• Bulletin of the Korean Chemical Society
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• v.22 no.11
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• pp.1192-1196
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• 2001

A diaphorase enzyme electrode for the catalytic reduction of NAD+ , the oxidized form of nicotinamide adenine dinucleotide, has been prepared. The enzyme layer grew spontaneously over an aminoethanethiol self assembled monolayer on a go ld plate electrode. The growth was accomplished by simply dipping the electrode covered by the aminoethanethiol monolayer into a solution containing both glutaraldehyde and diaphorase. We suggested that the glutaraldehyde as a cross-linking reagent was attached to the amino groups of the aminoethanethiol monolayer and the diaphorase enzyme molecules were bound to free aldehyde groups of the glutaraldehyde. Further attachments of the enzyme molecules over the bound enzyme molecules continued with the bridging of the glutaraldehyde. In frequency measurements with a quartz crystal microbalance, the frequency decrease was much more than it was for that of the enzyme monolayer formation, and an enzyme layer thicker than a monolayer was formed. The modified electrode was employed to reduce NAD+ , using diffusional methyl viologen as an electron transfer mediator. The NAD+ was electrocatalytically reduced, and the catalytic current was almost equivalent to that with the multilayered electrode of ten enzyme layers.

• Journal of Pharmaceutical Investigation
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• v.21 no.3
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• pp.143-147
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• 1991

${\alpha}-amylase$ was immobilized on the surface of polyethylene glycol(M.W. 2000) grafted polyurethane film using diisocyanate in an attemp to develop enzyme immobilized polymeric materials. The surface morphology of the modified polyurethane film was examined by SEM. Effects of pH and temperature on the activity of the immobilized ${\alpha}-amylase$ were investigated. The optimal pH range of the activity was $7.0{\sim}7.5.$ The immobilized ${\alpha}-amylase$ demonstrated high thermal stability and maintained consistent activity during long-term storage.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.2
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• pp.140-145
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• 2006

The behaviour of alginate immobilized and soluble watermelon (Citrullus vulgaris) urease in water miscible organic solvents like, acetonitrile, dimethylformamide (DMF), ethanol, methanol, and propanol is described. The organic solvents exhibited a concentration dependent inhibitory effect on both the immobilized and the soluble urease in the presence of urea. Pretreatment of soluble enzyme preparations with organic solvents in the absence of substrate for 10 min at $30^{\circ}C$ led to rapid loss in the activity, while similar pretreatment of immobilized urease with 50% (v/v) of ethanol, propanol, and acetonitrile was ineffective. Time-dependent inactivation of immobilized urease, both in the presence and in the absence of urea, revealed stability for longer duration of time even at very high concentration of organic solvents. The soluble enzyme, on the other hand, was rapidly inactivated even at fairly lower concentrations. The results suggest that the immobilization of watermelon urease in calcium alginate make it suitable for its application in organic media. The observations are discussed.

• Journal of Life Science
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• v.20 no.11
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• pp.1656-1661
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• 2010

Alkaline protease for the hydrolysis of egg yolk protein was immobilized on five carriers - Duolite A568, Celite R640, Dowex-1, Dowex 50W and Silica gel R60. Duolite A568 showed a maximum immobilization yield of 24.7%. Optimum pH for the free and immobilized enzyme was pH 8 and 9, respectively. However, no change was observed in optimum temperature ($50^{\circ}C$). Thermal stability was observed in immobilized enzymes compared to free enzymes. The immobilized enzyme retained 86% activity after 10 cycle operations in a repeated batch process. The effect of flow rate on the stability of enzyme activity in continuous packed-bed reactor was investigated. Lowering flow rate increased the stability of the immobilized enzyme. After 96 hr of continuous operation in a packed-bed reactor, the immobilized enzyme retained 83 and 61% activity when casein and egg yolk were used as a raw materials, respectively.

• KSBB Journal
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• v.28 no.4
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• pp.225-229
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• 2013

Bovine Carbonic anhydrase (BCA) was immobilized on a submicro-porous membrane through covalent immobilization. The immobilization was conducted on the porous membrane surface with the treatment of polyethyleneimine, glutaraldehyde, and the anhydrase, in sequence. The immobilization was confirmed using X-ray photon spectrometer. The pH values of carbon-dioxide saturated solution with buffer were monitored with respect to time to calculate the catalytic activities of hydration of carbon-dioxide for free and immobilized CA. The catalytic rate constant values for free CA, immobilized CA on polystyrene nanoparticles, and immobilized CA on a porous cellulose acetate membrane were 0.79, 0.67, and 0.56 $s^{-1}$, respectively. Reusability was studied up to 10 cycles of $CO_2$ sequestration. The activity for the CA immobilized on the membrane was kept to 95% after 10 cycles, and comparable to the CA on the nanoparticles. The stabilities for heat and storage were also investigated for the three cases. The results suggested that the CA immobilized the membrane had the least loss rate of the activity compared to the others. From this study, the porous membrane was feasible as a carrier for the CA immobilization in hydration and sequestration of carbon-dioxide.

• 한국신재생에너지학회:학술대회논문집
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• 2005.06a
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• pp.471-471
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• 2005

• Journal of dental hygiene science
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• v.2 no.2
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• pp.121-124
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• 2002

The fructan-producing enzyme, fructosyltransferase (FTF), was immobilized on the surface of 4 differents hydroxyapatites by simple incubation. Among the resins, the Type I (${20{\mu}m}$) hydroxyapatite gave the highest yield in terms of immobilization yield and enzyme activity. The fructan synthesis activity of the immobilized FTF was similar with that of the free enzyme, and were about 70-80% on fructose basis with substrate at 100~200 g/L. The binding time between the hydroxyapatite and FTF was terminated within 2 h, and the binding capacity of the Type I ($20{\mu}m$) hydroxyapatite was above 60 U of FTF/g of the resin. By immobilization, the FTF stability against proteolysis, was greatly improved by approximately 3-4 fold. Thus, the present study demonstrated that immobilization of FTF on the surface of the hydroxyapatite occurrs in simple reaction and the binding strength is strong enough to produce the fructan.

• Microbiology and Biotechnology Letters
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• v.48 no.4
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• pp.491-505
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• 2020

A newly isolated strain, Proteus vulgaris OR34, from olive mill waste was found to secrete an alkaline extracellular lipase at 11 U·ml-1 when cultivated on an optimized liquid medium. This lipase was purified 94.64-fold with a total yield of 9.11% and its maximal specific activity was shown to be 3232.58 and 1777.92 U·mg-1 when evaluated using the pH-stat technique at 55℃ and pH 9 and Tributyrin TC4 or olive oil as the substrate. The molecular mass of the pure OR34 lipase was estimated to be around 31 kDa, as revealed by SDS-PAGE and its substrate specificity was investigated using a variety of triglycerides. This assay revealed that OR34 lipase preferred short and medium chain fatty acids. In addition, this lipase was stable in the presence of high concentrations of bile salt (NaDC) and calcium ions appear not to be necessary for its activity. This lipase was inhibited by THL (Orlistat) which confirmed its identity as a serine enzyme. In addition, the immobilization of OR34 lipase by adsorption onto calcium carbonate increased its stability at higher temperatures and within a larger pH range. The immobilized lipase exhibited a high tolerance to organic solvents and retained 60% of its activity after 10 months of storage at 4℃. Finally, the OR34 lipase was applied in biodiesel synthesis via oleic acid mediated esterification of methanol when using hexane as solvent. The best conversion yield (67%) was obtained at 12 h and 40℃ using the immobilized enzyme and this enzyme could be reused for six cycles with the same efficiency.

• 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.

• Microbiology and Biotechnology Letters
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• v.18 no.1
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• pp.56-60
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• 1990

A bacterial strain of Brevibacterium sp. CHI was isolated from soil and used to produce an enzyme (nitrile hydratase) necessary for carrying out the bioconversion of acrylonitrile to acrylamide. Various immobilization methods and enzyme stability were investigated. The nitrile hydratase showed the maximum stability at pH 7 for the free cells. EDTA and phenyl methyl sulfonyl fluoride were selected as the protease inhibitor and the enzyme stability was evaluated by changing inhibitor concentration. Acrylamide beads were the best carriers among four carriers we tested in terms of stability and physicoehemical strength. The storage stability of the immobilized cells decreased with increasing acrylamide concentration of the gel phase at 4$^{\circ}C$, and was very low at acrylarnide concentration above 25%.