Elucidation of the catalytic mechanism of bacterial cell-surface-expressed chitinase ChiW and chitin oligosaccharide production by using the high transglycosylation activity of ChiW.
| Project/Area Number |
25850234
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Environmental agriculture(including landscape science)
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| Research Institution | Fukui Prefectural University |
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Principal Investigator |
Itoh Takafumi 福井県立大学, 生物資源学部, 准教授 (10402827)
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| Research Collaborator |
HIBI Takao 福井県立大学, 生物資源学部, 教授 (00285181)
KIMOTO Hisashi 福井県立大学, 生物資源学部, 教授 (70283166)
FUJII Yutaka 福井大学, 医学部, 教授 (80211522)
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| Project Period (FY) |
2013-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | Chitinase / Paenibacillus / Structural biology / chitin oligosaccharides / chitinase / structural biology / chitin / oligosaccharides |
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| Outline of Final Research Achievements |
The purposes of this study are to elucidate the degradation mechanism of the novel multi-modular surface-expressed chitinase, ChiW, of Paenibacillus sp. str. FPU-7 and to enhance the transglycosylation activity of the enzyme.
(1) The degradation mechanism of ChiW. The crystal structure of ChiW was determined at 2.0 angstrom resolution. The structural and functional analyses reveal that the enzyme is composed of six structural domains for efficient chitin degradation. (2) Enhancement of the transglycosylation activity of ChiW. The important residues for substrate binding were selected as the mutation sites. The mutant enzymes were prepared by site-specific mutagenesis. As the result, two mutant enzymes with higher transglycosylation activities (120-130%) were obtained. (3) Identification of P. FPU-7 proteins for chitin degradation. Proteomic analysis was carried on P. FPU-7 extracellular proteins. A number of chitinase and chitin-related proteins were identified thorough the analysis.
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Report
(4 results)
Research Products
(17 results)
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[Journal Article] Crystallization and preliminary X-ray analysis of the catalytic domains of Paenibacillus sp. strain FPU-7 cell-surface-expressed chitinase ChiW.2014
Author(s)
Itoh, T., Hibi, T., Sugimoto, I., Suzuki, F., Fujii, Y., Taketo, A. and Kimoto, H.
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Journal Title
Acta Crystallogr. F Struct. Biol. Commun.
Volume: 70
Issue: 3
Pages: 350-353
DOI
Related Report
Peer Reviewed
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[Journal Article] Cooperative degradation of chitin by extracellular and cell surface-expressed chitinases from Paenibacillus sp. strain FPU-7.2013
Author(s)
Itoh, T., Hibi, T., Fujii, Y., Sugimoto, I., Fujiwara, A., Suzuki, F., Iwasaki, Y., Kim, J-K., Taketo, A. and Kimoto, H.
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Journal Title
Appl. Environ. Microbiol.
Volume: 79
Issue: 23
Pages: 7482-7490
DOI
Related Report
Peer Reviewed
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[Presentation] Chitinases secreted by Paenibacillus sp. strain IK-5 form a huge complex.2013
Author(s)
Kimoto, H., Itoh, T., Hibi, T., Fujiwara, A., Fujii, Y., Tanaka, Y., Suzuki, F., Taketo, A., Kusaoke, H.
Organizer
10th Asia-Pacific Chitin & Chitosan Symposium
Place of Presentation
米子市, 米子コンベンションセンター BIG SHIP
Related Report
Invited
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[Presentation] Paenibacillus str. FPU-7由来キチナーゼ ChiWのN末端領域の機能解析2013
Author(s)
高宮 杏奈, 伊藤 貴文, 日ビ 隆雄, 大門 結花, 髙木 良樹, 仲上 真由, 鈴木 史子, 杉本 郁美, 藤井 豊, 武藤 明, 木元 久
Organizer
2013北陸合同バイオシンポジウム
Place of Presentation
七尾市, 国民宿舎能登小牧台 コンベンションホール「万葉」
Related Report
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