• KSBB Journal
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• v.13 no.4
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• pp.411-417
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• 1998

The possibility of application of membrane type immobilized adsorbent to the fed-batch or perfusion culture system with anchorage-independent cells as well as batch system was investigated. The improvement in cell density and cell viability due to the combination of immobilized adsorbent with each culture system was evaluated for the investigation, and the optimum culture system employing immobilized adsorbent system was suggested based on the results. It was observed that the system with immobilized adsorbent showed better cell growth and cell viability than that without immobilized adsorbent in every operation system of batch, fed-batch, and perfusion. In case of batch system, 200% improvement of maximum cell density was observed in the system where ammonium chloride was added on purpose. And 50% improvement of maximum cell density was observed in the fed-batch system where ammonium ion accumulates significantly, while small increase in maximum cell density was observed in the perfusion system where dilution of waste byproducts exists. Especially, the fed-batch system showed the most significant improvement on cell growth because both compensation of nutrient and removal of ammonium ion occurred simultaneously in the system. Therefore a combined system of immobilized adsorbent and fed-batch operation could be suggested as an optimum system with in situ removal of ammonium ion.

• Journal of applied mathematics & informatics
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• v.19 no.1_2
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• pp.203-214
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• 2005

In this study the optimal control of fed-batch glycerol fermentation is investigated based on an impulsive dynamical system. Considering the sudden increase of the glycerol and alkali in fed-batch culture of biodissimilation of glycerol to 1,3-propanediol, this paper proposes a non-linear impulsive system of fed-batch culture. The existence, uniqueness and regularity properties of piecewise solution for the system are proved. In view of the controllability of volumes of glycerol added to the reactor instantaneously, the paper constructs an optimal control model based on the nonlinear impulsive system and the existence of the optimal control is obtained. The control variables here are the moments and the sizes of jumps in the states at the discrete instants and the objective is to maximize the productivity of 1,3-propanediol over one cycle.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.2
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• pp.123-129
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• 2000

A high cell density culture system for the anchorage dependent CHO cells was developed based on the combination of in removal of ammonium ion and microcarrier culture system, and semi-fed-batch feeding of glucose and glutamine was employed to the developed culture system. The glass bead was selected as an optimum microcarrier in terms of cell growth. An ammonium ion selective zeolite, Phillipsite-Gismondine, was packed in a dialysis menium ion. The semi-fed-batch operation was employer to the novel culture system for the high density cell culture, and the results showed the cell growth was improved by 32% and tPA productivity by 250%.

• KSBB Journal
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• v.15 no.5
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• pp.489-496
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• 2000

Several culture systems including batch, two-stage CSTR, semi-fed batch, and two-stage cyclic fed-batch were investigated for the efficient production of the Fab fraction of PDC-E2 specific human monoclonal antibody using high cell density recombinant E. coli. A two-phase batch system and a two-stage continuous system were examined to overcome plasmid instability problems, by separating the growth and the production stages. The cell density and productivity of the two-stage continuous culture was better than that of the two-phase batch fermentation. In the two-stage continuous culture system with DO-stat, the cell growth and the productivity were superior to those of the system without the DO control. Also, almost total plasmid stability was maintained in the two-stage continuous culture system. Modified M9 medium was selected as an optimum feeding medium for the fed-batch process, and the optimum C/N ratio determined to be 2:3. The optimum feeding rate was $0.6g/\ell/hr$ for a constant feeding strategy in semi-fed batch system. When the feeding medium was fed by pulsing, it was observed that more frequent pulsing resulted in improved cell growth. The linear feeding method was the most efficient of the various feeding methods tested. Finally, high cell density culture using a two-stage cyclic fed batch system with pH-stat was tried because the linear feeding method showed limitations in terms of obtaining high cell densities, and a cell density of $54 g/\ell$ was achieved. It was concluded that the two-stage cyclic fed batch system was the most efficient system for high cell density culture of the systems tested. However, productivity improvements were lower than expected due to the extremely high accumulations of acetate, although the low levels of residual glucose were maintained.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.6
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• pp.753-757
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• 1999

The growth of rotifer, Brachionus rotundiformis was evaluated at different culture conditions. Rotifer was fed on condensed freshwater Chlorella. The productivity of rotifer in the high density culture system was compared to that of rotifer in the batch culture system, in which rotifer was fed on baker's yeast. The growth rate of rotifer increased as temperature increased in the culture system supplied with air or oxygen gas. The maximum density of rotifer in the culture systems supplied with air was in range of 16,300$\~$17,000 ind./ml at $24^{\circ}C$. In the culture systems supplied with oxygen gas, it ranged 26,300$\~$30,500 ind/ml at $28^{\circ}C$. When the concentration of dissolved oxygen in the culture system supplied with air reached to below 1 ppm or when the concentration of undissolved ammonia in the culture system supplied with oxygen gas reached 16.6$\~$22.6 ppm, the growth of rotifer decreased. When oxygen gas was supplied and pH was adjusted to 7, the maximum density of rotifer reached to 43,000 ind/ml at $32^{\circ}C$. The production costs for 10 billion rotifer in the high density culture and batch culture were 693,000 and 961,000 won, respectively. Therefore, it was concluded that the productivity of rotifer in the high density culture was higher than that in a batch culture.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.338-341
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• 2000

An internal membrane bioreactor system was employed for continuous succinic ac id production from glucose in order to prove its performance and practicality. Succinic acid-producing Anaerobiospirillum succiniciproducens required more $CO_2$ for the proper growth and succinic acid production in cell recycled continuous culture than in batch culture. The maximum productivity obtained in cell recycled continuous culture was about 3.3 g/L-h which was ca. 3.3 times higher than that obtained in batch culture.

• Journal of Microbiology and Biotechnology
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• v.16 no.5
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• pp.673-677
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• 2006

A perfusion culture of transgenic Nicotiana tabacum cell suspensions, transformed to express recombinant glucuronidase (GUS), was successfully performed in a 5-1 stirred tank bioreactor. With 0.1 $day^{-1}$ of perfusion rate, the maximum dry cell weight (DCW) reached to 29.5 g/l in 16 days, which was 2.1-fold higher than the obtained in batch culture (14.3 g/l). In terms of the production of GUS, the volumetric activity could be increased up to 12.8 U/ml by using perfusion, compared with 4.9 U/ml in batch culture. The specific GUS activities in both perfusion and batch cultures were maintained at similar levels, 200-400 U/g DCW. Consequently, a perfusion culture could be a good strategy for the enhanced production of recombinant proteins in a plant cell culture system.

• Journal of Microbiology and Biotechnology
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• v.19 no.12
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• pp.1603-1611
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• 2009

High-cell-density cultivation of yeast was investigated using the agricultural waste products corn steep liquor (CSL) and molasses. The Saccharomyces cerevisiae KV-25 cell mass was significantly dependent on the ratio between C and N sources. The concentrations of molasses and CSL in the culture medium were statistically optimized at 10.25% (v/v) and 16.87% (v/v), respectively, by response surface methodology (RSM). Batch culture in a 5-l stirred tank reactor using the optimized medium resulted in a cell mass production of 36.5 g/l. In the fed-batch culture, the feed phase was preceded by a batch phase using the optimized medium, and a very high dried-cell-mass yield of 187.63 g/l was successfully attained by feeding a mixture of 20% (v/v) molasses and 80% (v/v) CSL at a rate of 22 ml/h. In this system, the production of cell mass depended mainly on the agitation speed, the composition of the feed medium, and the glucose level in the medium, but only slightly on the aeration rate.

• KSBB Journal
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• v.8 no.5
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• pp.478-482
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• 1993

Perfusion culture was conducted in Celligen perfusion culture system using a self-constructed hybridoma cell and low serum medium. The culture system employed hollow fiber to separate cells from the culture broth. Maximum cell density of $2.1\times10^7$ ce11s/m1, 10 times higher than in batch culture, could be achieved. Concentration of monoclonal antibody (MAb) was 4 times higher and production rate at maximum feed rate was 9 times higher than in batch culture. Glucose concentration was very important for the cell growth and MAb production. When glucose concentration was below 1g/l, i. e. 0.5~0.9g/l, specific MAb production rate decreased but cell concentration still increased. As the glucose concentration goes above 1g/l, specific MAb production rate increased and remained at maximum value at more than 1.5g glucose/l. The maximum value of the specific Mab production rate was similar to that of batch culture.

• Journal of Microbiology and Biotechnology
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• v.10 no.3
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• pp.321-326
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• 2000

To examine the feasibility of the long-term production of the human granulocyte colony stimulating factor (hG-CSF) using the $_{L}-arabinose$ promoter system of Escherichia coli, flask relay culture and cyclic fed-batch culture were performed. In the flask relay culture, it was found that the pismid was maintained stably up to about 170 generations in an uninduced condition, whereby the cells could also maintain the capability of expressing hG-CSF expression were maintained stably up to at least 100 generations. In contrast, in the cyclid fed-batch culture, segregational plasmid instability was observed within about 4 generations after induction, even though the cell growth and hG-CSF production reached their maximum balues, 78.0 g/l of dry cell weight and 7.0 g/l of hG-CSF, respectively. It would appear that, when compared to the flask relay culture, the high-cell density and high-level expression of hG-CSF in the cyclic fed-batch cultrure led to the segregational plasmid instability; in other words, a severe metabolic burden existe on the cells due to the high-level expression of hG-CSF. Accordingly, based on these long-term cultures, the segregational and structural plasmid instability was observed and a strategy to overcome such problems could be designed.