Production of recombinant human granulocyte-colony-stimulating factor in high cell density yeast cultures |
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Authors: | Doo Suk Yang Cheon Soon Bae Jeewon Lee |
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Institution: | (1) Bioprocess Engineering Laboratory, Hanhyo Institutes of Technology, #461-6, Jeonmin-Dong, Yusong-Ku Taejon, South Korea |
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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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