• Title/Summary/Keyword: chemical immobilization

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Impact of Solvent pH on Direct Immobilization of Lysosome-Related Cell Organelle Extracts on TiO2 for Melanin Treatment

  • Bang, Seung Hyuck;Kim, Pil;Oh, Suk-Jung;Kim, Yang-Hoon;Min, Jiho
    • Journal of Microbiology and Biotechnology
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    • v.25 no.5
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    • pp.718-722
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    • 2015
  • Techniques for immobilizing effective enzymes on nanoparticles for stabilization of the activity of free enzymes have been developing as a pharmaceutical field. In this study, we examined the effect of three different pH conditions of phosphate buffer, as a dissolving solvent for lysosomal enzymes, on the direct immobilization of lysosomal enzymes extracted from Hen's egg white and Saccharomyces cerevisiae. Titanium(IV) oxide (TiO2) nanoparticles, which are extensively used in many research fields, were used in this study. The lysosomal enzymes immobilized on TiO2 under each pH condition were evaluated to maintain the specific activity of lysosomal enzymes, so that we can determine the degree of melanin treatment in lysosomal enzymes immobilized on TiO2. We found that the immobilization efficiency and melanin treatment activity in both lysosomal enzymes extracted from Hen's egg white and S. cerevisiae were the highest in an acidic condition of phosphate buffer (pH 4). However, the immobilization efficiency and melanin treatment activity were inversely proportional to the increase in pH under alkaline conditions. In addition, enhanced immobilization efficiency was shown in TiO2 pretreated with a divalent, positively charged ion, Ca2+, and the melanin treatment activity of immobilized lysosomal enzymes on TiO2 pretreated with Ca2+ was also increased. Therefore, this result suggests that the immobilization efficiency and melanin treatment activity of lysosomal enzymes can be enhanced according to the pH conditions of the dissolving solvent.

A Simple Method for Measuring the Immobilization Solids of Coating Colors Using an AA-CWR Water Retention Meter

  • Park, Chang-hak;Lee, Do-Ik;Margaret K. Joyce
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.34 no.5
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    • pp.39-48
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    • 2002
  • The water retention of coating colors can be accurately measured by devices such as an AA-GWR water retention meter whose principle of measurement Is based on pressure filtration of coatings under an externally applied air pressure over a certain period of time. It was hypothesized that such devices could be also used to determine the immobilization solids (IMS) of coating colors by determining a sudden drop in the rate of dewatering, that is, a sudden change in the drainage curves. To test this hypothesis, the immobilization solids of coating colors containing various thickeners and water retention additives at different levels were first accurately measured by a modified immobilization tester based on the well-known gloss drop method, and then their values were compared with those obtained by an AA-GWR water retention tester. They agreed very well and showed that the standard deviation is only 0.14% in the IMS points between both methods. This good agreement was not surprising because both test methods are based on the same end-point, that is, the immobilization solids point at which menisci begin to form at the coating surface. Theoretical considerations supporting this new method for measuring the immobilization solids of coating colors are presented and some recommendations for the test method are discussed. Also, the effect of various thickeners and water retention additives on the properties and printability of coated papers is discussed.

Biosorption of Heavy Metal Sons by Biomass of Marine Brown Algae in Cheju using Their immobilization Techniques: Biosorption of Copper by Undaria pinnatifida

  • Sang-Kyu Kam;Min-Gyu Lee
    • Journal of Environmental Science International
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    • v.1 no.2
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    • pp.157-166
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    • 1992
  • The biosorptlon perFormances of copper were Investigated by the immobilized biomass of nonliving marine brown algae Undaria pinnatifida by each of the Ca-alginate method(Ca-ALG), Ba-alginate method(Ba-ALG), polyethylene glycol method(PEG), and carrageenan method (CARR). The copper removal performance increased but the copper uptake decreased as the biomass amount was increased. However, the copper uptake by the immobilized biomass increased with increasing initial copper concentration. Among the immobilization methods, the copper uptake decreased in the following sequence: Ca-ALG > Ba-ALG > PEG > CARR. The pattern of copper uptake by the immobilized biomass fitted the Langmuir isotherm better than the Freundlich isotherm. Desorption of deposited copper with 0.05 ~0.5M HCI, resulted in no changes of the copper uptake capacity of the immobilized biomass by the immobilization methods except for PEG, through five subsequent biosorptioydesorption cycles. There was no damage to the immobilized biomass which retained its macroscopic appearance in repeated copper uptake/elution cycles.

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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.

An Immobilization of Extracellular Laccase to Humus-Iron Complex

  • Ginalska, Grazyna;Cho, Nam-Seok;Lobarzewski, Jerzy;Piccolo, Alessandro;Leonowicz, Andrzej
    • Journal of the Korean Wood Science and Technology
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    • v.29 no.3
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    • pp.104-111
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    • 2001
  • There are some evidence that active enzymatic proteins, e.g. fungal laccase, exist in the naturally occured soil humus. This study was performed to investigate the covalent binding of fungal laccase to the humic acid-iron complex, and to measure laccase activity of immobilized ones. Seven methods were adopted to form the covalent binding of fungal laccase with soil humic acids complexed with iron. Using these seven methods it was possible to change the dimension of spacer arm between laccase and support, and also to regulate the mode of covalent binding of this enzyme. The spacer arm was regulated from 2C to 11C. There was not observed any straight relationship between the spacer arm longitude and the laccase activity after immobilization, but the binding mode more effective than the former. Three out of the seven methods gave the high activity of immobilized laccase, and which active products of laccase immobilization was stable up to 10 days after the process. It is indicated that natural soil condition might be prevented the laccase activation by the toxic influence of some phenolic humic compounds. It was shown, for the first time, the possibilities to obtain the high activity of fungal laccase by binding to humic acids, and especially in complex with iron.

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Optimization of Catechol Production Using Immobilized Resting Cells of Pseudomonas putida in Aqueous/organic Two-phase System

  • Chae, Hee-Jeong;Yoo, Young-Je
    • Journal of Microbiology and Biotechnology
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    • v.7 no.5
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    • pp.345-351
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    • 1997
  • An aqueous/organic two-phase reaction system was applied to the production of catechol using immobilized resting cells of Pseudomonas putida CY 400. Water/ethyl ether system was used because of high partition coefficient of catechol and thus to reduce the product inhibition and degradation. Among the tested immobilization carriers, polyacrylamide gel gave the highest catechol productivity. The immobilization seemed to protect the cells against solvent toxicity. From the simulation of reaction conditions based on two-phase models, it was found that there was an optimum acetate concentration at fixed benzoate and cell concentrations for the catechol productivity. A lower phase volume ratio (lower fraction of organic phase) gave a higher productivity. However, the substrate conversion was low at low phase volume ratio.

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Hybridization by an Electrical Force and Electrochemical Genome Detection Using an Indicator-free DNA on a Microelectrode-array DNA Chip

  • Choi, Yong-Sung;Lee, Kyung-Sup;Park, Dae-Hee
    • Bulletin of the Korean Chemical Society
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    • v.26 no.3
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    • pp.379-383
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    • 2005
  • This research aims to develop DNA chip array without an indicator. We fabricated microelectrode array by photolithography technology. Several DNA probes were immobilized on an electrode. Then, indicator-free target DNA was hybridized by an electrical force and measured electrochemically. Cyclic-voltammograms (CVs) showed a difference between DNA probe and mismatched DNA in an anodic peak. Immobilization of probe DNA and hybridization of target DNA could be confirmed by fluorescent. This indicator-free DNA chip microarray resulted in the sequence-specific detection of the target DNA quantitatively ranging from $10^{-18}\;M\;to\;10^{-5}$ M in the buffer solution. This indicator-free DNA chip resulted in a sequence-specific detection of the target DNA.

Immunosensor for Detection of Escherichia coli O157:H7 Using Imaging Ellipsometry

  • Bae Young-Min;Park Kwang-Won;Oh Byung-Keun;Choi Jeong-Woo
    • Journal of Microbiology and Biotechnology
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    • v.16 no.8
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    • pp.1169-1173
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    • 2006
  • Imaging ellipsometry (IE) for detection of binding of Escherichia coli O157:H7 (E. coli O157:H7) to an immunosensor is reported. A protein G layer, chemically bound to a self-assembled layer of 11-mercaptoundecanoic acid (11-MUA), was adopted for immobilization of monoclonal antibody against E. coli O157:H7 (Mab). The immobilization of antibody was investigated using surface plasmon resonance. To fabricate antibody spots on a gold surface, protein G solution was spotted onto the gold surface modified with an 11-MUA layer, followed by immobilizing Mab on the protein G spot. Ellipsometric images of the protein G spot, the Mab spot, and Mab spots with binding of E. coli O157:H7 in various concentrations were acquired using the IE system. The change of mean optical intensity of the Mab spots in the ellipsometric images indicated that the lowest detection limit was $10^3$CFU/ml for E. coli O157:H7. Thus, IE can be applied to an immunosensor for detection of E. coli O157:H7 as a detection method with the advantages of allowing label-free detection, high sensitivity, and operational simplicity.