• Title/Summary/Keyword: Covalently

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Multianalyte Sensor Array using Capillary-Based Sample Introduction Fluidic Structure: Toward the Development of an "Electronic Tongue"

  • Sohn, Young-Soo;Anslyn, Eric V.;McDevitt, John T.;Shera, Jason B.;Neikirk, Dean P.
    • Journal of Sensor Science and Technology
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    • v.13 no.5
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    • pp.378-382
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    • 2004
  • A micromachined fluidic structure for the introduction of liquid samples into a chip-based sensor array composed of individually addressable polymeric microbeads has been developed. The structure consists of a separately attached cover glass, a single silicon chip having micromachined channels and microbead storage cavities, and a glass carver. In our sensor array, transduction occurs via colorimetric and fluorescence changes to receptors and indicator molecules that are covalently attached to termination sites on the polymeric microbeads. Data streams are acquired for each of the individual microbeads using a CCD. One of the key parts of the structure is a passive fluid introduction system driven only by capillary force. The velocity of penetration of a horizontal capillary for the device having a rectangular cross section has been derived, and it is quite similar to the Washburn Equation calculated for a pipe with a circular cross section having uniform radius. The test results show that this system is useful in a ${\mu}$-TAS and biomedical applications.

Subunit Organization of Bacterial Malonate Decarboxylases: The Smallest ${\delta}$ Subunit as an Acyl-Carrier Protein

  • Byun, Hye-Sin;Kim, Yu-Sam
    • BMB Reports
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    • v.30 no.2
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    • pp.132-137
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    • 1997
  • In order to compare molecular structure, malonate decarboxylases from Acinetobacter calcoaceticus, Pseudomonas fluorescens, and Pseudomonas putida aerobically grown on malonate, were purified by the method employing streptomycin sulfate treatment, chromatography with PBE 94 and ${\omega}-aminohexyl$ agarose. Molecular masses were estimated to be 185, 200, and 200 kDa, respectively. All malonate decarboxylases were multimeric enzymes consisting of four different subunits, $2{\alpha},\;1{\beta},\;1{\gamma},\;and\;1{\delta}$. The molecular masses of the Pseudomonas enzyme subunits were $65({\alpha})$, $33({\beta})$, $30({\gamma})$, and $11kDa({\delta})$; which are very similar to those, $65({\alpha})$, $32({\beta})$, $25({\gamma})$, and $11kDa({\delta})$ of Acinetobacter enzyme. The ${\delta}-subunit$ of the active form of the enzymes was acetylated. The acetyl group may form a thioester bond with the thiol group of the prosthetic group covalently linked to the enzyme. It suggests that such molecular organization is common in all malonate decarboxylases.

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Photo-grafting Dyeing of Wool Fabrics with ${\alpha}$-bromoacrylamide reactive dye (반응성 염료를 이용한 양모직물의 광그라프트 염색)

  • Dong, Yuanyuan;Jang, Jin-Ho
    • Proceedings of the Korean Society of Dyers and Finishers Conference
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    • 2011.03a
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    • pp.31-31
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    • 2011
  • Lanasol dyes containing ${\alpha}$-bromoacrylamide or ${\alpha},{\beta}$-dibromopropionylamide group are used for wool dyeing. They are normally applied to wool under pH 4.5 to 6.5 at $100^{\circ}C$. Although wool fabric can be dyed to obtain deep colour, high light and wet fastness, the dyeing processes need long dyeing time at high temperature, with salt addition, which inevitably causes environmental problems. Grafting is a modification method for textile where monomers are covalently bonded onto the polymer chain. It can be initiated by ozone, ${\gamma}$ rays, electron beams, plasma, corona discharge and UV irradiation. Coloration by UV-induced photografting exhibits several advantages such as fast reaction rate, energy saving, simple equipment, easy exploitation and environmentally friendliness. Also it requires much lower energy compared to the conventional dyeing and less damage to the substrate. In this study, a direct sequential UV-induced photografting onto wool fabrics was discussed. To understand the graft polymerization mechanism further, several characterization methods were used. Moreover, the effects of several principal factors on the graft photopolymerization were investigated. Furthermore, the colorfastness results were compared with conventional dyeing methods.

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Prediction of Ultra-High ON/OFF Ratio Nanoelectromechanical Switching from Covalently Bound $C_{60}$ Chains

  • Kim, Han Seul;Kim, Yong-Hoon
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.02a
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    • pp.645-645
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    • 2013
  • Applying a first-principles computational approach combining density-functional theory and matrix Green's function calculations, we have studied the effects [2+2] cycloaddition olligormerization of fullerene $C_{60}$ chains on their junction charge transport properties. Analyzing first the microscopic mechanism of the switching realized in recent scanning tunneling microscope (STM) experiments, we found that, in agreement with experimental conclusions, the device characteristics are not significantly affected by the changes in electronic structure of $C_{60}$ chains. It is further predicted that the switching characteristics will sensitively depend on the STM tip metal species and the associated energy level bending direction in the $C_{60}-STM$ tip vacuum gap. Considering infinite $C_{60}$ chains, however, we confirm that unbound $C_{60}$ chains with strong orbital hybridizations and band formation should in principle induce a much higher conductance state. We demonstrate that a nanoelectromechanical approach in which the $C_{60}-STM$ tip distance is maintained at short distances can achieve a metal-independent and drastically improved switching performance based on the intrinsically better electronic connectivity in the bound $C_{60}$ chains.

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Screening of Peptides Bound to Anthrax Protective Antigen by Phage Display

  • Kim, Joung-Mok;Park, Hye-Yeon;Choi, Kyoung-Jae;Jung, Hoe-Il;Han, Sung-Hwan;Lee, Jae-Seong;Park, Joon-Shik;Yoon, Moon-Young
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1784-1790
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    • 2006
  • Bacillus anthracis is a causative agent of anthrax. Anthrax toxins are composed of a protective antigen (PA), lethal factor (LF), and edema factor (EF), in which the PA is a central mediator for the delivery of the two enzymatic moieties LF and EF. Therefore, the PA has been an attractive target in the prevention and vaccinization for anthrax toxin. Recently, it has been reported that the molecule consisting of multiple copies of PA-binding peptide, covalently linked to a flexible polymer backbone, blocked intoxification of anthrax toxin in an animal model. In the present study, we have screened novel diverse peptides that bind to PA with a high affinity (picomolar range) from an M13 peptide display library and characterized the binding regions of the peptides. Our works provide a basis to develop novel potent inhibitors or diagnostic probes with a diverse polyvalence.

Inhibitory Mechanism of Novel Inhibitors of UDP-N-Acetylglucosamine Enolpyruvyl Transferase from Haemophilus influenzae

  • Jin, Bong-Suk;Han, Seong-Gu;Lee, Won-Kyu;Ryoo, Sung-Weon;Lee, Sang-Jae;Suh, Se-Won;Yu, Yeon-Gyu
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1582-1589
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    • 2009
  • Bacterial UDP-N-acetylglucosamine enolpyruvyl transferase (MurA) catalyzes the transfer of enolpyruvate from phosphoenolpyruvate (PEP) to uridine diphospho-N-acetylglucosamine (UNAG), which is the first step of bacterial cell wall synthesis. We identified thimerosal, thiram, and ebselen as effective inhibitors of Haemophilus influenzae MurA by screening a chemical library that consisted of a wide range of bioactive compounds. When MurA was preincubated with these inhibitors, their 50% inhibitory concentrations ($IC_{50}s$) were found to range from 0.1 to $0.7\;{\mu}M$. In particular, thimerosal suppressed the growth of several different Gram-negative bacteria such as Escherichia coli, Pseudomonas aeruginosa, and Salmonella typhimurium at a concentration range of $1-2\;{\mu}g/ml$. These inhibitors covalently modified the cysteine residue near the active site of MurA. This modification changed the open conformation of MurA to a more closed configuration, which may have prevented the necessary conformational change from occurring during the enzyme reaction.

Immunochemical Studies on Expression of Quinoproteins in Escherichia coli

  • Ryou, Chong-Suk;Kim, Jae-Beom;Kwon, Moo-Sik
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.95-98
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    • 2000
  • An immunochemical method has been develooped as the most sensitive tool for studying the expression of quinoproteins containing pyrroloquinoline qinone(PQQ) in E. coli. The PQQ was conjugated to bovine serum albumin (BSA), and the conjugant was purified by using a $KwikSep^{TM}$ dextran desalting column chromatography. The PQQ-BSA conjugant was immunized to rabbits, and the IgG fractions of the antisera were purified. The most sensitive antibody against PQQ-BSA conjugant recognized some nanogram quantity of the antigen on the blot, but had little cross reactivity with BSA. Using this batch of the antibody, all the immunochemical assays of quinoproteins in E. coli were preformed. Some six different PQQ-specfic spots were detected by Western blot analysis of the soluble proteins in E. coli were performed. Some six different PQQ-specific spots were detected by Western blot analysis of the soluble proteins in E. coli after two-dimensional gel electrophoresis. Their molecular weights on the blot were estimated to be about 100-, 90-, 72-, 58-, 52-, and 50kDa. Their pI values fell in the range from 4.8 to 5.5. These results stronly suggest that quinoproteins are present in E. coli, and that the protein moieties were covalently bound to PQQ.

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Chemically Modified Sepharose as Support for the Immobilization of Cholesterol Oxidase

  • Yang, Hailin;Chen, Yi;Xin, Yu;Zhang, Ling;Zhang, Yuran;Wang, Wu
    • Journal of Microbiology and Biotechnology
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    • v.23 no.9
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    • pp.1212-1220
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    • 2013
  • Because the cholesterol oxidase from Brevibacterium sp. M201008 was not as stable as the free enzyme form, it had been covalently immobilized onto chemically modified Sepharose particles via N-ethyl-N'-3-dimethylaminopropyl carbodiimide. The optimum immobilization conditions were determined, and the immobilized enzyme activity obtained was 12.01 U/g Sepharose-ethylenediamine. The immobilization of the enzyme was characterized by Fourier transform infrared spectroscopy. The immobilized enzyme exhibited the maximal activity at $35^{\circ}C$ and pH 7.5, which was unchanged compared with the free form. After being repeatedly used 20 times, the immobilized enzyme retained more than 40.43% of its original activity. The immobilized enzyme showed better operational stability, including wider thermal and pH ranges, and retained 62.87% activity after 20 days of storage at $4^{\circ}C$, which was longer than the free enzyme.

Characterization of Antibacterial activity and Synthesis of Sulfanilamide Polymer using Crosslinking Agent (가교제를 이용한 Sulfanilamide 중합체의 합성과 항균특성)

  • Kim, Jong-Woan;Yoon, Chul-Hun;Hwang, Sung-Kwy;Kong, Seung-Dae;Lee, Han-Seab
    • Journal of the Korean Applied Science and Technology
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    • v.17 no.1
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    • pp.37-42
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    • 2000
  • Drug delivery system(DDS) have been actively studied for the past twenty years. Dual action agents are unique chemical entities comprised of two different types of antibacterial compounds covalently linked together in a single molecule in such a way that both components are able to exert their bactericidal properties. In spite of the advent of the antibacterial agent the sulfa agents are the most widely used antibacterial agent today. In this study, new antibacterials derivative was synthesized using glutaraldehyde such as crosslinking agent for the purpose of dual-action as DDS study. Antibacterial activity of these new synthetic derivative between their structures and activities were examined by disc diffusion method. As a result, new synthetic derivative exhibited the broad antibacterial activities against Gram(+) and Gram(-) bacilli. Especially, the antibacterial effect of new synthetic derivative against Gram negative(Esherichia. coli) was much stronger than that against Gram positive.

The Surface Characteristics of Biomaterials Fixed with a Hydrophilic Membrane by Photochemical Reaction (광화학적 방법으로 친수성막이 고정화된 생체 재료의 표면특성 분석)

  • Kim, H.J.;Jeon, S.M.;Lee, K.B.;Min, B.G.
    • Proceedings of the KOSOMBE Conference
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    • v.1997 no.05
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    • pp.11-15
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    • 1997
  • We used the photoreactive poly(allylamie) (PPA) as the hydrophilic membrane to control the release of drug from polyurethane(PU). PPA was covalently bonded onto PU surface through the highly reactive nitrene intermediate upon UV light irradiation $(3.3mW/cm^2)$ at 254nm for 5min. Thus the release rate of rifampicin from PU surface was controlled. To know the characteristics of PU surface bonded with PPA, we measured the ATR-FTIR, ESCA, Static Contact Angle and SEM. From these, we suggest that PPA as a hydrophilic membrane is enable to control the release rate of a hydrophobic drug from polymer without the change of bulk property.

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