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High-level production of Fc-fused kringle domain in Pichia pastoris

  • Fermentation, Cell Culture and Bioengineering
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Recently, as a new non-immunoglobulin-based protein scaffold, a human kringle domain was successfully engineered toward biologically functional agonists and antagonists. In this study, the fed-batch cultivation conditions were optimized for enhanced production of an Fc-fused kringle domain (KD548-Fc) in Pichia pastoris. Fed-batch cultivations were performed in 5-l laboratory-scale bioreactors, and in order to find the optimal conditions for high-level production of KD548-Fc, several parameters including the initial carbon source (glycerol) concentration, temperature, and pH were investigated. When cells were cultivated at pH 4.0 and 25 °C with 9.5 % glycerol in the initial medium, the highest production yield (635 mg/l) was achieved with high productivity (7.2 mg/l/h). Furthermore, functional KD548-Fc was successfully purified from the culture broth using a simple purification procedure with high purity and recovery yield.

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Acknowledgments

This research was supported by the Converging Research Center Program (grant no. 2009-0082332) and Basic Science Research Program (grant no. 2011-0016235) through the National Research Foundation (NRF) of Korea, funded by the Ministry of Science, ICT, and Future Planning.

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Authors and Affiliations

  1. Department of Chemical and Biomolecular Engineering, KAIST, 335 Gwahagno, Yuseong-gu, Daejeon, 305-701, Republic of Korea

    Gu Min Jeong, Yong Jae Lee & Ki Jun Jeong

  2. Department of Molecular Science and Technology, Ajou University, San 5, Woncheon-dong, Yeongtong-gu, Suwon, 443-749, Republic of Korea

    Yong Sung Kim

  3. Institute for the BioCentury, KAIST, 335 Gwahagno, Yuseong-gu, Daejeon, 305-701, Republic of Korea

    Ki Jun Jeong

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  1. Gu Min Jeong
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  2. Yong Jae Lee
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Correspondence to Ki Jun Jeong.

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Jeong, G.M., Lee, Y.J., Kim, Y.S. et al. High-level production of Fc-fused kringle domain in Pichia pastoris . J Ind Microbiol Biotechnol 41, 989–996 (2014). https://doi.org/10.1007/s10295-014-1435-2

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  • DOI: https://doi.org/10.1007/s10295-014-1435-2

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