• Title/Summary/Keyword: semi-fed-batch

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Production of an Anticoagulant Hirudin by Fed-batch and Continuous Cell Recycle Fermentations Using Recombinant Saccharomyces cerevisiae (유가식과 세포재순환 연속공정을 이용한 항혈전제 hirudin의 생산)

  • 최치민;김명동;이상기;서진호
    • KSBB Journal
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    • v.13 no.4
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    • pp.456-460
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    • 1998
  • Fed-batch fermentations were carried out in order to improve the efficiency of hirudin production by recombinant Saccharomyces cerevisiae. A fed-batch fermentation done with the optimized semi-synthetic medium resulted in a maximum hirudin concentration of 342mg/$\ell$ by keeping a galactose concentrations between 10 and 30g/$\ell$ which corresponded to a 11.4-fold increase in hirudin concentration compared with the simple bach fermentation done with the same medium. Comparison of the chromatographic pattern of proteins in the growth medium clearly showed that the use of the semi-synthetic medium is more advantageous for separation of hirudin than the case o fusing the complex medium. Continuous cell recycle fermentation done at dilution rate of 0.1h-1 and an inlet galactose concentration of 100g/$\ell$ yielded a maximum hirudin productivity of 19.1mg hirudin/$\ell$$.$h.

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Development of High Density Mammalian CellCulture system for the Production of Tissue-Type Plasminogen Activator

  • Park, Byong-Gon;Chun, Joo-Mi;Lee, Chang-Jin;Chun, Gie-Taek;Kim, Ik-Hwan;Jeong, Yeon-Ho
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.2
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    • pp.123-129
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    • 2000
  • A high cell density culture system for the anchorage dependent CHO cells was developed based on the combination of in removal of ammonium ion and microcarrier culture system, and semi-fed-batch feeding of glucose and glutamine was employed to the developed culture system. The glass bead was selected as an optimum microcarrier in terms of cell growth. An ammonium ion selective zeolite, Phillipsite-Gismondine, was packed in a dialysis menium ion. The semi-fed-batch operation was employer to the novel culture system for the high density cell culture, and the results showed the cell growth was improved by 32% and tPA productivity by 250%.

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Development of an Immobilized Adsorbent for In Situ Removal of Ammonium Ion from Animal Cell Culture Media and Its Applications to Animal Cell Culture System : II. Application to Cell Culture System (동물세포 배양액으로부터 암모늄 이온의 동시제거를 위한 고정화 흡착제의 개발과 동물세포 배양 시스템에의 응용 : II. 세포배양 시스템에의 응용)

  • 박병곤;이해익;전계택;김익환;정연호
    • KSBB Journal
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    • v.13 no.4
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    • pp.411-417
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    • 1998
  • The possibility of application of membrane type immobilized adsorbent to the fed-batch or perfusion culture system with anchorage-independent cells as well as batch system was investigated. The improvement in cell density and cell viability due to the combination of immobilized adsorbent with each culture system was evaluated for the investigation, and the optimum culture system employing immobilized adsorbent system was suggested based on the results. It was observed that the system with immobilized adsorbent showed better cell growth and cell viability than that without immobilized adsorbent in every operation system of batch, fed-batch, and perfusion. In case of batch system, 200% improvement of maximum cell density was observed in the system where ammonium chloride was added on purpose. And 50% improvement of maximum cell density was observed in the fed-batch system where ammonium ion accumulates significantly, while small increase in maximum cell density was observed in the perfusion system where dilution of waste byproducts exists. Especially, the fed-batch system showed the most significant improvement on cell growth because both compensation of nutrient and removal of ammonium ion occurred simultaneously in the system. Therefore a combined system of immobilized adsorbent and fed-batch operation could be suggested as an optimum system with in situ removal of ammonium ion.

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Studies of Repeated Fed-Batch Fermentation of Cephalosporin C in an Immobilized Cell Bioreactor

  • Park, Hong-Je;Khang, Yong-Ho
    • Journal of Microbiology and Biotechnology
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    • v.5 no.4
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    • pp.229-233
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    • 1995
  • Acremonium chrysogenum was immobilized in ionotropic gel beads to develop semi-continuous production of cephalosporin C (CPC). Barium alginate beads were more stable than calcium alginate or strontium alginate beads in chemically defined media. The gel stability of Ba-alginate was further increased by cross-linking with polyethyleneimine (PEI). The presence of carboxymethyl cellulose inside Ba-alginate beads did not reduce mass transfer resistance. Ba-alginate microbeads that had little diffusion limitation increased CPC production rate 1.6 fold higher than that of normal beads. CPC fermentation with immobilized cells in Ba-alginate microbeads was performed continuously for 40 days by way of repeated fed-batch operations. Mathematical modeling was developed to describe the repeated fed-batch fermentation system. Results of the computer simulation agreed well with the experimental data, which made it possible to predict an optimal feeding rate that could maximize total CPC productions.

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Production of Pyruvate Dehydrogenase Complex-E2 Specific Human Monoclonal Antibody in Fed-batch Culture Systems with High Cell Density Recombinant Escherichia coli (고농도 재조합 대장균의 Fed-batch 배양 시스템을 이용한 Pyruvate Dehydrogenase Complex-E2 특이성 인간 모노클론 항체의 생산)

  • 이미숙;전주미;차상훈;정연호
    • KSBB Journal
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    • v.15 no.5
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    • pp.489-496
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    • 2000
  • Several culture systems including batch, two-stage CSTR, semi-fed batch, and two-stage cyclic fed-batch were investigated for the efficient production of the Fab fraction of PDC-E2 specific human monoclonal antibody using high cell density recombinant E. coli. A two-phase batch system and a two-stage continuous system were examined to overcome plasmid instability problems, by separating the growth and the production stages. The cell density and productivity of the two-stage continuous culture was better than that of the two-phase batch fermentation. In the two-stage continuous culture system with DO-stat, the cell growth and the productivity were superior to those of the system without the DO control. Also, almost total plasmid stability was maintained in the two-stage continuous culture system. Modified M9 medium was selected as an optimum feeding medium for the fed-batch process, and the optimum C/N ratio determined to be 2:3. The optimum feeding rate was $0.6g/\ell/hr$ for a constant feeding strategy in semi-fed batch system. When the feeding medium was fed by pulsing, it was observed that more frequent pulsing resulted in improved cell growth. The linear feeding method was the most efficient of the various feeding methods tested. Finally, high cell density culture using a two-stage cyclic fed batch system with pH-stat was tried because the linear feeding method showed limitations in terms of obtaining high cell densities, and a cell density of $54 g/\ell$ was achieved. It was concluded that the two-stage cyclic fed batch system was the most efficient system for high cell density culture of the systems tested. However, productivity improvements were lower than expected due to the extremely high accumulations of acetate, although the low levels of residual glucose were maintained.

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Lovastatin biosynthesis enhanced by thiamine in Aspergillus terreus

  • An, U-Seok;Han, Gyu-Beom
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.184-187
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    • 2002
  • Lovastatin is a cholesterol-lowering agent, which plays a role of an inhibitor of 3-hydroxy-3- methylglutaryl coenzyme A reductase (HMG-CoA). When thiamine was supplemented in 3L batch fermentation, the production of lovastatin was improved. At the same time, the levels of pyruvic acid and NAD(P)H were estimated in the course of the fermentation of A. terreus. For the high level production of lovastatin, semi fed-batch fermentation was performed. And the thiamine level was maintained to a concentration of 20 mg/L and glucose was supplied. The final dry cell weight was lowered by 30 % and final lovastatin concentration was increased by 33 %. Final lovastatin concentration of 3.3 g/L was achieved in 8 days.

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Potential Use of Microalgae Scenedesmus acuminatus for Tertiary Treatment of Animal Wastewater (축산폐수 고도처리를 위한 미세조류 Scenedesmus acuminatus의 이용 가능성)

  • Park, Ki-Young;Lim, Byung-Ran;Lee, Ki-Say;Lee, Soo-Koo
    • Journal of The Korean Society of Agricultural Engineers
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    • v.53 no.1
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    • pp.63-69
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    • 2011
  • The green algae Scenedesmus acuminatus was cultured in different media: animal wastewater and an artificial culture medium in order to evaluate potential use for tertiary treatment. The experiments were conducted with air flowrate 1~2 L/min at $28{\sim}30^{\circ}C$. The nitrogen and phosphorus showed very similar removal efficiencies (68~77 % and 69~80 % for nitrogen and phosphorus respectively). The optimal fed period was estimated as three days in the semi-continuous experiment. The effects of $CO_2$ (4.5 %) injection on nutrient uptake from animal wastewater (biological treatment effluent) were compared to an air injection under the same conditions of light and photoperiod. The uptake rates of nutrient with air injection were observed 0.009 gN/gChl-a/day, 0.028 gN/gChl-a/day and T-P 0.003 gP/gChl-a/day for nitrate, total nitrogen and phosphorus respectively. The rates were enhanced by addition of $CO_2$ to 0.026 gN/gChl-a/day, 0.076 gN/gChl-a/day and T-P 0.018 gP/gChl-a/day. This study establishes that $CO_2$ addition during nutrient deprivation of microalgal cells may accelerate tertiary wastewater treatment.

Efficient Use of Lactose for Production of the Soluble Recombinant Human Epidermal Growth Factor in Escherichia coli. (대장균에서 lactose를 이용한 수용성 재조합 인간 상피 세포 성장 인자의 생산)

  • 박세철;권태종;고인영;유광현
    • Microbiology and Biotechnology Letters
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    • v.26 no.1
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    • pp.61-67
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    • 1998
  • Recombinant human epidermal growth factor (rhEGF) was produced by E. coli BL2l (DE3) harboring a plasmid pYHB101. The production of rhEGF was 44.5 mg/L when the E. coli BL2l (pYHB101) was cultured at 27$^{\circ}C$ for 48 hr in the modified MBL medium containing 10 $\mu\textrm{g}$/L glucose with 10 $\mu\textrm{m}$ IPTG/lactose induction at 2 hr after inoculation. It was shown that lactose is able to induce the rhEGF expression of E. coli BL2l (pYHB101) with the same efficiency as IPTG. In the batch culture system, when induced with 10 $\mu\textrm{m}$ lactose, E. coli BL2l (pYHB101) produced maximum 45 mg/L of the rhEGF at 28 hr culture in the modified MBL medium containing 10 g/L glucose. In the semi-fed batch culture system, the volumetric yield was 160 mg/L when the culture was added with 0.5% (w/v) lactose and 0.25% (w/v) yeast extract in the late logarithmic phase and 94.3% of rhEGF was secreted as soluble form. However, when the culture was added with them in the early logarithmic phase, the volumetric yield was 120 mg/L and 20.9% of rhEGF was found in cytoplasmic insoluble aggregates. It was found that the addition time of lactose was important for production of soluble rhEGF from E. coli BL21 (pYHB101).

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Production of High Acetic Acid Vinegar Using Two Stage Fermentation (Two Stage 발효에 의한 고산도 식초 생산)

  • 이영철;이금용;김형찬;박기범;유익제;안평욱;최춘언;손세형
    • Microbiology and Biotechnology Letters
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    • v.20 no.6
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    • pp.663-667
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    • 1992
  • The production of vinegar containing 16.0-18.0% of acetic acid was examined in two stage fermentation consisting of semi-continuous and fed-batch type. The optimum conditions were obtained when the fermentation was carried out at agitation of 600 rpm, aeration of 0.1 vvm and temperature of $30^{\circ}C$. The initial and residual ethanol concentration in 1st stage were $50.0g/{\ell}$ and $5.0g/{\ell}$, respectively, and the ethanol concentration in 2nd stage was maintained from 5.0 to $10.0g/{\ell}$. The maximum productivity was 3.3 gll-hr and the acidity was 17.6% after the two days of acetic acid fermentation.

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A Study on the Substrate Removal Efficiency with Varing SRT in Anaerobic Treatment of High Organic Wastewater (고농도 유기성폐수의 혐기성처리시 SRT변화에 따른 유기물질 제거효율에 관한 연구)

  • Kim Byeong Ho;Kim Dong Min
    • Journal of environmental and Sanitary engineering
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    • v.4 no.1 s.6
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    • pp.43-52
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    • 1989
  • A synthetic wastewater composed of powdered baby food and inorganic nutrients was treated by five 3.6L anaerobic reactors in order to test the relationship between solids retention time (SRT) and media surface ratio, and the removal efficiencies of organic substrate. Of the five reactors, four were semi-continuously fed stirred-tank reactors and one continuously-stirred batch reactor. The influent was 7430mg/L in COD, 7120 mg/L $BOD_L$ and 6350mg/L in $BOD_5$, respectively. Operating temperature was $35{\pm}1^{\circ}C$ and pH in the range 6.9 to 7.2. In this experimental study it was found that a linear relationship existed, within the experiment range, between SRT and media surface ratio, and that SRT and removal efficiency increased with increasing media surface ratio. The substrate removal efficiencies were 82.7 to $88.2\%$ in COD, 82.9 to $88.4\%$ in $BOD_L$ and 83.3 to $88.7\%$ in $BOD_5$, respectively.

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