Orphan receptors and their ligands in Streptomyces secondary metabolism

2015 Fiscal Year Final Research Report

• 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
• Keywords: 放線菌 / 二次代謝 / オーファン受容体 / 未知リガンド

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.

Free Research Field

分子微生物学