• Title/Summary/Keyword: fed-batch cultivation

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Fermentation Studies on Pseudomonas aeruginosa Producing Antifungal Secondary Metabolite, PAFS. (항진균물질을 생합성하는 Pseudomonas aeruginosa의 배양생리적 특성 연구)

  • 송성기;윤권상;정용섭;전계택
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.52-59
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    • 2004
  • When both fructose and galactose were added to a production medium as carbon sources, the productivity of PAFS (Psedomonas Antifungal Substance) biosynthesized by Pseudomonas aeruginosa was observed to be reduced significantly due to the well-known phenomenon of catabolite repression. In order to overcome this phenomenon by use of fermentation bioprocess, fed-batch cultivation method was examined. In addition, a high producer mutant strain, AP-20 obtained by a rational screening method was tested for its productivity of PAFS in both batch and fed-batch fermentation processes. Notably fed-batch operation showed approximately 4 fold higher PAFS productivity than traditional batch operation process. It was appeared that galactose was utilized principally for the cell growth of Pseudomonas aeruginosa whereas large portion of fructose was used for the biosynthesis of PAFS. Furthermore it was observed that composition and feeding rate of production media should be optimized even in the fed-batch fermentation bioprocess. As an example, very slow feeding of carbon sources gave rather negative effect on the production of PAFS due to significant limitation of carbon and energy sources available for the producer microorganism.

An Optical-Density-Based Feedback Feeding Method for Ammonium Concentration Control in Spirulina platensis Cultivation

  • Bao, Yilu;Wen, Shumei;Cong, Wei;Wu, Xia;Ning, Zhengxiang
    • Journal of Microbiology and Biotechnology
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    • v.22 no.7
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    • pp.967-974
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    • 2012
  • Cultivation of Spirulina platensis using ammonium salts or wastewater containing ammonium as alternative nitrogen sources is considered as a commercial way to reduce the production cost. In this research, by analyzing the relationship between biomass production and ammonium-N consumption in the fed-batch culture of Spirulina platensis using ammonium bicarbonate as a nitrogen nutrient source, an online adaptive control strategy based on optical density (OD) measurements for controlling ammonium feeding was presented. The ammonium concentration was successfully controlled between the cell growth inhibitory and limiting concentrations using this OD-based feedback feeding method. As a result, the maximum biomass concentration (2.98 g/l), productivity (0.237 g/l d), nitrogen-to-cell conversion factor (7.32 gX/gN), and contents of protein (64.1%) and chlorophyll (13.4mg/g) obtained by using the OD-based feedback feeding method were higher than those using the constant and variable feeding methods. The OD-based feedback feeding method could be recognized as an applicable way to control ammonium feeding and a benefit for Spirulina platensis cultivations.

Enhancement of Alcohol Fermentation Yield by Adding the Extract of Dried Rehmannia glutinosa Liboschitz (건지황 추출물을 이용한 알콜 발효 수율 증진)

  • Ahn, Sang-Wook;Kim, Min-Hoe;Chung, Woo-Taek;Hwang, Baek;Seong, Nak-Sul;Lee, Hyeon-Yong
    • Korean Journal of Medicinal Crop Science
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    • v.8 no.4
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    • pp.351-361
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    • 2000
  • The juice extract of Rehmannia glutinosa Liboschitz was used to improve the productivity of ethanol in alcohol fermentation process using a 5 L fermentor under batch and fed-batch cultivations. For batch cultivation, both cell density and ethanol production were increased as the extract of R. glutinosa was increased, showing 11.8 (g/L) of maximum cell density and 0.092 (% /hr) of maximum alcohol productivity in adding 30% (v/v) of the extract. However, in adding more than 40% of the extract both cell growth and ethanol production were dropped. The cell growth was severely inhibited in 50% addition. It was found that fed-batch cultivation in adding 30% of the extract of R. glutinosa was an effective process than batch cultivation, yielding up to 30% cell growth and ethanol production. This ethanol productivity was also 30-40% higher than that obtained from a conventional alcohol fermentation. It can tell that dried R. glutinosa Liboschitz is to be used for both enhancing the yield of alcohol fermentation and utilizing biologically active substances possibly transported from R. glutinosa Liboschitz into fermented broth.

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Ammonia Production from Yeast Extract and Its Effect on Growth of the Hyperthermophilic Archaeon Sulfolobus solfataricus

  • Park, Chan-Beum;Lee, Sun-Bok
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.3 no.2
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    • pp.115-118
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    • 1998
  • Utilization of yeast extract and formation of byproduct metabolite were investigated for hyperthermophilic archaeon Sulfolobus solfataricus (DSH 1617). In both batch and fed-batch cultivations of S. solfataricus, maximal cell density, {{{{ { NH}`_{4 } ^{ +} }}} ion production and pH change were highly dependent on the ratio of yeast extract to glucose in the medium. Variation of {{{{ { NH}`_{4 } ^{ +} }}}} ion level was identified as a major cause of pH change during cultivation, and acidification of culture broth was attributed to consumption of {{{{ { NH}`_{4 } ^{ +} }}}} ions rather than formation of acid byproducts. It was also observed that increase of {{{{ { NH}`_{4 } ^{ +} }}}} ion concentrations in the medium resulted in greater degree of growth inhibition.

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High Cell Density Cultivation of Pseudomonas oleovorans for the Production of Poly(3-Hydroxyalkanoates)

  • Lee, Sang-Yup
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.1 no.1
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    • pp.51-53
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    • 1996
  • Fed-batch culture of Pseudomonas oleovorans was carried out for the production of medium-chain-length polyhydroxyalkanoates (MCL-PHAs) using octanoate as a carbon source. Octanoate and the salt solution containing ammounium sulfate and magnesium sulfate were intermittently fed in the course of fermentation. Cell mass and PHA concentrations of 42.8 and 16.8g/L, respectively, could be obtained in 40 h. The PHA content and the PHA productivity were 39.2% and 0.42 g PHA/L-h, respectively. The yields of cell mass and PHA were 0.71 g dry cell mass/g octanoate and 0.28g PHA/g octanoate, respectively. Therefore, octanoate can be used for the production of MCL-PHAs to a high concentration with high productivity.

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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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Fed-batch Cultivation of Escherichia coli YK537 (pAET-8) for Production of phoA Promoter-controlled Human Epidermal Growth Factor

  • Wang Yonggang;Du Peng;Gan Renbao;Li Zhimin;Ye Qin
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.2
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    • pp.149-154
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    • 2005
  • Secretion of the expressed heterologous proteins can reduce the stress to the host cells and is beneficial to their recovery and purification. In this study, fed-batch cultures of Escherichia coli YK537 (pAET-8) were conducted in a 5-L fermentor for the secretory production of human epidermal growth factor (hEGF) whose expression was under the control of alkaline phosphatase promoter. The effects of feeding of glucose and complex nitrogen sources on hEGF production were investigated. When the fed-batch culture was conducted in a chemically de-fined medium, the cell density was 9.68 g/L and the secreted hEGF was 44.7 mg/L in a period of 60 h. When a complex medium was used and glucose was added in pH-stat mode, the secreted hEGF was improved to 345 mg/L. When the culture was fed with glucose at a constant specific rate of $0.25\;gg^{-1}h^{-1}$, hEGF reached 514 mg/L. The effects of adding a solution containing yeast extract and tryptone were further studied. Different rate of the nitrogen source feeding resulted in different levels of phosphate and acetic acid formation, thus affected hEGF expression. At the optimal feeding rate, hEGF production achieved 686 mg/L.

Expression profile analysis of metabolism of Escherichia coli during high cell density cultivation using DNA chip

  • Yun, Seong-Ho;Lee, Sang-Yeop;Im, Geun-Bae
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.600-603
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    • 2000
  • DNA chip containing 207 E. coli genes related to important metabolisms such as (TCA cycle, glycolysis, fermentation and etc) were used to carry out a comprehensive investigation of the change in metabolism and physiology during high cell density culture of E. coli by fed-batch cultivation.

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Efficient Production of ε-Poly-L-Lysine by Streptomyces ahygroscopicus Using One-Stage pH Control Fed-Batch Fermentation Coupled with Nutrient Feeding

  • Liu, Sheng-Rong;Wu, Qing-Ping;Zhang, Ju-Mei;Mo, Shu-Ping
    • Journal of Microbiology and Biotechnology
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    • v.25 no.3
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    • pp.358-365
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    • 2015
  • ε-Poly-L-lysine (ε-PL) is a homopolymer of L-lysine molecules connected between the epsilon amino and alpha carboxyl groups. This polymer is currently used as a natural preservative in food. Insufficient biomass is a major problem in ε-PL fermentation. Here, to improve cell growth and ε-PL productivity, various nitrogen-rich nutrients were supplemented into flask cultures after 16 h cultivation, marking the onset of ε-PL biosynthesis. Yeast extract, soybean powder, corn powder, and beef extract significantly improved cell growth. In terms of ε-PL productivity, yeast extract at 0.5% (w/v) gave the maximum yield (2.24 g/l), 115.4% higher than the control (1.04 g/l), followed by soybean powder (1.86 g/l) at 1% (w/v) and corn powder (1.72 g/l) at 1% (w/v). However, supplementation with beef extract inhibited ε-PL production. The optimal time for supplementation for all nutrients examined was at 16 h cultivation. The kinetics of yeast-extract-supplemented cultures showed enhanced cell growth and production duration. Thus, the most commonly used two-stage pH control fed-batch fermentation method was modified by omitting the pH 5.0-controlled period, and coupling the procedure with nutrient feeding in the pH 3.9-controlled phase. Using this process, by continuously feeding 0.5 g/h of yeast extract, soybean powder, or corn powder into cultures in a 30 L fermenter, the final ε-PL titer reached 28.2 g/l, 23.7 g/l, and 21.4 g/l, respectively, 91.8%, 61.2%, and 45.6% higher than that of the control (14.7 g/l). This describes a promising option for the mass production of ε-PL.

Extracellular Overproduction of $\beta$-Cyclodextrin Glucanotransferase in a Recombinant E. coli Using Secretive Expression System

  • Lee, Kwang-Woo;Shin, Hyun-Dong;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.12 no.5
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    • pp.753-759
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    • 2002
  • $\beta$-Cyclodextrin glucanotransferase ($\beta$-CGTase) was overproduced extracellularly using recombinant E. coli by transforming the plasmid pECGT harboring a secretive signal peptide. The $\beta$-CGTase gene of alkalophilic Bacillus firmus var alkalophilus was inserted into the high expression vector pET20b(+) containing a secretive pelB signal peptide, and then transformed into E. coli BL2l(DE3)pLysS. The optimum culture conditions fer the overproduction of $\beta$-CGTase were determined to be TB medium containing 0.5% (w/v) soluble starch at post-induction temperature of $25^{\circ}C$. A significant amount of $\beta$-CGTase, up to 5.83 U/ml, which was nine times higher than that in the parent strain B. firmus var. alkalophilus, was overproduced in the extracellular compartment. A pH-stat fed-batch cultivation of the recombinant E. coli was also performed to achieve the secretive overproduction of $\beta$-CGTase at a high cell density, resulting in production of up to 21.6 U/ml of $\beta$-CGTase.