• Title/Summary/Keyword: Bioreactor Cultures

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Marked effect of Cuscuta on puerarin accumulation in cell cultures of Pueraria tuberosa grown in shake flasks and a bioreactor

  • Goyal, Shaily;Sharma, Varsha;Ramawat, Kishan G.
    • Plant Biotechnology Reports
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    • v.5 no.2
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    • pp.121-126
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    • 2011
  • Isoflavonoid production in cell cultures of Pueraria tuberosa as influenced by an angiospermic parasite, Cuscuta reflexa, was studied. During the time course, maximum isoflavonoid content was recorded when Cuscuta elicitor was added on day 15 of culture. Among various concentrations of elicitor tried, $1g\;l^{-1}$ of Cuscuta elicitor was found to be the most effective. The optimized elicitation conditions were used in vessels of varying capacity where maximum yield of ${\sim}91mg\;l^{-1}$ of isoflavonoid was recorded in a 2-l bioreactor which was about 19% higher than the control cultures. In this case, puerarin content increased up to $11mg\;l^{-1}$ which was 580% higher that the value recorded in the control cultures. In the bioreactor, 8 days of elicitation was optimal for the high accumulation of isoflavonoid, giving productivity of ${\sim}4mg\;l^{-1}\;day^{-1}$. The study showed persistent high isoflavonoid yield even during scale-up. Use of a preparation of Cuscuta reflexa as an elicitor is reported for the first time. The increase in isoflavonoid content was elicitor dose-dependent and can be explored to trigger high yields of isoflavonoid/secondary metabolites in production.

An Investigation Into the Relationship Between Metabolic Responses and Energy Regulation in Antibody-Producing Cell

  • Sun, Ya-Ting;Zhao, Liang;Ye, Zhao-Yang;Fan, Li;Liu, Xu-Ping;Tan, Wen-Song
    • Journal of Microbiology and Biotechnology
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    • v.23 no.11
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    • pp.1586-1597
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    • 2013
  • Energy-efficient metabolic responses were often noted in high-productive cultures. To better understand these metabolic responses, an investigation into the relationship between metabolic responses and energy regulation was conducted via a comparative analysis among cultures with different energy source supplies. Both glycolysis and glutaminolysis were studied through the kinetic analyses of major extracellular metabolites concerning the fast and slow cell growth stages, respectively, as well as the time-course profiles of intracellular metabolites. In three cultures showing distinct antibody productivities, the amino acid metabolism and energy state were further examined. Both the transition of lactate from production to consumption and steady intracellular pools of pyruvate and lactate were observed to be correlated with efficient energy regulation. In addition, an efficient utilization of amino acids as the replenishment for the TCA cycle was also found in the cultures with upregulated energy metabolism. It was further revealed that the inefficient energy regulation would cause low cell productivity based on the comparative analysis of cell growth and productivity in cultures having distinct energy regulation.

Improved guggulsterone production from sugars, precursors, and morphactin in cell cultures of Commiphora wightii grown in shake flasks and a bioreactor

  • Mathur, Meeta;Ramawat, K.G.
    • Plant Biotechnology Reports
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    • v.2 no.2
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    • pp.133-136
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    • 2008
  • Cell cultures of Commiphora wightii (Arnott.) Bhandari were grown in shake flasks and a bioreactor and an increase in guggulsterone accumulation up to $18{\mu}g\;l^{-1}$ was recorded in cells grown in the production medium containing a combination of sucrose:glucose (4% total), precursors (phenylalanine, pyruvic acid, xylose, and sodium acetate), morphactin, and 2iP. A yield of $10g\;l^{-1}$ biomass and ${\sim}200{\mu}g\;l^{-1}$ guggulsterone was recorded in a 3-l flask and in a 2-l stirred tank bioreactor compared with 6.6 g biomass and $67{\mu}g\;l^{-1}$ guggulsterone in 250-ml flasks. Increased vessel size was correlated with increased biomass and guggulsterone accumulation. 2iP alone was not effective for biomass and guggulsterone accumulation in cell cultures of C. wightii.

Basic Studies on Cultivation of Transformed Plant Tissue in Bioreactor (생물반응기에서 형질전환된 식물조직 배양을 위한 기초연구)

  • 박성화;박돈희
    • KSBB Journal
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    • v.5 no.3
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    • pp.207-214
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    • 1990
  • Growth properties of carrot hairy roots transformed by Agrobacterium rhizogenes were compared in flask and bioreactor. Oxygen transfer coefficient KLa was measured during the cultivation in bioreactor. In flask cultures, initially sucrose 30g/l was nearly exhausted after 20days. pH was dropped from initially 5.8 to 4.79 after 4 days, but it is stable after that time. Finally, after 28 days, hairy roots were grown about twelve times. In view of the results studied optimum conditions, hairy roots were maintained high growth rates in sucrose 50g/l, pH 5.8, total nitrogen 60mM. Also in bioreactor cultures, fixed stainless sieve in bottom and aerated 0.31 vvm, the results of cultivation by the use of sucrose 50g/l had grown about twenty-eight times and pH variations were liked in flask. As a results, growth rate of 1.756g fresh weight/day/g inoculum in bioreactor were higher about three times than 0.57g fresh weight/day/g inoculum in flask culture. KLa values were showed a tendency to decrease from 0.209 min-1 to 0.068 min-1.

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In Vitro Mass Propagation and Economic Effects of Bioreactor Culture in Ever-bearing Strawberry 'Goha' (Bioreactor를 이용한 사계성 딸기 기내대량증식과 경제성)

  • Lee, Jong-Nam;Kim, Hye-Jin;Kim, Ki-Deog;Kwon, Young-Seok;Im, Ju-Sung;Lim, Hak-Tae;Yeoung, Young-Rok
    • Horticultural Science & Technology
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    • v.28 no.5
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    • pp.845-849
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    • 2010
  • This study was conducted to compare various culture methods and evaluate economic feasibility of each method for mass propagation of new ever-bearing strawberry 'Goha'. Four different methods such as semi-solid culture, solid culture, liquid suspension culture and bioreactor culture were compared. The solid culture and bioreactor culture showed the shortest and longest root length, such as 3.6 cm and 8.3 cm, respectively. Fresh weights of plants cultured in bioreactor were 2,261 mg, which were heavier than those of cultures. Dry weights of plants cultured in bioreactor were the heavier compared to those in other cultures. The number of axillary bud developed in bioreactor was seven, but axillary bud was not developed in other cultures. Production cost through bioreactor culture was calculated to be 303 won per plant which was 542 won less than that of solid culture. As a result, we found that the bioreactor culture was the most cost effective culture method for in vitro mass propagation in new ever-bearing strawberry 'Goha'.

Effects of Elicitors on Scopolamine Production of Scopolia parviflora Nakai Adventitious Roots in Bubble Column Bioreactor

  • Jung, Hee-Young;Kim, Won-Jung;Kang, Seung-Mi;Park, Dong-Jin;Kang, Young-Min;Choi, Myung-Suk
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.5
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    • pp.378-383
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    • 2004
  • Scopolamine and hyoscyamine are important anticholinergic compounds. To increase the productivity, we have selected various elicitors and developed culture system using a bubble column bioreactor (BCB). As the same manner of elicitation in flask cultures, the elicitors were introduced into BCB cultures and the productivity was investigated. Except the bacterial elicitor of Staphyllococcus aureus, the elicitors inhibited hyoscyamine production. In scopolamine production, the elicitors revealed different responses from the results obtained in flask cultures. The elicitors of KCl and Candida albicans less increased the production than flask cultures. However, methyl jasmonate and S. aureus showed stronger positive effects on tropane alkaloid production. In particular, S. aureus was the most effective elicitor on scopolamine production and the elicitor resulted in the highly increased production, approximately 10 times higher than the control culture.

Adventitious root culture for secondary metabolite production in medicinal plants: A Review

  • Rahmat, Endang;Kang, Youngmin
    • Journal of Plant Biotechnology
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    • v.46 no.3
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    • pp.143-157
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    • 2019
  • Medicinal plants are high-value natural resources that have been used as precautionary drugs by many people globally. The increasing global demand for bioactive compounds from medicinal plants has led to the overexploitation of many valuable species. One widely used approach to overcome this problem is the use of adventitious root cultures as a propagation strategy. This review examines the scientific research published globally on the application of adventitious root cultures for many medicinal plants. Adventitious roots generated under aseptic environments in suitable phytohormone-augmented medium exhibit high growth rates and production of important secondary metabolites. Parameters such as medium properties and composition, growth hormone type, and elicitation strategies for in vitro grown adventitious roots of medicinal plants, are the main topics discussed in this review. We also examine current developments in bioreactor system cultivation for plant bioactive compounds using adventitious root cultures, a technology with possible commercial applications, via several studies on adventitious root culture of medicinal plants in which bioreactor systems play a role. In conclusion, the development of adventitious root cultures for medicinal plants is highly useful because of their capability for vegetative propagation and germplasm preservation.

Effects of Various Bioreactors on Growth and Ginsenoside Accumulation in Ginseng Adventitious Root Cultures(Panax ginseng C.A. Meyer) (다양한 생물반응기 형태가 인삼(Panax ginseng C.A. Meyer) 부정근의 생장과 Ginsenoside 생산에 미치는 영향)

  • Kim, Yun-Soo;Hahn, Eun-Joo;Paek, Kee-Yoeup
    • Journal of Plant Biotechnology
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    • v.31 no.3
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    • pp.249-253
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    • 2004
  • The type of air lift bioreactor affected the root growth in ginseng adventitious root cultures. Among bioreactors used in this experiment, bulb type bubble bioreactor (BU) was the best to increase root growth (41.92 g dry weight). The kLa value representing the oxygen transfer capacity from medium to explants (6.98 h$^{-1}$ ) in BU with 5 cm bubble column was higher than other bioreactors. On the other hand, cylindric tube bioreactor (CT) without bubble column resulted in minimum root growth (38.55 g dry weight) and kLa value (5.25 h$^{-1}$ ). Furthermore, the root growth (50.30 g dry weight) in BU with 10 cm bubble column more increased than 5 cm bubble column. However, the kLa value do not affected the secondary metabolite such as ginsenosides. These results show that the bubble column in air lift bioreactor increase kLa value and increased kLa value stimulate the growth of ginseng adventitious roots.

Kinetics of Cell Growth and Cyclosporin A Production by Tolypocladium inflatum when Scaling Up from Shake Flask to Bioreactor

  • El Enshasy, H.;Fattah, Y. Abdel;Atta, A.;Anwar, M.;Omar, H.;Magd, S. Abou El;Zahra, R. Abou
    • Journal of Microbiology and Biotechnology
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    • v.18 no.1
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    • pp.128-134
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    • 2008
  • The kinetics of cell growth and Cyclosporin A (Cyc A) production by Tolypocladium inflatum were studied in shake flasks and bioreactors under controlled and uncontrolled pH conditions. In the case of the shake flask, the production time was extended to 226 h and the maximal antibiotic concentration was 76 mg/l. When scaling up the cultivation process to a bioreactor level, the production time was reduced to only 70h with a significant increase in both the cell growth and the antibiotic production. The maximal dry cell weights in the case of the controlled pH and uncontrolled pH cultures in the bioreactor were 22.4g/l and 14.2g/l, respectively. The corresponding maximal dry cell weight values did not exceed 7.25g/l with the shake flask cultures. The maximal values for Cyc A production were 144.72 and 131.4 mg/l for the controlled and uncontrolled pH cultures, respectively. It is also worth noting that a significant reduction was observed in both the dry cell mass and the antibiotic concentration after the Cyc A production phase, whereas the highest rate of antibiotic degradation was observed in the stirred tank bioreactor with an uncontrolled pH. Morphological characterization of the micromorphological cell growth (mycelial/pellet forms) was also performed during cultivation in the bioreactor.

Importance of Strain Improvement and Control of Fungal cells Morphology for Enhanced Production of Protein-bound Polysaccharides(β-D-glucan) in Suspended Cultures of Phellinus linteus Mycelia (Phellinus linteus의 균사체 액상배양에서 단백다당체(β-D-glucan)의 생산성 향상을 위한 균주 개량과 배양형태 조절의 중요성)

  • Shin, Woo-Shik;Kwon, Yong Jung;Jeong, Yong-Seob;Chun, Gie-Taek
    • Korean Chemical Engineering Research
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    • v.47 no.2
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    • pp.220-229
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    • 2009
  • Strain improvement and morphology investigation in bioreactor cultures were undertaken in suspended cultures of Phellinus linteus mycelia for mass production of protein-bound polysaccharides(soluble ${\beta}$-D-glucan), a powerful immuno-stimulating agent. Phellineus sp. screened for this research was identified as Phellinus linteues through ITS rDNA sequencing method and blast search, demonstrating 99.7% similarity to other Phellinus linteus strains. Intensive strain improvement program was carried out by obtaining large amounts of protoplasts for the isolation of single cell colonies. Rapid and large screening of high-yielding producers was possible because large numbers of protoplasts ($1{\times}10^5{\sim}10^6\;protoplasts/ml$) formed using the banding filtration method with the cell wall-disrupting enzymes could be regenerated in relatively high regeneration frequency($10^{-2}{\sim}10^{-3}$) in the newly developed regeneration medium. It was demonstrated that the strains showing high performances in the protoplast regeneration and solid growth medium were able to produce 5.8~6.4%(w/w) of ${\beta}$-D-glucan and 13~15 g/L of biomass in stable manners in suspended shake-flask cultures of P. linteus mycelia. In addition, cell mass increase was observed to be the most important in order to enhance ${\beta}$-D-glucan productivity during the course of strain improvement program, since the amount of ${\beta}$-D-glucan extracted from the cell wall of P. linteus mycelia was almost constant on the unit biomass basis. Therefore we fully investigated the fungal cell morphology, generally known as one of the key factors affecting cell growth extent in the bioreactor cultures of mycelial fungal cells. It was found that, in order to obtain as high cell mass as possible in the final production bioreactor cultures, the producing cells should be proliferated in condensed filamentous forms in the growth cultures, and optimum amounts of these filamentous cells should be transferred as active inoculums to the production bioreactor. In this case, ideal morphologies consisting of compacted pellets less than 0.5mm in diameter were successfully induced in the production cultures, resulting in shorter period of lag phase, 1.5 fold higher specific cell growth rate and 3.3 fold increase in the final biomass production as compared to the parallel bioreactor cultures of different morphological forms. It was concluded that not only the high-yielding but also the good morphological characteristics led to the significantly higher biomass production and ${\beta}$-D-glucan productivity in the final production cultures.