Discovery of a biosynthetic gene cluster for an anti-tubercular agent its application for the establishment of overproduction system for the anti-tubercular agent and the production of anti-cancer drug-

• Project/Area Number: 22790068
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Biological pharmacy
• Research Institution: Hiroshima University
• Principal Investigator: KUMAGAI Takanori 広島大学, 大学院・医歯薬保健学研究院, 准教授 (70274058)
• Project Period (FY): 2010 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2012: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2011: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2010: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 抗結核薬 / D-サイクロセリン / 生合成 / 異種発現 / 放線菌 / サイクロセリン / 生合成遺伝子 / 異種生産 / 抗癌剤 / ヒドロキシ尿素 / 遺伝子破壊 / 遺伝子相補 / クローニング / 一酸化窒素合成酵素 / Streptomyces lavendulae

Research Abstract

In the present study, the biosynthetic pathway for the anti -tubercular agent, D-cycloserine (DCS), was elucidated by the gene disruption analysis of the DCS biosynthetic gene cluster in a DCS-producing Streptomyces lavendulae ATCC11924. In addition, DCS production system in Escherichia (E.) coli was established by introducing the DCS biosynthetic genes into E. coli. Furthermore, DcsA was found to be a novel heme protein involved in the biosynthesis of DCS.

Research Products

• [Journal Article] Establishment of an in vitroD-cycloserine-synthesizing system by using O-ureido-L-serine synthase and D-cycloserine synthetase found in the biosynthetic pathway (2013)
  • Author(s): Uda, N., Matoba, Y., Kumagai, T., Oda, K., Noda, M., and Sugiyama, M
  • Journal Title: Antimicrob. Agents Chemother Volume: 57 Issue: 6 Pages: 2603-2612
  • DOI: 10.1128/aac.02291-12
  • Peer Reviewed
• [Journal Article] Crystallographic study to determine the substrate specificity on an L-serine-acetylating enzyme found in the D-cycloserine biosynthetic pathway (2013)
  • Author(s): Oda, K., Matoba, Y., Kumagai, T., Noda, M., and Sugiyama, M
  • Journal Title: J. Bacteriol Volume: 195 Issue: 8 Pages: 1741-1749
  • DOI: 10.1128/jb.02085-12
  • Peer Reviewed
• [Journal Article] Heme protein and hydroxyarginase necessary for biosynthesis of D-cycloserine (2012)
  • Author(s): Kumagai, T., Takagi, K., Koyama, Y., Matoba, Y., Oda, K., Noda, M., and Sugiyama, M
  • Journal Title: Antimicrob. Agents Chemother Volume: 56 Issue: 7 Pages: 3682-3689
  • DOI: 10.1128/aac.00614-12
  • Peer Reviewed
• [Journal Article] The obesity and fatty liver are reduced by plant-derived Pediococcus pentosaceus in high fat diet-induced obese mice (2012)
  • Author(s): Zhao, X., et al
  • Journal Title: PLOS One Volume: 7 Issue: 2 Pages: e30696-e30696
  • DOI: 10.1371/journal.pone.0030696
  • Peer Reviewed
• [Journal Article] A molecular mechanism for copper transportation to tyrosinase that is assisted by a metallochaperone, caddie protein (2011)
  • Author(s): Matoba, Y., et al
  • Journal Title: The Journal of Biological Chemistry Volume: 286 Issue: 34 Pages: 30219-30231
  • DOI: 10.1074/jbc.m111.256818
  • Peer Reviewed
• [Journal Article] Structural evidence that puromycin hydrolase is a new type of aminopeptidase with a prolyl oligopeptidase family fold (2011)
  • Author(s): Matoba, Y., et al
  • Journal Title: Proteins Volume: 79 Issue: 10 Pages: 2999-3005
  • DOI: 10.1002/prot.23139
  • Peer Reviewed
• [Journal Article] Molecular cloning and heterologous expression of a biosynthetic gene cluster for the antitubercular agent D-cycloserine produced by Streptomyces lavendulae. Antimicrob (2010)
  • Author(s): Kumagai, T., Koyama, Y., Oda, K., Noda, M., Matoba, Y., and Sugiyama, M
  • Journal Title: Agents Chemother Volume: 54 Issue: 3 Pages: 1132-1139
  • DOI: 10.1128/aac.01226-09
  • Peer Reviewed
• [Journal Article] Molecular cloning and heterologous expression of a biosynthetic gene cluster for the antitubercular agent D-cycloserine produced by Streptomyces lavendulae (2010)
  • Author(s): Kumagai, T., et al.
  • Journal Title: Antimicrobial Agents and Chemotherapy Volume: 54 Pages: 1132-1139
  • Peer Reviewed
• [Presentation] 抗結核薬 D-サイクロセリンの生合成に関与する DcsA タンパク質の機能解析 (2012)
  • Author(s): 熊谷孝則, 高木紀抄, 的場康幸, 野田正文, 杉山政則
  • Organizer: 2012 年度日本放線菌学会大会
  • Place of Presentation: 東京
• [Presentation] D-サイクロセリン生合成に関わるL-セリン-O-アセチルトランスフェラーゼの基質特異性に関する構造生物学的研究 (2012)
  • Author(s): 小田康祐, 的場康幸, 熊谷孝則, 野田正文, 杉山政則
  • Organizer: 2012 年度日本放線菌学会大会
  • Place of Presentation: 東京
• [Presentation] D-サイクロセリン生合成に関わるO-ウレイドL-セリン合成酵素の基質特異性に関する構造生物学的研究 (2012)
  • Author(s): 宇田成利, 的場康幸, 小田康祐, 野田正文, 熊谷孝則, 杉山政則
  • Organizer: 2012 年度日本放線菌学会大会
  • Place of Presentation: 東京
• [Presentation] 抗結核薬D-サイクロセリンの生合成に関与するDcsAタンパク質の機能解析 (2012)
  • Author(s): 熊谷孝則, 高木紀抄, 的場康幸, 野田正文, 杉山政則
  • Organizer: 日本放線菌学会
  • Place of Presentation: 東京
• [Presentation] A hydroxylation reaction necessary in the biosynthesis of an antitubercular antibiotic D-cycloserine (2011)
  • Author(s): Kumagai, T., et al
  • Organizer: International Union of Microbiological Society 2011 Congress
  • Place of Presentation: 札幌市
  • Year and Date: 2011-09-08
• [Presentation] A hydroxylation reaction necessary in the biosynthesis of an antitubercular antibiotic D-cycloserine (2011)
  • Author(s): Kumagai, T., Takagi, K., Noda, M., Oda, K., Matoba, Y., and Sugiyama, M
  • Organizer: IUMS2011
  • Place of Presentation: Sapporo, Japan
• [Presentation] 生合成酵素を用いたD-サイクロセリンの試験管内合成 (2011)
  • Author(s): 宇田成利,的場康幸,小田康祐,野田正文,熊谷孝則,杉山政則
  • Organizer: 第52回日本生化学会中国・四国支部例会
  • Place of Presentation: 広島
• [Presentation] D-サイクロセリン生合成遺伝子クラスターのクローニングとその機能解析 (2011)
  • Author(s): 高木紀抄, 熊谷孝則,小山佑介,宇田成利,小田康祐,野田正文,的場康幸,杉山政則
  • Organizer: 第52回日本生化学会中国・四国支部例会
  • Place of Presentation: 広島
• [Presentation] 遺伝子破壊を用いたD-サイクロセリン生合成遺伝子クラスターの機能解析 (2010)
  • Author(s): 熊谷孝則
  • Organizer: 2010年度日本放線菌学会大会
  • Place of Presentation: 東京都
  • Year and Date: 2010-09-02
• [Presentation] 遺伝子破壊を用いたD-サイクロセリン生合成遺伝子クラスターの機能解析 (2010)
  • Author(s): 熊谷孝則,小山祐介,小田康祐,野田正文,的場康幸,杉山政則
  • Organizer: 2010 年度日本放線菌学会大会
  • Place of Presentation: 東京
• [Presentation] D-サイクロセリン生合成に関与する酵素群の機能解析 (2010)
  • Author(s): 宇田成利,的場康幸,小田康祐,野田正文,熊谷孝則,杉山政則
  • Organizer: 2010 年度日本放線菌学会大会
  • Place of Presentation: 東京