• KSBB Journal
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• v.14 no.2
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• pp.241-247
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• 1999

Experimental studies were carried out to optimize the culture conditions of Corynebacterium diphtheriae for the production of diphtheria toxin. A new media which does not contain any meat digest products was selected. The main ingredient of new medium was enzymatic digests of casein known as NZ-Case. In fermenter experiments, the toxin production was increased with the increase of cell growth. The optimum initial pH of media, air flow rate and agitation speed were 7.0, 0.22, vvm and 400 rpm, respectively. The contents of iron and calcium-phosphate precipitate were important for maximal cell growth and toxin production. The optimum concentration of iron was 0.3 mg/L and calcium-phosphate precipitate could serve in gradual supply of iron to maintain the optimal culture condition which is required for enhanced yield of toxin production. In potency test, the potency of toxoid from fermentor culture was higher than that from static culture. When diphtheria toxin is produced by fermentor culture, it is possible to produce higher levels of toxin and better toxoid quality in terms of safety, yield, productivity and immunity.

• Reproductive and Developmental Biology
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• v.36 no.1
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• pp.21-26
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• 2012

Suspension culture is a useful tool for culturing embryonic stem (ES) cells in large-scale, but the stability of pluripotency and karyotype has to be maintained $in$ $vitro$ for clinical application. Therefore, we investigated whether the chromosomal abnormality of ES cells was induced in suspension culture or not. The ES cells were cultured in suspension as a form of aggregate with or without mouse embryonic fibroblasts (MEFs), and 0 or 1,000 U/ml leukemia inhibitory factor (LIF) was treated to suspended ES cells. After culturing ES cells in suspension, their karyotype, DNA content, and properties of pluripotency and differentiation were evaluated. As a result, the formation of tetraploid ES cell population was significantly increased in suspension culture in which ES cells were co-cultured with both MEFs and LIF. Tetraploid ES cell population was also generated when ES cells were cultured alone in suspension regardless of the existence of LIF. On the other hand, the formation of tetraploid ES cell population was not detected in LIF-free condition, in which MEFs were included. The origin of tetraploid ES cell population was turned out to be E14 ES cells and not MEFs by microsatellite analysis and the basic properties of them were still maintained despite ploidy-conversion to tetraploidy. Furthermore, we identified the ploidy shift from tetraploidy to near-triploidy as tetraploid ES cells were differentiated spontaneously. From these results, we demonstrated that suspension culture system could induce ploidy-conversion generating tetraploid ES cell population. Moreover, optimization of suspension culture system may make possible mass-production of ES cells.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.268-272
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• 2003

Experiments were carried out to investigate the selection of industrial medium and optimization of fermentation process for the production of erythritol by Candida magnoliae SR101. In the batch fermentation, light steep water(LSW) was the best nitrogen source for the industrial production of erythritol. For the optimization of culture condition, the batch culture was performed. When the concentration of LSW was 65 mL/L in the defined medium containing 250 g/L of glucose, 44% of erythritol yield with 110 g/L of erythritol concentration and 0.66 g/L-hr of productivity, respectively were obtained. Two-stage fed-batch culture was performed to improve the volumetric productivity of erythritol. High density cell culture in the growth stage was performed by batch type with 100 g/L glucose and 500 mL/L LSW concentration, respectively. The cell yield was 0.72 g-cell/g-glucose. Productivity of erythritol was increased and concentration of organic acids such as gluconic acid and acetic acid were decreased when initial pH of 6.5 controlled by 28% ammonia water For increasing yield of erythritol, glucose concentration in the production stage was tested. 37% of total erythritol yield with 186 g/L of erythritol concentration and 1.66 g/L-hr of erythritol productivity were obtained when 820 g of glucose powder was directly added for making up 450 g/L of glucose at production stage.

• Journal of Embryo Transfer
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• v.28 no.4
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• pp.373-379
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• 2013

Spermatogonial stem cells (SSCs) developed into sperms through spermatogenesis have been utilized as a useful tool in the field of regenerative medicine and infertility. However, a small number of highly qualified SSCs are resided in the seminiferous tubule of testis, resulted in developing effective in-vitro culture system of SSCs for solving simultaneously quantitative and qualitative problems. Presently, SSCs can be enriched on testicular stromal cells (TSCs), but there are no systematic researches about TSC culture. Therefore, we tried to optimize culture condition of TSCs derived from mouse with different strains. For these, proliferation and viability were measured and compared by culturing ICR outbred or DBA/2 inbred mouse-derived TSCs at 35 or $37^{\circ}C$. In case of ICR strain, primary TSCs cultured at $37^{\circ}C$ showed significantly higher proliferation and viability than those at $35^{\circ}C$ and significant increase of proliferation and viability in sub-passaged TSCs was detected in the $35^{\circ}C$ culture condition. Moreover, sub-passage of primary TSCs at $35^{\circ}C$ induced no significant effects on proliferation and viability. In contrast, in case of DBA/2 strain, significantly improved proliferation were detected in the primary TSCs cultured at $35^{\circ}C$, which showed no significant difference in the viability, compared to those at $37^{\circ}C$. Furthermore, sub-passaged TSCs cultured at $37^{\circ}C$ showed no significant differences in proliferation and viability, compared to those at $35^{\circ}C$. However, with significant decrease of proliferation induced by sub-passage of primary TSCs at $35^{\circ}C$, no significant effects on proliferation and viability were resulted from sub-passage of primary TSCs at $37^{\circ}C$. From these results, culture temperature of primary TSCs derived from outbred and inbred strain of mouse could be separately optimized in primary culture and subculture.

• Journal of Microbiology and Biotechnology
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• v.10 no.3
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• pp.367-374
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• 2000

The composition of dissolved gases and nutrients in a liquid medium were determined for establishment of the optimum conditions for in vitro culture of Helicobacter pylori. A microaerobic condition facored by the organism was prepared by adjusting the partial pressure of the gas, agitation speed, and viscosity of the medium. The gaseous concentrations were controlled by utilizing CampyPak Plus that reduced oxygen while augmenting carbon dioxide. Agitation of the broth facilitated the oxygen transfer to the cells, yet inhibited the growth at high rates. An increase of viscosity in the medium repressed the culture although this variable was relatively insignificant. The chemical constituents of the liquid broth were examined to establish an economic model for H. pylori cultivation. The microbe required a neutral pH for optimum growth, and yet was also able to proliferate in an acidic condition, presumably by releasing the acidity-modulating enzyme, urease. Cyclodextrin and casamino acid were investigated as growth enhancers in place of serum, while yeast extract unexpectedly inhibited the cells. A low concentration of glucose, the unique carbon source for the organism, increased the cell density, yet high concentrations resulted in an adverse effect. Under optimally dissolved gas conditions, the cell concentration in brucella broth supplemented with serum substitutes and glucose reached $1.6{\times}10^8$ viable cells/ml which was approximately 50% higher than that obtained in the liquid medium added with only cyclodextrin or serum.

• KSBB Journal
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• v.25 no.1
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• pp.79-84
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• 2010

In the present study, we isolated a new bacterial strain producing invertase (EC 3.2.1.26) and determined optimized culture condition in flask culture. The strain was identified as Bacilus flexus determined by the 16S rDNA sequencing method. The invertase was produced only in the sucrose medium as the sole carbon source. Potassium nitrate was an adequate nitrogen source for enzyme production, whereas meat peptone showed the highest bacterial growth. Enzyme production was increased about 2-fold when $MgSO_4\cdot7H_2O$ was supplemented to the growth media. The optimum temperature was found to be $30^{\circ}C$ for both enzyme production and bacterial growth. Invertase exhibited pH optima in the range 5.0-6.0 and have a temperature optimum at $40^{\circ}C$, similarly to other invertases found from different microbial sources. Several mineral ions (K and Fe) stimulated the invertase activity, whereas some bioelements (Ag, Mg, and Mn) inhibited enzyme activity. Under the optimized culture condition, the maximum enzyme production (over 250 units/mL) was achieved at 20 h. To the best of our knowledge, this is the first time to report on invertase production by Bacilus flexus.

• KSBB Journal
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• v.30 no.1
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• pp.33-37
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• 2015

Recently, several health benefits of rice wine, makgeolli, were known due to the interest on the traditional Korean liquor and the researches on the rice wine are increasing. Organic acids produced during the process of rice wine fermentation play important roles in the taste and flavor. In this study, we have examined the optimal conditions for lactic acid production in rice koji as a carbon source. Skim milk was also used as a supplementary ingredient for the optimization of lactic acid fermentation. Bacterial growth of Lactobacillus sakei was monitored under this condition. The pH, acidity of the culture and the ethanol tolerance of this bacterium were also tested. Through these experiments, we were able to optimize the growth condition of lactic acid bacteria by the addition of skim milk. This was also able to affect the change of pH, acidity, sugar concentration and alcohol tolerance, which might contribute to the improvement of the quality of rice wine. The optimal condition for the growth was 2 days with 10% (w/v) of skim milk concentration. With these results, it was confirmed that rice koji was an effective carbon source for the growth of lactic acid bacteria.

• Journal of Life Science
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• v.13 no.5
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• pp.705-711
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• 2003

The gellan was produced by Pseudomonas elodea under aerobic condition. In this study, the effects of inoculum size, carbon sources and concentration, nitrogen source, and C/N ratio on the cell growth and the production of gellan were evaluated. The maximum growth of P. elodea and gellan production was obtained at 5% (v/v) of inoculum size and glucose showed best results among 9 carbon sources tested. The maximum specific yield of 2.22 and productivity of $0.03 g/\ell$h were obtained at 1.0% (w/v) of glucose. The maximum gellan production was obtained at medium without ammonium nitrate. This indicates that nitrogen limitation is essential for the production of gellan. The highest cell and gellan production were obtained at 20 of C/N ratio.

• Microbiology and Biotechnology Letters
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• v.34 no.3
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• pp.235-243
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• 2006

The study was performed to identification of producing microbe Non-Cariogenicity Sugar (NCS; Fuc($1{\to}4$) gaINAc($2{\to}6$)NeuAc) with anti-caries activity, and to optimization of production condition. A typical strain which produced the NCS was identified alkalophilic Bacillus sp. S-1013 through the results of morphological, biochemical and chemotaxonomic characteristics and 16S rDNA sequencing. The optimal medium composition for the maximal production of the NCS from alkalophilic Bacillus sp. S-1013 was as follow: soluble starch 30 g, dextrin 15 g, yeast extract 5 g, peptone 10 g, $K_{2}HPO_4$ 2 g in a liter of distilled water. Optimal temperature and pH were 25 and 11.0, respectively. The highest production of NCS was shown 60 hrs cultivation using the optimal medium, and then NCS productivity and dry cell weight of culture broth increased 4.24 and 2.67 time than initial medium, respectively.

• Culinary science and hospitality research
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• v.23 no.4
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• pp.81-92
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• 2017

This research was studied to optimize the recipe of rice pound cake with two concentrations of Euonymus alatus and grape seed oil, using central composite design (CCD). The mixing condition of rice pound cake was optimized by subjecting it to sensory evaluation and mechanical and physicochemical analysis, using response surface methodology (RSM). The results of the mechanical and physicochemical analysis showed significant values for color (lightness, redness, yellowness), texture (hardness, springiness, chewiness, gumminess, cohesiveness), loss rate, volume, specific volume, sweetness, saltiness, moisture content and pH (p<0.05). The results of the sensory evaluation showed significant values for color, flavor, taste, softness, appearance and overall quality (p<0.05). As a result, the optimized compounding ratio was found to be 4.28 g of Euonymus alatus and 33.18 g of grape seed oil.