were analyzed and compared with those of the normal tissues by two-dimensional NMR spectroscopy. NMR data were analyzed with the help of the metabolome database and the statistics software. Cancerous tissues showed significantly altered metabolic profiles as compared to the normal tissues. Among such metabolites, the concentrations of taurine, glutamate, choline were notably increased in the cancerous tissues of most patients, and those of glucose, malate, and glycerol were decreased. Changes in individual metabolites varied significantly from patient to patient, but the combination of such changes could be used to distinguish cancerous tissues from normal ones, which could be done by PCA analysis. The traditional chemometric analysis was also performed using AMIX software. By comparing those two results, the analysis via $^1H-^{13}C$ HSQC spectra proved to be more robust and effective in assessing and classifying global metabolic profiles of the colorectal tissues.

NMR Spectroscopic Assessment of the Structure and Dynamic Properties of an Amphibian Antimicrobial Peptide (Gaegurin 4) Bound to SDS Micelles

  • Park, Sang-Ho;Son, Woo-Sung;Kim, Yong-Jin;Kwon, Ae-Ran;Lee, Bong-Jin
    • BMB Reports
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    • v.40 no.2
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    • pp.261-269
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    • 2007
  • The structure and dynamics of a 37-residue antimicrobial peptide gaegurin 4 (GGN4) isolated from the skin of the native Korean frog, Rana rugosa, was determined in SDS micelles by NMR spectroscopy. The solution structure of the peptide in SDS micelles was determined from 352 NOE-derived distance constraints and 22 backbone torsion angle constraints. Dynamic properties for the amide backbone were characterized by $^1H-^{15}N $heteronuclear NOE experiments. The structural study revealed two amphipathic helices spanning residues 2-10 and 16-32 and that the helices were connected by a flexible loop. An intraresidue disulfide bridge was formed between residues Cys31 and Cys37 near the C-terminus. The loop region (11-15) connecting the two helices are were slightly more flexible than these helices themselves. From the fact that since there is no contact NOEs between two helices, it is implied that the GGN4 peptide shows an independent motion of both helices which has an angle of about $ 60^{\circ}-120^{\circ}$ from each other.

Metabolic Profiling of Urine Samples from Colorectal Cancer Patients Before and After Surgical Treatments

  • Chae, Young-Kee;Kang, Woo-Young;Kim, Seong-Hwan;Joo, Jong-Eun;Han, Joon-Kil;Hong, Boo-Whan
    • Journal of the Korean Magnetic Resonance Society
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    • v.14 no.1
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    • pp.28-37
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    • 2010
  • Metabolites of urine samples from 6 colorectal cancer patients were analyzed by two-dimensional NMR spectroscopy, where the samples were collected before and after the surgical treatments per patient. NMR data were analyzed with the help of the metabolome database and the statistics software. Urine samples before and after the treatments showed significantly different metabolic profiles from each other. We were able to compare 10 different metabolites. Most of the assigned metabolites of every patient showed a tendency of increase after the surgery except for a few cases. The amount of changes in individual metabolites varied significantly from patient to patient, but the combination of such changes could be used to distinguish the condition before the surgery from after, which could be done by PCA analysis. The analysis via $^{1}H-^{13}C$ HSQC spectra proved to be applicable in assessing and classifying global metabolic profiles of the urines from colorectal cancer patients.

NMR study of the interaction of T4 Endonuclease V with DNA

  • Lee, Bong-Jin;Im, Hoo-Kang;Hyungmi Lihm;Yu, Jun-Suk
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.80-80
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    • 1995
  • T4 Endonuclease V (Mw 16,000) acts as a repair enzyme for UV induced pyrimidine dimers in DNA. Many researchers have studied the biochemical characteristics of the enzyme. However the precise action mechanism of T4 endo V has not fully elucidated yet. In our laboratory NMR spectroscopy technique is being used for the structural study of T4 endo V. Because of its low temperature stability and high content of ${\alpha}$-helix, the conventional $^1$H NMR technique was inapplicable. Therefore we utilized stable isotope labeling technique and so far prepared about 10 amino acid specific labeled proteins. The HSQC spectra of amino acid specific labeled proteins will help us to interpret the triple resonance 3D, 4D data which are under processing, We also studied the behaviors of specific amino acid residues whose roles might be critical. When the enzyme labeled by $\^$15/N-Thr was mixed with the substrate oligonucleotide (semispecific -TT- sequence), one crosspeak in its HSQC spectrum was completely desappeared, which means that one of seven Thr residues is in the binding site of the enzyme with DNA, This result is well consistent with previous report that implicated the Thr 2 residue in the activity of the enzyme. Similar studies were carried on the behaviors of Arg and Tyr residues.

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Detection of electromagnetic interference shielding effect of Hanji mixed with carbon nanotubes using nuclear magnetic resonance techniques

  • Byun, Young Seok;Chae, Shin Ae;Park, Geun Yeong;Lee, Haeseong;Han, Oc Hee
    • Carbon letters
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    • v.27
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    • pp.90-97
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    • 2018
  • Electromagnetic interference (EMI) shielding is an important issue in modern daily life due to the increasing prevalence of electronic devices and their compact design. This study estimated EMI-shielding effect (EMI-SE) of small ($8-14{\times}17mm$) Hanji (Korean traditional paper) doped with carbon nanotubes (CNTs) and compared to Hanji without CNT using $^2H$ (92.1 MHz) and $^{23}Na$ (158.7 MHz) nuclear magnetic resonance (NMR) peak area data obtained from 1 M NaCl in $D_2O$ samples in capillary tubes that were wrapped in the Hanji samples. The simpler method of using the variation of reflected power and tuning frequency by inserting the sample into an NMR coil was also tested at 242.9, 158.7, and 92.1 MHz. Overall, EMI shielding was relatively more effective at the higher frequencies. Our results validated that NMR methods to be useful to evaluate EMI-SE, particularly for small, flexible shielding materials, and demonstrated that EMI shielding by absorption is dominant in Hanji mixed with CNT.

The High Resolution NMR Solution Structure of Monocyte Chemoattractant Protein-3

  • Kwon Do-Yoon;Lee Duck-Yeon;Sykes Brian D.;Kim Key-Sun
    • Journal of the Korean Magnetic Resonance Society
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    • v.9 no.2
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    • pp.74-92
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    • 2005
  • The high resolution solution structure of MCP-3 was determined using multinuclear, multidimensional NMR spectroscopy with an expressed and $^{13}C-\;and\;^{15}N-labeled$ protein. The MCP-3 has a typical chemokine fold including 3 anti-parallel $\beta-sheets$, and a C-terminal helix, but it exists as a monomer in solution under the conditions where the structure was determined (2 mM, pH 5.1 at $30^{\circ}C$). Based on the structure and the amino acid sequence compared to other chemokines we propose that Ile20 and Leu25 in MCP-3 play key roles in the formation of N-loop (residues between the $2^{nd}$ cysteine and the I sheet) which has been implicated as a determinant of chemokine specificity. Additional receptor binding surface is supplied by the 40s loop (residues between the 2 and the 3 sheet) and the binding interface of the acidic N-terminal region of chemokine receptor to MCP-3 would resemble the dimerization interface of CC type dimer.

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Effect of Substituted Trifluoromethyl Groups on Thermal and Mechanical Properties of Fluorine-containing Epoxy Resin

  • Heo, Gun-Young;Park, Soo-Jin
    • Macromolecular Research
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    • v.17 no.11
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    • pp.870-873
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    • 2009
  • In this study, 2-diglycidylether of benzotrifluoride (2-DGEBTF) and 4-diglycidylether of benzotrifluoride (4-DGEBTF) epoxy resins, which contained fluorine groups in the main chain, were synthesized. The resins were characterized by FTIR, $^1H$ NMR, $^{13}C$ NMR and $^{19}F$ NMR spectroscopy. The 2-DGEBTF and 4-DGEBTF epoxy resins were cured with triethylene tetramine (TETA), and the effect of the fluorine group on the synthesized epoxy resin on the cure behavior, thermal, and mechanical properties was investigated. The 2-DGEBTF/TETA system was more reactive than the 4-DGEBTF/TETA system, whereas the thermal stability factor i.e., the decomposition activation energy ($E_d$), of 4-DGEBTF/TETA was higher than that of 2-DGEBTF/TETA. These results can be explained by the decrease in cross-linking density and decomposition of the short side chains, resulting in the $CF_3$ group at the para position. However, the $K_{IC}$ value of 4-DGEBTF/TETA was higher than that of 2-DGEBTF/TETA. This was attributed to the increase in flexibility in the epoxy backbone, resulting in a difference in steric hindrance and polarlizability.

Isolation and Structural Determination of a Complete Set of Intact Bacteriochlorophyll-d Homologs and Isomers from Green Sulfur Bacterium Chlorobium vibrioforme and Their Aggregation Properties in Hydrophobic Solvents

  • Mizoguchi, Tadashi;Saga, Yoshitaka;Tamiaki, Hitoshi
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.344-346
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    • 2002
  • Eight intact bacteriochlorophyll (BChl)-d homologs and isomers were isolated from a strain of green sulfur bacterium Chlorobium vibrioforme. All the molecular structures of the BChl-d components were fully determined by a combination of mass spectrometry and $^1$H-NMR spectroscopy. The aggregation behavior of the isomerically pure BChls-d in hydrophobic organic solvents was examined with respect to the stereoisomeric configuration at the C3$^1$ position as well as the bulkiness of the C8 and C12 side-chains by using electronic- absorption spectroscopy.

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