• 제목/요약/키워드: three-dimensional characterization

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Thermus aquaticus YT-1의 내열성 프로테아제 aqualysin I의 구조와 특징 (Characterization of aqualysin I structure(a thermophilic alkaline Serine protease) of Thermus aquaticus YT-1)

  • 권석태
    • Applied Biological Chemistry
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    • 제31권3호
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    • pp.274-283
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    • 1988
  • Aqualysin I is an alkaline serine protease which is secretet into the culture medium by Thermus aquaticus YT-1, an extreme thermophile. Aqualysin I was purified, and its partial amino acid sequence was determined. The gene encoding aqualysin I was cloned into E. coli using synthetic oligodeoxyribonucleotides as hybridization probes. The nucleotide sequence of the cloned DNA was determined. The primary structure of aqualysin I, deduced from the nucleotide sequenc, agreed with the determid amino acid sequences, including the $NH_2-$ and COOH terminal sequence of the tryptides derived from aqualysin I. Aqualysin I comprised 281 amino acid residues and its molecular mass was determined to be 28350. On alignment of the whole amino acid sequence, aqualysin I showed high sequence homology with the subtilisin type serine protease, and 43% identity with proteinase K, 37-30% with subtilisins and 34% with thermitase. Extremely high sequence identity was observed in the regions containing the active-site residues, corresponding to Asp32, His64 and Ser221 of subtilisin BPN'. Aqualysin I contains two disulfide bonds, Cys67-Cys99 and Cys163-Cys194, and these disulfide bonds seem to contribute to the heat stability of the enzyme. The determined positions of the twe disulfide bonds of aqualysin I agreed with those predicted previously on the basis of computer graphics of the crystallographic data for subtilisin BPN'. Therefore, these findings sugests that the three-dimensional structure of aqualysin I is similar to that of subtilisin BPN' Aqualysin I is produced as a lage precursor, which contains $NH_2-$ and COOH- terminal portions besides the mature protease sequence.

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집누에(Bombyx mori)와 멧누에(Bombyx mandayina)의 종간교잡에 있어서 란각구조 및 Chorion 단백질 (Morphological and Biochemical Characterization of the Chorion in Interspecific Hybrid Between Bombyx mori and Bombyx mandarina)

  • 김종길;노시갑
    • 한국잠사곤충학회지
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    • 제36권1호
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    • pp.30-36
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    • 1994
  • 집누에와 멧누에의 종간교잡에 의한 란각구조 및 chorion 단백질의 변이와 양 종간의 유연관계를 검토했다. 1. 교잡종 란의 측면부의 란각표면구조는 양 종이 모두 집누에와 유사한 그물상구조를 나타내지만 각각의 교잡 모체쪽의 구조와 유사했다. 2. 교잡종에서의 란각단면구조는 멧누에의 구조와 유사했으며 최외층인 덮개층은 측면부에서 주변부로 갈수록 더욱 뚜렷이 관찰되었다. 3. Chorion 단백질의 성분분석 결과 등전점이 4.0-6.5 분자량 10-50 kd 사이에 대부분의 성분이 검출되었으며 교잡종이 chorion 단백질성분은 영동 pattern, 성분수, 등전점 및 분자량 등에 있어서 교잡모체의 chorion 단백질과 유사했다.

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Synthesis and Characterization of Novel pH-Sensitive Hydrogels Containing Ibuprofen Pen dents for Colon-Specific Drug Delivery

  • Mahkam, Mehrdad;Poorgholy, Nahid;Vakhshouri, Laleh
    • Macromolecular Research
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    • 제17권9호
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    • pp.709-713
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    • 2009
  • The aim of this study was to develop novel intestinal specific drug delivery systems with pH sensitive swelling and drug release properties. The carboxyl group of ibuprofen was converted to a vinyl ester group by reacting ibuprofen and vinyl acetate as an acylating agent in the presence of catalyst. The glucose-6-acrylate-1, 2, 3, 4-tetraacetate (GATA) monomer was prepared under mild conditions. Cubane-1, 4-dicarboxylic acid (CDA) linked to two 2-hydroxyethyl methacrylate (HEMA) group was used as the crosslinking agent (CA). Methacrylic-type polymeric prodrugs were synthesized by the free radical copolymerization of methacrylic acid, vinyl ester derivative of ibuprofen (VIP) and GATA in the presence of cubane cross linking agent. The structure of VIP was characterized and confirmed by FTIR, $^1H$ NMR and $^{13}C$ NMR spectroscopy. The composition of the cross-linked three-dimensional polymers was determined by FTIR spectroscopy. The hydrolysis of drug polymer conjugates was carried out in cel-lophane membrane dialysis bags, and the in vitro release profiles were established separately in enzyme-free simulated gastric and intestinal fluids (SGF, pH 1 and SIF, pH 7.4). The detection of a hydrolysis solution by UV spectroscopy at selected intervals showed that the drug can be released by hydrolysis of the ester bond between the drug and polymer backbone at a low rate. Drug release studies showed that increasing the MAA content in the copolymer enhances the rate of hydrolysis in SIP. These results suggest that these polymeric prodrugs can be useful for the release of ibuprofen in controlled release systems.

Heterologous Expression and Characterization of a Thermostable Exo-β-D-Glucosaminidase from Aspergillus oryzae

  • Wu, Dingxin;Wang, Linchun;Li, Yuwei;Zhao, Shumiao;Peng, Nan;Liang, Yunxiang
    • Journal of Microbiology and Biotechnology
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    • 제26권2호
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    • pp.347-355
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    • 2016
  • An exo-β-D-glucosaminidase (AorCsxA) from Aspergillus oryzae FL402 was heterologously expressed and purified. The deduced amino acid sequence indicated that AorCsxA belonged to glycoside hydrolase family 2. AorCsxA digested colloid chitosan into glucosamine but not into chitosan oligosaccharides, demonstrating exo-β-D-glucosaminidase (CsxA) activity. AorCsxA exhibited optimal activity at pH 5.5 and 50℃; however, the enzyme expressed in Pichia pastoris (PpAorCsxA) showed much stronger thermostability at 50℃ than that expressed in Escherichia coli (EcAorCsxA), which may be related to glycosylation. AorCsxA activity was inhibited by EDTA and most of the tested metal ions. A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of Vmax and kcat values with those of the wild-type enzyme. The three-dimensional structure suggested the tightened pocket at the active site of F769W enabled efficient substrate binding. The AorCsxA gene was heterologously expressed in P. pastoris, and one transformant was found to produce 222 U/ml activity during the high-cell-density fermentation. This AorCsxA-overexpressing P. pastoris strain is feasible for large-scale production of AorCsxA.

Foundation Design the 151 story Incheon Tower in Reclamation Area

  • Abdelrazaq, Ahmad;Badelow, Frances;Kim, Sung-Ho;Park, Yung-Ho
    • 한국지반공학회:학술대회논문집
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    • 한국지반공학회 2009년도 세계 도시지반공학 심포지엄
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    • pp.157-171
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    • 2009
  • A 151 storey super high-rise building located in an area of reclaimed land constructed over soft marine clay in Songdo, Korea is currently under design. This paper describes the design process of the foundation system of the supertall tower, which is required to support the large building vertical and lateral loads and to restrain the horizontal displacement due to wind and seismic forces. The behaviour of the foundation system due to these loads and foundation stiffness influence the design of the building super structure, displacement of the tower, as well as the raft foundation design. Therefore, the design takes in account the interactions between soil, foundation and super structure, so as to achieve a safe and efficient building performance. The site lies entirely within an area of reclamation underlain by up to 20m of soft to firm marine silty clay, which overlies residual soil and a profile of weathered rock. The nature of the foundation rock materials are highly complex and are interpreted as possible roof pendant metamorphic rocks, which within about 50m from the surface have been affected by weathering which has reduced their strength. The presence of closely spaced joints, sheared and crushed zones within the rock has resulted in deeper areas of weathering of over 80m present within the building footprint. The foundation design process described includes the initial stages of geotechnical site characterization using the results of investigation boreholes and geotechnical parameter selection, and a series of detailed two- and three-dimensional numerical analysis for the Tower foundation comprising over 172 bored piles of varying length. The effect of the overall foundation stiffness and rotation under wind and seismic load is also discussed since the foundation rotation has a direct impact on the overall displacement of the tower.

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결정형이 생체이용률에 미치는 영향 (Effect of Crystal Form on Bioavailability)

  • 손영택
    • Journal of Pharmaceutical Investigation
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    • 제34권6호
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    • pp.443-452
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    • 2004
  • Habit is the description of the outer appearance of a crystal. If the environment of a growing crystal affects its external shape without changing its internal structure, a different habit results. Crystal habit and the internal structure of a drug can affect bulk and physicochemical properties, which range from flowability to chemical stability. A polymorph is a solid crystalline phase of a given compound resulting from the possibility of at least two different arrangements of the molecules of that compound in the solid state. Chemical stability and solubility changes due to polymorphism can have an impact on a drug's bioavailability and its development program. During crystallization from a solution, crystals separating may consist of a pure component or be a molecular compound. Solvates are molecular complexes that have incorporated the crystallizing solvent molecule in their lattice. When the solvent incorporated in the solvate is water, it is called a hydrate. To distinguish solvates from polymorphs, which are not molecular compounds, the term pseudopolymorph is used. Identification of possible hydrate compounds is important since their aqueous solubilities can be significantly less than their anhydrous forms. Conversion of an anhydrous compound to a hydrate within the dosage form may reduce the dissolution rate and extent of drug absorption. An amorphous solid may be treated as a supercooled liquid in which the arrangement of molecules is random. Amorphous solids lack the three-dimensional long-range order found in crystalline solids. Since amorphous forms are usually of higher thermodynamic energy than corresponding crystalline forms, solubilities as well as dissolution rates are generally greater. A study on crystal form includes characterization of (l)crystal habit, (2)polymorphism, (3)pseudopolymorphism, (4)amorphous solid.

Characterization of the v-cath Gene of Bombyx mori Nuclear Polyhedrosis Virus K1

  • Lee, Kwang Sik;Li, Jianhong;Je, Yeon Ho;Woo, Soo Dong;Sohn, Hung Dae;Jin, Byung Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • 제9권2호
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    • pp.217-223
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    • 2004
  • A cathepsin L-like cysteine protease, v-cath, encoded by the baculovirus has been shown to playa role in host liquefaction. We have identified a v-cath gene in the silkworm virus, Bombyx mori nuclear polyhedrosis virus (BmNPV) K1 strain. The 969 bp v-cath has an open reading frame of 323 amino acids. A putative cleavage site and catalytic sites were conserved in BmNPV-K1 v-cath. The predicted three-dimensional structure of BmNPV-K1 v-cath revealed that the overall fold of BmNPV-K1 v-cath is similar to that of other proteases of the papain family. The deduced amino acid sequence of BmNPV-K1 v-cath showed 98% and 97% protein sequence identity to BmNPV T3 strain and to Autographa californica nuclear polyhedrosis virus, respectively. The BmNPV-K1 v-cath differed at 4 amino acid positions from BmNPV T3. The v-cath gene in BmNPV-K1 genome is located on the EcoRV 6 kb and XhoI 9 kb fragments. Northern hybridization analysis of BmNPV K1 v-cath gene revealed that it is expressed late in infection.

콘빔형 소동물용 전산화단층촬영(CT) 장치의 성능평가 (Characterization of Cone-beam Computed Tomography System for Small Animal)

  • 강형근;천권수
    • 한국방사선학회논문지
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    • 제4권1호
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    • pp.19-24
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    • 2010
  • 콘빔형 소동물용 CT 장치는 바이오-의료분야의 기초 실험에 활발히 응용되고 있다. $10{\mu}m$의 직경을 가진 텅스텐 와이어(wire)와 물 팬텀을 이용하여 비교적 간단한 방법으로 소동물용 CT 장치를 평가할 수 있는 방법을 제시하였다. X선 튜브가 안정적으로 장시간 운전 가능한 조건에서 $1^{\circ}$ 간격으로 360개의 투영영상을 획득하고, 영상재구성을 통하여 슬라이스 영상 및 3차원 영상을 얻었다. 콘빔형 소동물용 CT 장치는 배율 1.07배 위치에서 13.78lp/mm(공간해상도 $36.2{\mu}m$)를 나타내었고, 80kV의 관전압에서 CNR 10.33 및 S/N 5.87을 보였다.

다양한 지구통계기법의 지하매질 예측능 및 적용성 비교연구 (Comparative Analysis of Subsurface Estimation Ability and Applicability Based on Various Geostatistical Model)

  • 안정우;정진아;박은규
    • 한국지하수토양환경학회지:지하수토양환경
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    • 제19권4호
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    • pp.31-44
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    • 2014
  • In the present study, a few of recently developed geostatistical models are comparatively studied. The models are two-point statistics based sequential indicator simulation (SISIM) and generalized coupled Markov chain (GCMC), multi-point statistics single normal equation simulation (SNESIM), and object based model of FLUVSIM (fluvial simulation) that predicts structures of target object from the provided geometric information. Out of the models, SNESIM and FLUVSIM require additional information other than conditioning data such as training map and geometry, respectively, which generally claim demanding additional resources. For the comparative studies, three-dimensional fluvial reservoir model is developed considering the genetic information and the samples, as input data for the models, are acquired by mimicking realistic sampling (i.e. random sampling). For SNESIM and FLUVSIM, additional training map and the geometry data are synthesized based on the same information used for the objective model. For the comparisons of the predictabilities of the models, two different measures are employed. In the first measure, the ensemble probability maps of the models are developed from multiple realizations, which are compared in depth to the objective model. In the second measure, the developed realizations are converted to hydrogeologic properties and the groundwater flow simulation results are compared to that of the objective model. From the comparisons, it is found that the predictability of GCMC outperforms the other models in terms of the first measure. On the other hand, in terms of the second measure, the both predictabilities of GCMC and SNESIM are outstanding out of the considered models. The excellences of GCMC model in the comparisons may attribute to the incorporations of directional non-stationarity and the non-linear prediction structure. From the results, it is concluded that the various geostatistical models need to be comprehensively considered and comparatively analyzed for appropriate characterizations.

Tribological Performance of Ni-Cr Composite Coating Sprayed onto AISI 4340 (SNCM439) Steel by High Velocity Oxygen Fuel

  • Umarov, Rakhmatjon;Pyun, Young-Sik;Amanov, Auezhan
    • Tribology and Lubricants
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    • 제34권6호
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    • pp.217-225
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    • 2018
  • In this study, we spray a Ni-Cr composite powder onto AISI 4340 steel using the high velocity oxygen fuel method. We subsequently subject the Ni-Cr coating (as-sprayed) to ultrasonic nanocrystal surface modification (UNSM) process to improve the tribological performance. This study aims at increasing the wear resistance and durability of the Ni-Cr coating by altering the surface integrity and microstructure via the UNSM process. The UNSM process reduces the surface roughness of the as-sprayed coating by about 64%, which is explained by observing the elimination of high peaks and valleys and filling up micro-pores. Furthermore, a change in the microstructure of the coating due to continuous high-frequency strikes to the surface by a tip can lead to an increase in hardness from about 48 to 60 HRC. Furthermore, we investigate the characterization of the friction and wear behavior of Ni-Cr coating by a ball-on-disc tribometer in the dry conditions. We determine that after the UNSM process, there is a significant reduction in the friction coefficient of the as-sprayed coating from approximately 1.1 to 0.75. This is owing to the increased hardness and smoothed surface roughness. In addition, we investigate the surface morphology and wear track of the coatings before and after the UNSM process using a scanning electron microscope, energy dispersive spectrometer, and three-dimensional laser scanning microscope. We observe that the wear track of the Ni-Cr coating after the UNSM process is lower than that of the as-sprayed one. Thus, we confirm that the UNSM process has a significant influence on the improvement of the tribological performance of the Ni-Cr composite coating.