• Title/Summary/Keyword: fab E

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Structure Determination of Antifungal KRF-001 Produced by Bacillus subtilis subsp. krictiensis (Bacillus subtilis subsp. krictiensis가 생산하는 항진균 물질 KRF-001의 구조 결정)

  • 김성기;이남경;정태숙;김영국;최진자;복성해
    • Microbiology and Biotechnology Letters
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    • v.19 no.6
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    • pp.598-603
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    • 1991
  • An antifungal mixture of six members (component A to F), KRF-001 produced by Bacillzts subtilis subsp. krictiensis was isolated from the fermentation broth. Molecular weight of component A to F was determined by FAB-MS to be 1042, 1056, 1056, 1070, 1070 and 1084 respectively. Various instrumental analyses (amino acid analysis, GC-MS, $^1H-NMR, ^1HH$ COSY NMR) revealed that the mixture was a homologous cyclic peptide composed of each one mole of glutamine, proline, tyrosine, serine, unusual $\beta$-amino acid and three moles of asparagine. The structural differences of component A to F were found in carbon number and terminal structure of the unusual $\beta$-amino acid. After determination of the sequence and stereochemistry of those amino acids, the tentative structure of KRF-001 was determined.

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An Adaptive Dispatching Architecture for Constructing a Factory Operating System of Semiconductor Fabrication : Focused on Machines with Setup Times (반도체 Fab의 생산운영시스템 구축을 위한 상황적응형 디스패칭 방법론 : 공정전환시간이 있는 장비를 중심으로)

  • Jeong, Keun-Chae
    • IE interfaces
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    • v.22 no.1
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    • pp.73-84
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    • 2009
  • In this paper, we propose a dispatching algorithm for constructing a Factory Operating System (FOS) which can operate semiconductor fabrication factories more efficiently and effectively. We first define ten dispatching criteria and propose two methods to apply the defined dispatching criteria sequentially and simultaneously (i.e. fixed dispatching architecture). However the fixed type methods cannot apply the criteria adaptively by considering changes in the semiconductor fabrication factories. To overcome this type of weakness, an adaptive dispatching architecture is proposed for applying the dispatching criteria dynamically based on the factory status. The status can be determined by combining evaluation results from the following three status criteria; target movement, workload balance, and utilization rate. Results from the shop floor in past few periods showed that the proposed methodology gives a good performance with respect to the productivity, workload balance, and machine utilization. We can expect that the proposed adaptive dispatching architecture will be used as a useful tool for operating semiconductor fabrication factories more efficiently and effectively.

Characterization of Extremely Hydrophobic Immunostimulatory Lipoidal Peptides by Matrix Assisted Laser Desorption Ionization Mass Spectrometry

  • 장정석;이성택;장윤석
    • Bulletin of the Korean Chemical Society
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    • v.17 no.11
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    • pp.1036-1039
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    • 1996
  • Synthetic lipoidal peptides based on viral protein sequences have been prepared. These peptides contain an N-palmitoyl group at the N-terminal residue, which is a modified cysteine, containing a S-[2,3-bis(acyloxy)-(2-R,S)-propyl] moiety. When this residue (Pam3Cys) is at the N-terminus of a synthetic peptide, it acts as potent immunoadjuvant to enhance both IgM and IgG antibody responses to the attached peptide. Conventional analytical procedures (e.g., Edman degradation and amino acid analysis) are either not applicable due to the N-terminal modification, or do not provide confirmation of the intact structure. Chromatographic analysis is also hindered by the tendency of these lipoidal Pam3Cys peptides to form large aggregates, and in some cases to be permanently adsorbed on reversed phase columns. We have applied several mass spectrometric techniques, including fast atom bombardment (FAB), electrospray ionization (ESI) and matrix assisted laser desorption ionization (MALDI) to characterize the intact structures of a number of different Pam3Cys synthetic peptides. The MALDI-MS has been found to be the most sensitive for the analysis of the structure of Pam3Cys peptides.

Detection of Sugar Process Contamination Using Dextran Binding hages Produced by Batch Fermentation of Escherichia coli (대장균의 회분식 발효에 의해 생산된 덱스트란 결합 파아지를 활용한 설탕 제조공정 오염 검출)

  • Kim, Du-Woon
    • Food Science and Preservation
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    • v.15 no.5
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    • pp.617-621
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    • 2008
  • Sequential passes through $Sephadex^{TM}$ columns were used to select phages that displays ligands for dextran ($\alpha$-1,6 linked linear chains) from a phage antibody library. Those phages that bound to the $Sephadex^{TM}$ in each iteration were replicated in E. coli. A phage preparation isolated on the third round selection produced 5.4 nephelos turbidity units (NTU) in a dextran specific immunonephelometric assay, a 2.2 fold higher value than the phage preparation from the first round selection. This phage gave $72\;{\pm}\;10$ normalized intensity (N.I.) in a dip-stick assay against high molecular size dextran (T2000, $2\;{\times}\;10^6) and significantly lower color ($30\;{\pm}\;6$ N.I.) against low molecular size dextran (T10, $10^4$). The presence of an Fab insert in each of these phages was confirmed using a $\beta-galactosidase linked assay and polymerase chain reaction.

Three New Flavonoids Of Spatholobus suberectus

  • Yoon, Jeong-Seon;Lim, Song-Won;Yang , Hye-Kyeong;Ahn , Tae-Kyeong;Kim, Eun-Sil;Sung, Sang-Hyun;Kim, Young-Choong
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.202.1-202.1
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    • 2003
  • Six compounds were isolated from the 90% MeOH fraction of the vine stem of Spatholobus suberectus Dunn (Leguminosae) using silica gel, reverse phase column chromatography and RP-HPLC. Structures of compounds 1-6 were elucidated by spectroscopic parameters of IR, E1 MS, FAB MS, lD-NMR and 2D-NMR spectrum and identified as pseudobaptigenin (1), genistein (2), (2R)-7-hydroxy-6-methoxyflavanone (3), (3R,4R)-2",4"-dihydroxy-6,7-methylenedioxy-isoflavan-4-ol (4), (3R,4R)-7,2" -dihydroxy- 4"-methoxyisoflavan-4-ol (5), sativan (6), respectively. Compounds 3, 4 and 5 have been newly reported in nature.

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Generation and Characterization of Monoclonal Antibodies to the Ogawa Lipopolysaccharide of Vibrio cholerae O1 from Phage-Displayed Human Synthetic Fab Library

  • Kim, Dain;Hong, Jisu;Choi, Yoonjoo;Han, Jemin;Kim, Sangkyu;Jo, Gyunghee;Yoon, Jun-Yeol;Chae, Heesu;Yoon, Hyeseon;Lee, Chankyu;Hong, Hyo Jeong
    • Journal of Microbiology and Biotechnology
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    • v.30 no.11
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    • pp.1760-1768
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    • 2020
  • Vibrio cholerae, cause of the life-threatening diarrheal disease cholera, can be divided into different serogroups based on the structure of its lipopolysaccharide (LPS), which consists of lipid-A, core-polysaccharide and O-antigen polysaccharide (O-PS). The O1 serogroup, the predominant cause of cholera, includes two major serotypes, Inaba and Ogawa. These serotypes are differentiated by the presence of a single 2-O-methyl group in the upstream terminal perosamine of the Ogawa O-PS, which is absent in the Inaba O-PS. To ensure the consistent quality and efficacy of the current cholera vaccines, accurate measurement and characterization of each of these two serotypes is highly important. In this study, we efficiently screened a phage-displayed human synthetic Fab library by bio-panning against Ogawa LPS and finally selected three unique mAbs (D9, E11, and F7) that specifically react with Ogawa LPS. The mAbs bound to Vibrio cholerae vaccine in a dose-dependent fashion. Sequence and structure analyses of antibody paratopes suggest that IgG D9 might have the same fine specificity as that of the murine mAbs, which were shown to bind to the upstream terminal perosamine of Ogawa O-PS, whereas IgGs F7 and E11 showed some different characteristics in the paratopes. To our knowledge, this study is the first to demonstrate the generation of Ogawa-specific mAbs using phage display technology. The mAbs will be useful for identification and quantification of Ogawa LPS in multivalent V. cholerae vaccines.

Recent Progress of Ti3Ci2Tix MXene Electrode Based Self-Healing Application (Ti3Ci2Tix MXene 기반 전극 소재의 자가 치유 적용 기술 개발 동향)

  • Jun Sang Choi;Seung-Boo Jung;Jong-Woong Kim
    • Journal of the Microelectronics and Packaging Society
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    • v.30 no.3
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    • pp.20-34
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    • 2023
  • Single or multi-layered two-dimensional (2D) materials, with thicknesses in the order of a few nanometers, have garnered substantial attention across diverse research domains owing to their distinct properties, including electrical conductivity, flexibility, and optical transparency. These materials are frequently subjected to repetitive mechanical actions in applications like electronic skin (E-Skin) and smart textiles. Moreover, they are often exposed to external factors like temperature, humidity, and pressure, which can lead to a deterioration in component durability and lifespan. Consequently, significant research efforts are directed towards developing self-healing properties in these components. Notably, recent investigations have revealed promising outcomes in the field of self-healing composite materials, with Ti3Ci2Tix MXene being a prominent component among the myriad of available 2D materials. In this paper, we aim to introduce various synthesis methods and characteristics of Ti3Ci2Tix MXene, followed by an exploration of self-healing application technologies based on Ti3Ci2Tix MXene.

Expression of a Functional zipFv Antibody Fragment and Its Fusions with Alkaline Phosphatase in the Cytoplasm of an Escherichia coli

  • Hur, Byung-Ung;Choi, Hyo-Jung;Yoon, Jae-Bong;Cha, Sang-Hoon
    • IMMUNE NETWORK
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    • v.10 no.2
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    • pp.35-45
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    • 2010
  • Background: Expression of recombinant antibodies and their derivatives fused with other functional molecules such as alkaline phosphatase in Escherichia coli is important in the development of molecular diagnostic reagents for biomedical research. Methods: We investigated the possibility of applying a well-known Fos-Jun zipper to dimerize $V_H$ and $V_L$ fragments originated from the Fab clone (SP 112) that recognizes pyruvate dehydrogenase complex-E2 (PDC-E2), and demonstrated that the functional zipFv-112 and its alkaline phosphatase fusion molecules (zipFv-AP) can be produced in the cytoplasm of Origami(DE3) trxB gor mutant E. coli strain. Results: The zipFv-AP fusion molecules exhibited higher antigen-binding signals than the zipFv up to a 10-fold under the same experimental conditions. However, conformation of the zipFv-AP seemed to be influenced by the location of an AP domain at the C-terminus of $V_H$ or $V_L$ domain [zipFv-112(H-AP) or zipFv-112(L-AP)], and inclusion of an AraC DNA binding domain at the C-terminus of VH of the zipFv-112(L-AP), termed zipFv-112(H-AD/L-AP), was also beneficial. Cytoplasmic co-expression of disulfide-binding isomerase C (DsbC) helped proper folding of the zipFv-112(H-AD/L-AP) but not significantly. Conclusion: We believe that our zipFv constructs may serve as an excellent antibody format bi-functional antibody fragments that can be produced stably in the cytoplasm of E. coli.

Production of Pyruvate Dehydrogenase Complex-E2 Specific Human Monoclonal Antibody in Fed-batch Culture Systems with High Cell Density Recombinant Escherichia coli (고농도 재조합 대장균의 Fed-batch 배양 시스템을 이용한 Pyruvate Dehydrogenase Complex-E2 특이성 인간 모노클론 항체의 생산)

  • 이미숙;전주미;차상훈;정연호
    • KSBB Journal
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    • v.15 no.5
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    • pp.489-496
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    • 2000
  • Several culture systems including batch, two-stage CSTR, semi-fed batch, and two-stage cyclic fed-batch were investigated for the efficient production of the Fab fraction of PDC-E2 specific human monoclonal antibody using high cell density recombinant E. coli. A two-phase batch system and a two-stage continuous system were examined to overcome plasmid instability problems, by separating the growth and the production stages. The cell density and productivity of the two-stage continuous culture was better than that of the two-phase batch fermentation. In the two-stage continuous culture system with DO-stat, the cell growth and the productivity were superior to those of the system without the DO control. Also, almost total plasmid stability was maintained in the two-stage continuous culture system. Modified M9 medium was selected as an optimum feeding medium for the fed-batch process, and the optimum C/N ratio determined to be 2:3. The optimum feeding rate was $0.6g/\ell/hr$ for a constant feeding strategy in semi-fed batch system. When the feeding medium was fed by pulsing, it was observed that more frequent pulsing resulted in improved cell growth. The linear feeding method was the most efficient of the various feeding methods tested. Finally, high cell density culture using a two-stage cyclic fed batch system with pH-stat was tried because the linear feeding method showed limitations in terms of obtaining high cell densities, and a cell density of $54 g/\ell$ was achieved. It was concluded that the two-stage cyclic fed batch system was the most efficient system for high cell density culture of the systems tested. However, productivity improvements were lower than expected due to the extremely high accumulations of acetate, although the low levels of residual glucose were maintained.

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Toxicity Monitoring of Endocrine Disrupting Chemicals (EDCs) Using Freeze-dried Recombinant Bioluminescent Bacteria

  • Kim, Sung-Woo;Park, Sue-Hyung;Jiho Min;Gu, Man-Bock
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.6
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    • pp.395-399
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    • 2000
  • Five different freeze-dried recombinant bioluminescent bacteria were used for the detection of cellular stresses caused by endocrine disrupting chemicals. These strains were DPD2794 (recA::luxCDABE), which is sensitive to DNA damage, DPD2540 (fabA::luxCDABE), sensitive to cellular membrane damage, DPD2511 (katG::luxCDABE), sensitive to oxidative damage, and TV1061 (grpE::luxCDABE), sensitive to protein damage. GC2, which emits bioluminescence constitutively, was also used in this study. The toxicity of several chemicals was measured using GC2. Damage caused by known endocrine disrupting chemicals, such as nonyl phenol, bisphenol A, and styrene, was detected and classified according to toxicity mode, while others, such as phathalate and DDT, were not detected with the bacteria. These results suggest that endocrine disrupting chemicals are toxic in bacteria, and do not act via an estrogenic effect, and that toxicity monitoring and classification of some endocrine disrupting chemicals may be possible in the field using these freeze-dried recombinant bioluminescent bacteria.

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