• KSBB Journal
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• v.15 no.3
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• pp.247-253
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• 2000

This study was aimed to enhance polycyclic aromatic hydrocarbon(PAHS) biodegradation rate by repeated-batch treatment using immobilized cells of Phanerochaete chrysosporium. In the repeated-batch operations with 30 mg/L of pyrene the maximum degradation rate was 6.58 mg/L day. As the number of batches increased the concentration of immobilized cells significantly decreased and the degradation rate and specific acitivity gradually increased to a maximum value and then decreased. To have PAH degradation activity and cell mass recovered one batch of cultivation using the growth medium instead of the PAH-degrading medium was carried in the course of repeated-batch operations. This maximum degradation rates of pyrene and anthracene were 4.29 and 4.46 mg/L$.$day respectively. Overall the rate of PAH degradation could be enhanced 2.5-30 folds by using immobilized cells compared to the case of using suspended cells.

• Journal of Microbiology and Biotechnology
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• v.21 no.9
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• pp.972-978
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• 2011

In this study, we investigated the performance of an immobilized ${\beta}$-galactosidase inclusion bodies-containing Escherichia coli cell reactor, where the cells were immobilized in alginate beads, which were then used in repeated-batch operations for the hydrolysis of o-nitrophenyl-${\beta}$-D-galactoside or lactose over the long-term. In particular, in the Tris buffer system, disintegration of the alginate beads was not observed during the operation, which was observed for the phosphate buffer system. The o-nitrophenyl-${\beta}$-D-galactoside hydrolysis was operated successfully up to about 80 h, and the runs were successfully repeated at least eight times. In addition, hydrolysis of lactose was successfully carried out up to 240 h. Using Western blotting analyses, it was verified that the ${\beta}$-galactosidase inclusion bodies were sustained in the alginate beads during the repeated-batch operations. Consequently, we experimentally verified that ${\beta}$-galactosidase inclusion bodies-containing Escherichia coli cells could be used in a repeated-batch reactor as a biocatalyst for the hydrolysis of o-nitrophenyl-${\beta}$-D-galactoside or lactose. It is probable that this approach can be applied to enzymatic synthesis reactions for other biotechnology applications, particularly reactions that require long-term and stable operation.

• Journal of Microbiology and Biotechnology
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• v.5 no.4
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• pp.229-233
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• 1995

Acremonium chrysogenum was immobilized in ionotropic gel beads to develop semi-continuous production of cephalosporin C (CPC). Barium alginate beads were more stable than calcium alginate or strontium alginate beads in chemically defined media. The gel stability of Ba-alginate was further increased by cross-linking with polyethyleneimine (PEI). The presence of carboxymethyl cellulose inside Ba-alginate beads did not reduce mass transfer resistance. Ba-alginate microbeads that had little diffusion limitation increased CPC production rate 1.6 fold higher than that of normal beads. CPC fermentation with immobilized cells in Ba-alginate microbeads was performed continuously for 40 days by way of repeated fed-batch operations. Mathematical modeling was developed to describe the repeated fed-batch fermentation system. Results of the computer simulation agreed well with the experimental data, which made it possible to predict an optimal feeding rate that could maximize total CPC productions.

• Journal of Microbiology and Biotechnology
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• v.22 no.11
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• pp.1486-1493
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• 2012

In this study, we investigated whether galactooligosaccharide (GOS) can be stably and steadily synthesized using immobilized ${\beta}$-galactosidase (${\beta}$-gal) inclusion body (IB)-containing E. coli cells during long-term repeated-batch operation. To improve the operational stability of this enzyme reactor system, immobilized E. coli cells were crosslinked with glutaraldehyde (GA) after immobilization of the E. coli. When we treated with 2% GA for E. coli crosslinking, GOS production continued to an elapsed time of 576 h, in which seven batch runs were operated consecutively. GOS production ranged from 51.6 to 78.5 g/l ($71.2{\pm}10.5$ g/l, n = 7) during those batch operations. In contrast, when we crosslinked E. coli with 4% GA, GOS production ranged from 31.5 to 64.0 g/l ($52.3{\pm}10.8$, n = 4), and only four consecutive batch runs were operated. Although we did not use an industrial ${\beta}$-gal for GOS production, in which a thermophile is used routinely, this represents the longest operation time for GOS production using E. coli ${\beta}$-gal. Improved stability and durability of the cell immobilization system were achieved using the crosslinking protocol. This strategy could be directly applied to other microbial enzyme reactor systems using cell immobilization to extend the operation time and/or improve the reactor system stability.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.410-413
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• 2001

In this study a repeated batch production of lactic acid was investigated for the purpose of reducing amount of yeast extract dosage. First batch operation was performed with nutrient rich media(glucose 100-150 g/${\ell}$, and high yeast extract contents; 15 g/${\ell}$), and subsequential batch operations were performed with nutrient poor media(glucose 100-150 g/${\ell}$, and reduced yeast extract contents; 4-5 g/${\ell}$). Volumetric productivities(6.03-6.20 g/${\ell}{\cdot}h$) and substrate conversion rate(94-98%) of eve η subsequential batch operation were maintained with the same level of first batch operation(6.19 g/${\ell}{\cdot}h$, and 99%, respectively), when concentrations of glucose and yeast extract were 100, and 4 g/${\ell}$ respectively.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.6
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• pp.544-549
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• 2006

The performance of immobilized fungal cells on celite beads for the production of gibberrelic acid was investigated in flasks and 7-L stirred-tank reactor. Repeated incubations of immobilized fungal cells increased cell concentrations and volumetric productivity. The maximum volumetric productivity obtained in the immobilized-cell culture was 3-fold greater than that in suspended-cell culture. The concentration of cotton seed flour (CSF), among the various nutrients supplied, most significantly influenced productivity and operational stability. Notably, insoluble components in CSF were found to be essential for production. CSF at 6 g/L with 60 g/L glucose was found to be optimal for gibberellic acid production and stable operation by preventing excessive cell growth.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.6
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• pp.367-370
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• 2002

Candida cylindracea lipase was immobilized by adsorption on acid washed glass beads. It was observed that protein loading of the support depends on the size of the particle, with smaller particle containing higher amount of protein per unit weight. Initial reaction rate linearly varied up to enzyme concentration of 17.25 U/mL. Amount of free fatty acids produced was linearly proportional up to the enzyme loading of 1650 $\mu$g/g of bead. Achievement of chemical equilibrium took longer time in the case of less protein loading. Degree of hydrolysis was found to decrease in second and third consecutive batch operations on repeated use of immobilized lipase.

• The Korean Journal of Mycology
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• v.39 no.3
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• pp.243-248
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• 2011

We studied the repeated-batch process for the bioethanol production from the hydrolysate of Ulva pertusa Kjellman using yeast Pichia stipitis, which is able to assimilate C6- and C5-monosaccharides. During 180-hour operations, the repeated-batch process was carried out stably, and the average bioethanol concentration reached 11.9 g/L from about 30 g/L of reducing sugar in the hydrolysate. Meanwhile, the bioethanol yields, based on the reducing sugar and the quantitative TLC analysis, were 0.40 and 0.37, respectively, which corresponded to 78.4% and 72.5% of theoretical value, respectively. Throughout the quantitative process analysis, it was also demonstrated that 39.67 g-bioethanol could be produced from 1 kg of dried Ulva pertusa Kjellman. In this study, we verified that the bioethanol production from the hydrolysate of Ulva pertusa Kjellman was feasible using a yeast Pichia stipitis, particularly during the repeated-batch operation.

• KSBB Journal
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• v.14 no.5
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• pp.585-592
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• 1999

This study was aimed to enhance the Fe(II) oxidation rate using immobilized cells of Thiobacillus ferroxidans. For this purpose, a medium for the minimization of jarosite formation was developed first. Secondly, cell immobilization in celite beads was carried out. And then, repeated-batch and continuous operatons of Fe(II) oxidation by using immobilization cells were performed. In a series of flask cultures, three types of media were tested: media with a much lower salt concentration than that of the 9K medium; media which contained different nitrogen sources from that of the 9K medium, that is $(NH_4)_2HPO_4$, $NH_4Cl and HNO$_3$; media which contained $(NH_4)_2HPO_4$ as nitrogen and phosphate source, but without $K_2HPO_4$ as nitrogen and phosphate source in the 9K medium. As a result, the M16 medium which contained 3 g/L of $(NH_4)_2HPO_4$ as nitrogen and phosphate source was found to be the optimal one. It sustained good cell growth allowing no jarosite formation. In the repeated-batch operations, the rate of Fe(II) oxidation gradually increased to reach a maximum value as the batch was repeated. As a result of repeated-batch operations. a maximum Fe(II) oxidation rate was 2.33 g/L . h. In the continuous operations, the iron oxidation rate could be increased to 2.14 g/L .h at a dilution rate of 0.25 $h^{-1}$ which is greater than the maximum specific growth rate (0.12 $h^{-1}$) of the bacteria.

• 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.