• Journal of Applied Biological Chemistry
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• 제43권2호
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• pp.109-111
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• 2000

Effect of the two-stage operation and cell concentration on indirubin production was investigated using bioreactor culture of Polygonum tinctorium. Two-stage culture was operated successfully for 110 days without any adverse effects on continuous indirubin production. Maximum indirubin concentration was found to be at 80 mg/bioreactor. Initial cell concentration significantly affected indirubin production. The indirubin production at 29.2% PCV was improved by 845%, compared to that at 5% PCV. For high-density bioreactor culture of P. tinctorium, a maximum production rate of 10.2 mg indirubin/L day was obtained. Indirubin recovery for bioreactor operation was also examined using XAD-2, XAD-4, XAD-7, and solid silicon. XAD-4 was 1.6-fold more effective than that for solid silicon in indirubin recovery.

• Journal of Plant Biotechnology
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• 제3권1호
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• pp.1-5
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• 2001

Traditional culture technology of medicinal plants mainly depends on the field culture, which has many problems. With progress of modern culture technology, it has become possible to produce valuable secondary metabolites from medicinal plants. In this paper, we discuss about the pigment and saikosaponin production from too medicinal plants, Carthamus tinctorius and Bupleurum falcatum, through bioreactor culture system. A two-stage bioreactor culture system was established for the production of yellow and red pigments and saikosaponins by cell suspension cultures of Carthamus tinctorius and Bupleurum falcatum. In Carthamus tinctorius, balloon type airlift bioreactors and column type airlift bioreactors were employed for the tell culture and for the pigment production, respectively. The greatest pigment production was obtained on White medium supplemented with 4 mg/L kinetin, high levels of sucrose concentration and photosynthetic photon flux. In Bupleurum falcatum, adventitious roots were cultured in balloon type airlift bioreactors and the root growth was greatest on SH medium containing 5 mg/L IBA and 0.2 mg/L kinetin. HPLC analysis showed that the contents of main active saikosaponins a, c, and d in adventitious roots were almost the same as those in field cultured root.

• 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.

• Journal of Plant Biotechnology
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• 제5권1호
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• pp.1-6
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• 2003

The adventitious root of Panax ginseng C.A. Meyer is regarded as an efficient alternative to cell culture or hairy root culture for biomass production due to its fast growth and stable metabolite production. To determine optimal culture conditions for the bioreactor culture of ginseng roots, experiments have been conducted on physical and chemical factors such as bioreactor type, dissolved oxygen, gas supply, aeration, medium type, macro- and micro-elements, medium supplement during culture period, sucrose concentration, osmotic agents, medium pH and light. Elicitation is a key step to increase ginsenoside accumulation in the adventitious roots but biomass growth is severely inhibited by elicitor treatment. To obtain high ginsenoside content with avoiding biomass decrease, we applied two-stage bioreactor culture system. Ginseng adventitious roots were cultured for 40 days to maximize biomass increase followed by elicitation for 7 days to enhance ginsenoside accumulation. We also experimented on types and concentrations of jasmonate to determine optimal elicitation methods. In this paper, we discussed several factors affecting the root propagation and ginsenoside accumulation. Based on the results obtained from previous experiments we have established large-scale bioreactor system (1 ton-10 ton) for the efficient production of ginseng adventitious roots and bioactive compounds including ginsenoside. Still, experiments are on going in our laboratory to determine other bioactive compounds having effects on diet, high blood pressure, DPPH elimination and increasing memories.

• 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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• 제12권4호
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• pp.617-621
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• 2002

In order to optimize the carrot hairy root culture for SOD production, a fed-batch culture of hairy roots was performed in a bioreactor. Maximum SOD activity was obtained when the hairy roots were transferred to the MS medium containing 110 g/1 concentration of sucrose. By controlling the sucrose concentration (70 g/1 sucrose for growth and 110 g/1 sucrose far production, respectively) In a two-stage fed-batch culture, 29 g/1 of the hairy roots was obtained based on the final dry mass. The volumetrically determined SOD activity and productivity in the fed-batch culture were about 6 times higher than those from the flask culture containing sucrose at 30 g/1 concentration.

• Biotechnology and Bioprocess Engineering:BBE
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• 제7권3호
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• pp.138-149
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• 2002

Plant cell culture provides a viable alternative over whole plant cultivation for the production of secondary metabolites. In order to successfully cultivate the plant cells at large scale, several engineering parameters such as, cell aggregation, mixing, aeration, and shear sensitivity are taken into account for selection of a suitable bioreactor. The media ingredients, their concentrations and the environmental factors are optimized for maximal synthesis of a desired metabolite. Increased productivity in a bioreactor can be achieved by selection of a proper cultivation strategy (batch, fed-batch, two-stage etc.), feeding of metabolic precursors and extraction of intracellular metabolites. Proper understanding and rigorous analysis of these parameters would pave the way towards the successful commercialization of plant cell bioprocesses.

• Journal of Microbiology and Biotechnology
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• 제31권10호
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• pp.1430-1437
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• 2021

Cronobacter sakazakii is an opportunistic pathogenic bacterium found in powdered infant formula and is fatal to neonates. Antibiotic resistance has emerged owing to overuse of antibiotics. Therefore, demand for high-yield bacteriophages as an alternative to antibiotics has increased. Accordingly, we developed a modified mass-production method for bacteriophages by introducing a two-stage self-cycling (TSSC) process, which yielded high-concentration bacteriophage solutions by replenishing the nutritional medium at the beginning of each process, without additional challenge. pH of the culture medium was monitored in real-time during C. sakazakii growth and bacteriophage CS01 propagation, and the changes in various parameters were assessed. The pH of the culture medium dropped to 5.8 when the host bacteria reached the early log phase (OD₅₄₀ = 0.3). After challenge, it decreased to 4.65 and then recovered to 4.94; therefore, we set the optimum pH to challenge the phage at 5.8 and that to harvest the phage at 4.94. We then compared phage production during the TSSC process in jar-type bioreactors and the batch culture process in shaker flasks. In the same volume of LB medium, the concentration of the phage titer solution obtained with the TSSC process was 24 times higher than that obtained with the batch culture process. Moreover, we stably obtained high concentrations of bacteriophage solutions for three cycles with the TSSC process. Overall, this modified TSSC process could simplify large-scale production of bacteriophage CS01 and reduce the unit cost of phage titer solution. These results could contribute to curing infants infected with antibiotic-resistant C. sakazakii.

• Journal of Plant Biotechnology
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• 제1권1호
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• pp.1-7
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• 1999

Large-scale cultures of plant cell, tissue, and organ have been achieved by using BTBB. When different sized BTBBs (5 L, 20 L, 100 L, 300 L, and 500 L) were tested for the culture of yew cells (Taxus cuspidata Sieb. et Zucc.), cell growth increment reached to 94.5% in SCV after 24 days of culture with 30% of inoculation cell density. However, there were some variations in the production of taxol and its derivatives among the BTBBs of different size. Approximate 4 ㎎/l of taxol and 84 ㎎/l of total taxanes were obtained by using a 500L BTBB after 6 weeks of culture. With a 20L BTBB, about 20,000 cuttings of virus-free potatoes (cv. Dejima) could be obtained by inoculating 128 explants and maintaining 8 weeks under 16 hr light illumination. The frequency of ex vitro rooting of the cuttings revealed as more than 99% under 30% shade. By incorporating two-stage culture process consisting of multiple bulblet formation in solid medium and bulblet development in liquid medium, mass propagation of lily through bioreactor seemed to be possible. In the case of 'Marcopolo', the growth of mini-bulblets in BTBB was nearly 10 folds faster than that of the solid medium. Time course study revealed that maximum MAR yield of ginseng (Panax ginseng C. A. Meyer) in a 5 L and 20 L BTBB after 8 weeks of culture was 500 g and 2.2 ㎏, respectively. By cutting the MAR once and/or twice during the culture, the yield of root biomass could be increased more than 50% in fresh weight at the time of harvest. With initial inoculum of 500 g of sliced MAR in a 500 L BTBB, 74.8 ㎏ of adventitious root mass was obtained after 8 weeks of culture. The average content of total ginseng saponin obtained from small-scale and/or pilotscale BTBBs was approximately 1% per gram dry weight. Based on our results, we suggest that large-scale cultures of plant cell, tissue, and organ using BTBB system should be quite a feasible approach when compared with conventional method of tissue culture.

• KSBB Journal
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• 제8권5호
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• pp.495-503
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• 1993

페퍼민트 세포의 회분배양에 의한 monoterpenoid 풍미성분의 생산 가능성과 세포의 생육 및 정유생산 특성을 알아보기 위해 캘러스로부터 얻은 모배양액 을 기포반응기 형태의 생물반응기에 접종하여 배양 하였다. 회분배양 중 접종량, abiotic stress, yeast elicitor, 그리고 2단 배양 등의 배양조건이 세포생육 과 박하정유 생성 빛 조성에 마치는 영향을 조사하 였으며 배지의 sucrose 농도와 세포생육의 kinetics 를 분석하였다. 접종량을 탈리하여 배양한 결과 2.0 %(PCV)를 접종한 경우가 반응기에서 배양이 가장 적당하였는데 배양 10일 만에 5.8g/P 의 세포생육을 얻었으며 0.109g/P 의 박하정유가 생성되였다. Abi-otic stress의 영향을 보기 위해 16시간은 $27^{\circ}C$ 에서 명상태로,8시간은 암상태로 하고 10'C로 온도를 낮추어 배양한 결과 세포의 생육은 떨어졌으나 박하정유 생성량은 0.546g/P 로 높은 수율을 보여 주었다. 사용한 Lin-Staba 배 지 에 100mg/P 의 yeast extract를 elicitor로 첨가했을 때 세포의 생육과 정유 의 생성량이 높았으며 유일하게 menthol이 정유 중 22.5%의 높은 농도로 생성되었다. 배지의 당농도를 1/2로 줄이고 $27^{\circ}C$ , 명조건으로 6일간 배양한 후 1OOmg/P 의 yeast elicitor를 첨가하여 8시간을 암 상태와 lOoe로 저온처리를 실시하여 6일간 배양한 결과 8일 이후에는 세포의 생육이 감소하였고 정유 생성량은 증가하지 않았으며 menthol도 생생되지 않았다. Dry cell yield는 O.38g dry cell/g sugar이 였으며 specific growth rate은 O.25day-1 이었다. 회분배양에 의해 박하정유가 생성됨을 알 수 있었으 냐 menthol이 생성되지 않고 전구체인 pulegone과 p piperitone이 축적되였다. 그러나 특이하게도 yeast elicitor를 첨가한 경우 menthol이 22.5%로 생성되 었는데 이제까지 박하세포 배양에 의해 생성된 men­t thol 함량으로는 최대치를 보여주는 결과이였다.