• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.219-222
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• 2000

Physiological changes of Escherichia coli during the fed-batch fermentation process were characterized in this study. Overall cellular protein samples prepared at the different stage of fermentation were separated by 2-dimensional gel electrophoresis (2-DE), and differently expressed 15 proteins, Phosphotransferase enzyme I, GroEL, Trigger factor, ${\beta}$ subunit of ATP synthase, Transcriptional regulator KDGR, Phosphoglycerate mutase 1, Inorganic pyrophosphatase, Serine Hydroxymethyl-transferase, ${\alpha}$ subunit of RNA polymerase, Elongation factor Tu, Elongation factor Ts, Tyrosine-tRNA ligase, DnaK suppressor protein, Transcriptional elongation factor, 30S ribosomal protein S6 were identified using matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS). When bacterial cells grow to high cell density, and IPTG-inducible heterologous protein is produced, expression level of overall cellular proteins was decreased. According to their functions in the cell, identified proteins were classified into three groups, proteins involved in transport process, small-molecule metabolism, and synthesis and modification of macromolecules.

• KSBB Journal
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• v.8 no.5
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• pp.473-477
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• 1993

Up to now, 10% Fetal Bovine Serum(FBS(V/V)) was added to basal medium for the cultivation of hybridoma. For the cultivation of hybridoma cell line, CH07E02, against colon cancer, serum concentration was reduced to 3% FBS without influence on cell growth and maximum cell concentration. By the addition of cell growth promoting substances-insulin (I), pyruvate (P), oxaloacetate(O), Pluronic F-68(P) and 2-mercaptoethanol(2-ME)-to 1% FBS medium, a cell density higher than that with 1% FBS medium alone was achieved. FBS 3% medium was replaced by very cheap 2% Calf Serum (CS) medium without influence on cell growth rate and concentration. Cells grew vigorously in 0.5% CS＋IPOP medium. This composition was used during suspension culture and exhibited good viability and high specific growth rate.

• Journal of Microbiology and Biotechnology
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• v.21 no.2
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• pp.204-211
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• 2011

To yield high concentrations of protein expressed by genetically modified Escherichia coli, it is important that the bacterial strains are cultivated to high cell density in industrial bioprocesses. Since the expressed target protein is mostly accumulated inside the E. coli cells, the cellular product formation can be directly correlated to the bacterial biomass concentration. The typical way to determine this concentration is to sample offline. Such manual sampling, however, wastes time and is not efficient for acquiring direct feedback to control a fedbatch fermentation. An E. coli K12-derived strain was cultivated to high cell density in a pressurized stirred bioreactor on a pilot scale, by detecting biomass concentration online using a capacitance probe. This E. coli strain was grown in pure minimal medium using two carbon sources (glucose and glycerol). By applying exponential feeding profiles corresponding to a constant specific growth rate, the E. coli culture grew under carbon-limited conditions to minimize overflow metabolites. A high linearity was found between capacitance and biomass concentration, whereby up to 85 g/L dry cell weight was measured. To validate the viability of the culture, the oxygen transfer rate (OTR) was determined online, yielding maximum values of 0.69 mol/l/h and 0.98mol/l/h by using glucose and glycerol as carbon sources, respectively. Consequently, online monitoring of biomass using a capacitance probe provides direct and fast information about the viable E. coli biomass generated under aerobic fermentation conditions at elevated headspace pressures.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.290-293
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• 2001

In this study, media optimization by statistically designed experiments stimulated an increase in cell growth of Bacillus sp. during batch cultivation. Plackett-Surman design method selected 3 components among 7 components of production medium. Box-Behnken design method calculated the optimum concentration of selected components by Plackett-Surman design. In the optimized medium, viable cell number increased 2 times. Addition of antifoam effected the cell growth depending on the type of antifoam Vegetable oil, are a carbon source and an antifoam. increased cell growth and controlled foaming

• KSBB Journal
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• v.14 no.3
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• pp.264-272
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• 1999

Benzoylformate was converted to benzaldehyde by whole cell enzyme from Pseudomonas putida KCTC 1751. We investigated the effect of the composition of the growth medium on th accumulation of benzoylformate decarboxylase in the microbial cell. We prepared a calcium alginate capsule containing Pseudomonas putida cells to develop a reusable whole cell enzyme. Pseudomonas putida cells were inoculated in the capsule and cultured in M1 medium for 1 day followed by cultivation in M3 medium for 3 days. The dry cell density reached 77.75 g/L on the basis of the inner volume of the capsule. The specific activity of encapsulated whole cell benzoylformate decarboxylase was half as high as that of free whole cell enzyme. The activity of encapsulated whole cell benzoylformate decarboxylase was half as high as that of free whole cell enzyme. The activity of encapsulated whole cell benzoylformate decarboxylase decreased 20 % after use for 20 batches and 40% after use for 30 batches. The dry cell density reduced about 10 % after 30 trials.

• 한국생물공학회:학술대회논문집
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• 2005.10a
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• pp.312-315
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• 2005

A process for efficient recycle fed-batch culture was carried out to increase cell mass and spore production by Lactic acid bacteria isolated from Kimchi. A large quantity of cell mass obtained by feeding concentration of sugar in recycle fed-batch culture. When the high density of salt was created that the cell mass was come-down. In this study, cultured in different feeding concentration of sugar conditions. Lactic acid bacteria by recycle fed-batch culture was investigated in 2L working volume of fermenter, obtained the maximum cell mass was 15.17g/L.

• KSBB Journal
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• v.25 no.5
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• pp.471-477
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• 2010

In order to improve bio-ethanol productivity by various cultivation methods in this paper, the culture modes using food wastes, such as batch culture, high-cell-density fermentation, SSF (simultaneous saccharification and fermentation) by fill & draw, continuous culture by fill & draw were performed and their productivities were compared. SSFs by fill & draw were performed by continuous decompression using 1 L evaporator system, and by 10 L bioreactor without decompression. In addition, the continuous cultures by fill & draw mode using SFW (saccharafied food wastes) medium were performed by changes of 40% culture broth with intervals of 12 h (0.03 $h^{-1}$), 6 h (0.07 $h^{-1}$), 3 h (0.13 $h^{-1}$). Consequently, productivities of bio-ethanol were 2.52 g/L-h and 1.30 g/L-h in batch culture and high- cell-density fermentation, respectively. The productivities of SSF by fill & draw showed 2.24 g/L-h and 2.03 g/L-h in continuous decompression with 1 L evaporator and 10 L bioreactor without decompression, respectively. Also, the productivities in continuous culture by fill & draw modes showed 2.02 g/L-h, 4.07 g/L-h and 6.25 g/L-h by medium change with intervals of 12 h, 6 h, and 3 h, respectively. In conclusion, the highest ethanol productivity was obtained in the continuous culture mode by fill & draw with dilution rate of 0.13 $h^{-1}$.

• Microbiology and Biotechnology Letters
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• v.45 no.3
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• pp.185-199
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• 2017

Methane, which is emitted from natural and anthropogenic sources, is a representative greenhouse gas for global warming. Methanotrophs are widespread in the environment and play an important role in the biological oxidation of methane via methane monooxygenases (MMOs), key enzymes for methane oxidation with broad substrate specificity. Methanotrophs have attracted attention as multifunctional bacteria with promising applications in biological methane mitigation technology and environmental bioremediation. In this review, we have summarized current knowledge regarding the biodiversity of methanotrophs, catalytic properties of MMOs, and high-cell density cultivation technology. In addition, we have reviewed the recent advances in biological methane mitigation technologies using methanotrophs in field-scale systems as well as in lab-scale bioreactors. We have also surveyed information on the dynamics of the methanotrophic community in biological systems and discussed the various challenges pertaining to methanotroph-related biotechnological innovation, such as identification of suitable methanotrophic strains with better and/or novel metabolic activity, development of high-cell density mass cultivation technology, and the microbial consortium (methanotrophs and non-methanotrophs consortium) design and control technology.

• KSBB Journal
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• v.5 no.3
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• pp.307-313
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• 1990

A microcomputer-aided fermentation system was constructed for high density fed-batch culture using dissolved oxygen(DO) as a substrate feeding indicator. DO signal was processed prior to aquisition to computer. Agitation speed and oxygen flow rate was changed stepwisely to maintain DO value at a constant level. Agitation speed was controlled by the output signal of D/A converter. Oxygen flow rate was controlled by a flow rate control valve connected to a stepping motor. Substrate was fed with a feeding pump operated by the abrupt increase of DO signal. Methylobacillus sp. SK1 was cultivated to test the system and 16.53g/l of cell density was obtained after 10 hr.

• Applied Chemistry for Engineering
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• v.28 no.3
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• pp.306-312
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• 2017

Changes in the cell growth and lipid accumulation of marine microalga Dunaliella tertiolecta were investigated in response to the combination of different stress factors including the variation of iron supply as a primary stress factor and different options in light irradiation and $CO_2$ supply as a secondary stress factor. High or limited Fe conditions could act as a stress for lipid synthesis. As a secondary stress factor, non-$CO_2$ condition was good for lipid accumulation, but the overall cell growth was sacrificed significantly after a long-time cultivation. Dark condition as a secondary stress factor also favored lipid accumulation and the extent of cell density reduction at the early period in the dark was small compared to other stress conditions. The two-stage cultivation strategy was necessary to maximize lipid production because tendencies of the cell growth and lipid content were not identical under the chosen stress condition. The first stage was for preparing a high cell density under the normal growth-favoring condition and the second stage was the stress condition to induce lipid accumulation in a short time. The short-term (12 h) incubation under the 5X Fe (3.25 mg/L) and dark conditions resulted in the best lipid productivity of 1.44 g/L/d providing 2 g/L inoculum at the second stage.