• Title/Summary/Keyword: sol-gel immobilization

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Improved Immobilized Enzyme Systems Using Spherical Micro Silica Sol-Gel Enzyme Beads

  • Lee, Chang-Won;Yi, Song-Se;Kim, Ju-Han;Lee, Yoon-Sik;Kim, Byung-Gee
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.4
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    • pp.277-281
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    • 2006
  • Spherical micro silica sol-gel immobilized enzyme beads were prepared in an emulsion system using cyclohexanone and Triton-X 114. The beads were used for the in situ immobilization of transaminase, trypsin, and lipase. Immobilization during the sol to gel phase transition was investigated to determine the effect of the emulsifying solvents, surfactants, and mixing process on the formation of spherical micro sol-gel enzyme beads and their catalytic activity. The different combinations of sol-gel precursors affected both activity and the stability of the enzymes, which suggests that each enzyme has a unique preference for the silica gel matrix dependent upon the characteristics of the precursors. The resulting enzyme-entrapped micronsized beads were characterized and utilized for several enzyme reaction cycles. These results indicated improved stability compared to the conventional crushed form silica sol-gel immobilized enzyme systems.

Characterization of carbon dioxide sensitive fluorescence dye immobilized on the sol-gel

  • Sohn, Ok-Jae;Lam, Tuan-Hung;Rhee, Jong-Il
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.478-481
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    • 2005
  • In this study optical sensing membrane was developed for the queantification of dissolved carbon dioxide in micro-bioreactor using an immobilized 8-hydroxypyrene-1,3,6-trisulfonic acid trisodium salt (HPTS). For the immobilization of HPTS sol-gel was synthesied by using 3-glycidoxypropyl-dimethoxymethylsiline and tetraethyl orthosilicate.

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Investigation of Direct and Mediated Electron Transfer of Laccase-Based Biocathode

  • Jamshidinia, Zhila;Mashayekhimazar, Fariba;Ahmadi, Masomeh;Molaeirad, Ahmad;Alijanianzadeh, Mahdi;Janfaza, Sajad
    • Journal of Electrochemical Science and Technology
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    • v.8 no.2
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    • pp.87-95
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    • 2017
  • Enzymatic fuel cells are promising low cost, compact and flexible energy resources. The basis of enzymatic fuel cells is transfer of electron from enzyme to the electrode surface and vice versa. Electron transfer is done either by direct or mediated electron transfer (DET/MET), each one having its own advantages and disadvantages. In this study, the DET and MET of laccase-based biocathodes are compared with each other. The DET of laccase enzyme has been studied using two methods; assemble of needle-like carbon nanotubes (CNTs) on the electrode, and CNTs/Nafion polymer. MET of laccase enzyme also is done by use of ceramic electrode containing, ABTS (2,2'-azino-bis [3-ethylbenzthiazoline-6-sulphonic acid]) /sol-gel. Cyclic voltammetric results of DET showed a pair of well-defined redox peaks at $200{\mu}A$ and $170{\mu}A$ in a solution containing 5and $10{\mu}M$ o-dianisidine as a substrate for needle-like assembled CNTs and CNTs-Nafion composite respectively. In MET method using sol-gel/ABTS, the maximum redox peak was $14{\mu}A$ in the presence of 15 M solution o-dianisidine as substrate. The cyclic voltammetric results showed that laccase immobilization on needle-like assembled CNTs or CNTs-Nafion is more efficient than the sol-gel/ABTS electrode. Therefore, the expressed methods can be used to fabricate biocathode of biofuel cells or laccase based biosensors.

Immobilization of Phospholipase C by sol-gel method

  • Sohn, Ok-Jae;Yim, Yong-Sik;Jung, Jin-Su;Lee, Yong-Jun;Lee, Hye-Na;Rhee, Jong-Il
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.120-122
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    • 2005
  • Phospholipases C (PLCs) from B. cereus 318 and recombinant Pichia pastoris were immobilized on sol-gel coated glass beads. The pH and temperature on immobilized PLC activity were investigated. Operational and storage stability of the immobilized PLCs was measured by spectro- photometric assay. The PLCs immobilized on sol-gel coated glass beads were photographed by scanning electron microscope (SEM).

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Electrogenerated Chemiluminescence Sensor Based on Tris(2,2'-bipyridyl) ruthenium(II) Immobilized in the Composite Film of Multi-walled Carbon Nanotube/Sol-gel Zinc oxide/Nafion

  • Choi, Eun-Jung;Kang, Chang-Hoon;Choi, Han-Nim;Lee, Won-Yong
    • Bulletin of the Korean Chemical Society
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    • v.30 no.10
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    • pp.2387-2392
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    • 2009
  • A composite film of multi-walled carbon nanotube (MWCNT)/sol-gel-derived zinc oxide(ZnO)/Nafion has been utilized as an efficient immobilization matrix for the construction of a highly sensitive and stable tris(2,2'-bipyridyl) ruthenium(II) (Ru(${bpy)_3}^{2+})$ electrogenerated chemiluminescence (ECL) sensor. The electrochemical and ECL behaviors of Ru(${bpy)_3}^{2+})$ ion-exchanged into the composite film were strongly dependent upon the sol-gel preparation condition, the amount of MWCNT incorporated into the ZnO/Nafion composite film, and the buffer solution pH. The synergistic effect of MWCNTs and ZnO in the composite films increased not only the sensitivity but also the long-term stability of the ECL sensor. The present ECL sensor based on the MWCNT/ZnO/Nafion gave a linear response ($R^2$ = 0.999) for tripropylamine concentration from 500 nM to 1.0 mM with a remarkable detection limit (S/N = 3) of 15 nM. The present ECL sensor showed outstanding long-term stability (94% initial signal retained for 5 weeks). Since the present ECL sensor exhibits large response towards NADH, it could be applied as a transduction platform for the ECL biosensor in which the NADH is produced from the dehydrogenase-based enzymatic reaction in the presence of NA$D^+$ cofactor.

Recombinant S-Layer Proteins of Lactobacillus brevis Mediating Antibody Adhesion to Calf Intestine Alleviated Neonatal Diarrhea Syndrome

  • Khang, Yong-Ho;Park, Hee-Young;Jeong, Yoo-Seok;Kim, Jung-Ae;Kim, Young-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.19 no.5
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    • pp.511-519
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    • 2009
  • A chimeric gene encoding enhanced green fluorescent protein (EGFP) and a S-layer protein from Lactobacillus brevis KCTC3102, and/or two copies of the Fe-binding Z-domain, a synthetic analog of the B-domain of protein A, was constructed and expressed in Escherichia coli BL21(DE3). The S-layer fusion proteins produced in a 500-1 fermentor were likely to be stable in the range of pH 5 to 8 and $0^{\circ}C$ to $40^{\circ}C$. Their adhesive property enabled an easy and rapid immobilization of enzymes or antibodies on solid materials such as plastics, glass, sol-gel films, and intestinal epithelial cells. Owing to their affinity towards intestinal cells and immunoglobulin G, the S-layer fusion proteins enabled the adhesion of antibodies to human epithelial cells. In addition, feeding a mixture of the S-layer fusion proteins and antibodies against neonatal calf diarrhea (coronavirus, rotavirus, Escherichia coli, and Salmonella typhimurium) to Hanwoo calves resulted in 100% prevention of neonatal calf diarrhea syndrome (p<0.01), whereas feeding antibodies only resulted in 56% prevention.

Development of Optical Fiber Glucose and Lactate Biosensors for Bioprocess Monitoring (생물공정 모니터링을 위한 광섬유 포도당 및 젖산 센서의 개발)

  • Jung, Chang Hwan;Sohn, Ok-Jae;Rhee, Jong Il
    • KSBB Journal
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    • v.32 no.1
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    • pp.35-45
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    • 2017
  • In this work the optical fiber glucose and lactate biosensors were developed by using fluorescent dye and enzyme immobilized on the end tip of an optical fiber. 3-Glycidyloxypropyl)methyldiethoxysilane (GPTMS), (3-Aminopropyl) trimethoxysilane (APTMS) and Methyltrimethoxysilane (MTMS) were used to immobilize glucose oxidase (GOD), lactate oxidase (LOD) and ruthenium(II) complex (tris(4,7-diphenyl-1,10-phenanthroline) ruthenium(II), $Ru(dpp)_3^{2+}$) as oxygen sensitive fluorescent dye. MTMS sol-gel was an excellent supporting material for the immobilization of $Ru(dpp)_3^{2+}$, GOD, and LOD on the optical fiber. Storage stability of the optical fiber glucose sensor was kept constant over 20 days, while the optical fiber lactate sensor had constant storage stability over 17 days. The optical fiber glucose and lactate biosensors also maintained good operational stability for 20 hours and 14 hours, respectively. The activities of the immobilized enzymes were most excellent at pH 7 and at $25^{\circ}C$. On-line monitoring of glucose and lactate in a simulated process was performed with the optical fiber glucose and lactate biosensors. On-line monitoring results were agreed with those of off-line data measured with high performance liquid chromatography (HPLC).

Surface Modification of Nano Porous Silica Particle for Enzyme Immobilization (효소 고정화를 위안 실리카 나노세공 입자의 표면개질)

  • Cho, Hyung-Min;Kim, Jong-Kil;Kim, Ho-Kun;Lee, Eun-Kyu
    • KSBB Journal
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    • v.21 no.5
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    • pp.360-365
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    • 2006
  • The objectives of this study were to develop nano-pore silica particles and to modify its surface for use as an enzyme immobilization matrix. Sol-gel reaction was used to produce silica particles of various nano pore sizes with hydroxyl groups on their surfaces. The surface was modified with aldehyde that was confirmed by fluorescence imaging. Trypsin was covalently immobilized by reductive amination. Surface density of the immobilized trypsin was ca. $350{\mu}g/m^2$, which was approximately 17- and 35-fold higher than those from the surfaces with hydroxyl and amine group, respectively. About 90% of the initial enzyme activity was maintained after the 12th use of repeated use. When compared with the commercial matrices, the nano-pore silica particle was superior in terms of immobilization yield and specific activity. This study suggests the nano porous silica particles can be used as enzyme immobilization matrix for industrial applications.