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

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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.

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Yim, .S., Jeong, .K., Chang, .H. et al. High-level secretory production of human granulocyte-colony stimulating factor by fed-batch culture of recombinant Escherichia coli . Bioprocess Biosyst Eng 24, 249–254 (2001). https://doi.org/10.1007/s004490100267

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  • DOI: https://doi.org/10.1007/s004490100267

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