• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.5
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• pp.383-388
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• 2004

The conversion of a cellulose-producing cell ($Cel^+$) from Gluconacetobacter hansenii PJK (KCTC 10505 BP) to a non-cellulose-producing cell ($Cel^-$) was investigated by measuring the colony forming unit (CFU). This was achieved in a shaking flask with three slanted baffles, which exerted a strong shear stress. The addition of organic acid, such as glutamic acid and acetic acid, induced the conversion of microbial cells from a wild type to $Cel^-$ mutants in a flask culture. The supplementation of $1\%$ ethanol to the medium containing an organic acid depressed the con-version of the microbial cells to $Cel^-$ mutants in a conventional flask without slanted baffles. The addition of ethanol to the medium containing an organic acid; however, accelerated the conversion of microbial cells in the flask with slanted baffles. The $Cel^+$ cells from the agitated culture were not easily converted into $Cel^-$ mutants on the additions of organic acid and ethanol to a flask without Slanted baffles, but some portion of the $Cel^+$ cells were converted to $Cel^-$ mutants in a flask with slanted baffles. The conversion ratio of $Cel^+$ cells to $Cel^-$ mutants was strongly re-lated to the production of bacterial cellulose independently from the cell growth.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.253-257
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• 2003

This study was executed to investgate the effects of medium components on liquid culture in the flask culture of Coriolus versicolor. This work was focused on raising the mycelial growth and the polysaccharide production. In order to optimize the medium, different carbon and nitrogen sources were investgated. Glucose and yeast extract were chosen for the production of mycelia and polysaccharide as carbon and nitrogen sources, respectively, in the flask culture. For the mycelia growth and polysaccharide production, the medium contained glucose 20g/L, yeast extract 6g/L, $KH_2PO_4$ 0.46g/L, $MgSO_4.7H_2O$ 0.5g/L. The liquid culture conditions for the mycelial growth were $27^{\circ}C$, 200rpm and working volume 100mL using 250mL flask.

• KSBB Journal
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• v.8 no.4
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• pp.382-389
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• 1993

In order to elucidate the effect of acetate ion on the growth of Escherichia coli, flask and fermentor cultures were performed using M9 and LB media. Acetic acid was secreted at a higher rate under the conditions of high glucose concentration as well as of richer medium, i. e., LB broth. The pH in flask culture could not be controlled as i fermentor and pH decreased with the formation of acetic acid. The inhibition effect of acetic acid was pronounced at a lower pH, and the effective inhibitory concentrations of acetic acid were 2.0g/l for LB flask culture, 4.0g/l for M9 flask culture, and 8.0g/l for M9 fermentor culture. Dialysis flask culture was designed to slowly provide E coli cells with glucose. Solid LB agar was layered under LB liquid medium with the variation of agar concentration and solid volume, The increase in the solid portion in the total volume(agar＋liquid) resulted in the increase of the final cell concentration. This can be ascribed to the fact that the larger solid phase behaves like a larger reservoir for glucose and controls the growth of E. coli with a controlled rate.

• Journal of Life Science
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• v.12 no.5
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• pp.577-583
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• 2002

The growth kinetics and the production of $\beta$-glucuronidase from transgenic tobacco's suspension culture was investigated in the flask culture and a 2.5 L bubble column reactor. The growth of bubble column reactor was similar to that of flask culture. However, in the bubble column reactor, the production of $\beta$ -glucuronidase reached 2850 U/mg (85-fold higher than that of flask culture). In both case, the production level of $\beta$ -glucuronidase was fluctuated, which was resulted from periodical degradation of the protein. Sucrose is important component in plant culture medium. Twice addition of sucrose in bubble column reactor could not improve cell growth, since other components in a medium were already depleted. However, the addition of sugar decreased cell size, which facilitated the operation of bioreactor. The production of $\beta$ -glucuronidase was continuously increased, however final concentration of $\beta$ -glucuronidase was similar to that without sucrose addition.

• KSBB Journal
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• v.11 no.2
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• pp.254-261
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• 1996

Attenuated varicella-zoster virus (VZV) was propagated in human lung fibroblast (HLF) cells Among media tested in this work, DMEM was the best medium for the growth of HLF cells. Because HLF was a normal finite cell line, cell growth late was dependent on the age of HLF cells. When the population doubling level (PDL) was higher than 46, apoptosis of HLF cells started and cell growth rate decreased. The optimum temperature for the cell growth and virus propagation in the T-flask culture was $37^{\circ}C$. In a microcarrier culture system in which Cytodex-3 was used for the VZV propagation in spinner vessels, the yield of plaque forming cells was lower than that in the T-flask culture. The relatively high shear environment near microcarriers was thought to cause the low yield of VZV in the microcarrier culture system.

• KSBB Journal
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• v.13 no.3
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• pp.231-237
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• 1998

An attenuated Japanese encephalitis virus (JEV) clone SA-14-14-2(Vero) was obtained through successive adaptation of a primary cell adapted strain, SA-14-14-2(PDK) to Vero cell, a continuous line of monkey kidney cells. The virus titer reached above the 107 plaque forming unit (pfu) per mL of culture supernatant with 3 passages in Vero cells and was maintained close to this level in the further passages. The optimum temperature for the virus growth was $35^{\circ}C$. Growth pattern of the virus indicated that optimum time for the virus harvest is 4 days post infection and the virus accomplished rapid initial propagation even in medium containing no serum supplement. The roller bottle (RB) system and the spinner flask (SF) system using micro-carrier (Cytodex-1) for the JEV cultivation were explored. When RB, SF, and T-flask culture system were compared, there was no significant difference in virus yield. Furthermore, the results indicated that virus could be harvested multiple times from 3 days to 9 days post infection; neither severe cytopathic effect (CPE) in the infected cells nor the decrease in the titer was observed on duration of 9 days.

• The Korean Journal of Mycology
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• v.27 no.1 s.88
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• pp.1-9
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• 1999

For the practical using of liquid spawn we carried out selection test of nutrient sources, cultural methods and cultural apparatus for liquid spawn production of oyster mushroom(ASTI 2001, ASTI 2018, ASTI 2072, ASTI 2016, ASTI 2070, ASTI 2180, ASTI 2183, ASTI 2042). The optimal temperature and pH range for mycelial growth of Pleurotus species were $25^{\circ}C\;to\;30^{\circ}C$ and 5.5 to 6.5, respectively. The effect of carbon sources, nitrogen sources and mineral salts on the mycelial growth was studied using petridish culture. Generally, the disaccharides and polysaccharides showed good effect for mycelial growth of Pleurotus species, and the polysaccharides were superior to the other classes of carbon sources for mycelial growth. For the mycelial growth of the 8 oyster mushroom stains, soybean flour was superior to the other kinds of nitrogen sources. On the other hand, addition of mineral salts did not affect, and even poor under certain mineral salts, the mycelial growth of the 8 oyster mushroom stains. The brown sugar selected out the carbon source of the agricultural medium. Also the soybean flour selected out the nitrogen source of agricultural medium. In the medium selection, we selected out agricultural optimum medium composed of brown sugar 3%, soybean flour 0.3%, potassium phosphate 0.05%, magnesium sulfate 0.05%. Under the 250 ml erlenmeyer culture, the effects of such factors as the inoculum rate, the working volume, cultural method and flask shapes on the mycelial growth were examined. The optimal inoculum rate and working volume on the mycelial growth of oyster mushroom was 2 mycelial disk (diameter 6 mm) and 100 ml, respectively. The shake flask culture was enhanced the mycelial growth than at the erlenmeyer flask. Pulp form growth of mycelium in the erlenmeyer flask culture was obtained in the culture with glass rod of length 50 mm, diameter 10 mm.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.235-238
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• 2000

The production of polysaccharide from Paecilomyces japonica was studied in the shake flask culture. For the cell growth and the polysaccharide production, the optimum synthetic medium was glucose peptone(YMP) medium. The flask culture conditions for the polysaccharide production were $27^{\circ}C$ and 200rpm with the initial pH 9 for 8days cultivation.

• KSBB Journal
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• v.19 no.3
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• pp.241-244
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• 2004

An automated flow injection analysis apparatus manufactured economically was used in the automatic measurement of E. col growth in shake-flask culture of nutrient broth. The detailed measurement of whole growth was possible at intervals of 10 minutes by the automatic measurement system which adopted the sterilized nutrient broth as a carrier solution. Using distilled water as a carrier solution showed less accurate results than nutrient broth.

• Journal of Plant Biotechnology
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• v.5 no.3
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• pp.187-191
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• 2003

This paper reported the establishment of mass production system of adventitious roots of Eleutherococcus sessiliflorus through the shake flask and bio-reactor culture. Induction of adventitious roots was started from the explants of germinated somatic embryos on half-strength Murashing and Skoog (MS) solid medium. The frequency of adventitious root formation was better in the explants comprising the basal hypocotyl parts than root explants alone. Among the different auxins tested (NAA, IBA and IAA), frequency of adventitious root induction was highest on medium with 0.5 mg/L NAA, and produced $16.3\pm1.9$ roots per explant. In shake-flask culture, deletion of $NH_4NO_3$ of MS medium was effective for induction of adventitious root compared with both full and half-strength MS media. Fresh weight increase of induced adventitious roots was performed well in medium with 0.5 mg/L IBA. When adventitious roots produced in shake-flask culture were transferred to 10-liter bioreactor, 5.5 times of fresh weight increase was gained after one month of culture. HPLC analysis revealed that the amount of eleutheroside E and E1 was higher in in vitro cultured adventitious roots than the 3 year-old field cultivated root barks of Eleutherococcus sessiliflorus. The content of eltutheroside B was much lower in adventitious roots than that of field cultivated one.