• Title/Summary/Keyword: Fed-batch

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The Optimization of Expression System for Recombinant Protein Production by Pichia pastoris and Hansenula polymorphs (유전자 재조합 단백질 생산에 있어서 Pichia pastoris와 Hansenula polymorpha를 이용한 최적 발현 방법 개발)

  • 강환구;전희진;김재호
    • KSBB Journal
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    • v.15 no.2
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    • pp.174-180
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    • 2000
  • Pichia pastoris and Hansenula polymorpha, the methylotrophic yeasts have been widely used as a host for the production of e eudaryotic proteins due to the advantages related to their inherited characters. This paper describes the method to enhance t the productivity of recombinant proteins by P. pastoris and H. po$\psi$morpha. In the production of recombinant proteins using a f fed batch fermentation system, the effects of specific growth rate on the specific expression rate of re$\infty$mbinant proteins w were studied. In both species, the expression system of recombinant proteins using the fed batch fermentation was optimezed.

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Removal of Simultaneously Biological Organic, Nitrogen, and Phosphorus Removal in Sequencing Batch Reactors using Night-soil (연속회분식 반응기(Sequencing Batch Reactor)를 이용한 분뇨중 유기물과 질소 및 인의 동시제거)

  • 한기백;박동근
    • Journal of Environmental Science International
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    • v.6 no.6
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    • pp.697-709
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    • 1997
  • Sequencing Batch Reactor(SBR) experiments for organics and nutrients removal have been conducted to find an optimum anaerobic/anoxic/aerobic cycling time and evaluate the applicability of oxidation-reduction potential(ORP) as a process control parameter. In this study, a 61 bench-scale plant was used and fed with night-soil wastewater in K city which contained TCODcr : 10, 680 mg/l, TBm : 6, 893 mg/l, $NH_4^+-N$ : 1, 609 mg/l, $PO_4^{3-}-P$ : 602 mg/l on average. The cycling time In SBRs was adjusted at 12 hours and 24 hours, and then certainly included anaerobic, aerobic and inoxic conditions. Also, for each cycling time, we performed 3 series of experiment simultaneously which was set up 10 days, 20 days and 30 days as SRT From the experimental results, the optimum cycling time for biological nutrient removal with nlght-soil wastewater was respctively 3hrs, 5hrs, 3hrs(anaerobic-aerobic-anoxic), Nitrogen removal efficiency was 77.9%, 77.9%, 81.7% for each SRT, respectively. When external carbon source was fed in the anoxic phase, ORP-bending point indicating nitrate break point appeared clearly and nitrogen removal efficiency increased as 96.5%, 97.1%, 98.9%. Phosphate removal efficiency was 59.8%, 64.571, 68.6% for each SRT. Also, we finded the applicability of ORP as a process control parameter in SBRs.

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1,2-Propanediol Production by Using Saccharomyces cerevisiae M3G3 (Saccharomyces cerevisiae M3G3를 이용한 1,2-Propanediol의 생산 최적화)

  • Koo, Ja-Ryong;DaSilva, Nancy A.;Yun, Hyun-Shik
    • KSBB Journal
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    • v.26 no.5
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    • pp.439-442
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    • 2011
  • 1,2-propanediol (1,2-PD) is a commodity chemical that is currently produced from petrochemical derivatives. Saccharomyces cerevisiae is well characterized and a successful industrial microorganism to enable the improvement of the 1,2-propanediol production by metabolic engineering. A recombinant S. cerevisiae M3G3 was used to produce 1,2-propanediol. S. cerevisiae M3G3 is the diploid strain that contains 3 copies of mgs (methylglyoxal synthase) and gldA (glycerol dehydrogenase). S. cerevisiae M3G3 was cultivated at various culture conditions by changing culture temperature, glucose concentration, and inducer concentration. Also the effect of induction time was studied to optimize the production of 1,2-propanediol. Batch and fed-batch cultivation of S. cerevisiae M3G3 was performed by using a 5 L jar fermenter. The highest concentration of 1,2-propanediol in batch cultivation was 0.86 g/L and it was further improved to 1.33 g/L in fed-batch cultivation.

Ethanol Fermentation of Corn Starch by a Recombinant Saccharomyces cerevisiae Having Glucoamylase and $\alpha$-Amylase Activities

  • Lee, Dae-Hee;Park, Jong-Soo;Ha, Jung-Uk;Lee, Seung-Cheol;Hwang, Yong-Il
    • Preventive Nutrition and Food Science
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    • v.6 no.4
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    • pp.206-210
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    • 2001
  • Starch is an abundant resource in plant biomass, and it should be hydrolyzed enzymatically into fermentable sugars for ethanol fermentation. A genetic recombinant yeast, Saccharomyces cerevisiae GA-7458, was constructed by integrating the structural gene of both $\alpha$-amylase from Bacillus stearothermophilus and the gene (STA1) encoding glucoamylase from S. diastaticus into the chromosome of S. cerevisiae SH7458. The recombinant yeast showed active enzymatic activities of $\alpha$-amylase and glucoamylase. The productivity of ethanol fermentation from the pH-controlled batch culture (pH 5.5) was 2.6 times greater than that of the pH-uncontrolled batch culture. Moreover, in a fed-batch culture, more ethanol was produced (13.2 g/L), and the production yield was 0.38 with 2% of corn starch. Importantly, the integrated plasmids were fully maintained during ethanol fermentation.

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Two-Stage Fed-Batch Culture of Candida magnoliae for the Production of Erythritol using an Industrial Medium (산업용 배지를 이용한 Candida magnoliae의 2단계 유가식 배양에서 에리스리톨의 생산)

  • 박선영;서진호;유연우
    • KSBB Journal
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    • v.18 no.4
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    • pp.249-254
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    • 2003
  • Experiments were carried out to select an industrial nitrogen source and optimize erythritol production by Candida magnoliae in fed-batch culture. Among the industrial nitrogen sources tested, light steep water (LSW) was found to be the best nitrogen source for producing erythritol, based on erythritol yield and raw material price. The maximum erythritol concentration obtained a 131.6 g/L, with a 52.6% yield and 0.52 g/L-hr productivity from a 250 g/L glucose and 43.3 mL/L LSW in batch culture. Two-stage fed-batch culture was chosen to improve the volumetric productivity and the yield of erythritol. High cell density culture in cell growth stage was achieved by batch type culture containing 100 g/L glucose and 500 mL/L LSW. The cell concentration was 71.0 g/L after 23 hours of culture. Erythritol productivity was decreased by increasing glucose concentration in the production stage. But 37.3% of the maximum erythritol yield was obtained with 185.5 g/L of erythritol and 1.66 g/L-hr of productivity when 820 g of glucose powder was directly added to a concentration of 450 g/L glucose in production stage.

Control of Both Foam and Dissolved Oxygen in the Presence of a Surfactant for Production of $\beta$-Carotene in Blakeslea trispora

  • Kim, Seon-Won;Lee, In-Young;Jeong, Jae-Cheol;Lee, Jung-Heon;Park, Young-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.548-553
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    • 1999
  • A production of $\beta-Carotene$was attempted in a fed-batch culture of Blakeslea trispora by controlling both foam and dissolved oxygen in the presence of surfactant, Span 20. Results obtained from the shake flask cultures indicated that a high concentration of dissolved oxygen was needed for both cell growth and $\beta-Carotene$ synthesis, and the optimal concentration of glucose was found to be in the range of 50-100 g/l. In order to maintain the dissolved oxygen concentration level at higher than 50% of air saturation, pure oxygen was automatically sparged into the medium with air. Foam was controlled by bypassing air from the submerged aeration to the headspace in response to the foam that was caused by Span 20. High agitation speed was found to be detrimental to the cell growth due to shear damage, even though it provided sufficient dissolved oxygen. On the other hand, a low aeration speed caused stagnant regions in the fermentor because of improper mixing. Thus, for the fed-batch operation, agitation speed was increased gradually from 300 to 700 rpm to prevent cell damage at the initial stage of fermentation and to give efficient mixing for a viscous culture broth as the culture proceeded. By controlling dissolved oxygen and foam, a high concentration of $\beta-Carotene$otene (1,190 mg/l) was obtained in 6 days of the fed-batch culture of B. trispora with 2.5% of the dry cell weight, which was approximately 5 times higher than that of the batch cultures.

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Production of Mycelia and Water Soluble Polysaccharides from Submerged Culture of Lentinus lepideus in Bioreactor (생물반응기를 이용한 잣버섯(Lentinus lepideus)의 균사체 및 수용성 다당체 생산특성)

  • Ahn, Jin-Kwon;Ka, Kang-Hyeon;Lee, Wi-Young
    • The Korean Journal of Mycology
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    • v.35 no.1
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    • pp.37-42
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    • 2007
  • The mushroom Lentinus lepideus was used to produce mycelial as well as soluble polysaccharides in bioreactor cultures. To determine optimal submerged culture conditions, both growth characteristics and water soluble polysaccharides production were compared among four different types of bioreactor and culture conditions. For the production of mycelial biomass, the following bioreactors were proven to be effective in decreasing order: an external-loop type air-lift bioreactor (ETAB; 7g/l), a balloon type air bubble bioreactor (BTBB; 6.2g/l), a stirrer type bioreactor (STB; 6g/l), and a column type air bubble bioreactor (CTBB; 5g/l). Maxiaml production of water soluble exopolysaccharides (EPS; 0.62g/l) and endopolysaccharides (PPS; 7.7%) could also be obtained from BTBB. The mycelial biomass increased with increase in glucose concentration from 15g/l to 75g/l in the media. In contrast, PPS contents in the cells decreased with increase in glucose concentration in the media, showing the highest PPS content (7%) at 15g/l. Among different medium feeding types, fed-batch culture based on concentration control in media (10g/l) produced higher mycelia than fed-batch culture based on volume control of media (5.8g/l) or batch culture (3.4g/l). EPS production was also higher in fed-batch culture based on medium concentration control than that in other feeding types.

Optimal Strategy for Ethanol Production in Repeated Fed-batch Operation Using Flocculent Sacchromyces cerevisiae (응집성 Sacchromyces cerevisiae 를 이용한 반복 유가식 ethanol 생산에서의 최적 운전전략)

  • Lee, Sang-Eun;Yeon, Ji-Hyeon;Seo, Yong-Chang;Kang, Do-Hyung;Lee, Hyeon-Yong;Jung, Kyung-Hwan
    • KSBB Journal
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    • v.25 no.2
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    • pp.179-186
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    • 2010
  • We investigated the optimal strategy for ethanol production using flocculent Sacchromyces cerevisiae ATCC 96581. Considering the characteristic of flocculent yeast, a repeated fed-batch ethanol fermentation was designed, in which non-sterile glucose powder was fed every 12 hours and, after cell flocculation, new feeding medium was exchanged every 24 or 36 hours. We particularly compared this fermentation process with those when cell flocculation was not carried out. Finally, the maximal total ethanol production was 825 g-ethanol during 120 hours, in which the time interval of withdrawal-fill of feeding medium was 24 hours and cell flocculation was carried out.