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Bioprocess and Biosystems Engineering

, Volume 24, Issue 4, pp 249–254 | Cite as

High-level secretory production of human granulocyte-colony stimulating factor by fed-batch culture of recombinant Escherichia coli

  •  S. Yim
  •  K. Jeong
  •  H. Chang
  •  S. Lee

Abstract.

Secretory production of human granulocyte colony-stimulating factor fusion protein (hG-CSF) by fed-batch culture of Escherichia coli was investigated in both 2.5-L and 30-L fermentors. To develop a fed-batch culture condition that allows efficient production of hG-CSF, different feeding strategies including pH-stat, exponential and constant feeding were examined. Among these, the constant feeding strategy (0.228 g glucose×min–1) and the exponential feeding that supports a low specific growth rate (µ=0.116 h–1) resulted in the best hG-CSF production. Under these conditions, 4.4 g×L–1 of hG-CSF was produced. The effect of induction time on the protein production was also investigated. For the fed-batch cultures carried out with the pH-stat and exponential feeding strategies, induction at higher cell density (late-exponential phase) resulted in more hG-CSF production compared with induction at lower cell density (early to mid-exponential phase). The constant feeding strategy that supported best hG-CSF production was applied to the scale-up production of hG-CSF in 30 L of fermentor. The maximum dry cell weight and hG-CSF concentration of 51.7 and 4.2 g×L–1, respectively, was obtained.

Keywords

Induction Time Feeding Strategy Secretory Production Lower Cell Density Exponential Feeding 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag 2001

Authors and Affiliations

  •  S. Yim
    • 2
  •  K. Jeong
    • 1
  •  H. Chang
    • 2
  •  S. Lee
    • 1
  1. 1.Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical Engineering and BioProcess Engineering Research Center, Korea Advanced Institute of Science and Technology, 373-1 Kusong-dong, Yusong-gu, Taejon 305-701, Korea
  2. 2.Department of Chemical Engineering and BioProcess Engineering Research Center, Korea Advanced Institute of Science and Technology, 373-1 Kusong-dong, Yusong-gu, Taejon 305-701, Korea

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