• KSBB Journal
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• v.15 no.2
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• pp.139-144
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• 2000

Automatic addition of glucose and ammonium to an alcohol distille깨 wastewater and the control of them at low $\infty$ncentration were tried to efficiently increase the cell concentration of a baker's yeast c비tivated in that wastewater. Added g glucose was indirectly controlled to less than 116 mg/L by a method which used DO as control parameter. Ammonium was a automatically measured and controlled within the range of 7.0~27.7 mM by a homemade on-line system which adopted FIA a as measurement method. Maximum specific growth rate and biomass yield to glucose were $0.21 hr^{-1}$ and about 0.78 g/g, w which were significantly increased values in contrast to those of an experiment without ammonium control. Biomass yield to a ammonium was 11.3 gIg. Cell cone엉ntration could be increased from 2.6 g/L to 18.5 g/L by the add ion of glucose and a ammonium.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.65-68
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• 2000

To understand the antioxidative mechanism in plant cell cultures, we investigated the levels of antioxidant enzymes and low molecular antioxidants in 100 cell lines derived from different plant species. SOD and POD activities in plant cell lines were significantly higher than intact plants. The cell lines from sweet potato (Ipomoea batatas) and cassava (Manihot esculeanta) showed the highest POD and SOD activities, respectively, suggesting that the cell cultures of sweet potato and cassava are good biomaterials for the mass production and molecular study of antioxidant enzymes. The average ascorbate content in plant cell lines was several hundred times lower than intact plants, whereas the glutathione content was 2-3 times higher than plants. Interestingly, the ratio of reduced and oxidized ascorbate and glutathione was different from plant species. In conclusion, the results strongly suggest that plant cell cultures are good biomaterials for the study of antioxidative mechanism and the production of useful components including antioxidants.

• Proceedings of the Korean Society for Bio-Environment Control Conference
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• 2001.11a
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• pp.174-175
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• 2001

미나리 묘를 대량으로 생산하는데 있어서 체세포배를 이용하는 방법이 매우 유리하다. 체세포배는 계절에 영향을 받지 않고 좁은 공간에서 한꺼번에 많은 량의 묘를 생산할 수 있다. 또한 생물반응기 등을 이용하여 대량 생산할 수 있으면 실생묘를 대체할 수 있을 것으로 생각된다. 배양기 내의 공기 환경의 영향은 삼각플라스크를 이용한 소규모 배양에서는 문제가 되지 않았지만 배양기가 커짐에 따라 공기 환경의 변화도 커져 체세포배의 발생과 발아에 영향을 미친다. 체세포배를 배양하는 기간은 배발생과 발아로 나누어, 세포에서 배가 발생하는 처음 2주 동안에는 밀폐 환경, 즉 환기가 거의 되지 않는 환경이 유리하지만 이후의 발아 동안에는 산소를 많이 요구한다. 삼각플라스크를 이용한 소규모 배양에서는 aluminum foil을 플라스크의 뚜껑으로 하여, 배지를 교체할 때를 제외하고는 배발생과 발아가 진행되는 전 배양 기간 동안 이를 유지한다. 배가 발생하는 초기 2주의 경우는 이 밀폐 환경이 발아를 촉진시킨다. 그러나 배가 발아하여 정상 식물체로 발달하는데는 유리하지 않다. 체세포배는 발아할 때 산소를 많이 필요로 하며 에틸렌이 많이 축적되면 발아율이 낮아지며, 적절한 공기 환경이 주어지면 체세포배 발아가 동조화되어 균일한 묘를 얻을 수 있고 수확시기도 예측할 수 있다. 본 실험에서는 배가 발아하는 기간 동안 플라스크 내의 공기 환경을 다르게 하기 위해 플라스크를 막는 뚜껑의 소재를 달리하여 배양한 후 발아율을 측정하였다. 또한 가장 효율적인 환기량을 구명하기 위하여 인위적으로 플라스크에 공기를 넣어 강제 환기시키는 실험을 수행하였다. 미나리 cell은 처음 8일까지는 생장을 하지 않다가 이후 급속히 생장을 시작하여 35일 정도까지는 생장을 하다가 다시 생장이 둔화되었다. 밀폐시킨 삼각플라스크에서 자라는 Cell은 상태도 좋지 않고 전반적인 증식량도 적었다. Cell은 환기정도에 민감한 것으로 판단되며 삼각플라스크에서 약 35일 정도의 생장 주기를 가지는 것으로 사료된다. 배양 3주까지는 플라스틱 뚜껑으로 밀폐시킨 bottle에서 가장 많은 체세포배를 얻었다. Air filter를 달아 2일 마다 신선한 공기를 넣어 주었을 때는 배의 발달이 많이 늦어져 배양 3주째에 다른 처리보다 배의 수가 훨씬 적었다. 체세포배가 발달하는 동안에는 산소를 많이 요구하지 않으나 성숙하는 동안에는 산소를 많이 요구하는 것으로 생각된다.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.88-88
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• 2003

본 연구는 배란후 48시간째 회수된 난소로부터 황체세포를 체외배양 후 progesterone 분비에 대한 LH, IGF-1 및 TGF$\beta$첨가효과에 대해 검토하였다. 축산기술연구소에서 사육중인 돼지(체중 145$\pm$kg) 12두로부터 발정을 유도시켜 배란후 약 48시간째 도축하여 난소를 회수하였다. 회수된 난소로부터 황체를 분리하여 세절한 후 0.25% collagenase용액(0.025mg DNase, 50mM EDTA, 50mM Dithiothreitol)으로 37$^{\circ}C$의 진탕수조에서 30분간 배양하여 황체세포를 분리 회수하였다. 회수된 황체세포는 D-MEM 용액(GIBCO, 10% FCS와 antibiotics 첨가)으로 2회 세정하여 1$\times$$10^{6}$live cell/$m\ell$이 되도록 희석한 후 24 well culture plate(Coming, New Tork 14831)에 분주하여 $CO_2$ 배양기($CO_2$: 5%)에서 12시간 간격으로 배양액을 회수하였으며 48시간까지 배양하였다. 배양액 1$m\ell$당 LH, IGF-I 및 TGF$\beta$1을 단독 혹은 공배양하여 회수된 배양액내의 progesterone 농도를 RIA법으로 분석하였다. 돼지황체세포는 배양후 24시간째에 높은 농도의 progesterone이 측정되었으며 LH를 첨가한 대조구에 비해 유의적으로 높은 progesterone이 측정되었다. 또한 IGF-1 과 TGF$\beta$1을 첨가한 군에서도 높은 농도의 progesterone이 측정되었다 그러나 TGF$\beta$ 혹은 IGF-I 단독으로는 대조구의 결과와 큰 차이가 없었다 따라서 돼지 황체세포에서의 progesterone 분비는 TGF$\beta$ 및 IGF-I 그리고 LH의 황체세포의 progesterone 분비기능을 촉진하는 것으로 나타났다.

• 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.21 no.2
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• pp.103-109
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• 2006

For the study of Hericium erinaceum as a useful functional foods and materials, liquid cultivation under two different bioreactors(air-lift fermenter and jar fermenter) which was not studied systematically until now, was conducted as a method of mass cultivation for H. erinaceum. A batch cultivation in an air-lift fermenter and a jar fermenter was examined for enhancing the productivity because of small amounts of mycelial weight and slow growth in case of a liquid culture for H. erinaceum. We found that air lift fermenter system was more effective than jar fermenter for mycelial production of H. erinaceum, and mycelial morphology was a critical factor of the growth. By scale-up and cultivation based on morphological analysis, the conditions for mass production with 30 L and 500 L jar fermenter was 200 and 150 rpm of agitation speed at 1 vvm of aeration rate, respectively, and mycelial dry weight under these conditions was enhanced to about $13{\sim}14g/L$.

• Korean Journal of Plant Resources
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• v.21 no.5
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• pp.368-373
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• 2008

In order to investigate the effects of low temperature pretreatment of floral bud and plant growth regulators on anther-derived callus and shoot differentiation, anthers were cultured on 1/2 MS medium supplemented with 2,4-D, NAA, BA and TDZ. This plant depends on the plant growth regulators, for these anthers couldn't respond on 1/2 MS medium without plant growth regulators. 2,4-D was a prerequisite substance in this experiment, especially 52.6% of callus formation on MS medium with 2.0mg/L 2,4-D alone. However, the optimum medium was on 1/2 MS medium with 0.1 mg/L 2,4-D and 1.0mg/L BA for continuous growth and shoot differentiation from the anther. Calli derived from on MS medium with 2.0mg/L 2,4-D transferred to the 1/2MS medium with TDZ and BA. TDZ were less superior to BA, only one anther could produce shoot on MS media with 1.0mg/L TDZ. On the other hand, when the calli transferred to the medium with 3.0mg/L BA, adventitious shoots were proliferated, subsequently, regenerated shoots elongated from the embryogenic calli. After floral buds of one week before anthesis were incubated at $5^{\circ}C$ refrigerator for eight or fifteen days, anthers seperated from floral buds were cultured on 1/2MS medium supplemented with 0.1mg/L 2,4-D and 1.0mg/L BA. Callusing and shoot differentiation on anthers from treated at $5^{\circ}C$ for eight days were more effective than those of fifteen days or control.

• Korean Journal of Breeding Science
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• v.40 no.1
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• pp.31-38
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• 2008

This experiment was carried out to improve the anther culture efficiency of barley (Hordeum vulgare L.). Callus induction rates from anther cultures of the five domestic naked barley and four unhulled varieties ranged from 0 to 5.6%, and plant regeneration rate to callus was 30.4% in the donor plants grown in a greenhouse during winter, among which the green plant regeneration rates ranged from 0 to 4.4%. Plant regeneration rate was 30.4% in the donor plants grown in a greenhouse during winter, whereas 21.3% in the normal field condition in spring. In addition, callus induction rates were 19.2% in plants grown in a normal field and 7.2% in drought-stressed condition, respectively. Being Considered the anther culture efficiency affected by the sampling time, the optimum sampling stage of anthers was 3~4 days before heading when the length between the 1st and 2nd auricles reaches 5 to 10 cm and at the uninucleate of pollen which the tip of the 2nd auricle aligns with the middle of panicle in the leaf sheath. Best callus induction rates came from the anthers stored at $4^{\circ}C$ for 3 weeks in a 10 to 15 cm diameter polyethylene bag with 5 to 10 panicles and Duwonchapssalbori and Saessalbori showed the higher induction rate of 4.8% and 1.7%, respectively.

• Korean journal of food and cookery science
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• v.18 no.1
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• pp.43-50
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• 2002

This study was carried out to evaluate the quality attributes of frozen soy yogurts prepared by freezine soy yogurts, which are made of different types of Bifidobacteria (B. bifidum, B.breve, B. infantis) and oligosaccharides (fructooligosaccharides, galactooligosaccharides, isomaltooligosaccharides) containing $\alpha$-chymotrypsin treated soy protein isolate were evaluated in terms of overrun, melt-down quality, changes in the total number of Bifidobacteria after freezing, and sensory evaluation. The quality attributes of soy yogurts were also evaluated in terms of changes in the number of viable cells of Bifidobacteria in soy yogurts after incubation at 37$\^{C}$, pH 3.0 for 90 min, water holding capacity, and viscosity. The overrun of frozen soy yogurts fermented by B. bifidum showed the hiehest value but those fermented by B. infantis showed the lowest, while the melt-down quality of soy yogurts were vice versa. The total numbers of Bifidobacteria after freezing for 30 min in ice cream maker showed more than 10$\^$9/ CFU/ml. In sensory evaluation, all $\alpha$-chymotrypsin treated frozen soy yogsurt showed little beany flavor. In sour, sweet, and bitter tastes and mouth feel, the frozen soy yogurts fermented by B. bifidum evaluated better but those fermented by B. infantis evaluated worse. Also in the overall quality, the frozen soy yogurts fermented by B. bifidum were evaluated desirable but those fermented by B. infantis were evaluated undesirable. The water holding capacity and viscosity of soy yogurts fermented by B. bifidum showed the highest values but those fermented by B. infantis showed the lowest values. The total numbers of Bifidobacteria of all soy yogurts decreased from 10$\^$9/ CFU/ml to 10$\^$8/ CFU/ml after incubation at 37$\^{C}$, pH 3.0 for 90 min.

• Microbiology and Biotechnology Letters
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• v.6 no.2
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• pp.75-80
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• 1978

This study was conducted to investigate the condition of enhancing glutathione content of yeast. Rhodotorula glutinis in various kind of yeasts produced high content of glutathione and dry cell by cultivating for 48 hours at 30'C and pH 6.0 on reciprocal shaker. In order to enhance the glutathione content, as 0.7％ of amino acid was applied into the medium. Glutathione was produced high for 36 hours cultivation. When glutamic acid, cysteine and glycine composing the glutathione were added, glutathione content increased to 219 $\mu\textrm{g}$/ml. However the control showed to 73 $\mu\textrm{g}$/ml.