| Project/Area Number |
18560749
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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 |
Biofunction/Bioprocess
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| Research Institution | Niigata University |
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Principal Investigator |
TANIGUCHI Masayuki Niigata University, Institute of Science and Technology, Professor (00163634)
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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 |
¥3,980,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥480,000)
Fiscal Year 2007: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2006: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | Antimicrobial protein / Rice protein / Antimicrobial activity / Protein purification / Periodontal disease / システインプロテアーゼ / 歯周病菌 / プロテアーゼ阻害剤 |
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| Research Abstract |
In this study, the preparation of cysteine protease inhibitors (oryzacystatins: Ocs) as a recombinant protein and their properties were investigated. The screening of antimicrobial proteins from rice and rice bran against Porphyromonas gingivalis was also studied. The results obtained in this study were summarized as follows.
1. Five types of recombinant-OCs were expressed in Escherichia coli BL21 as the GST or His-tag fusion protein and purified by the conventional method. The inhibitory activities of OCs for several cysteine proteases were estimated.
2. To measure sensitively antimicrobial activity, turbidity assay, ATP-chemical luminescence assay and fluorescence assay using DNA-binding reagent were developed and their performances were compared.
3. The proteins inhibitory to proteases (Arg- and Lys-gingipains) from P. gingivalis were detected in the extracts of rice and rice bran.
4. The proteins of MW 1-16 kDa obtained from the extract of rice bran have antimicrobial activity against P. gingivalis. The chemical structure of the proteins purified by two-dimensional electrohoresis was analyzed by MALDI-TOF-MS. The several proteins in the purified samples were identified as candidates of antimicrobial proteins.
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