• Korean Journal of Microbiology
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• v.37 no.1
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• pp.92-99
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• 2001

High-cell-density cultivation of Ralstonia eutopha KHB 8862 by fed-batch fermentation in a 200 l pilot plant was carried out for the mass production of poly(3-hydroxybutyrate) (PHB). After 80 h of cultivation, the dry cell weight (DCW), PHB concentration, and PHB yield from fructose syrup reached 168 g/l, 74%DCW, and 0.27 (w/w), respectively, resulting in a productivity of 1.6 g of PHB/L/h. Based on these results, the PHB production cost from bacterial fermentation was analyzed and economic evaluation was performed. In the case of new investment being implemented or not, the production cost of PHB was US$ 3.15/kg and US$ 2.41/kg, respectively. PHB productivity and PHB yield on a carbon substrate were both important factors to be optimized. The increase of PHB yield on a carbon sources significantly decreased the PHB production cost but the increase in productivity had a relatively slight effect on the decrease in PHB production cost because the cost of carbon sources (37%) for PHB was larger in proportion to total cost than the depreciation cost (17%). These results suggest that the increased PHB yield from carbon sources and the development of new cheaper substrates would be more effective in decreasing PHB production cost than the increase in productivity. It was demonstrated that PHB is not in competition with consumable plastics such as PET in present market. Therefore, it is essential to lower production cost to be used as a bulk product and desirable to develop new application fields for PHB such as biomedical and cosmeceuticals.

• Korean Journal of Microbiology
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• v.35 no.4
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• pp.302-306
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• 1999

Production of biomass by fed-batch culture of Saccharomyces cerevisiae Hansen CBS5926, which is used to treat intestinal disorders, was investigated using ethanol as the sole carbon source. Ethanol was a better carbon source than glucose for high cell density culture of the st-rain since it could decrease the frequency of contamination while increasing the efficiency and final productivity of the fermentation process. Under optimal conditions, 38 g/ℓ of dry cell weight with $2.2{\times}10^{9}$ cfu/㎖ of maximum viable cell count was achieved after 72h cultivation. Freeze-drying of the cultured yeast cells resulted in severe reduction of viability. Of the freeze-drying protectants tested, 20% sucrose and 30% lactose were most effective for the preservation of yeast cells with a viability level of 16.3%. A combination of skim milk and lactose with 20% sucrose(w/v) exerted no synergistic influence upo the viability of the cells during cryopreservation by freeze-drying.

• Journal of Microbiology and Biotechnology
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• v.30 no.1
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• pp.136-145
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• 2020

Chlorella sp. HS2, which previously showed excellent performance in phototrophic cultivation and has tolerance for wide ranges of salinity, pH, and temperature, was cultivated heterotrophically. However, this conventional medium has been newly optimized based on a composition analysis using elemental analysis and ICP-OES. In addition, in order to maintain a favorable dissolved oxygen level, stepwise elevation of revolutions per minute was adopted. These optimizations led to 40 and 13% increases in the biomass and lipid productivity, respectively (7.0 and 2.25 g l^-1d^-1 each). To increase the lipid content even further, 12 h heat shock at 50℃ was applied and this enhanced the biomass and lipid productivity up to 4 and 17% respectively (7.3 and 2.64 g l^-1d^-1, each) relative to the optimized conditions above, and the values were 17 and 14% higher than ordinary lipid-accumulating N-limitation (6.2 and 2.31 g l^-1d^-1). On this basis, heat shock was successfully adopted in novel Chlorella sp. HS2 cultivation as a lipid inducer for the first time. Considering its fast and cost-effective characteristics, heat shock will enhance the overall microalgal biofuel production process.

• KSBB Journal
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• v.7 no.4
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• pp.296-301
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• 1992

A Pseudomonas fluorescens was selected from mushrooms and studied in both batch and fed-batch cultures in order to get maximal biomass concentration. P. fluorescens is an aerobic bacterium and antagonistic to Pseudomonas tolaasii which causes blotch disease on the mushroom cap. P fluarescens and P. tolaasii were identified by Gram staining, gelatin liquefaction, oxidase test, etc. and were characterized by pigment production, temperature sensitivity, salt tolerance and rapid pitting test, etc., Celts of P. fluorescens well in medium containing 30g/L of glucose, whereas the growth was inhibited at the glucose levels at higher than 30g/L. The highest values of specific growth rate and productivity were obtained when using 10g/1 of yeast extract. Optimum concentrations of $NH_4Cl$ and ${(NH_4Cl)}_2SO_4$ for culture were found to be 1.0g/L and 0.1g/L respectively. Optimum concentration of $MgSO_4{\cdot}7H_2O$ used as a sulfursource was 1.0g/L. It was also found that the cell concentrations reached the maximum level when grown on the medium containing 1.0g/L of $KH_2PO_4$ and 0.1g/L of $CaCl_2$. Also, the optimum culture conditions were $30^{\circ}C$ and pH 6.0. Cultivation of P. fluarescens at high dissolved oxygen (DO) concentration led to a decrease of bacterial productivity in batch culture. Maximum productivity was achieved at 40% DO concentration.

• Journal of Life Science
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• v.14 no.2
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• pp.235-238
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• 2004

The possibility for developing multistage culture system to cultivate larvae by computer simulation with basic experiments was investigated. This culture system was composed of 3 stages. At the 1st stage, Chlorella sp. were cultivated and at 2nd stage Chlorella sp. were supplied to rotifer (Brachionus plicatilis), and rotifer were supplied to larvae at the 3rd stage. In this study, Chlorella sp. were cultivated by batch culture to search for the possibility of continuous feeding to rotifer at 2 nd stage. The maximum specific growth rate ($\mu$$_{max}$) of Chlorella sp. at the logarithmic phase was 0.56 [1/day]. Rotifer was cultivated by fed-batch culture at the feeding rate of Chlorella sp., $10^3$, $10^4$, $10^{5}$ and $10^{6}$ [cells/rotiferㆍh] to search for the relation between the feeding rate of Chlorella sp. and the growth rate of rotifer. As the results, the minimum feeding rate of Chlorella sp. was 2.8 ${\times}$ $10^4$ [cells/rotiferㆍday] in the multistage culture system, then the change of rotifer concentration at 2nd stage was simulated by computer. The required amount of rotifer for the growth of larvae was also increased as the growth of larvae. On the 9th day of the culture, the rotifer uptake rate of larvae was 250 [cells/rotiferㆍday]. Based on these basic experiments and results, It was suggested that the possibility of multistage culture system to cultivate larvae with continuous feeding of Chlorella sp. and rotifer.r.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.5
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• pp.269-278
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• 2003

Gibberellic acid (GA$_3$) is a commercially important plant growth hormone, which is gaining much more attention all over the world due to its effective use in agriculture and brewing industry. Industrially it is produced by submerged fermentation technique using Ascomycetous fungus Gibberella fujikuroi. Solid state and immobilized cell fermentation techniques had also been developed as an alternative to obtain higher yield of GA$_3$. This review summarizes the problems of GA$_3$ fermentation such as production of co-secondary metabolites along with GA$_3$, substrate inhibition and degradation of GA$_3$ to biologically inert compound gibberellenic acid, which limits the yield of GA$_3$ in the fermentation medium. These problems can be overcome by various bioprocessing strategies e.g. two - stage and fed batch cultivation processes. Further research on bioreactor operation strategies such as continuous and / or extractive fermentation with or without cell recycle / retention system need to be investigated for improvement in yield and productivity. Down stream processing for GA$_3$ isolation is also a challenge and procedures available for the same have been critically evaluated.

• KSBB Journal
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• v.15 no.2
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• pp.134-138
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• 2000

Addition of carbon and nitrogen source to an alcohol distillery wastewater was tried to increase the cell concentration of a b baker's yeast cultivated in that wastewater. Carbon was found to be primary limiting nutrient and nitrogen secondary limiting o one. Glucose addition increased the cell concentration 1.3 times higher than no addition, and both glucose and $(NH_4)_2S0_4$ a addition did 5.8 times. A fed-batch cultivation by the automatic addition of glucose and ammonium was executed. Added g glu$\infty$se was automatically controlled to low concentration by a method using DO as control parameter. Ammonium was a automatically added as NH40H used as pH $\infty$ntrol agent after initiating glucose addition. By this simple cultivation method t the cell concentration $\infty$내d be efficiently increased from 2.6g/L to 12.0g/L, and maximum specific growth rate and biomass y yield to glu$\infty$se were $0.18hr^{-1}$ and about 0.54g/g respectively. By increasing cell concentration, COD of the wastewater m media could be additionally reduced by about 22%.

• KSBB Journal
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• v.19 no.3
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• pp.192-199
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• 2004

In this work the production of glutathione (GSH) by yeast Saccharomyces cerevisiae and the monitoring of the process were studied. In shaking culture the production of GSH was high at initial pH value of 4 and at temperature of 30$^{\circ}C$. But when L-cysteine was added to the culture medium at the beginning of the cultivation, the productivity of GSH was low. In case 0,5% (v/v) of L-cysteine, glycine and glutamic acid were introduced to the culture medium in the exponential cell growth phase, high concentration of GSH (about 90 mg/L) was produced in the bioreactor. A fed-batch operation with stepwise glucose feeding strategy allowed to produce 102 mg/L of GSH. The cultivation processes were on-line monitored by a 2-dimensional fluorescence sensor. A few off-line data such as cell growth, cystein concentration, phosphate concentration and GSH productivity could be well correlated to the fluorescence intensity of some combinations of excitation and emission wavelengths.

• KSBB Journal
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• v.9 no.5
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• pp.518-524
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• 1994

Serum free medium was developed to cultivate Human Embryonic Kidney cells with Cytodex III microcarriers. The cells normally attached and spreaded on the microcarriers in serum free medium, and grew well by bead to bead processes. 85% of attachment yield was obtained on Cytodex III in this medium, compared to about 93% in 1% serum containing medium. About 90% of the attached HEK cells spreaded after 6 hours of post-attachment periods on the surface of microcarrier. Maximum cell density and scu-PA concentration in a serum free medium were $9.1{\times}10^5$ cells/ml and 1790 IU/ml, respectively, with fed-batch cultivation. Maximum cell density and scu-PA concentration in this medium with perfusion cultivation were $2.5{\times}10^6$ cells/ml and 1820 IU/ml, respectively. The conversion of single chain urokinase type plasminogen activator (scu-PA) into two chain type plasminogen activator (tc-PA) was less than 5% in a serum free medium compared to about 10% in 0.5% serum containing medium.

• Applied Biological Chemistry
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• v.36 no.6
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• pp.488-494
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• 1993

A photosynthetic bacterium strain P17 having high growth rate and assimilating ability of organic acids was isolated from several soil samples, which was identified as Rhodospirillum rubrum. Cultural conditions of the strain P17 were examined for the production of starter culture used in the treatment of organic waste water. The addition of organic acids mixture as carbon source containing 0.2% Na-acetate, 0.1% Na-propionate and 0.2% Na-lactate and 0.1% of yeast extract as growth factor stimulated the cell growth. The maximal cell production was obtained at $30^{\circ}C$, pH 7.0, 2,500 lux of illumination and $50{\sim}100\;rpm$ of agitation. Under the optimal conditions of batch and fed-batch culture systems in a Jar fermentor, 5.17 g/l and 7.93 g/l of cells were obtained after S days of cultivation, respectively. In continuous culture system, the cell productivity was 0.206 g/l/h at a dilution rate of 0.21 $h^{-1}$. When R. rubrum P17 was cultivated in a soybean curd waste water, initial COD level(3,240 mg/l) of the waste water was reduced to 250 mg/l after 4 days of cultivation.