| Project/Area Number |
01580158
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
物質生物化学
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| Research Institution | Kyoto Institute of Technology |
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Principal Investigator |
HARA Saburo Kyoto Institute of Technology Faculty of Engineering and Design Chemistry and Materials Technology Professor, 工芸学部, 教授 (00028193)
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| Project Period (FY) |
1989 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
|
| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1991: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1990: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1989: ¥900,000 (Direct Cost: ¥900,000)
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| Keywords | Serine proteinase inhibitor / Squash family inhibitor / Molecular evolution / 分子進化 / 一次構造 / カンピョウ / ニガウリ |
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| Research Abstract |
Squash family inhibitors which are included in cucurbitaceae seeds are composed of only 28 amino acid residues. Therefore, we could draw a phylogenetic tree simply by tracing substitutions of amino acids among amino acid sequences of inhibitors.
For this purpose, many kinds of cucurbitaceous seeds were investigated for their proteinase inhibitory activities. From the results, bitter gourd, bottle gourd, gourd and oriental pickling melon were choosed for the purification of inhibitors. By the investigation of the purification method, it was clarified that acetone precipitation and chromatographies on ion-exchangers and reversed phase matrices were the simple and convenient method for the purification of squash family inhibitors.
Total 12 inhibitors were isolated from 4 kinds of cucurbitaceous seeds (4 inhibitors from bitter gourd, 2 from bottle gourd, 3 froin gourd and 3 from oriental pickling inelon) and their amino acid sequences were determined as follows.
Bitter gourd
MCTI-I <ERRCPRILKQCKRDSDCPGECICMAHGFCG
MCTI-II RRCPRIWMECKRDSDCMAQCICVD-GFOG
MCTI-III <ERGCPRILKQCKQDSDCPGECICMAHGFCG
MCEI-I RICPLIWMECKRDSDCLAQCICVD-GHCG
Bottle gourd
LLDTI-I <ERRCPRIYMECKHDSDCLADCVCLEHGICG
LLDTI-II RRCPRIYMECKHDSDCLADCVCLEHGICG
Gourd
LLTI-I <ERRCPRIYMECKHDSDCLADCVCLEHGICGG
LLTI-II RRCPRIYMECKHDSDCLADCVCLEHGICGG
LLTI-III ERRCPRIYMECKHDSDCLADCVCLEHGICGG
Oriental pickling melon
CMCTI-I MCPKILMKCKQDSDCLLDCVCKE-GFCG
CMCTI-II RMCPKILMKCKQDSDCLLDCVCKE-GFCG
CMCTI-III <ERMCPKILMKCKQDSDCLLDCVCKE-GFCG
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