• Journal of the Korean Wood Science and Technology
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• 제17권3호
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• pp.45-51
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• 1989

A major problem in the enzymatic hydrolysis of lignocellulosic substrates is the very strong bonding of cellulase to lignin and even cellulose in the hydrolysis residues. This phenomenon inhibits recycle of the cellulase which is a major expense of the enzymatic hydrolysis process. In this paper, autohydrolyzed wood was delignified by two-stage with a 0.3% Na OH extraction and oxygen-alkali bleaching and was subjected to enzymatic hydrolysis with cellulase. Also, an improved almost quantitative recycle process of cellulase enzyme was discussed. In enzyme recovery by affinity method. the first recycling showed relatively high hydrolysis rate of 97.4%. Even at the third recycle. hydrolysis rate was 86.7 percents. In the case of cellulase recovery by ultrafiltration method, first 2 recycling treatments resulted very high hydrolysis rate(97.0-97.7%). Even the third recycling showed about 94.2%. Authoydrolysis of oak wood followed by 2-stage delignification with alkali and oxygen-alkali produced a substrate for enzymatic hydrolysis that allowed almost quantitative recycle of cellulase.

• Journal of the Korean Wood Science and Technology
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• 제19권1호
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• pp.14-21
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• 1991

Steam-exploded woods were delignified by two-stage with a 0.3% NaOH extraction and oxygen-alkali bleaching and were subjected to the enzymatic hydrolysis with cellulase enzyme. Also, an improved almost quantitative recycle process of cellulase enzyme was discussed. In enzyme recovery by affinity method, The first recycling showed relatively high hydrolysis rate of 96.4%. Even at the third recycle, hydrolysis rate was 87.0 percents. In the case of cellulase recovery by ultrafiltration method, first 2 recycling treatments resulted in very high hydrolysis rates, 96.8% and 95.0%, respectively. Even the third recycling showed about 93.6%. Steam-explosion treatment of oak wood followed by 2-stage delignification with alkali and oxygen-alkali produced a excellant substrate for the enzymatic hydrolysis that allowed almost quantitative recycle of cellulase.

• Journal of Microbiology and Biotechnology
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• 제22권1호
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• pp.92-99
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• 2012

The optimal physical factors affecting enzyme production in an airlift fermenter have not been studied so far. Therefore, the physical parameters such as aeration rate, pH, and temperature affecting PLA-degrading enzyme production by Actinomadura keratinilytica strain T16-1 in a 3 l airlift fermenter were investigated. The response surface methodology (RSM) was used to optimize PLA-degrading enzyme production by implementing the central composite design. The optimal conditions for higher production of PLA-degrading enzyme were aeration rate of 0.43 vvm, pH of 6.85, and temperature at $46^{\circ}C$. Under these conditions, the model predicted a PLA-degrading activity of 254 U/ml. Verification of the optimization showed that PLA-degrading enzyme production of 257 U/ml was observed after 3 days cultivation under the optimal conditions in a 3 l airlift fermenter. The production under the optimized condition in the airlift fermenter was higher than un-optimized condition by 1.7 folds and 12 folds with un-optimized medium or condition in shake flasks. This is the first report on the optimization of environmental conditions for improvement of PLA-degrading enzyme production in a 3 l airlift fermenter by using a statistical analysis method. Moreover, the crude PLA-degrading enzyme could be adsorbed to the substrate and degraded PLA powder to produce lactic acid as degradation products. Therefore, this incident indicates that PLA-degrading enzyme produced by Actinomadura keratinilytica NBRC 104111 strain T16-1 has a potential to degrade PLA to lactic acid as a monomer and can be used for the recycle of PLA polymer.

• KSBB Journal
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• 제25권5호
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• pp.453-458
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• 2010

본 연구에서는 효소의 재활용성과 안정성을 확보하기 위해 자석으로 분리가 가능한 polyaniline nanofiber를 개발하였다. 개발된 고정화 효소는 상온에서 8일 동안 90% 이상의 활성도를 보유하였으며 온도가 높은 $55^{\circ}C$에서는 60% 이상의 활성도를 보유하여 안정성의 유지현상을 보였다. 개발된 고정화 효소는 자석으로 분리가 가능하였으며 이 효소를 이용하여 curdlan, agarose, cellulose, 및 미역을 분해한 결과 포도당을 생산하였으며 curdlan을 분해시킨 경우에는 분해 속도가 1.2 g/L/h로 나타나 다른 다당류에 비해 3-10배 이상 빠른 속도를 나타내었다. 고정화 효소를 반복하여 사용하는 경우 10번 반복 사용했을 때 75% 이상의 활성도를 유지하는 것으로 측정되었다. 젖은 미역 줄기를 10 g/L를 분해하기 위하여 5 mg의 고정화 효소를 사용한 결과 24시간 만에 1 g/L의 glucose를 생산하였다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제7권5호
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• pp.268-274
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• 2002

Enzymatic hydrolysis of waste office paper was evaluated using three commercial cellulases, Acremonium cellulase, Meicelase, and Cellulosin T2. Varying the enzyme loading from 1 to 10% (w/w) conversion of waste office paper to reducing sugar was investigated. The conversion increased with the increase in the enzyme loading: in the case of enzyme loading of 10% (w/w), Acremonium cellulase yielded 79%conversion of waste office paper, which was 17% higher compared to Meicelase, 13% higher than that of Cellulosin T2. Empirical model for the conversion (%) of waste office paper to re-ducing sugar (x) was derived from experimental results as follow, x = $kE^{m}t^{(aE+b)}$ where k, m, a, and b de-note empirical constants. E indicates initial enzyme concentration.

• Journal of Microbiology and Biotechnology
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• 제27권2호
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• pp.289-296
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• 2017

Lysine decarboxylase (CadA) converts ${\small{L}}-lysine$ into cadaverine (1,5-pentanediamine), which is an important platform chemical with many industrial applications. Although there have been many efforts to produce cadaverine through the soluble CadA enzyme or Escherichia coli whole cells overexpressing the CadA enzyme, there have been few reports concerning the immobilization of the CadA enzyme. Here, we have prepared a cross-linked enzyme aggregate (CLEA) of E. coli CadA and performed bioconversion using $CadA^{CLEA}$. $CadA^{free}$ and $CadA^{CLEA}$ were characterized for their enzymatic properties. The optimum temperatures of $CadA^{free}$ and $CadA^{CLEA}$ were $60^{\circ}C$ and $55^{\circ}C$, respectively. The thermostability of $CadA^{CLEA}$ was significantly higher than that of $CadA^{free}$. The optimum pH of both enzymes was 6.0. $CadA^{free}$ could not be recovered after use, whereas $CadA^{CLEA}$ was rapidly recovered and the residual activity was 53% after the $10^{th}$ recycle. These results demonstrate that $CadA^{CLEA}$ can be used as a potential catalyst for efficient production of cadaverine.

• 한국미생물·생명공학회지
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• 제19권2호
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• pp.171-178
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• 1991

The two-stage enzyme reactor, packed with cyclodextrin glucanotransferase (CGTase) immobilized on Amberite IRA 900, coupled with ultrafiltration membrane was investigated for continuous production of cyclodextrin (CD). 5% (w/v) of soluble starch was partially cyclized, in the 0.1 l first-stage immobilized enzyme reactor, up to CD conversion yield of 10% (w/w) at retention time of 0.56hr and 1.5 units of immobilized CGTase/1g of carrier. In the second stage main immobilized enzyme reactor capacity of 1.5 l, the maximum CD conversion yield of 39% (w/v) was achieved at retention time of 2.8hr and 0.47 unit of CGTase/1 g of carrier. Unreacted residual dextrin was fractionated with ultrafiltration membrane, and then, recycled into the second-stage main bioreactor to increase the CD conversion yield. The most suitable membrane size and the volume concentration ratio (concentrate: filterate) for recycling of unreacted residual dextrin were found to be 5K dalton and 4:6, respectively. CD conversion yield was increased about 3~4% upon co-immobilization of pulluanase along with CGTase. Spent Amberite IRA 900 can be reutilized consecutively more than 3 times for immobilization of CGTase after regeneration.

• Journal of Microbiology and Biotechnology
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• 제25권8호
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• pp.1291-1298
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• 2015

Five different polymer nanofibers, namely, polyaniline nanofiber (PANI), magnetically separable polyaniline nanofiber (PAMP), magnetically separable DEAE cellulose fiber (DEAE), magnetically separable CM cellulose fiber (CM), and polystyrene nanofiber (PSNF), have been used for the immobilization of lactase (E.C. 3.2.1.23). Except for CM and PSNF, three polymers showed great properties. The catalytic activities (k_cat) of the free, PANI, PAMP, and magnetic DEAE-cellulose were determined to be 4.0, 2.05, 0.59, and 0.042 mM/min·mg protein, respectively. The lactase immobilized on DEAE, PANI, and PAMP showed improved stability and recyclability. PANI- and PAMP-lactase showed only a 0-3% decrease in activity after 3 months of vigorous shaking conditions (200 rpm) and at room temperature (25℃). PANI-, PAMP-, and DEAE-lactase showed a high percentage of conversion (100%, 47%, and 12%) after a 1 h lactose hydrolysis reaction. The residual activities of PANI-, PAMP-, and DEAE-lactase after 10 times of recycling were 98%, 96%, and 97%, respectively.

• 생명과학회지
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• 제17권9호통권89호
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• pp.1260-1265
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• 2007

자석으로 분리가 가능한 ${\alpha}-Amylase$가 코팅된 나노고분자를 제조하여 녹말의 분해공정에 활용하였다. 본 연구에서 개발된 고정화 효소의 안정성은 크게 증가하여 상온에서 200rpm으로 교반하면서 보관한 경우 30일 동안에 92.7%의 활성도를 유지하였다 . 고정화 효소를 자석을 이용하여 재사용한 경우 10 회 동안 사용했을 경우 95.2%의 활성도 회수율을 보 여 효소의 재사용 가능성을 확인시켜 주었다 . 고정화 효소 0.5mg을 사용하여 녹말 분해 공정 에 활용하였을 때 2 ml의 40 g/l 녹말 용액을 40분만에 완전히 분해 시켰다, 이러한 고정화 효소를 사용하여 연속 효소반응기를 개발하여 녹말 분해공정에 활용한 결과 체류시간을 1시간으로 하였을 때 녹말 30 g/l를 76% 분해시켜 산업적으로 활용 가능성을 보여 주었다.

• Journal of Microbiology and Biotechnology
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• 제31권3호
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• pp.419-428
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• 2021

To efficiently recycle GH78 thermostable rhamnosidase (TpeRha) and easily separate it from the reaction mixture and furtherly improve the enzyme properties, the magnetic particle Fe3O4-SiO2-NH2-Cellu-ZIF8 (FSNcZ8) was prepared by modifying Fe3O4-NH2 with tetraethyl silicate (TEOS), microcrystalline cellulose and zinc nitrate hexahydrate. FSNcZ8 displayed better magnetic stability and higher-temperature stability than unmodified Fe3O4-NH2 (FN), and it was used to adsorb and immobilize TpeRha from Thermotoga petrophilea 13995. As for properties, FSNcZ8-TpeRha showed optimal reaction temperature and pH of 90℃ and 5.0, while its highest activity approached 714 U/g. In addition, FSNcZ8-TpeRha had better higher-temperature stability than FN. After incubation at 80℃ for 3 h, the residual enzyme activities of FSNcZ8-TpeRha, FN-TpeRha and free enzyme were 93.5%, 63.32%, and 62.77%, respectively. The organic solvent tolerance and the monosaccharides tolerance of FSNcZ8-TpeRha, compared with free TpeRha, were greatly improved. Using naringin (1 mmol/l) as the substrate, the optimal conversion conditions were as follows: FSNcZ8-TpeRha concentration was 6 U/ml; induction temperature was 80℃; the pH was 5.5; induction time was 30 min, and the yield of products was the same as free enzyme. After repeating the reaction 10 times, the conversion of naringin remained above 80%, showing great improvement of the catalytic efficiency and repeated utilization of the immobilized α-L-rhamnosidase.