• Title/Summary/Keyword: High density culture

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Continuous high cell density culture of Anaerobiospirillum succiniciproducens with membrane filtration for the production of succinic acid

  • Lee, Pyeong-Cheon;Lee, U-Gi;Lee, Sang-Yeop;Jang, Ho-Nam
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.338-341
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    • 2000
  • An internal membrane bioreactor system was employed for continuous succinic ac id production from glucose in order to prove its performance and practicality. Succinic acid-producing Anaerobiospirillum succiniciproducens required more $CO_2$ for the proper growth and succinic acid production in cell recycled continuous culture than in batch culture. The maximum productivity obtained in cell recycled continuous culture was about 3.3 g/L-h which was ca. 3.3 times higher than that obtained in batch culture.

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고정상세포분리기의 개발 및 Cyclosporin A 생산을 위한 고정화 연속배양공정에의 적용

  • Lee, Tae-Ho;Park, Sung-Kwan;Chang, Yong-Keun;Chun, Gie-Taek
    • Microbiology and Biotechnology Letters
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    • v.24 no.6
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    • pp.717-725
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    • 1996
  • We have developed an efficient immobilized cell separator for continuous operation of immobilized fungal cell cultures, and applied this separator to actual fermentation process for the production of cyclosporin A (CyA), a powerful immunosuppressant. In the experiments employing highly viscous polymer (carboxymethyl cellulose) solution, the decantor showed good separating performances at high solution viscosites and fast dilution rates. Air duct and cylindrical separator installed inside the decantor turned out to play key roles for the efficient separation of the immobilized cells. By installing the decantor in an immobilized perfusion reactor system (IPRS), continuous immobilized culture was stably carried out even at high dilution rate for a long period, leading to high productivities of free cells and CyA. Almost no immobilized biomass existed in effuluent stream of the IPRS, demonstrating the effectiveness of the decan- tor system for a long-term continuous fermentation. It was noteworthy that we could obtain these results despite of the unfavorable fermentation conditions, i.e., reduced density of the biosupports caused by overgrowth of cells inside the bead particles and existence of high density of suspended fungal cells (10g/l) in the fermentation broth.

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Calculation of Light Penetration Depth in Photobioreactors

  • Lee, Choul-Gyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.4 no.1
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    • pp.78-81
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    • 1999
  • Light penetration depth in high-density Chlorella cultures can be successfully estimated by Beer-Lambert's law. The efficiency of light energy absorption algal cultures was so high that algal cells near the illuminating surface shade the cells deep in the culture. To exploit the potential of high-density algal cultures, this mutual shading should be eliminated or minimized. However, providing more light energy will not ease the situation and it will simply drop the overall light utilization efficiency.

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Production of a Platelet Aggregation Inhibitor, Salmosin, by High Cell Density Fermentation of Recombinant Escherichia coli

  • Seo, Myung-Ji;Choi, Hak-Jong;Chung, Kwang-Hoe;Pyun, Yu-Ryang
    • Journal of Microbiology and Biotechnology
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    • v.21 no.10
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    • pp.1053-1056
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    • 2011
  • Optimal conditions for a high cell density fermentation were investigated in a recombinant Escherichia coli producing salmosin, a platelet aggregation inhibitor. The optimized carbon and nitrogen sources were glycerol 10 g/l, yeast extract 30 g/l, and bacto-tryptone 10 g/l, yielding the dry cell weight (DCW) of 10.61 g/l in a 500 ml flask culture. The late-stage induction with 1% L-arabinose in a 5 l jar fermentor showed the highest DCW of 65.70 g/l after 27 h of the fed-batch fermentation. Around 2,200 mg/l of the protein was expressed as an inclusion body that was then refolded to obtain the active salmosin of 96 mg/l. We also confirmed the inhibitory activity against platelet aggregation of the active salmosin from the high cell density fermentation.

High Density Cultivation of Methylobacillus sp. SK1 in Fed-Batch System (Methylobacillus sp. SK1의 고농도 유가배양)

  • 이형춘;이계호김시욱
    • KSBB Journal
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    • v.5 no.3
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    • pp.269-277
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    • 1990
  • Methylobacillus sp. SK1, an obligate methylotroph, was cultivated in a fed-batch culture using DO as a methanol feeding indicator with a micro computer-aided control system. While 2.07g/l of cell density was obtained after 13 hr in the batch culture (initial methanol concentration: 1.0%(v/v)),45.3g/l of cell density was obtained after 17 hr by feeding methanol and metal ions in the fed-batch culture with oxygen supply. The high-density biomass was obtained in short cultuivation time by fed-batch culture with feedback control, and consequently the biomass productivity was significantly increased. It was mainly due to extension of logarithmic growth period by methanol feeding without methanol inhibition and intensive aeration without DO limitation with microcomputer-aided control system.

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Effect of Low Temperature Preservation and Cell Density on Metabolic Function in a Bioartificial Live

  • Park, Yueng-Guen;Takehiko Tosha;Satoshi Fujita;Boru Zhu;Hiroo Iwata;Ryu, Hwa-Won
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.1
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    • pp.41-46
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    • 2003
  • Difficulties associated with bioartificial liver (BAL) preservation limit not only the commercialization of BAL, but also its clinical trials. In this study, the possibility of cold preservation of BAL cartridges containing porcine hepatocytes was examined at 4$^{\circ}C$. In an in vitro perfusion culture System, BAL cartridges maintained cytochrome P450 metabolic function for at least 50 days. However, all BAL cartridges completely lost their ammonia eliminating ability when stored at 4$^{\circ}C$. We a1so studied the effect of cell density on the maintenance of BAL liver function In a highly differentiated and healthy state. As expected, BALs containing a larger number of hepatocytes demonstrated higher metabolic functions. When metabolic functions were compared per gram of hepatotytes, no large differences were observed between devices containing different densities of hepatocytes. Decreased cell density did not Successfully prolong BAL function. The viability and function of isolated hepatotytes highly depend on the culture conditions, such as cell density, substrata, culture media, and additives to the culture media. Perfusion culture of BAL cartridges at 4$^{\circ}C$ gave a promosing result with respect to the maintenance of P450 activity. However, as indicated by the rapid loss of ammonia metabolic activity, many factors still remain to be optimized for preservation of BAL keeping high metabolic functions for a longer time.

High Density Culture of KA112 Hybridoma and Effect of Glucose Concentration on MAb Productivity (하이브리도마의 고농도 배양과 포도당 농도가 MAb 생산성에 미치는 영향)

  • 박상재;최차용
    • KSBB Journal
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    • v.8 no.5
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    • pp.478-482
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    • 1993
  • Perfusion culture was conducted in Celligen perfusion culture system using a self-constructed hybridoma cell and low serum medium. The culture system employed hollow fiber to separate cells from the culture broth. Maximum cell density of $2.1\times10^7$ ce11s/m1, 10 times higher than in batch culture, could be achieved. Concentration of monoclonal antibody (MAb) was 4 times higher and production rate at maximum feed rate was 9 times higher than in batch culture. Glucose concentration was very important for the cell growth and MAb production. When glucose concentration was below 1g/l, i. e. 0.5~0.9g/l, specific MAb production rate decreased but cell concentration still increased. As the glucose concentration goes above 1g/l, specific MAb production rate increased and remained at maximum value at more than 1.5g glucose/l. The maximum value of the specific Mab production rate was similar to that of batch culture.

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Development of Curdlan Separation Process with Density Gradient Centrfugation (Density Gradient를 이용한 식품소재를 커들란의 분리공정개발)

  • 김봉영;이중헌
    • KSBB Journal
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    • v.16 no.5
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    • pp.523-525
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    • 2001
  • Curdlan is one biopolymer composed of ${\beta}$1,3-glucan and dissolved in a alkali solution but formed salt under neutral or acid condition. It was produced by Agrobactrium species and the separation process is necessary to make pure curdlan from the culture broth. The pH swing separation method was as feasible separation process using solubility changes with the pH difference. however, this method requires a lot of acid and alkali solution also produces a lot of waste. Therefore, an efficient process which could save energy and minimize toxic waste was developed. A density gradient separation process was developed in this research. High density sucrose solution was used as a separation agent. Curdlan was separated from the culture broth when the density of the sucrose solution was 1.15 g/L. Since the curdlan was produced on the surface of cell wall. the pre-treatment of culture broth was necessary. Curdlan recovery yield was increased up to 83% with the homogenization of the culture broth and further increased up to 87% with the treatment of alkai-acid solution.

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Effects of biological Factors on In Vitro Production of Hanwoo Embryos (한우 난포란 유래 배반포의 체외생산을 위한 생물학적 요인들의 영향)

  • 박흠대;김재영;주재홍;공건오;윤산현;공일근;이상민;이상진;송해범
    • Journal of Embryo Transfer
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    • v.15 no.2
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    • pp.129-136
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    • 2000
  • This study was carried out to investigate the effect of biological factors on the in vitro production(IVP) of bovine oocytes for development of simple culture methods and medium. Oocytes from the slaughterhouse ovaries were matured and fertilized using general protocol and this study was examined if there were necessary to co-culture, media change, media type and embryo density. This results were as follows: 1. The development rate according to co-culture with cumulus cells and non co-culture as drop culture was not significantly different in cleavage (68.9 vs 71.7%), 8-cell stage (41.2 vs 44.1%) and blastocyst stage (12.2 vs 13.8%), respectively (p<0.05) 2. The blastocyst development rates in YS and CRIaa were higher than that in TCM199 (12.4, 10.4$ vs 3.7%), but the cleavage (69.0, 77.8 and 61.0%) and 8-cell stage (31.7, 37.0 and 35.7%) development accoring to YS, TCM199 and CRIaa ws not significantly different, respectively (p<0.05). 3. There was no significantly different in cleavage (62.6, 59.5 and 61.2%), 8-cell(34.7, 37.9 and 34.0%) and blastocyst (9.5, 11.6 and 12.8%) development among medium change time as control, Group I and Group II, respectively (p<0.05). 4. Blastocyst formation of 8-cell stage according to embryo density was not significantly different in 1, 10 and 25 embryos (27.3, 22.5 and 34.0%), respectively (p<0.05). These results indicated that simple culture system could reduce bovine IVP embryos as drop culture as non co-culture system, high density embryo (25 embryos/50 $\mu$1 drop). YS defined medium and no medium change for whole culture period, although other biological factors need to examine in order to produce efficient IVP bovine embryos.

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