• Korean Journal of Food Science and Technology
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• v.8 no.4
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• pp.253-259
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• 1976

Two kinds of sepharose-bound pronases were successfully prepared. The immobilized pronase, directly coupled to cyanogen-bromide activated sepharose, retains 22.6% of original specific activity against casein. However, ${\omega}-aminoalkyl$ sepharose immobilized pronases, in which extension arms of ${\omega}-aminoalkyl$ group $(number\;of\;-CH_2-\;is\;8,\;10,\;and\;12)$ are used, retain almost 100% of original specific activity. Studies of enzyme stability, pH dependence, temperature dependence, and Km values are presented.

• Applied Biological Chemistry
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• v.26 no.4
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• pp.222-230
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• 1983

This study was designed to develop a process for the immobilization of xylose isomerase(D-xylose ketol isomerase, EC 5.3.1.5) from Streptomyces griseolus previously isolated by the authors and its application on a pilot plant scale for the production of high fructose corn syrup. The biomass which has endo-excreted xylose isomerase was homogenized under a pressure of $500kg/cm^2$ and 90.8% of the enzyme recovery of the native activity was obtained as compared to 54.7% recovery by the lysozyme treatment. Ionic bonding method was adopted for the enzyme immobilization due to its many reported merits. It was found that the porous resins such as Diaion HP 20, Duolite A-7, Amberlite IRA 93 and 94 were effective in immobilizing the enzyme. In addition, it was disclosed that the regeneration form of $BO_4--$ is effective for Amberlite IRA 93 and $HCO_3-$ for Diaion HP 20. Optimal immobilization condition for Amberlite IRA 93 was pH 8.0 and $55^{\circ}C$ yielding 80.6% of immobilization. Activity decay test showed half life of the immobilized enzyme with Amberlite IRA 93 was more than 24 days at $65^{\circ}C$. The carrier was evaluated to be resuable and its result showed the relative immobilization yields were 98.2, 93.3, 90.7 and 87.5%, respectively at second, third, forth and fifth rebinding test of the enzyme on Amberlite IRA 93. Optimal temperature of the immobilized enzyme was slightly lowered and the range widened to $60\sim70^{\circ}C$, while optimal pH moved toward $8.0\sim8.3$ in its isomerization reaction. The trial production result of high fructose corn syrup in pilot scale immobilization showed that one liter of immobilized xylose isomerase (350 IXIU/ml-R) is capable producing about 293l high fructose corn syrup(75% dry substance) in 30 days.

• Korean Journal of Food Science and Technology
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• v.22 no.4
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• pp.426-433
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• 1990

The conditions for immobilization of the partially purified ${\beta}-galactosidase$ form Bacillus subtilis HP4 and the properties of the immobilized enzyme have been investigated. The crude enzyme precipitated with cold acetone was purified about 68-fold through DEAE-cellulose and sephadex G-100 chromatography and its recovery was 19.9% The optimal conditions for Immobilization of enzyme were obtained in 2%(w/v) sodium alginate, 15%(v/v) enzyme solution and 2%(w/v) calcium chloride, and also the optimal stirring thme was 2 hours on the above conditions. The optimum temperature and pH values for immobilized enzyme were $55^{\circ}C$ and 6.5, respectively. Its residual activity was show 25% after heat treatment for an hour at $65^{\circ}C$, and found its high stability in pH 6.0 to 8.0. The enzyme activity was not affected b)· EDTA, 2-mercaptoethanol, KCN, protective agents, and other methal ions except Hg ion and Cu ion. The $K_m\;and\;V_{max}$ values of the immobilized enzyme on ONPG were $1.82{\times}10^{-2}M\;and\;3.57{\times}10^{-8}mole/min$, whereas those on lactose were $2.94{\times}10^{-2}M\;and\;1.68{\times}10^{-7} mole/min$, respectively. The remained enzyme activity for the immobilized enzyme was 95%t of original activity after storage of 40 days at $4^{\circ}C$, and when reused for 5 times was 81%. When skim milk(4.8% lactose) and 5% lactose solution were reacted with the immobilized enzyme(250 units/g) of lactose were 51% and 43%, respectively.

• Applied Biological Chemistry
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• v.39 no.5
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• pp.333-337
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• 1996

For continuous production of galactooligosaccharides(GOS), $\beta-galactosidase$ with h1gh transgalactosylation activity from Bacillus sp. A 4442 was Immobilized onto $Diaion^{TM}$ HPA 75(styrene-divinylbenzene resin). The parameters influencing enzyme immobilization were scrutinized in order to maximize immobilization yield while minimizing enzyme inactivation. The optimum conditions turned out to be: Tris buffer concentration 30 mM, pH 8.0, contact time at room temperature 3 hr, and enzyme loading 25 mg protein/g resin. Both the thermal stability and the operational stability of immobilized enzyme were markedly enchanced by the treatment with 0.5% glutaraldehyde as a cross-linker. Under the experimental conditions established, the yield of ${\beta}-galactosidase$ immobilization was 40% or more and the activity of the immobilized enzyme ca. 200 U/g resin. When a packed-bed reactor was employed to continuously convert lactose to GOS, the specific production, which refers to as the amount of commercially valuable GOS produced by a unit amount of immobilized ${\beta}-galactosidase$, was found to be ca. 300 g GOS/g carrier.

• Analytical Science and Technology
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• v.23 no.2
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• pp.172-178
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• 2010

Glutathione S-transferase is known to play a crucial role in detoxification in many cases. To develop a herbicide detection biosensor, we in this study attempted to immobilize glutathione S-transferase enzyme on solid supports, polystyrene and agarose, and Na-alginate. These matrixes were attractive materials for the construction of biosensors and might also have utility for the production of immobilized enzyme bioreactors. We also compared the activities of glutathione-S-transferase immobilized OsGSTF3 and free OsGSTF3. The specific activity of the free enzyme in solution was 3.3 higher than the immobilized enzyme. These results suggest that 50% of the enzyme was bound with the catalytic site in polystyrene-alkylamine bead and immobilized enzymes showed 80% remaining activity until 3 times reuse.

• Current Research on Agriculture and Life Sciences
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• v.6
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• pp.157-162
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• 1988

In order to study of practical purpose of immobilized Mucor spp L42 milk clotting enzyme on activated succimylamino-propyl glass beads with glutaraldehyde in continuous curd coagulation, acidified milk(pH5.6, $8^{\circ}C$) was treated through reactor packed with immobilized beads, and warmed at $30^{\circ}C$ and allowed to coagulation for the determination of enzyme stability, deactivation of milk clotting ability by continuous reaction, the beads treatment conditions, and contact time of milk and beads in reactors. The results obtained were summarized as follow ; 1) After 3 month's storage, activity of immobilized Mucor spp L42 milk clotting enzyme in 0.2M phosphate buffer(pH 4.6) with 0.06% sodium azide was only 80% of initial activity. 2) Milk clotting activity of the beads was decreased by continuouse exposure on acidified skim milk. Nitrogen accumulation on the beads paralled loss of the activity in initial reaction stage. 3) After 6 hours continuous treatment of the beads at 60 sec/ml surface time, the milk-clotting activity of the beads was about 70% of initial activity. 4) Bead reactor and shaking bed reactor were more effective than column reactor on continuouse skim milk coagulation.

• KSBB Journal
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• v.10 no.4
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• pp.468-474
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• 1995

The disposable glucose bio-sensor using composite of CTA and PCL membrane was developed for measurement of glucose. The most effective membrane was composed of CTA/PCL(80/20, w/w) and glutaraldehyde one-step immobilization method ($10{\mu}m$ thickness) for glucose sensor gave the best result among various methods, considering oxygen permeability and electronic sensitivity. A scanning electron micrograph of the cross-section of a typical asymmetric CTA/PCL composite membrane showed that the membrane fused with a dense layer covered with a GOD-glutaraldehyde. Glucose oxidase immoblilized on the membrane showed the linearity between difference of absolute amperometric values and glucose concentrations within 7mM when the GOD immobilized electrode was used. About 35% of activity was remained after 8 days when the tyrosinase was immobilized on CTA/PCL (80/20) membrane.

• Journal of Life Science
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• v.17 no.9 s.89
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• pp.1260-1265
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• 2007

Magnetically separable enzyme-coated nanofibers were developed for the hydrolysis of starch. Stability of ${\alpha}-amylase-coated$ nanofiber was greatly improved and its residual activity was maintained over 92.7% after 32 days incubation at room temperature and under shaking conditions (200 rpm). The recovery of enzyme was high and enzyme activity after 10 recycle was 95.2% of its original activity. Developed enzyme-coated nanofibers were used for the hydrolysis of starch. When 0.5 mg of magnetically separable enzyme nanofibers was used, 40 g/l of starch (2 ml) was completely degraded within 40 min. The continuous enzyme reactor was developed and used for starch hydrolysis and 76% of starch (30 g/l) was hydrolyzed with 1 hr residence time.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.1
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• pp.83-87
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• 1999

Immobilization of amyloglucosidase (AMG) with chitosan microbead and its possible applications were evaluated. The diameter of chitosan inicrobead was about 1.2 mm and the optimum enzyme concentration for immobilization was 6 mg/ml. The relative activity of the immobilized enzyme was $97.8\%$ at pH 4.2 and $55^{\circ}C$ and the optimum condition for the immobilized enwme was the same to that of free enzyme. In case of temperature above $30^{\circ}C$, the activity of the immobilized enzyme was a little higher than that of free enzyme. The enzyme activities of both free and immobilized were stable for 6 months when stored at $35^{\circ}C$. The optimum temperatures of both enzymes for saccharification of the dextrinized starch were $55^{\circ}C$ while the relative activity of the immobilized enzlme was $62.6\%$.

• KSBB Journal
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• v.4 no.3
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• pp.229-234
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• 1989

NAD+ analogs, 8-( 6-aminohexyl) aminonicotinamide adenine dinucleotide and N6-[(6- aminohewl)-carbamoylmethyl]- NAD+, were imobilized on bovine caseins by the action of hansglutaminase. It appears that NAD+ analogs bind with $\alpha$S1-and $\beta$-caseins through formation of the r-glutamylamine bond between the amino groups attached to the hexyl chains in NAD+ analogs and the glutaminyl residues in caseins. The NAD+ analogs immobilized on the caseins were enzymatically reducible by alcohol dehydrogenase. $\beta$-Casein was more useful carrier than the $\alpha$S1-casein and 8-substituted NAD+ analog was more effective than N6-substituted one in immobilization. Michaelis constant of 8-substituted NAD+ analog immobilized on $\beta$-casein in alcohol dehydrogenase reaction was similar to that of free from of NAD+ and that of NAD+ analog. Immobilized NAD+ was much more stable at alkaline pH than free NAD+ and its analog while maximum velocity was reduced to 31% of the free NAD+ analog. The coenzyme casein conjugated was recovered almost completely in casein precipitated by calcium.