• Microbiology and Biotechnology Letters
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• v.9 no.3
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• pp.165-171
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• 1981

The penicillin amidase from Escherichia coli (ATCC 9637) was immobilized by entrappment in gelatin and DEAE-cellulose mixture cross-linked with glutaraldehyde, and the kinetics in a differential column reactor was studied. The optimal operating condition of a differential reactor was reasonably met when the enzyme loading was 1g, and 30 mM substrate solution in 0.1 M phosphate buffer (pH 8.0) was fed at flow rate 4$m\ell$/min and 4$0^{\circ}C$. The optimal pH and temperature were found to be 8.0 and 55$^{\circ}C$, respectively. The Michaelis-Menten constant was 4.8 mM while the maximum velocity was 308 units/g of the immobilized enzyme under the condition of the differential reactor. The effect of substrate inhibition disappeared in the immobilized enzyme preparation. The differential reactor was proved to be good for studying the true kinetics since the pH drop and the external diffusional resistance could be eliminated.

• Korean Journal of Food Science and Technology
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• v.12 no.3
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• pp.176-181
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• 1980

Yeast invertase was immobilized on the 2 kinds of matrices : one is an indirectly coupled enzyme to the cyanogen bromide activated Sepharose by using ${\omega}-aminohexyl$ group as an extension arm, and the other is a tightly adsorbed enzyme on the modified hydrophobic cellulose derivative which has a phenoxyacetyl group as a linkage. The enzyme preparation coupled on Sepharose retained 26.0% of the original activity against sucrose as a substrate, while the preparation immobilized on phenoxyacetyl cellulose retained 72.9% . The immobilized invertase preparation on ${\omega}-aminohexyl$ Sepharose showed the optimal pH 4.5, optimal temperature $60^{\circ}C$, activation energy $5,941\;cal/mole{\cdot}deg$ and Km' 22.2 mM against sucrose, while the preparation adsorbed on phenoxyacetyl cellulose showed the optimal pH 4.0, optimal temperature $60^{\circ}C$, activation energy $7,769\;cal/mole{\cdot}deg$ and Km' 69.9 mM.

• Journal of the Korean Wood Science and Technology
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• v.38 no.3
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• pp.251-261
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• 2010

Cellulase from Formiptosis pinicola KMJ812 is an efficient cellulose degradation enzyme complex, especially with a high ${\beta}$-glucosidase activity. In this study, the change in enzymatic characteristics by immobilization and the reduction of immobilized enzyme activity by repeated usages were evaluated using cellulases from F. pinicola KMJ812. Among tested four resins, Duolite A568 resin had the best enzyme activity yield with 61.7% cellulase activity and 64.4% ${\beta}$- glucosidase activity during the cellulase immobilization. The best reaction temperature was $55^{\circ}C$ for both cellulase and ${\beta}$-glucosidase activities which were higher than the unimmobilized soluble cellulases. The best reaction pH was 4.0 for cellulase activity which was a little more basic than a soluble form and 4.5 for ${\beta}$-glucosidase activity. The immobilized cellulase activity was remained 98% of the beginning activity after 72 h incubation at $50^{\circ}C$ and 50% of the beginning activity after eight times usage at $50^{\circ}C$.

• KSBB Journal
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• v.5 no.2
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• pp.107-112
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• 1990

The kinetic characteristics of an invertase immobilized covalently on porous silica have been appraised for the applicability of porous silica to immobilization supports of enzymes. The invertase was covalently bound with glutaraldehyde on 3-aminopropyltriethoxy-silane-activated porous silica to give a maximum loading of 120mg invertase per 1 gram of dry silica and 26.9 to 70.2% retention of original activity. The porous structure of silica seems to be suitable for enzyme immobilization, judging from the observed results of high immobilization capacity and comparably satisfactory retention of enzyme activity.

• 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.

• Applied Biological Chemistry
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• v.27 no.2
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• pp.112-118
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• 1984

Inverase was entrapped in calcium alginate gel. The immobilized beads had excellent uniform size and configuration. To screen the optimal conditions possessing high enzyme activity, effects of sodium alginate concentration, $CaCl_2$ concentration, and the incubation time of beads in $CaCl_2$ solution during the immobilization procedure were investigated. Immobilized beads prepared from the optimal conditions had 18.8 units per ml of gel, which is equivalent to 68% of the activity of soluble enzyme. Several kinetic parameters were determined: Km value. 143 mM; optimum pH, 4.5; optimum temperature, $50^{\circ}C$. Enzyme loading capacity was 150 mg per 20 ml gel. No significant decrease in sugar conversion was observed during the column operation for 6 days.

• Microbiology and Biotechnology Letters
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• v.31 no.4
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• pp.368-376
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• 2003

Cyclodextrin glucanotransferase (CGTase) from Paenibacillus sp. JB-13 was immobilized on various carriers by several immobilization methods such as ionic binding, covalent linkage and ultrafiltration to improve the process performance. The ultrafiltration and covalent linkage with CNBr-activated sepharose 4B were found as the best method for immobilization of CGTase. The ability of CGTase immobilization onto CNBr-activated sepharose 4B was as high as 18,000 units/g resin when the conditions was as follows: contact time 9 hrs at $37^{\circ}C$, pH 6.0, 100 nm and enzyme loading 24,000 units/g resin. The optimum conditions for production of 2-O-$\alpha$-D-Glucopyranosyl L-Ascorbic acid by immobilized CGTase turned out to be: pH 5.0, temperature $37^{\circ}C$, 20% substrate solution containing 8% (w/v) of soluble starch and 12% (w/v) of L-ascorbic acid sodium salt, 100 rpm, far 25 hrs and with 800 units of immobilized CGTase/ml substrate solution. Moreover the CGTase activity could be stably maintained for 8 times of repetitive reactions after removing products by ultrafiltration through YM 10 membrane.

• KSBB Journal
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• v.25 no.5
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• pp.453-458
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• 2010

The magnetically separable polyaniline nanofiber enzymes were developed for the recycle of enzyme and enhanced enzyme stability. The stability of enzyme was maintained over 90% for 8 days under room temperature and vigorous shaking conditions (200 rpm). The residual activity of immobilized enzyme was over 60% after 8 days incubation at $55^{\circ}C$. Glucose was produced from various polysaccharides, agarose, curdlan, cellulose, and sea weed, using magnetically separable immobilized enzyme. Glucose production rate with curdlan was 1.2 g/(l h) and showed high decomposition rate due to high mass transfer. After 10 times recycle, the residual activity of immobilized enzyme was over 75%. 1 g/L of glucose was produced with 5 mg of immobilized enzymes.

• 한국신재생에너지학회:학술대회논문집
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• 2005.11a
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• pp.267-275
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• 2005

차세대 재생산성 에너지로 각광을 받고 있는 바이오디젤은 현재 주로 알칼리촉매를 이용하는 화학공정으로 생산하고 있으나 고에너지 요구성이며 대규모 생산시 폐수발생 등 환경오염 유발요인이 있기 때문에 친환경 생물공정의 필요성이 대두되고 있다. 생물촉매 리파제(lipase)를 이용하는 친환경 생물공정은 화학공정에 비해 다양한 장점을 제공하고 있으나 고가의 효소생산 비용문제로 실용화에 어려움이 있다. 따라서 본 연구에서는 저비용의 생물학적 바이오디젤 생산 시스템 구축을 위해 고활성의 효소 개발, 경제적 재조합 대량생산, 반복 재사용을 위한 효소고정화 등을 통해 고효율의 생산반응계를 개발하였다. 우선 바이오디젤 생산공정에 적합한 리파제로서 CalB(Lipase B of Candida antarctica)를 선택하고 분자 진화기술을 이용하여 효소활성을 17배 향상시킨 CalB14를 개발하였다. CalB14를 효모 발현시스템을 이용하여 경제적 대량생산하기 위해 단백질분비를 획기적으로 개선할 수 있는 맞춤형 분비융합합인자기술(TFP technology)을 이용하여 재조합 CalB를 2 grams/liter 수준으로 분비생산하였다. 생산된 효소를 반복 재사용이 가능하도록 다양한 레진에 고정화하였고 최적의 바이오디젤 전환반응용 고정화효소를 개발하였다. 고정화효소를 효율적으로 재사용하기 위해 바이오디젤 생산용 고정상반응기(packed-bed reactor)를 제작하였으며 기질을 12시간내에 95% 이상 바이오디젤로 수십회 이상 반복전환할 수 있는 경제적인 생물학적 바이오디젤 전환 시스템을 구축하였다.

• Korean Chemical Engineering Research
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• v.50 no.2
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• pp.257-263
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• 2012

Biodiesel is a renewable energy which is nontoxic and acting as a replacement for conventional diesel which derived from fossil fuel. Classified biodiesel producing way such as acid, base, supercritical and enzyme methods, this study focused on eco-friendly production of biodiesel using supercritical and immobilized enzyme process. Assuming a plant with a production rate of 10,000 tons a year, a PRO II simulator program was used to simulate the product conversion rate and total energy consumption. The product conversion in supercritical process and immobilized enzyme was found to be 91.17% (including 0.9% glycerol) and 93.18% (including 1.0% glycerol) respectively. The result shows that the efficiency of immobilized enzyme process is higher compared to supercritical process but having lower end product purity. From the energy consumption point of view, supercritical process consume about 8.9 MW while immobilized enzyme process consume much lower energy which is 3.9 MW. Consequently, this study certifies that energy consumption of supercritical process is 2.3 times higher than immobilized enzyme process.