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Enhanced protein secretion from multiprotease-deficient fission yeast by modification of its vacuolar protein sorting pathway

  • Applied Genetics and Molecular Biotechnology
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Abstract

Previously, we achieved approximately 30-fold enhanced secretion of the protease-sensitive model protein human growth hormone (hGH) by multiple gene deletion of seven obstructive proteases in the fission yeast Schizosaccharomyces pombe. However, intracellular retention of secretory hGH was found in the resultant multiprotease-deficient strains. As a solution, genetic modification of the intracellular trafficking pathway that is related to intracellular retention of hGH was attempted on a protease octuple deletant strain. Vacuolar accumulation of the intracellularly retained hGH was identified by secretory expression of hGH fused with EGFP, and three vacuolar protein sorting (vps)-deficient strains, vps10Δ, vps22Δ, and vps34Δ, were determined on account of their hGH secretion efficiency. The mutant vps10Δ was found to be effective for hGH secretion, which suggested a role for vps10 in the vacuolar accumulation of the intracellularly retained hGH. Finally, vps10 deletion was performed on the protease octuple deletant strain, which led to an approximately 2-fold increase in hGH secretion. This indicated the possible application of secretory-pathway modification and multiple protease deletion for improving heterologous protein secretion from the fission yeast S. pombe.

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Acknowledgments

This study was carried out as part of the project ‘The Development of Basic Technologies for Advanced Production Methods Using Microorganism Functions of New Industrial Science and Technology Frontiers’, by the Ministry of Economy, Trade and Industry (METI), and entrusted by New Energy and Industrial Technology Development Organization (NEDO) of Japan.

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Authors and Affiliations

  1. R&D Group, ASPEX Division, Research Center, Asahi Glass Co., Ltd., 1150 Hazawa-cho, Kanagawa-ku, Yokohama, 221-8755, Japan

    Alimjan Idiris, Hideki Tohda, Mayumi Sasaki, Katsunori Okada, Hiromichi Kumagai & Yuko Giga-Hama

  2. Laboratory of Applied Microbiology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Fukuoka, 812-8581, Japan

    Kaoru Takegawa

Authors
  1. Alimjan Idiris
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  2. Hideki Tohda
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  3. Mayumi Sasaki
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  4. Katsunori Okada
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  5. Hiromichi Kumagai
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  6. Yuko Giga-Hama
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  7. Kaoru Takegawa
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Correspondence to Alimjan Idiris.

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Idiris, A., Tohda, H., Sasaki, M. et al. Enhanced protein secretion from multiprotease-deficient fission yeast by modification of its vacuolar protein sorting pathway. Appl Microbiol Biotechnol 85, 667–677 (2010). https://doi.org/10.1007/s00253-009-2151-0

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  • DOI: https://doi.org/10.1007/s00253-009-2151-0

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