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Production of recombinant human granulocyte-colony-stimulating factor in high cell density yeast cultures

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Abstract

The recombinant human granulocyte-colony-stimulating factor (rhG-CSF) was synthesized in a fusion protein using a GAL1-10 UAS in recombinant Saccharomyces cerevisiae and the intracellular KEX2 cleavage led excretion of mature rhG-CSF into the extracellular culture broth. The recombinant yeast growth in fed-batch cultures was controlled by precise computer-aided medium feed. The optimal C/N ratio in preinduction (glucose/Casamino acids) and post-induction (galactose/yeast extract) feed media was determined at 3 and 2, respectively. The final rhG-CSF and cell concentration was more than 60 mg/L and 70 g/L, respectively, with around 90% plasmid stability and negligible ethanol accumulation. Comparing the cell growth between the hG-CSF + and hG-CSF - recombinant strains shows that the cloned gene product does not hamper the host cell growth.

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Authors and Affiliations

  1. Bioprocess Engineering Laboratory, Hanhyo Institutes of Technology, #461-6, Jeonmin-Dong, Yusong-Ku Taejon, South Korea

    Doo Suk Yang, Cheon Soon Bae & Jeewon Lee

Authors
  1. Doo Suk Yang
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  2. Cheon Soon Bae
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  3. Jeewon Lee
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Suk Yang, D., Soon Bae, C. & Lee, J. Production of recombinant human granulocyte-colony-stimulating factor in high cell density yeast cultures. Biotechnology Letters 19, 655–659 (1997). https://doi.org/10.1023/A:1018338815187

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  • DOI: https://doi.org/10.1023/A:1018338815187

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