| Project/Area Number |
23656522
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Biofunction/Bioprocess
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| Research Institution | Nagoya University |
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Principal Investigator |
NAKANO Hideo 名古屋大学, 生命農学研究科, 教授 (00237348)
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| Project Period (FY) |
2011 – 2012
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| Project Status |
Completed (Fiscal Year 2012)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2012: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2011: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
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| Keywords | 生物機能工学 / 微生物 / 分泌生産 / シグナルペプチド / 酵素 / 酵母 / ゲノムライブラリー |
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| Research Abstract |
The secretion level of foreign protein in recombinant microbe is strongly dependent onthe combination of signal peptides (SPs) and the target protein; therefore optimization of SP sequence to each target protein is a crucial step to maximize the productivity of the secreted protein in heterogenous expression both in prokaryote and eukaryote.In this research, we have developed a novel method to adjust SP for each target protein utilizing the library of endogenous signal peptides of host cells, named signal peptide optimization tool (SPOT), which can easily make a library of SP fused to a target protein without any interfacial sequence. As a model system, beta-galactosidase from Aspergillus oryzae was used as a target protein for the secretion from Saccharomyces secrevisiae, a typical eukaryotic expression host, and demonstrated that several SPs giving a larger amount of secreted protein from the recombinant yeast were found by the screening of relatively a small number of independent clones. In addition, a correlation between the secretion efficiency and the D-score of each signal peptide analyzed by Signal P software was found, suggesting the use of the software would be helpful to increase the quality of the library. The developed method can be used for variety of proteins in a variety of host cells.
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