| Project/Area Number |
61480419
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
小児・社会系歯学
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| Research Institution | The University of Tokushima |
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Principal Investigator |
NAKAMURA Ryo The University of Tokushima, Professor, 歯学部, 教授 (30034169)
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| Co-Investigator(Kenkyū-buntansha) |
SHIMADA Junko The University of Tokushima, Research Associate, 歯学部, 教務職員 (10170945)
HINODE Daisuke The University of Tokushima, Research Associate, 歯学部, 助手 (70189801)
ENDO Junko The University of Tokushima, Research Associate, 歯学部, 助手 (20176796)
MAEHARA Reiko The University of Tokushima, Lecturer, 歯学部附属病院, 講師 (30181605)
SATO Makoto The University of Tokushima, Associate Professor, 歯学部, 助教授 (10126229)
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| Project Period (FY) |
1986 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥5,700,000 (Direct Cost: ¥5,700,000)
Fiscal Year 1988: ¥200,000 (Direct Cost: ¥200,000)
Fiscal Year 1987: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1986: ¥4,000,000 (Direct Cost: ¥4,000,000)
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| Keywords | Bacteroides gingivalis / Collagenase inhibitor / Periodontal disease / 大豆 / バクテロイデス / ジンジバリス / コラゲナーゼ活性阻害剤 / 大豆タンパク |
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| Research Abstract |
Our previous paper have shown that Bacteroides gingivalis produces SH dependent collagenolytic enzyme and that the activity is inhibited by the extract of soy-bean meal. The purpose of this study is to describe the isolation, partial purification and some caracterization of this substance.
Dried soy-beans were pulverized to pass a 20 mesh sieve with a coffee mill. Two percent soy-bean extract in 10 mM tris-HCl buffer, pH 7.4 including 0.1 M NaCl, was centrifuged at 10,000 Xg. And the supernatant was usedv as crude soy-bean extract. The inhibitory substance in the crude extract was further purified by the procedures including deae-cellulose column chromatography and gel filtration on sepharose CL-6B, and the active fraction for the inhibition of the collagenolytic enzyme from B gingivalis was collected, concentrated and termed as soy-bean collagenase inhibitory substance(SCI). The crude extract contained soy-bean trypsin inhibitor (STI), and inhibited the trypsin and collagenolytic enzyme of B. gingivalis, but no inhibition was observed for clostridium histolyticum enzyme and the protease of B. gingivalis. However, the SCI was successfully separated from STI by these procedures. The estimated molecular weight of SCI by gel filtration was approximately 700 K dalton, but separated into several subunits which have molecular weight of less than 100 K dalton by the treatment with sodium dodecyl sulfate. The sTI was inactivated by the heat treatment in an autoclave for 20 min at 15 lbs/in^2, while SCI was stable for the same treatment. This substance was very effective in preveinting human gingival fibroblast cells (HGF) from cytopathogenecity of the collagenolytic enzyme of B. gingivalis. These results suggest that this collagenolytic enzyme inhitor isolated from soy-beans maintains normal turnover of gingival cells and prevents from the developement of periodontal disease.
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