• KSBB Journal
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• v.6 no.1
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• pp.85-90
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• 1991

Experiments of alcohol fermentation of the yeast,K. fragi1is CBS 1555 were performed to obtain the following results. In these experiments, the initial concentrations of sugar which was composed of inulin and fructose as weight ratio of one to one were 30, 50, 75, 100 and 150g/l and the initial densities of the microorganism were less than 0.5g/l, 10g/1 to 15g/1, and 50g/l. The functional relationship among specific growthrates, sugar concentrations, and alcohol concentrations could be expressed by Aiba-shoda equation and the specific growth rate represented the trend that decreased with increase in the initial concentration of the microorganism. Also, $\mu$max and Ks of Monod's equation could be expressed as the function of initial cell concentration like the following equations. $\mu$max=0.8-0.008X Ks=0.54X+8 In the region that sugar, alcohol and cell concentrations were 10g/1 to 120g/l, 0g/l to 60g/l and 0.5g/l to 50g/l respectively, the differences between the experimental values and the calculated ones for specific growth rate approached to 40% with respect to experimental values at the worst cases, but in most cases, those were distributed in the range of less than 20%.

• KSBB Journal
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• v.17 no.3
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• pp.307-310
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• 2002

In order to obtain high density seed cells for biofiltration, we studied batch and fed-batch culture of P. putida. Studies were carried out to find optimum fermentation conditions such as pH, concentration of glucose and agitation speed. Specific growth rate of P. putida was dependent on agitation speed and a high rpm of 300 was necessary to carry out the efficient aerobic growth of P. putida. Specific growth rate was highest at pH 7. Feeding glucose and yeast extract continuously at the initial growth phase was the most effective way to get high cell density of P. putida.

• KSBB Journal
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• v.13 no.3
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• pp.268-271
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• 1998

Bacteriorhodopsin in the purple membrane (PM) of halobacteria has recently been attracting much attention to be used as a component of molecular electron device and optical computers. In order to increase the productivity of bacteriorhodopsin in high cell density cultures of Halobacterium halobium R1, an internal membrane cell-retention bioreactor system was employed. As a result, the production of cell mass at OD660 of 12 and of bacteriorhodopsin at 125-130 mg/L were obtained using the internal membrane bioreactor system at a dilution rate of 0.066 hr-1. The productivity achieved by the internal membrane system (0.7 mg/L$.$hr) was 3.5-fold higher than that obtained by the corresponding batch cultivations (0.2 mg/L$.$hr).

• KSBB Journal
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• v.13 no.6
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• pp.681-686
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• 1998

Effect of glucose feeding strategy and initial concentration of glucose on Serratia marcescens ATCC 27117 in high cell density fed-batch fermentation was investigated. The final biomasses in batch, constant feeding, constant and exponentially feeding strategy at glucose starvation condition in fed-batch were 1.40, 5,07, 6,93 and 7.60 g/L at 40, 41, 24 and 40 hrs, respectively. Productivities of biomass were 0.035, 0.124, 0.289 and 0.190 g/L$.$h, respectively. As a result, constant feeding strategy at starvation condition was 1.5∼8.6 times higher than other strategies. The relationship between dissolved oxygen and glucose feeding times was good identified in exponential feeding strategy and constant feeding strategy at starvation condition. And high cell density cultivation was obtained when minimal media was used.

• Microbiology and Biotechnology Letters
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• v.23 no.5
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• pp.588-592
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• 1995

Fed-batch fermentation was used to produce the high concentrations of poly-$\beta $-hydroxybutyrate (PHB) and poly-$\beta $-(hydroxybutyrate-co-hydroxyvalerate) (PHB/V). Specific growth rate ($\mu $), yield of cell from glucose (Y$_{x/s}$) were calculated from the two samples in 3 to 5 hours of interval and they were reflected on the determination of glucose feeding rate to maintain the glucose concentration at around 10 g/l in the culture broth. PHB was accumulated after the nitrogen became limited at 60 g/l of dry cell weight by changing ammonia water to 4N-NaOH solution. As results, the final dry cell weight (DCW) of 170 g/l, PHB of 115 g/l were obtained in 50 hours and the overall productivity was 2.4 g/l$\cdot $h. After PHB accumulation, cosubstrate of glucose and propionic acid (PA) was fed to accumulate PHB/V. But, PA feeding rate was decreased from 3 g/l$\cdot $h to 1 g/l$\cdot $h to prevent PA from accumulating to high level in the broth, which is very inhibitory to the cells. As results, DCW, PHB and PHV were 147.5 g/l, 90 g/l and 8 mole % of hydroxyvalerate, respectively.

• Journal of Microbiology and Biotechnology
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• v.26 no.2
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• pp.347-355
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• 2016

An exo-β-D-glucosaminidase (AorCsxA) from Aspergillus oryzae FL402 was heterologously expressed and purified. The deduced amino acid sequence indicated that AorCsxA belonged to glycoside hydrolase family 2. AorCsxA digested colloid chitosan into glucosamine but not into chitosan oligosaccharides, demonstrating exo-β-D-glucosaminidase (CsxA) activity. AorCsxA exhibited optimal activity at pH 5.5 and 50℃; however, the enzyme expressed in Pichia pastoris (PpAorCsxA) showed much stronger thermostability at 50℃ than that expressed in Escherichia coli (EcAorCsxA), which may be related to glycosylation. AorCsxA activity was inhibited by EDTA and most of the tested metal ions. A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of V_max and k_cat values with those of the wild-type enzyme. The three-dimensional structure suggested the tightened pocket at the active site of F769W enabled efficient substrate binding. The AorCsxA gene was heterologously expressed in P. pastoris, and one transformant was found to produce 222 U/ml activity during the high-cell-density fermentation. This AorCsxA-overexpressing P. pastoris strain is feasible for large-scale production of AorCsxA.

• 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.14 no.1
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• pp.90-96
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• 2004

A synthetic defined medium, fortified with amino acids, was developed for the stable production of the kringle fragments of human apolipoprotein(a) (apo(a)), rhLK68. Using a complex rich medium containing yeast extract and a high-cell-density fed-batch culture, the expression level of rhLK68 reached 17% of the total cellular protein, which corresponded to $5\;g\;l^{-1}$ of the culture. To replace the complex media with chemically defined media, several amino acids that positively affect cell growth and gene expression were chosen by a statistical method. The various combinations of the selected amino acids were tested for its fortifying effect on a minimal defined medium. When glutamine only was added, the overall expression level of rhLK68 reached 93% of the complex rich medium increasing the specific expression level by 22.4% and decreasing the cell growth by 24%. Moreover, the addition of glutamine resulted in a 2-fold increase in the concentration of rhLK68 in the culture broth, compared with the minimal defined medium. The synthetic defined media developed in this study could be generally applied to high-cell-density cultures of the recombinant Escherichia coli BL21(DE3), especially for the production of therapeutic proteins that require a strict quality control of the culture media and fermentation processes.

• KSBB Journal
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• v.7 no.4
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• pp.247-251
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• 1992

The effect of biomass concentration of Saccharomyces cerevisiae ATCC 24858 on specific growth rates, biomass yields, ethanol yields and productivity in the batch fermentation of rotary shaker was investigated. The specific growth rate decreased according to the increase in the biomass density and finally became zero at a biomass concentration, 55g/L. The ethanol yield $Y_{p/s}$ represented a constant value, 0.43, regardless of the change of biomass concentrations. However, the biomass yield $Y_{x/s}$ showed a trend to diminish in values with augmentation of biomass density and ultimately to reach zero at 55g/L of biomass concentration. The ethanol productivity increased linearly with biomass concentration so that, in case of initial sugar concentration, 170g/L, the productivity for 55g/L of biomass density rose up to 30g/L$\cdot$hr for all the batch fermentations. And also the ethanol concentration inhibiting completely the growth was verified 95g/L by applying experimental data to Luong's equation.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.233-236
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• 2000

Bacillus thuringiensis (Bt) is the most widely used microbial insecticide in the biological control market. Cultivation of the microorganism to high cell densities offers potential for enhancing the rate of formation as well as the concentration of the desired products In the fermentation broths in bioreactor. With this objective, we developed the new bioreactor incorporating ceramic membrane module for the retention of cell mass. Cell yield and spore formation of Bacillus thuringiensis was improved markedly by adopting this new bioreactor based on glucose -limited feeding operation. It was possible to grow the cell and the heat-resistant spore to above $1.2\;{\times}\;10^{10}\;CFU/ml$ density. With glucose-limited operation, we studied the growth behavior of Bacillus thuringiensis during the cell retention culture. Linear growth of Bacillus thuringiensis was observed under glucose-limited culture, which matched well with simple mathematical model of cell retention culture.