Orphan receptors and their ligands in Streptomyces secondary metabolism

• Project/Area Number: 24310157
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Living organism molecular science
• Research Institution: Osaka University
• Principal Investigator: Nihira Takuya 大阪大学, 生物工学国際交流センター, 教授 (70144441)
• Co-Investigator(Kenkyū-buntansha): KITANI Shigeru 大阪大学, 生物工学国際交流センター, 准教授 (10379117) ; KINOSHITA Hiroshi 大阪大学, 生物工学国際交流センター, 助手 (20294035)
• Project Period (FY): 2012-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥19,240,000 (Direct Cost: ¥14,800,000、Indirect Cost: ¥4,440,000); Fiscal Year 2015: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000); Fiscal Year 2014: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000); Fiscal Year 2013: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000); Fiscal Year 2012: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000)
• Keywords: 放線菌 / 二次代謝 / オーファン受容体 / 未知リガンド / オーファンリセプター / TetR型制御因子 / 生理活性物質 / オーファンレセプター / 未同定リガンド / 信号伝達 / 低分子リガンド

Outline of Final Research Achievements

Using Streptomyces avermitilis which produces avermectin as important anthelmintic and insecticidal drugs, among 125 TetR family regulators and 60 two-component sensor kinases, ligand-responsing regulators were selected based on their transcriptional patterns during the cultivation. The selected regulators were overexpressed in E. coli and the recombinant regulators were purified using His-tag affinity chromatography. The His-tagged recombinant regulators were used as the affinity proteins, the cognate ligands were trapped from the culture broth in the affinity column, selectively eluted and purified. The purified ligand were analyzed by High-resolution NMR, LC, LC-MS/MS to decide the corresponding structures.

Research Products

• [Journal Article] Characterization of a gamma-butyrolactone synthetase gene homologue (stcA) involved in bafilomycin production and aerial mycelium formation in Streptomyces sp. SBI034 (2016)
  • Author(s): Intra, B., Nihira, T. and Panbangred, W.
  • Journal Title: Appl Microbiol Biotechnol Volume: 100 Issue: 6 Pages: 2049-2070
  • DOI: 10.1007/s00253-015-7142-8
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Regulation of production of the blue pigment indigoidine by the pseudo γ-butyrolactone receptor FarR2 in Streptomyces lavendulae FRI-5. (2016)
  • Author(s): 2)Kurniawan, YN., Kitani, S., Iida, A., Maeda, A., Nijeholt, J. L., Lee, YK. and Nihira, T.
  • Journal Title: J. Biosci. Bioeng. Volume: 121 Issue: 4 Pages: 372-379
  • DOI: 10.1016/j.jbiosc.2015.08.013
  • NAID: 40020812927
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] 29-Deoxymaklamicin, a new maklamicin analogue produced by a genetically engineered strain of Micromonospora sp. NBRC 110955. (2015)
  • Author(s): 3)Daduang, R., Kitani, S., Sudoh, Y., Pait, I., Thamchaipenet, A., Ikeda, H., Igarashi, Y. and Nihira, T.
  • Journal Title: J. Biosci. Bioeng. Volume: 120 Issue: 6 Pages: 608-613
  • DOI: 10.1016/j.jbiosc.2015.04.004
  • NAID: 110010022641
  • Peer Reviewed
• [Journal Article] Characterization of the biosynthetic gene cluster for maklamicin, a spirotetronate-class antibiotic of the endophytic Micromonospora sp. NBRC 110955. (2015)
  • Author(s): 4)Daduang, R., Kitani, S., Hashimoto, J., Thamchaipenet, A., Igarashi, Y., Ikeda, H. and Nihira, T.
  • Journal Title: Microbiol Res Volume: 180 Pages: 30-39
  • DOI: 10.1016/j.micres.2015.07.003
  • Peer Reviewed
• [Journal Article] SdrA: a new DeoR-family regulator involved in morphological development and antibiotic production in Streptomyces avermitilis. (2013)
  • Author(s): Ulanova, D., Kitani, S., Fukusaki, E. and Nihira, T.
  • Journal Title: Appl. Environ. Microbiol. Volume: 79 Issue: 24 Pages: 79161-7921
  • DOI: 10.1128/aem.02843-13
  • Peer Reviewed