• Korean Chemical Engineering Research
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• v.46 no.6
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• pp.1100-1106
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• 2008

In this study, we presented rapid and simple fabrication method of functionalized surface on various substrates as a universal platform for the selective immobilization of cells. The functionalized surface was achieved by using deposition of polyelectrolyte such as poly(allyamine hydrochloride) (PAH), poly(diallyldimethyl ammonium chloride) (PDAC), poly(4-ammonium styrene sulfonic acid) (PSS), poly(acrylic acid) (PAA) and fabrication of poly(ethylene glycol) (PEG) microstructure through micro-molding in capillaries (MIMIC) technique on each glass, poly(methyl methacrylate) (PMMA), polystyrene (PS) and poly(dimethyl siloxane) (PDMS) substrate. The polyelectrolyte multilayer provides adhesion force via strong electrostatic attraction between cell and surface. On the other hand, PEG microstructures also lead to prevent non-specific binding of cells because of physical and biological barrier. The characteristic of each modified surface was examined by using static contact angle measurement. The modified surface onto several substrates provides appropriate environment for cellular adhesion, which is essential technology for cell patterning with high yield and viability in the micropatterning technology. The proposed method is reproducible, convenient and rapid. In addition, the fabrication process is environmentally friendly process due to the no use of harsh solvent. It can be applied to the fabrication of biological sensor, biomolecules patterning, microelectronics devices, screening system, and study of cell-surface interaction.

• Journal of the Korean Electrochemical Society
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• v.2 no.1
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• pp.31-37
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• 1999

Copper-containig enzyme, laccase (Rhus vernicifera) was immobilized onto gold electrode using self-assembly technique and its surface properties and catalytic activities were examined. Laccase is an oxidoreductase capable to oxidize diphenols or diamines by 4-electron reduction of molecular oxygen without superoxide or peroxide intermediates. The electrode surface were modified by $\beta-mercaptopropionate$ to have a net negative charge in neutral solution and positively charged laccase (pI=9) was immobilized by electrostatic interaction. The successful immobilization was confirmed by cyclic voltammograms which showed typical surface-confined shapes and behaviors. The amount of charge to reduce the surface was similar to the charge calculated assuming the surface being covered by monolayer. The activity of the immobilized enzyme was tested by the capbility of oxidizing a substrate, ABTS (2,2-azine-bis-(3-ethylbenzthioline-6-sulfonic acid) and it was maintained for $2\~3$ days at $4^{\circ}C$. The immobilzed laccase showed about $10\~15\%$ activity compared to that in solution. The laccase-modified electrode showed the activity of elefoocatalytic reduction of oxygen in the presence of mediator, $Fe(CN)_6^{3-}$ The addtion of azide which is an inhibitor of laccase compeletly eliminated the catalytic current.

• Microbiology and Biotechnology Letters
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• v.21 no.4
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• pp.373-380
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• 1993

When the cells of yeast K35 were immobilized in Ca-alginate gel, cell concentration and viability decreased as alginate concentration increased. Considering the results, 2% (w/v) Ca-alginate concentration would be suitable. Among various concentrations of additives and cross-lin-king agent, the addition of 1.67% (w/v) of bentonite together with 0.33% (v/v) of glutaraldehyde (ABG bead) resulted in the highest ethanol production of 1.8%(w/v), using YPD medium containing 2% glucose. ABG bead seemed to be more resistant to phosphate ion than Ca-alginate bead. 0.33%(w/v) of phosphate was a proper concentration for the ethanol production by ABG bead. Scanning electron microscopic observation depicted that the immobilized cells on the bead surface were coated by alginate gel and that the cells in the internal bead were cross-linked with alginate matrix. When repeated-batch culture was performed with ABG bead for 40 days in a packed-bed reactor, ethanol concentration of about 90~110 g/l-gel was maintained. Cell viability was maintained around 70%, and outgrowing cell concentration was below 6.3% of total cell concentration. Consequently, the results showed that ABG head was a potential carrier for continuous production of ethanol compared to conventional Ca-alginate bead.

• Journal of Microbiology and Biotechnology
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• v.24 no.10
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• pp.1382-1388
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• 2014

Proteus vulgaris K80 lipase was expressed in Escherichia coli BL21 (DE3) cells and immobilized on amine-terminated magnetic microparticles (Mag-MPs). The immobilization yield and activity retention were 84.15% and 7.87%, respectively. A homology model of lipase K80 was constructed using P. mirabilis lipase as the template. Many lysine residues were located on the protein surface, remote from active sites. The biochemical characteristics of immobilized lipase K80 were compared with the soluble free form of lipase K80. The optimum temperature of K80-Mag-MPs was $60^{\circ}C$, which was $20^{\circ}C$ higher than that of the soluble form. K80-Mag-MPs also tended to be more stable than the soluble form at elevated temperatures and a broad range of pH. K80-Mag-MP maintained its stable form at up to $40^{\circ}C$ and in a pH range of 5.0-10.0, whereas soluble K80 maintained its activity up to $35^{\circ}C$ and pH 6.0-10.0. K80-Mag-MPs had broader substrate specificity compared with that of soluble K80. K80-Mag-MPs showed about 80% residual relative activity after five recovery trials. These results indicate the potential benefit of K80-Mag-MPs as a biocatalyst in various industries.

• Journal of Microbiology and Biotechnology
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• v.15 no.2
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• pp.234-238
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• 2005

Glucose oxidase was immobilized on the carboxylated multi-wall carbon nanotubes (MWNT-COOHs) in the presence of a coulping reagent, 1-ethy1-3-(3-dimethylaminopropy1) carbodiimide. Significant amounts of glucose oxidase were also immobilized on MWNT-COOHs without the coupling reagent. Various conditions for the immobilization of glucose oxidase were optimized. Optimal pH for the maximal activity of the immobilized glucose oxidase shifted to 7 from the optimal pH of 6 for the maximal activity of free enzyme due to the carboxy1 groups on the surface of MWNT-COOHs. An electrode of graphite rod with a diameter of 6 mm was fabricated using the immobilized glucose oxidase. The cyclic voltammetry study of the enzyme electrode revealed that the oxidation of glucose and subsequent transfer of electrons from the oxidation of glucose to the electrode were possible by the immobilized glucose oxidase without a mediator, implying that the enzyme electrode can be utilized for the development of biofuel cells.

• Genomics & Informatics
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• v.4 no.3
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• pp.133-136
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• 2006

The adsorption of proteins on the surface of glass slides is essential for construction of protein chips. Previously, we prepared a nickel-coated plate by the spin-coating method for immobilization of His-tagged proteins. In order to know whether the structural factor is responsible for the immobilization of His-tagged proteins to the nickel-coated glass slide, we executed a series of experiments. First we purified a His-tagged protein after expressing the vector in E. coli BL21 (DE3). Then we obtained the unfolding curve for the His-tagged protein by using guanidine hydrochloride. Fractions unfolded were monitored by internal fluorescence spectroscopy. The ${\Delta}G_{H20}$ for unfolding was $2.27kcal/mol{/pm}0.52$. Then we tested if unfolded His-tagged proteins can be adsorbed to the nickel-coated plate, comparing with $Ni^{2+}-NTA$ (nitrilotriacetic acid) beads. Whereas unfolded His-tagged proteins were adsorbed to $Ni^{2+}-NTA$ beads, they did not bind to the nickel-coated plate. In conclusion, a structural factor is likely to be an important factor for constructing the protein chips, when His-tagged proteins will immobilize to the nickel-coated slides.

• Bulletin of the Korean Chemical Society
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• v.31 no.4
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• pp.905-909
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• 2010

Magnetic beads (Dynabeads$^{(R)}$) embedded in ~1 micron size polystyrene beads bearing surface carboxylic acid groups were modified with aminobenzyl ethylenediaminetetraacetic acid (ABEDTA) to concentrate or separate metal ions using pH gradients on micro and nano scales. The immobilization of ABEDTA was achieved by amide formation. The presence of the metal chelating functional group in the fully deprotonated form was confirmed by FT-IR. The chelation efficiency of beads was tested by determining metal ions in supernatant using GFAAS when pH gradients from 3 to 7. Mixtures of Cu and Mg and of Cd and Mn (at 10 ng/mL of metal) were separated as the difference in formation constant with the functional group of ABEDTA. The separation was repeated twice with relative standard deviation of <18%. A polydimethylsiloxane (PDMS) microchip column packed with EDTA-coated magnetic beads was optimized to concentrate metal ion for practical applications by eluting a Cu solution of micro scale at pH 3.

• Membrane and Water Treatment
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• v.10 no.3
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• pp.245-250
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• 2019

Prussian blue (PB) is well known for its excellent $Cs^+$ ions adsorption capacity. Due to the high dispersibility of PB in aqueous phase, composite materials imbedding PB in supporting materials have been introduced as a solution. However, building PB particles inside porous supporting materials is still difficult, as PB particles are not fully formed and elute out to water. In this study, we suggest layer-by-layer (LBL) assembly to provide better immobilization of PB on supporting materials of poly vinyl alcohol sponge (PVA) and cellulose filter (CF). Three different PB attachment methods, ex-situ/in-situ/LBL assembly, were evaluated using PB leaching test as well as $Cs^+$ adsorption test. Changes of surface functionality and morphology during PB composite preparation protocols were monitored through Fourier transform infrared spectroscopy and scanning electron microscopy. The results indicate that LBL assembly led to better PB attachment on supporting materials, bringing less eluting PB particles in aqueous phase compared to other synthesis methodologies, such as ex-situ and in-situ synthesis. By enhancing the stability of the adsorbent, adsorption capacity of PVA-PB with LBL improved nine times and that of CF-PB improved over 20 times. Therefore, the results suggest that LBL assembly offers a better orientation for growing PB particles on porous supporting materials.

• Environmental Engineering Research
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• v.24 no.1
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• pp.173-179
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• 2019

Prussian blue (PB) is known to be an effective material for radioactive cesium adsorption, but its nano-range size make it difficult to be applied for contaminated water remediation. In this study, a simple and versatile approach to immobilize PB in the supporting matrix via surface functionalization was investigated. The commercially available poly vinyl alcohol (PVA) sponge was functionalized by acrylic acid (AA) to change its major functional group from hydroxyl to carboxylic, which provides a stronger ionic bond with PB. The amount of AA added was optimized by evaluating the weight change rate and iron(III) ion adsorption test. The FTIR results revealed the surface functional group changing to a carboxyl group. The surface functionalization enhanced the attachment of PB, which minimized the leaching out of PB. The $Cs^+$ adsorption capacity significantly increased due to surface functionalization from 1.762 to 5.675 mg/g. These findings showed the excellent potential of the PB-PAA-PVA sponge as a cesium adsorbent as well as a versatile approach for various supporting materials containing the hydroxyl functional group.

• Korean Journal of Food Science and Technology
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• v.31 no.1
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• pp.1-6
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• 1999

A piezoelectric (PZ) biosensor system detecting Salmonella spp. was developed. The system consisted of an oscillator, a frequency counter and an antibody-immobilized quartz crystal. An anti-Salmonella antibody was immobilized on one gold. surface of the quartz crystal with protein A. Salmonella detection was made by measuring resonant frequency shift owing to a mass change by specific binding of microbial cells to the gold surface of the PZ crystal. The PZ antibody sensor was operated optimally at 0.1M phosphate buffer, pH 7.2 and $35^{\circ}C$. The sensor was quite specific to Salmonella spp. The obtained frequency shift was correlated with the Salmonella concentration in the range of $10^5{\sim}10^6\;CFU/mL$. The frequency shift increased further by addition of polystyrene beads. The Salmonella detection which is indicated by a steady-state microbial adsorption to the quartz crystal was accomplished within 50min.