| Project/Area Number |
18580134
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Food science
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| Research Institution | Akita Prefectural Agriculture, Forestry and Fisheries Research Center |
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Principal Investigator |
TAKAHASHI Saori Akita Prefectural Agriculture, Forestry and Fisheries Research Center, Akita Research Institute for Food and Brewing, Chief Researcher (10142184)
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| Co-Investigator(Kenkyū-buntansha) |
HORI Kazuyuki Akita Prefectural Agriculture, Forestry, and Fisheries Research Center, Akita Research Institute for Food and Brewing, Senior Researcher (50181516)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,850,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥450,000)
Fiscal Year 2007: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2006: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | renin / renin-angiotensin system / synthetic substrate / inhibitor / insect cells / expression / soybean / legumes / アスパルティックプロテアーゼ / 結合タンパク質 / エピメラーゼ / ヌクレオチド / ATP |
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| Research Abstract |
The renin-angiotensin-aldosterone system (RAS) plays an essential role in blood pressure control in animals. Renin catalyzes the liberation of angiotensin I from plasma substrate angiotensinogen. The produced decapeptide angiotensin I is an inactive peptide and activated by angiotensin converting enzyme (ACE). ACE cleaves C-terminal dipeptide from angiotensin I. The produced angiotensin II raises blood pressure by vasoconstriction as well as stimulation of the synthesis and release of aldosterone. Thus, renin is a key enzyme of RAS. ACE has been used as a target enzyme in RAS for screening inhibitors because of its simple assay method; however, renin is a rate-limiting enzyme in RAS, so it was not used because the measurement is very complicated. In the present study, we expressed recombinant human renin in Escherichia coil cells, refolded and activated by trypsin. Using recombinant human renin as a target enzyme, we screened the renin inhibitory activity in fermented soybean paste (miso) and found that soybean, the major ingredient of miso, had renin inhibitory activity. Moreover, we expressed recombinant human renin in insect Sf-9 cells and found that mature active renin accumulated in the late stage of cultivation. Using the medium, we simply purified recombinant human renin and characterized various properties of it. Our method should provide a large quantity of recombinant human renin using the baculovirus expression system.
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