• 제목/요약/키워드: Covalent Bond

검색결과 83건 처리시간 0.026초

활성화된 Sepharose Gels에 공유결합으로 고정화된 Urokinase를 이용한 융합단백질 절단반응 (Fusion Protein Cleavage by Urokinase Covalentley Immobilized to Activated Sepharose Gels)

  • 서창우;강관엽;이효실;안상점;이은규
    • KSBB Journal
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    • 제15권1호
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    • pp.42-48
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    • 2000
  • 본 연구에서는 고정화 UK를 이용한 융합단백질의 절단방응에 대해 UK의 고정화, 고정화 UK의 특성과 절단방응, 절단반응 후의 분리정제 그리고 고정화 UK으 재생에 대해 실험하였다. 고정화 수율은 99% 이상이였고 고정화 후의 효소활성은 80%를 유지하였다. 융합단백질 전단반응에서 액상 UK와 고정화 UK를 이용한 회분식 반응 모두 약 70%의 절단반응을 얻었고, 특히 고정화 UK의 사용시 부반응이 매우 낮은 이점이 있었다. 컬럼식 절단반응에서는 기절의 주입속도에 따라 절단수율은 크게 변화하였다. 최적의 유속은 50%의 절단수율을 얻은 1 bed volume/h로 설정하였다. 고정화 효소반응의 이점인 안정성과 반복사용 측면에서는 액상 UK 대비 고정화 UK가 높은 열안정성을 보였고 낮은 pH에서는 10% 이상 높은 활성을 유지하였다. 반복사용을 위해 6M GuHCl을 사용하여 인위적으로 풀림, 재접힘을 한 경우 98%의 활성을 얻음으로 타당성이 있음을 제시하였다. 또한 목적 단백질의 분리를 위하여 산침전 후 expanded bed adsorption 크로마토그래피를 이용함으로써 연속화된 고수율의 정제공정을 가능하게 하였다. 이러한 고정화 UK를 이용한 절단방응 및 정제시스템을 구축함으로써 융합단백지의 생산공정에 매우 유용하게 사용될 것으로 생각되어진다.

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Vicinal Oxime 리간드의 새로운 구리(II) 착물에 대한 합성, 특성 및 ESR 연구 (Synthesis, Characterization and ESR Studies of New Copper(II) Complexes of Vicinal Oxime Ligands)

  • El-Tabl, Abdou S.;Shakdofa, Mohamad M.E.;El-Seidy, Ahmed M.A.
    • 대한화학회지
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    • 제55권4호
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    • pp.603-611
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    • 2011
  • Ethoxylacetyl oxime 리간드 [HL, (1) 및 $H_2L^1$, (3)]와 아세트산 구리(II) 일수화물을 반응시켜 팔면체 및 사각평면착물을 각각 얻었다. 이들 착물을 원소분석, IR 및 UV-Vis 스펙트럼, 자기수자율, 전기전도도 및 ESR 스펙트럼으로 조사하였다. DMF에서 이들 착물의 몰전기전도도 측정한 결과 비이온성을 보였다. 실온 및 77 K에서 [$(L)_2Cu(H_2O)_2$], (2) 착물의 ESR 스펙트럼은 배위결합성을 갖는 축대칭($d_{x2-y2}$)의 특성을 보이며, 또한 전형적인 쌍극자 상호작용에 의해 큰 띠나비를 갖는다. 그러나 착물 [$(L^1)Cu$], (4)는 고체상태에서 큰 띠넓임으로 인해 초미세 갈라짐이 관찰되지 않았는데, 이는 구리(II) 결합자리들 사이의 스핀-교환 상호작용에 의한 것으로 확인되었다. 실온에서 도포된 구리(II) 착물의 스펙트럼은 배위된 질소원자로 인해 극초미세 갈라짐을 보였다. 또한 이 물질은 공유 결합성을 갖는 축형태($d_{x2-y2}$)의 특성을 보이고, 구리(II) 이온의 환경은 기본적으로 사각-평면 배열을 갖는다. 77 K의 얼어있는 메탄올에서 [$(L_1)Cu$], (4)의 스펙트럼은 이합체 화합물의 삼중항 상태의 특성을 보이며, 두 구리(II) 이온 중심 사이의 거리를 계산한 결과 4.8 이다.

Investigation of the Copper (Cu) Binding Site on the Amyloid beta 1-16 (Aβ16) Monomer and Dimer Using Collision-induced Dissociation with Electrospray Ionization Tandem Mass Spectrometry

  • Ji Won Jang;Jin Yeong Lim;Seo Yeon Kim;Jin Se Kim;Ho-Tae Kim
    • Mass Spectrometry Letters
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    • 제14권4호
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    • pp.153-159
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    • 2023
  • The copper ion, Cu(II), binding sites for amyloid fragment Aβ1-16 (=Aβ16 ) were investigated to explain the biological activity difference in the Aβ16 aggregation process. The [M+Cu+(z-2)H]z+ (z = 2, 3 and 4, M = Aβ16 monomer) and [D+Cu+(z-2)H]z+ (z = 3 and 5, D = Aβ16 dimer) structures were investigated using electrospray ionization (ESI) mass spectrometry (MS) and tandem mass spectrometry (MS/MS). Fragment ions of the [M+Cu+(z-2)H]z+ and [D+Cu+(z-2)H]z+ complexes were observed using collision-induced dissociation MS/MS. Three different fragmentation patterns (fragment "a", "b", and "y" ion series) were observed in the MS/MS spectrum of the (Aβ16 monomer or dimer-Cu) complex, with the "b" and "y" ion series regularly observed. The "a" ion series was not observed in the MS/MS spectrum of the [M+Cu+2H]4+ complex. In the non-covalent bond dissociation process, the [D+Cu+3H]5+ complex separated into three components ([M+Cu+H]3+, M3+, and M2+), and the [M+Cu]2+ subunit was not observed. The {M + fragment ion of [M+Cu+H]3+} fragmentation pattern was observed during the covalent bond dissociation of the [D+Cu +3H]5+ complex. The {M + [M+Cu+H]3+} complex geometry was assumed to be stable in the [D+Cu+3H]5+ complex. The {M + fragment ion of [M+Cu]2+} fragmentation pattern was also observed in the MS/MS spectrum of the [D+Cu+H]3+ complex. The {M + [y9+Cu]1+} fragment ion was the characteristic fragment ion. The [D+Cu+H]3+ and [D+Cu+3H]5+ complexes were likely to form a monomer-monomer-Cu (M-M-Cu) structure instead of a monomer-Cu-monomer (M-Cu-M) structure.

Theoretical Studies on Electronic Structure and Absorption Spectrum of Prototypical Technetium-Diphosphonate Complex 99mTc-MDP

  • Qiu, Ling;Lin, Jian-Guo;Gong, Xue-Dong;Ju, Xue-Hai;Luo, Shi-Neng
    • Bulletin of the Korean Chemical Society
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    • 제32권7호
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    • pp.2358-2368
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    • 2011
  • Density functional theory (DFT) and time-dependent density functional theory (TDDFT) calculations, employing the B3LYP method and the LANL2DZ, 6-31G$^*$(LANL2DZ for Tc), 6-31G$^*$(cc-pVDZ-pp for Tc) and DGDZVP basis sets, have been performed to investigate the electronic structures and absorption spectra of the technetium-99m-labeled methylenediphosphonate ($^{99m}Tc$-MDP) complex of the simplest diphosphonate ligand. The bonding situations and natural bond orbital compositions were studied by the Mulliken population analysis (MPA) and natural bond orbital (NBO) analysis. The results indicate that the ${\sigma}$ and ${\pi}$ contributions to the Tc-O bonds are strongly polarized towards the oxygen atoms and the ionic contribution to the Tc-O bonding is larger than the covalent contribution. The electronic transitions investigated by TDDFT calculations and molecular orbital analyses show that the origin of all absorption bands is ascribed to the ligand-to-metal charge transfer (LMCT) character. The solvent effect on the electronic structures and absorption spectra has also been studied by performing DFT and TDDFT calculations at the B3LYP/6-31G$^*$(cc-pVDZ-pp for Tc) level with the integral equation formalism polarized continuum model (IEFPCM) in different media. It is found that the absorption spectra display blue shift in different extents with the increase of solvent polarity.

Influence of Surface Morphology and Substrate on Thermal Stability and Desorption Behavior of Octanethiol Self-Assembled Monolayers

  • ;강훈구;;;노재근
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2012년도 제43회 하계 정기 학술대회 초록집
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    • pp.219-219
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    • 2012
  • The formation and thermal desorption behaviors of octanethiol (OT) SAMs on single crystalline Au (111) and polycrystalline Au, Ag, and Cu substrates were examined by X-ray photoelectron microscopy (XPS), thermal desorption spectroscopy (TDS), and contact angle (CA) measurements. XPS and CA measurements revealed that the adsorption of octanethiol (OT) molecules on these metals led to the formation of chemisorbed self-assembled monolayers (SAMs). Three main desorption fragments for dioctyl disulfide (C8SSC8+, dimer), octanethiolate (C8S+), and octanethiol (C8SH+) were monitored using TDS to understand the effects of surface morphology and the nature of metal substrates on the thermal desorption behavior of alkanethiols. TDS measurements showed that a sharp dimer peak with a very strong intensity on single crystalline Au (111) surface was dominantly observed at 370 K, whereas a broad peak on the polycrystalline Au surface was observed at 405 K. On the other hand, desorption behaviors of octanethiolates and octanethiols were quite similar. We concluded that substrate morphology strongly affects the dimerization process of alkanethiolates on Au surfaces. We also found that desorption intensity of the dimer is in the order of Au>>Ag>Cu, suggesting that the dimerization process occurs efficiently when the sulfur-metal bond has a more covalent character (Au) rather than an ionic character (Ag and Cu).

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Preparation and Characterizations of C60/Polystyrene Composite Particle Containing Pristine C60 Clusters

  • Kim, Jung-Woon;Kim, Kun-Ji;Park, Soo-Yeon;Jeong, Kwang-Un;Lee, Myong-Hoon
    • Bulletin of the Korean Chemical Society
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    • 제33권9호
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    • pp.2966-2970
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    • 2012
  • Fullerene/polystyrene ($C_{60}$/PS) nano particle was prepared by using emulsion polymerization. Styrene and fullerene were emulsified in aqueous media in the presence of poly(N-vinyl pyridine) as an emulsion stabilizer, and polymerization was initiated by water soluble radical initiator, potassium persulfate. The obtained nano particles have an average diameter in the range of 400-500 nm. The fullerene contents in the nano particle can be controlled up to 15 wt % by varying the feed ratio, which was confirmed by themogravimetric analysis (TGA) and elemental analysis (EA). The structure and morphologies of the $C_{60}$/PS nano particles were examined by various analytical techniques such as dynamic light scattering (DLS), scanning electron microscope (SEM), transmission electron microscope (TEM), electron diffraction (ED) pattern, X-ray powder diffraction (XRD), and UV spectroscopy. Unlike conventional $C_{60}$/PS particles initiated by organic free radical initiators, in which the fullerene is copolymerized forming a covalent bond with styrene monomer, the prepared $C_{60}$/PS nano particles contain pristine fullerene as secondary particles homogeneously distributed in the polystyrene matrix.

Dyeing Properties of Nylon Fabric with Reactive Dyes Having Different Reactive Group

  • Lee, Byung-Sun;Lee, Jin-Ah;Lee, Kwon-Sun;Park, Jong-Ho;Kim, Sung-Dong
    • 한국염색가공학회지
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    • 제18권5호
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    • pp.1-7
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    • 2006
  • The dyeing properties of the several reactive dyes possessing different types of the reactive group on nylon 6 fabric have been investigated. Dyeing of nylon fiber with commercial reactive dyes was dependent on the dye bath pH. As pH increased, the extent of exhaustion decreased due to the deprotonation of the amino end group. But, the extent of fixation increased because of the enhanced reactivity with nylon fiber. It was found that all reactive dyes could be applied to nylon 6 reasonably at pH 5-6. But, the conflicting requirements of dye exhaustion and covalent bond formation limited the extent of fixation efficiency. The best method to improve the extent of the fixation efficiency of the reactive dyes was the conversion of sulphatoethylsulphone group to vinylsulphone group before dyeing with trisodium phosphate. The converted Supra type reactive dye gave the highest extent of fixation efficiency, which was about 90%.

Vinylsulfone Activated Agarose 에 Endo- 및 Exoinulinase의 고정화 (Immobilization of Endo- and Exoinulinase on Vinylsulfone Activated Agarose)

  • 한상배;송근섭;정용섭;손희숙;우순자;엄태봉
    • 한국미생물·생명공학회지
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    • 제20권1호
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    • pp.20-24
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    • 1992
  • Inulinase의 효율적인 재사용을 위하여 vinylsulfone activated agarose에 endo- 및 exoinulinase를 고정화시켰다. Gram gel당 exoinulinase는 400U, endoinulinase는 80U까지 고정화시킬 수가 있었고 열안정성은 exoinulinase 에서 증가되었다. 두 고정화 효소의 혼합비율에 따른 synergistic effect는 endo/exo가 0.5-0.1일 대 가장 컸으며, synergistic effect는 혼합되지 않은 상태의 고정화 효소에 비해 그 활성이 약 1.7배 증가하였다. 두 고정화 효소의 최적 pH는 4.4-5.0 범위이었으며 operational stability는 batch reactor에서 20번 반복된 실험결과 어떠한 효소활성의 감소도 보이지 않았다.

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포도당 센서의 제작을 위한 고정화 방법의 전기화학적 결정 (Electrochemical Determination of Immobilization Technique for Glucose Sensor Fabrication)

  • 정태훈;홍석인;노봉수;정용섭;윤정원;김태진
    • KSBB Journal
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    • 제13권1호
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    • pp.52-57
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    • 1998
  • The present work proposes a simple electrochemical method applicable to any immobilization processes of oxidase using a Clark type oxygen electrode as a base transducer. The present work suggests an optimal immobilization technique among three different methods of glucose oxidase(GOD) onto one side of $37[\mu}$mthick blend membranes, composed o 80% of cellulose triacetate and 20% of polycaprolactone, on the basis of the maximum Michaelis-Menten parameter(Vm) determined by either steady state or transient analyses. The electrode system was made of disk type gold cathode(4mm diameter) and Ag/AgCl anode. One side of the blend membrane was in contact with the cathode surface while the other side was immobilized with GOD either in covalent-bond or cross-linked forms, the latter being covered by $25{\mu}$m thick dialysis membrane of cellulose acetate. The resultant current density was on-line monitored by a potentiostat while glucose level was varied from 1 to 20 mM. The present study shows that direct cross-linking of GOD with glutaraldehyde was mostly preferred for fabrication of glucose sensor, on the basis of resultant kinetic parameters from either steady state or transient analyses.

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고압연소 소결(HPCS)법에 의한 탄화티타늄(TiC)의 합성 및 소결 (Simultaneous Synthesis and Sintering of Titanium Carbide by HPCS(High Pressure-Self Combustion Sintering))

  • 김지헌;최상욱;조원승;조동수;오장환
    • 한국세라믹학회지
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    • 제34권5호
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    • pp.473-482
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    • 1997
  • Titanium carbide(TiC) has a poor sinterability due to the strong covalent bond. Thus, it is generally fabricated by either hot pressing or pressureless-sintering at elevated temperature by the addition of sintering aids such as nickel(Ni), molybdenum(Mo) and cobalt(Co). However, these sintering methods have the following disadvantages; (1) the complicated process, (2) the high energy consumption, and (3) the possibility of leaving inevitable impurities in the product, etc. In order to reduce above disadvantages, we investigated the optimum conditions under which dense titanium carbide bodies could be synthesized and sintered simultaneously by high pressure self-combustion sintering(HPCS) method. This method makes good use of the explosive high energy from spontaneous exothermic reaction between titanium and carbon. The optimum conditions for the nearly full-densification were as follows; (1) The densification of sintered body becomes high by increasing the pressing pressure from 400kgf/$\textrm{cm}^2$ upto 1200 kgf/$\textrm{cm}^2$. (2) Instead of adding the coarse graphite or activated carbon, the fine particles of carbon black should be added as a carbon source. (3) The optimum molar ratio of carbon to titanium (C/Ti) was unity. In reality, titanium carbide body which were prepared under optimum conditions had relatively dense textures with the apparent porosity of 0.5% and the relative density of 98%.

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