Biosynthetic enzymes for streptothricin group antibiotic

• Project/Area Number: 15K14724
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Bioorganic chemistry
• Research Institution: Fukui Prefectural University
• Principal Investigator: Hamano Yoshimitsu 福井県立大学, 生物資源学部, 教授 (50372834)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2016: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: ペプチド結合 / tRNA / 生合成 / 二次代謝産物 / ペプチド

Outline of Final Research Achievements

The antibiotic streptothricin (ST) possesses an amino sugar bound to an L-β-lysine (β-Lys) residue via a peptide bond. The peptide-bond formation has been shown to be catalyzed by a nonribosomal peptide synthetase (NRPS) during ST biosynthesis. The focus of this study is the closely related ST analogue BD-12, which carries a glycine-derived side chain rather than a β-Lys residue. In the biosynthetic studies of BD-12, we revealed that the peptide bond between the amino sugar and the glycine residue is catalyzed by a Fem-like enzyme (Orf 11) in a tRNA-dependent manner rather than by an NRPS; Orf 11 utilizes Gly-tRNA as a substrate.
From a genome database, we further identified the enzyme (Sba18) homologous to Orf11 and investigated the substrate specificity. Interestingly, Sba18 was found to accept Ala-tRNA and Ser-tRNA as well as Gly-tRNA, producing new ST-related compounds which carry alanine and serine side chains.

Research Products

• [Journal Article] tRNA-dependent aminoacylation of an amino sugar intermediate in the biosynthesis of a streptothricin-related antibiotic (2016)
  • Author(s): Maruyama, C., Niikura, H., Izumikawa, M., Hashimoto, J., Shin-ya, K., Komatsu, M., Ikeda, H., Kuroda, M., Sekizuka, T., Ishikawa, J., Hamano, M.
  • Journal Title: Appl. Environ. Microbiol. Volume: 82 Issue: 12 Pages: 3640-3648
  • DOI: 10.1128/aem.00725-16
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] ストレプトスリシン類縁生合成遺伝子群に見出したAla-tRNAAla依存型ペプチド合成酵素の機能解析 (2016)
  • Author(s): 松田貫暉，丸山千登勢，橋本絢子，新家一男，濱野吉十
  • Organizer: 2016年度日本生物工学会大会
  • Place of Presentation: 富山国際会議場（富山市）
  • Year and Date: 2016-09-28
• [Presentation] 抗生物質BD-12生合成に関与するN-メチル基転移酵素の機能解析 (2016)
  • Author(s): 新倉春香，丸山千登勢，新家一男，濱野吉十
  • Organizer: 2016年度日本生物工学会大会
  • Place of Presentation: 富山国際会議場（富山市）
  • Year and Date: 2016-09-28
• [Presentation] ストレプトスリシン類縁生合成遺伝子群に見出したAla-tRNAAla依存型ペプチド合成酵素の機能解析 (2016)
  • Author(s): 松田貫暉，丸山千登勢，橋本絢子，新家一男，濱野吉十
  • Organizer: 2016年度日本放線菌学会大会
  • Place of Presentation: 東京大学農学部（東京都文京区）
  • Year and Date: 2016-09-08
• [Presentation] BD-12生合成におけるN-formimidoyl基転移酵素の酵素学的諸性質 (2016)
  • Author(s): 新倉春香，丸山千登勢，泉川美穂，新家一男，濱野吉十
  • Organizer: 日本農芸化学会2016年度大会
  • Place of Presentation: 札幌市コンベンションセンター（北海道、札幌市）
  • Year and Date: 2016-03-27
• [Presentation] ストレプトスリシン類縁生合成遺伝子群に見出したAla-tRNA依存型ペプチド合成酵素の機能解析 (2016)
  • Author(s): 松田貫暉，丸山千登勢，橋本絢子，新家一男，濱野吉十
  • Organizer: 日本農芸化学会・中部支部・第177回例会
  • Place of Presentation: 名古屋大学農学部（名古屋市）
• [Presentation] 抗生物質BD-12生合成遺伝子の機能解析 (2015)
  • Author(s): 新倉春香, 丸山千登勢, 泉川美穂, 石川淳, 池田治生, 新家一男, 濱野吉十
  • Organizer: 2015年度日本生物工学会大会
  • Place of Presentation: 城山観光ホテル（鹿児島県、鹿児島市）
  • Year and Date: 2015-10-26
• [Presentation] 抗生物質BD-12生合成遺伝子の機能解析 (2015)
  • Author(s): 新倉春香，丸山千登勢，泉川美穂，石川淳，池田治生，新家一男，濱野吉十
  • Organizer: 2015年度日本放線菌学会大会
  • Place of Presentation: 富山国際会議場（富山県、富山市）
  • Year and Date: 2015-09-07