| Project/Area Number |
23380050
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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 | Kyushu University |
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Principal Investigator |
SONOMOTO Kenji 九州大学, (連合)農学研究科(研究院), 教授 (10154717)
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| Co-Investigator(Kenkyū-buntansha) |
KOHDA Daisuke 九州大学, 生体防御医学研究所, 教授 (80186618)
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| Project Period (FY) |
2011-04-01 – 2014-03-31
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| Project Status |
Completed (Fiscal Year 2013)
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| Budget Amount *help |
¥18,850,000 (Direct Cost: ¥14,500,000、Indirect Cost: ¥4,350,000)
Fiscal Year 2013: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2012: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000)
Fiscal Year 2011: ¥8,450,000 (Direct Cost: ¥6,500,000、Indirect Cost: ¥1,950,000)
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| Keywords | ランチビオティック / 異常アミノ酸 / nukacin ISK-1 / ペプチドデザイン / 翻訳後修飾 / 脱水・環化 / 基質認識ドメイン / リーダーペプチド / 異常アミノ酸形成酵素NukM / 結晶化 / ペプチドの機能改変 / プロタミン / 表面プラズモン共鳴 |
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| Research Abstract |
Nukacin ISK-1 is a lantibiotic produced by Staphylococcus warneri ISK-1. Ribosomally synthesized precursor peptide, NukA undergoes introduction of unusual amino acids by a single modification enzyme, NukM, and the leader sequence is removed and the mature peptide is secreted by ABC transporter NukT. NukM was divided into two domains, NukMN and NukMC, based on a prediction that each domain is responsible for dehydration (phosphorylation and further phosphate elimination) and cyclization, respectively. The in vitro reconstitution of NukM, NukMN, and NukMC revealed that NukM can fully modify NukA, and NukMN could partially phosphorylate and dehydrate it. A series of NukT mutants were constructed and their transport activity in vivo and peptidase activity in vitro were investigated. It was suggested that peptidase activity of NukT depends on ATP hydrolysis, and the N-terminal peptidase domain of NukT may cooperatively function with the C-terminal ATP-binding domain.
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