• 한국약용작물학회지
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• 제12권5호
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• pp.372-377
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• 2004

Scopolia parviflora Nakai, a rare and endangered species, is the sole plant producing tropane alkaloids (TA) among the Korean native species. In order to enhance TA productivity the SP72 root line was selected by screening 100 of root line, and the optimal culture media for root growth and TA production were investigated with the SP72 roots. Based on the several media, SH and 2B5 medium were determined as growth medium and White and NN medium as production medium. Among the four combinations of two-stage culture, 2BN (2B5 as growth medium plus NN as production medium) showed more enhanced root growth and TA production as compared with production media of White and NN medium and growth media of SH and 2B5 medium, respectively. However, bubble column bioreactor (BCB) cultures applying two-stage culture did not reveal the effective results despite of the each successful operation of two-stage culture in conical flasks and BCB cultures.

• 한국미생물·생명공학회지
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• 제2권1호
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• pp.29-35
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• 1974

We have studied the applicability of the principles and inherent advantages of the two-stage dontinuous uclture technique to an enzyme process for the purpose of improving and optimizing the productivity of 3-ketosteroid-delta-1-dehydrogenase. By using a two-stage continuous culture system, the growth st ageand enzyme produdtion stage are separated. In each stage an optimal set of toperaing conditions was determined, and this was tested for feasibility for the period of 10 days. During this period, at least 70％ of the maximum enzyme productivity could be maintained. The important design parameters studied are: (1) optimal specific growth rate in the first stage which corresponds to the maximal cell productivity, (2) the optimal dilution rate in the second stage which in turn determines the size of second stage fermentor and the mean residence time of cells in the second stage, (3) cell concentration in both stages, add (4) the specific enzyme productivity and enzyme productivity of the second stage. In addition, by using two-stage continuous culture system we have been able to reduce or eliminate the effect of catabolite repression due to high medium concentration and the adverse effect of the solvent used to dissolve the inducer. We have found the balance between the opposing effects of induction and repression in the second stage judging from the observation that the enzyme productivity goes through a maximum.

• International Journal of Advanced Culture Technology
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• 제7권4호
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• pp.96-103
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• 2019

In this paper, we propose two-stage multichannel architecture for oyster product management system, called cloud stage and agent stage. There are two communication channels at each stage. In cloud stage, the embedded system in the smart scale communicates with the server through two channels, Ethernet or 3G/LTE mobile communication. In agent stage, PCs and smart phones called agents communicate with the server also through Internet and 3G/LTE mobile communication. Compared with previous system in which the amount of the oyster produced in oyster workplaces could be monitored only at the console of only one oyster main server, developed system makes it possible to monitor the amount of produced oyster at several PCs (or smart phones). In addition to the amount of oysters produced at all oyster workplaces the environment of oyster workplaces such as temperature and humidity can be monitored on agents to judge the freshness. Two-stage architecture with multiple channels makes it possible to monitor the amount of oyster product and environment of the oyster workplace at any place in real time.

• KSBB Journal
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• 제15권5호
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• pp.489-496
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• 2000

고농도 유전자 재조합 대장균을 이용하여 pyruvate dehy-drogenase complex-E2 특이성 인간 모노클론 항체의 Fab 부분을 효율적으로 생산하기 위해 회분식, 이단 연속식, 반 유 가식, two-stage cyclic fed-batch 동 여러 가지 배양 방법이 조사되었다. 먼저 플라스미드 안정성 문제를 극복하기 위해 growth stage와 production stage를 분리하는 two-phase 회분식 배양과 이단 연속식 시스템을 시도하였다 그 결과 two-phase 회분식 배양보다는 이단 연속식 배양에서의 세포농도와 항체 생산성이 우수하였다 또한 이단 연속식 배양에서의 세포 성 장과 항처l 생산성은 용존산소를 제어한 경우가 그렇지 않은 경우보다 월등하게 높았다. 그리고 plasmid 안정성에 있어서 는 실험기간 내에 거의 100%를 유지하여 높은 안정도를 보 여주었다. 유가식 공정에 적합한 공급 배지로 변형된 M9 배 지가 최적배지로 선정되었고 이 배지 중 최적의 CjN 비율을 조사한 결파 2:3으로 결정되었다. 반 유가식 시스템에서 constant feeding 전략을 사용할 경우 최적 공급속도는 $0.6g/\ell/hr$이었다. 또한 pulse에 의해 공급배지를 공급할 경우에는 총 공급 량이 같을 경우 소량으로 자주 공급해 주는 것이 공급배지를 한꺼번에 많은 양을 공급해주는 것 보다 바람직하였다. 여러 가지 feeding 전략을 조사해 본 결과 linear feeding 방법이 가장 효과적이었다. 하지만 linear feeding 방법마저도 고농도 세포배양에 한계가 있었기 때문에 pH-stat 방법을 이용한 two-stage cyclic fed-batch 시스템을 시도하여 $54 g/\ell$의 세포 를 얻을 수 있었다. 따라서 이 방법이 일단 생산성 향상을 위한 세포의 고농도 배양에는 조사한 여러 배양 시스템 중에 가장 효율적인 시스템임올 알 수 있었다 하지만 이 시스템 에서 포도당을 낮은 level로 유지할 수 있었으나, 초산의 과도한 축적으로 항체 생산성의 향상은 예상에 비해 크지 않았다.

• Journal of Microbiology and Biotechnology
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• 제11권5호
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• pp.739-748
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• 2001

A self-organizing fuzzy logic controller using a genetic algorithm is described, which controlled the glucose concentration for the enhancement of flavonoid production in a fed-batch cultivation of Scutellaria baicalensis G. plant cells. The substrate feeding strategy in a fed-batch culture was to increase the flavonoid production by using the proposed kinetic model. For the two-stage culture, the substrate feeding strategy consisted of a first period with 28 g/I of glucose to promote cell growth, followed by a second period with 5 g/I of glucose to promote flavonoid production. A simple fuzzy logic controller and the self-organizing fuzzy logic controller using a genetic algorithm was constructed to control the glucose concentration in a fed-batch culture. The designed fuzzy logic controllers were applied to maintain the glucose concentration at given set-points of the two-stage culture in fed-batch cultivation. The experimental results showed that the self-organizing fuzzy logic controller improved the controller\`s performance, compared with that of the simple fuzzy logic controller. The specific production yield and productivity of flavonoids in the two-stage culture were higher than those in the batch culture.

• 생약학회지
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• 제17권1호
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• pp.7-11
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• 1986

The young leaf of Perilla species was cultured by two stage culture system using the medium containing mevalonic acid lactone. The growth rate and productivity of essential oil of callus were increased. The essential oil from intact plant and callus was also analysed. Sesquiterpene hydrocarbons and one sesquiterpene alcohol were identified in essential oils of callus.

• 한국수산과학회지
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• 제50권2호
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• pp.160-168
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• 2017

Optimizing the conditions for stem cell culture is an essential prerequisite for the efficient utilization of stem cells. In the culture of fish embryonic stem cells (ESCs) or ESC-like cells, embryo extracts are important for stable growth, but there is no rule for determining the developmental stage of the embryos used to obtain extracts. Therefore, this study investigated the effects of the developmental stage of extract donor embryos on the culture of Oryzias dancena ESC-like cells. O. dancena ESC-like cells were cultured in different media containing each of four types of embryo extract depending on the developmental stage of the extract donor embryos. Growth, morphology, colony-forming ability, alkaline phosphatase (AP) activity, and embryoid body (EB) formation of the cells were investigated. While the developmental stage of the extract donor embryos did not influence the growth, morphology, AP activity, or EB formation of ESC-like cells, colony-forming ability was affected and the pattern of the effects differed completely between the two ESC-like cells investigated. These results suggest that the developmental stage of extract donor embryos should be selected carefully for the culture of ESC-like cells, according to the research purpose and type of cell line.

• Journal of Microbiology and Biotechnology
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• 제14권5호
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• pp.932-937
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• 2004

In situ extraction by organic solvent was studied in order to improve the recovery yield of hydrocarbon from the culture of Botryococcus braunii, a green colonial microalga. When the solvent mixture of octanol as an extractive solvent and n-octane as a biocompatible solvent was added to a two-phase column, the algal growth was seriously inhibited, even at a low concentration of polar octanol. Therefore, a two-stage cell-recycle extraction process was proposed to improve the contact area between the organic phase and the aqueous phase. The hydrocarbon recovery with in situ cell-recycle extraction showed a three-fold increase (57% of cell) in yield over that with two-phase extraction. In addition, over 60% of the hydrocarbon could be recovered without serious cell damage by downstream separation when this process was applied to the culture broth after batch fermentation.

• 한국미생물·생명공학회지
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• 제20권6호
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• pp.668-676
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• 1992

Alcaligenes eutrophus에 의한 poly-Beta-hydroxybuty-rate(PHB) 생산을 최적화하기 위해서 1단계에서는 성장을 최대화하고, 2단계에서는 생산을 최대화하는 이단 유가식 배양을 채택하였다. 성장단계에서 탄소원과 질소원의 최적농도는 회분식 배양 모델로부터 결정되었는데, 각기 16.64g/l와 0.54g/l이었다. 생산단계에서 질소원의 경우, 유가식 배양실험을 통해 0.07g/l가 최적인 것으로 나타났다. 탄소원의 최적농도는 모사로부터 결정되었고 이때 최적 전환시간도 함께 결정되었다. 생산공정의 제어를 위하여 시간지연을 고려한 적응비례 되먹임제어를 이용한 결과, 탄소원 농도 및 질소원농도를 최적의 상태로 유지시킬 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제24권6호
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• pp.781-787
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• 2014

2-Ketogluconic acid production by Klebsiella pneumoniae is a pH-dependent process, strictly proceeding under acidic conditions. Unfortunately, cell growth is inhibited by acidic conditions, resulting in low productivity of 2-ketogluconic acid. To overcome this deficiency, a two-stage fermentation strategy was exploited in the current study. During the first stage, the culture was maintained at neutral pH, favoring cell growth. During the second stage, the culture pH was switched to acidic conditions favoring 2-ketogluconic acid accumulation. Culture parameters, including switching time, dissolved oxygen levels, pH, and temperature were optimized for the fed-batch fermentation. Characteristics of glucose dehydrogenase and gluconate dehydrogenase were revealed in vitro, and the optimal pHs of the two enzymes coincided with the optimum culture pH. Under optimum conditions, a total of 186 g/l 2-ketogluconic acid was produced at 26 h, and the conversion ratio was 0.98 mol/mol. This fermentation strategy has successfully overcome the mismatch between optimum parameters required for cell growth and 2-ketogluconic acid accumulation, and this result has the highest productivity and conversion ratio of 2-ketogluconic and produced by microorganism.