• Title/Summary/Keyword: covalent cross-linking

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Involvement of Electrostatic Interactions between the Components of Toluene Dioxygenase from Pseudomonas putida Fl

  • Lee, Kyoung
    • Journal of Microbiology and Biotechnology
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    • v.8 no.4
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    • pp.416-421
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    • 1998
  • A carboxyl group modifier, l-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) was used to study the interactions between three components of toluene dioxygenase (TDO) from Pseudomonas putida FI. $Ferredoxin_{TOL}$ activity was increased by the treatment with EDC; however, the activity was rapidly declined in the prolonged incubation. In covalent cross-linking experiments with EDC, $Ferredoxin_{TOL}$ made a one-to-one complex with $Reductase_{TOL}$ or the large subunit of $ISP_{TOL}$. These results provide evidence for the involvement of electrostatic interactions in the TDO electron transfer system.

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Free-Standing Langmuir-Blodgett Films of Maleic Acid-Vinyl Ether Copolymers across 1 μm Pores

  • 이범종;최기선;권영수
    • Bulletin of the Korean Chemical Society
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    • v.16 no.12
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    • pp.1167-1172
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    • 1995
  • A coverage of about 1 μm-sized pores of a membrane filter by four monolayers of maleic acids copolymers and poly(allylamine) (PAA) was attained by Langmuir-Blodgett (LB) technique through a covalent cross-linking followed a polyion complexation at the air-water interface. The copolymers were prepared to have side chains of hydrocarbon tail, carboxyl, and/or oligoether in the repeat unit. The surface pressure-area isotherms showed that the monolayers on an aqueous PAA have more expanded area than on pure water. The monolayers were transferable on a calcium fluoride substrate and a fluorocarbon membrane filter as Y deposition type, and the resulting LB films were characterized by FT-IR spectroscopy and scanning electron microscopy. A polymer network produced through interchain amide formation was confirmed in as-deposited films. The films were heat-treated in order to complete the cross-linking. SEM observation of the heat-treated film on a porous membrane filter showed that the four layer film was sufficiently stable to cover the filter pore of about 1 μm. Immersion of the film in water or in chloroform did not cause any change in its appearance on SEM and in FT-IR spectra.

Evaluation of Immobilization Methods for Cyclodextrin Glucanotransferase and Characterization of its Enzymatic Properties

  • Lee, Sang-Ho;Shin, Hyun-Dong;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.1 no.1
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    • pp.54-62
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    • 1991
  • Cyclodextrin glucanotransferase(CGTase) derived from Bacillus macerans was immobilized by (1) covalent linkage on chitosan and chitin with glutaraldehyde, (2) adsorption on DEAE-cellulose and Amberite IRA 900 after succinylation, and (3) entrapment on alginate and polyacrylamide by cross linking. Adsorption on Amberite IRA 900 and covalent linking on chitosan were identified to be the most suitable immobilization methods considering the yield of activity and stability of immobilized CGTase. The enzymatic properties of immobilized CGTase were investigated and compared with those of the soluble CGTase. Thermal stability of CGTase immobilized on chitosan was increased from 50 to $55^{\circ}C$, and the optimum temperature of CGTase immobilized on Amberite IRA 900 was shifted from 55 to $50^{\circ}C$. The effect of molecular size of soluble starch (substrate) on immobilized CGTase investigated using partially liquefied substrates with different dextrose equivalent(DE). Cyclodextrin(CD) conversion yield augmented according to the increase of DE level for immobilized CGTase on Amberite IRA 900. CD conversion yield of partially cyclized starch with soluble CGTase was higher compared with liquefied one with ${\alpha}-amylase$.

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The Relation Among the Linear Energy Transfer and Changes of Polyphenylene Sulfide Surface by ion Implantation (이온주입에 의한 고분자(Polyphenylene Sulfide)표면 특성 변화와 선에너지전달(Pineal Energy Transfer)과의 관계)

  • Lee, Jae S.;Kim, Bo-Young;Lee, Jae-Hyung
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.18 no.5
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    • pp.407-413
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    • 2005
  • Ion implantation provides a unique way to modify the mechanical, optical and electrical properties of polymer by depositing the energy of ions in the material on the atomic scale. Implantation of ions into the polymers generally leads to a radiation damage, which, in many cases, modifies the properties of the surface and bulk of the material. These modifications result from the changes of the chemical structure caused in their turn by changing the chemical bonding when the incident ions cut the polymer chains, breaks covalent bonds, promotes cross-linking, and liberates certain volatile species. We studied the relation among the linear energy transfer (LET) and changes of surface microstructure and surface resistivity on PPS material using the high current ion implantation technology The surface resistivity of nitrogen implanted PPS decreased to $10^{7}{\Omega}/cm^{2}$ due to the chain scission, cross linking, ${\pi}$ electron creation and mobility increase. In this case, the surface conductivity depend on the 1-dimensional hopping mechanism.

Electrochemical and Mechanical Characteristics of Covalently Cross-Linked SPEEK Polymer Electrolyte Membrane for Water Electrolysis (수전해용 공유가교 SPEEK 고분자 전해질 막의 전기 화학적 및 기계적 특성)

  • Kim, Kyung-Eon;Jang, In-Young;Kweon, Oh-Hwan;Hwang, Yong-Koo;Moon, Sang-Bong;Kang, An-Soo
    • Transactions of the Korean hydrogen and new energy society
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    • v.18 no.4
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    • pp.391-398
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    • 2007
  • The covalently cross-linked sulfonated polyetheretherketone (CL-SPEEK) membrane was prepared by four-step synthesis of sulfonation-sulfochlorination, partial reduction, lithiation, and cross-linking, and its electrochemical and mechanical properties were investigated for water electrolysis application. The prepared ion exchange membranes showed good electrochemical and mechanical properties; proton conductivity of 0.116 S/cm at $80^{\circ}C$, water uptake of 44.6%, ion exchange capacity of 1.75 meq/g-dry-memb., tensile strength of 64.25 MPa and elongation of 61.11%. The membrane electrode assembly (MEA) with homemade membranes were prepared by non-equilibrium impregnation-reduction (I-R) method. Especially, the electrochemical surface area (ESA) and roughness factor of CL-SPEEK electrolyte by cyclic voltammetry method were 23.46 $m^2/g$ and 307.3 $cm^2-Pt/cm^2$, respectively. The prepared MEA was used in the unit cell of water electrolysis and the cell voltage was 1.81 V at 1 A/$cm^2$ and $80^{\circ}C$, with platinum loadings of 1.31 mg/$cm^2$.

Permeation Property of Ionomer Film with New Multifunctional Ionic Site (다관능기를 도입한 아이오노머 필름의 기체투과 특성)

  • Lee, Bo-Mi;Jeong, Sam-Bong;Nam, Sang-Yong
    • Korean Journal of Materials Research
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    • v.22 no.5
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    • pp.227-236
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    • 2012
  • Ionomer is a thermoplastic that is composed of covalent bonds and ionic bonds. It is possible to use this material in processes such as injection molding or extrusion molding due to the material's high oil resistance, weatherproof characteristics, and shock resistance. In this study, a new ionomer having a multifunctional group was prepared by a stepwise neutralization system with the addition of acidic and salt additives. In step I, to increase the contents of the multifunctional group and the acid degree in ethylene acrylic acid (EAA), MGA was added to the ionomer resin (EAA). A new ionomer was prepared via the traditional preparation method of the ionic cross-linking process. In step II, metal salt was added to the mixture of EAA and MGA. The extrusion process was performed using a twin extruder (L/D = 40, size : ${\varphi}30$). Ionomer film was prepared for evaluation of gas permeability by using the compression molding process. The degree of neutralized and ionic cross-linked new ionomer was confirmed by FT-IR and XRD analysis. In order to estimate the neutralization of the new ionomer film, various properties such as gas permeation and mechanical properties were measured. The physical strength and anti-scratch property of the new ionomer were improved with increase of the neutralization degree. The gas barrier property of the new ionomer was improved through the introduction of an ionic site. Also, the ionic degree of cross-linking and gas barrier property of the ionomer membrane prepared by stepwise neutralization were increased.

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

  • 정태훈;홍석인;노봉수;정용섭;윤정원;김태진
    • KSBB Journal
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    • v.13 no.1
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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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Immobilization of $\beta$-glucosidase and properties of Immobilized Enzyme ($\beta$-glucosidase의 고정화와 효소 반응특성)

  • 정의준;이상호이용현
    • KSBB Journal
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    • v.5 no.2
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    • pp.141-149
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    • 1990
  • $\beta$-glucosidase derived from Aspergillus niger was immobilized by (1) covalent linkage on chitin and chitosan with glutaraldehyde, (2) adsorption on DEAE-cellulose and Amberite IRA93 after succinylation, and (3) entrapment on alginate and polyacrylamide gels with various cross linking agents. The retention yield of $\beta$-glucosidase immobilized on chitosan was 31.5% and operational stability was 69% after continuous operation at column reactor(5$0^{\circ}C$ at pH 4.8) for 15 days. The retention yield and operational stability were 24.7% and 60% respectively, in adsorption on Amberite IRA 93. On the other hand, the entrapment method by alginate and polyacrylamide gel was identified to be not appropriate due to the continuous elution of inlmobilized $\beta$-glucosidase. Optimum conditions for the immobilization on chitosan were also studied with optimum pH of 4.8 and glutaraldehyde concentration of 0.4%(w/v). The properties and stability of immobilized $\beta$-glucosidase are also investigted. The conversion yield of cellobiose to glucose was also analyzed using the column type enzyme reactor to evaluate the effectiveness of immobilized enzyme.

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Advanced Glycation End Products and Diabetic Complications

  • Singh, Varun Parkash;Bali, Anjana;Singh, Nirmal;Jaggi, Amteshwar Singh
    • The Korean Journal of Physiology and Pharmacology
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    • v.18 no.1
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    • pp.1-14
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    • 2014
  • During long standing hyperglycaemic state in diabetes mellitus, glucose forms covalent adducts with the plasma proteins through a non-enzymatic process known as glycation. Protein glycation and formation of advanced glycation end products (AGEs) play an important role in the pathogenesis of diabetic complications like retinopathy, nephropathy, neuropathy, cardiomyopathy along with some other diseases such as rheumatoid arthritis, osteoporosis and aging. Glycation of proteins interferes with their normal functions by disrupting molecular conformation, altering enzymatic activity, and interfering with receptor functioning. AGEs form intra- and extracellular cross linking not only with proteins, but with some other endogenous key molecules including lipids and nucleic acids to contribute in the development of diabetic complications. Recent studies suggest that AGEs interact with plasma membrane localized receptors for AGEs (RAGE) to alter intracellular signaling, gene expression, release of pro-inflammatory molecules and free radicals. The present review discusses the glycation of plasma proteins such as albumin, fibrinogen, globulins and collagen to form different types of AGEs. Furthermore, the role of AGEs in the pathogenesis of diabetic complications including retinopathy, cataract, neuropathy, nephropathy and cardiomyopathy is also discussed.

The Covalent Immobilization of Biomolecules to Polymer Surface by Deep-UV Lithography Using N-Hydroxysuccinimidyl Azidobenzoate

  • Kim, H.J.;Shim, J.H.;Kim, J.H.;Kim, J.;Min, B.G.
    • Proceedings of the KOSOMBE Conference
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    • v.1997 no.05
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    • pp.47-50
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    • 1997
  • We synthesized N-Hydroxysuccinimidyl (NHS) azidobenzoate as a cross-linking reagent for immobilization of peptide onto the solid surface. Thin polystyrene(PS) films spin-coated with a NHS azidobenzoate solution were exposed with ultraviolet light at 245nm$(3.3mW/cm^2)$ for 5 min. The NHS active ester groups became covalently attached to the polymer via photogenerated, highly reactive nitrene intermediates derived from NHS azidobenzoate. Using this technique, it is demonstrated that well-defined surface regions can be functionalized with a minimum observable feature size of 1mm for UV exposure. Through reaction of this functionalized PS surface with primary amine-containing biomolecules, biological molecule had been immobilized on the polymer surface.

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