| Project/Area Number |
15380060
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied microbiology
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| Research Institution | Mie University |
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Principal Investigator |
SAKKA Kazuo Mie University, Faculty of Bioresources, Professor, 生物資源学部, 教授 (20154031)
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| Co-Investigator(Kenkyū-buntansha) |
KIMURA Tetsuya Mie University, Faculty of Bioresources, Associate Professor, 生物資源学部, 助教授 (00281080)
OHMIYA Kunio Mie University, Faculty of Bioresources, Professor, 生物資源学部, 教授 (60023488)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥14,100,000 (Direct Cost: ¥14,100,000)
Fiscal Year 2004: ¥4,200,000 (Direct Cost: ¥4,200,000)
Fiscal Year 2003: ¥9,900,000 (Direct Cost: ¥9,900,000)
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| Keywords | Secondary cell wall polymers / Peotidoglycan / SLH Modules / Clostridium josui / Clostridium stercorarium / Clostridium thermocellum / SLHドメイン / 嫌気性細菌 / 細胞表層タンパク質 / 2次細胞壁 / 細胞壁 / キシラナーゼ / モジュラー酵素 / クロストリジウム |
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| Research Abstract |
1.Secondary cell wall polymers(SCWP) and peptidoglycans were prepared from 3 clostridia, C.josui, C.stercorarium and C.thermocellum
2.SLH module polypeptides from C.josui xylanase Xyn10A, C.stercorarium xylanase Xyn10B and C.thermocellum cell surface protein SdbA were produced in Escherichia coli and purified.
3.Interactions between these polypeptides and various cell wall fractions were investigated. As a result, it was found that the SLH module polypeptides from C.josui Xyn10A and C.stercorarium Xyn10B bound to SCWP but that the SLH module polypeptide from C.thermocellum SdbA unexpectedly bound to peptidoglycan.
4.Binding assay using peptidoglycan fraction prepared from E.coli confirmed that the SLH module polypeptide from C.thermocellum SdbA had an affinity for peptidoglycan although it is generally believed that SLH modules bind to SCWP. The SLH module polypeptide from C.thermocellum SdbA should be applicable for cell surface engineering in bacteria lacking SCWP.
5.SCWP of C.josui, C.stercorarium and C.thermocellum were hydrolyzed with HC1 and constituent amino sugars were qualitatively analyzed. N-Acetylglucosamine and N-acetylmannosamine were detected as constituents for C.stercorarium SCWP and N-acetylgalactosamine as well as these amino sugars were identified for C.josui and C.thermocellum SCWP, suggesting that SCWP of these two bacteria include an unidentified structure.
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