Journal
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Volume 85, Issue 3, Pages 667-677Publisher
SPRINGER
DOI: 10.1007/s00253-009-2151-0
Keywords
Fission yeast; Heterologous protein secretion; Protease; Human growth hormone; Vacuolar protein sorting; vps10
Categories
Funding
- The Development of Basic Technologies for Advanced Production Methods Using Microorganism Functions of New Industrial Science and Technology Frontiers
- Ministry of Economy, Trade and Industry (METI)
- Energy and Industrial Technology Development Organization (NEDO) of Japan
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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, vps10 Delta, vps22 Delta, and vps34 Delta, were determined on account of their hGH secretion efficiency. The mutant vps10 Delta 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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