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Title page for etd-0814104-005713


URN etd-0814104-005713 Statistics This thesis had been viewed 2155 times. Download 9 times.
Author Chung-Yu Hung
Author's Email Address don0284@yahoo.com.tw
Department Bioengineering
Year 2003 Semester 2
Degree Master Type of Document Master's Thesis
Language zh-TW.Big5 Chinese Page Count 135
Title Studies on the Fermentation Properties and the Metabolic products during continuous Fermentation by Escherichia coli BL21(DE3)W/ pET20-LacZ
Keyword
  • E. coli
  • continuous fermentation
  • continuous fermentation
  • E. coli
  • Abstract The present study employed E. coli BL21 (DE3) and recombinant E. coli BL21 (DE3) w/pET20-LacZ to investigate extra-cellular and intra-cellular metabolic products in the continuous fermentation. The results could be used for understanding the metabolic flux of nutrients in the medium. Four dilution rates (0.57, 0.4, 0.23, 0.13 h-1) were performed in the continuous fermentation. The results revealed that glucose consumption rates (g/DCW/h) in the steady state for E. coli BL21 (DE3) w/pET20-LacZ were higher than those for E. coli BL21 (DE3). This indicated recombinant E. coli BL21 (DE3) w/pET20-LacZ consumed more glucose because of the burden of the plasmid. The plasmid of E. coli BL21 (DE3) w/pET20-LacZ was stable after performing continuous fermentation for 25 working volume without ampicillin. IPTG was introduced in the steady state of the continuous fermentation for E. coli BL21 (DE3) w/pET20-LacZ. The cell densities in terms of OD600 decreased from 2.85 and 3.14 to 2.12 and 3.02 respectively for the dilution rate of 0.4 and 0.57. The results revealed that the specific growth rate decreased because of nutrient was consumed for the production of recombinant protein. Metabolites from glycolysis and TCA cycle are analysed. Accumulation of oxaloacetate in-cell and medium at high dilution rate suggest over-supply of nutrient. For the amino acids, large amount of tryptophan, alanine, tyrosine, and histidine were consumed from the medium. However, the cell needed to convert nutrients to phenylalanine, valine, serine, and threonine that were not exited in the medium.
    Advisor Committee
  • Kow-Jen Duan - advisor
  • none - co-chair
  • none - co-chair
  • Files indicate in-campus access only
    Date of Defense 2004-07-26 Date of Submission 2004-08-14


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