Development of the comprehensive system for useful secondary metabolism and bioconversion of useful compounds

2010 Fiscal Year Final Research Report

• Project/Area Number: 20310122
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied genomics
• Research Institution: Kitasato University
• Principal Investigator: IKEDA Haruo Kitasato University, 大学院・感染制御科学府, 教授 (90159632)
• Co-Investigator(Kenkyū-buntansha): TAKAMATSU Satoshi 日本大学, 薬学部, 准教授 (30236351) ; KOMATSU Mamoru 北里大学, 大学院・感染制御科学府, 助教 (40414057)
• Project Period (FY): 2008 – 2010
• Keywords: 産業微生物ゲノム / 物質生産 / 染色体再構成 / 二次代謝産物

Research Abstract

Genetic analysis of production process in secondary metabolites reveals their biosynthetic pathways and their regulatory networks for gene expression. These results reflect on not only basic research but also application to the industrial production process. But almost Streptomyces microorganisms that produce valuable secondary metabolites including antibiotics cannot be applied to molecular genetic approaches (DNA transformation, gene targeting and so on). In this study, we have developed a versatile host for the production of exogenous secondary metabolites using efficient heterologous gene expression system. An industrial anthelmintic antibiotic, avermectin, producer S. avermitilis was used as this model and its chromosome has been engineered for heterologous expression of gene cluster for secondary metabolite biosynthesis. The results indicate that the versatile host constructed is useful for the heterologous expression of many gene clusters for secondary metabolite biosynthesis.

Research Products

• [Journal Article] Genome mining in Streptomyces. Elucidation of the role of Baeyer-Villiger monooxygenases and non-heme iron-dependent dehydrogenase/oxygenases in the final steps of the biosynthesis of pentalenolactone and neopentalenolactone. (2011)
  • Author(s): Seo, M-J. Zhu, D. Endo, S. Ikeda, H. Cane, D.E.
  • Journal Title: Biochemistry 50 Pages: 1739-1754
  • Peer Reviewed
• [Journal Article] Genome mining in Streptomyces. Discovery of an unprecedented P450-catalyzed oxidative rearrangement that is the final step in the biosynthesis of pentalenolactone. (2011)
  • Author(s): Zhu, D. Seo, M-J. Ikeda, H. Cane, D.E.
  • Journal Title: J.Am.Chem.Soc. 133 Pages: 2128-2131
  • Peer Reviewed
• [Journal Article] Characterization of a silent sesquiterpenoid biosynthetic pathway in Streptomyces avermitilis controlling epi-isozizaene and albaflavenone biosynthesis and isolation of a new oxidized epi-isozizaene metabolite. (2011)
  • Author(s): Takamatsu, S. Lin, X. Nara, A. Komatsu, M. Cane, D.E. Ikeda, H.
  • Journal Title: Microb.Biotechnol. 4 Pages: 184-191
  • Peer Reviewed
• [Journal Article] Biosynthesis of 2-methylisoborneol in Cyanobacteria. (2011)
  • Author(s): Giglio, S. Chou, W.K.W. Ikeda, H. Cane, D.E. Monis, T.
  • Journal Title: Environ.Sci.Technol. 45 Pages: 992-998
  • Peer Reviewed
• [Journal Article] Pentalenic acid is a shunt metabolite in the biosynthesis of the pentalenolactone family of metabolites : Hydroxylation of 1-deoxypentalenic acid mediated by CYP105D7 (SAV_7469) of Streptomyces avermitilis. (2011)
  • Author(s): Takamatsu, S. Lin, X. Fushinobu, S. Shoun, H, Komatsu, M. Cane, D.E. Ikeda, H.
  • Journal Title: J.Antibiot. 64 Pages: 65-71
  • Peer Reviewed
• [Journal Article] Genome sequence of Kitasatospora setae NBRC 14216T : An evolutionary snapshot of the family Streptomycetaceae. (2010)
  • Author(s): Ichikawa, N. Oguchi, A. Ikeda, H. Ishikawa, J. Kitani, S. Watanabe, Y. Nakamura, S. Katano, Y. Kishi, E. Sasagawa, M. Ankai, A. Fukui, S. Hashimoto, Y. Kamata, S. Otoguro, M. Nihira, T. Horinouchi, S. Ohnishi, Y. Hayakawa, M. Kuzuyama, T. Arisawa, A. Nomoto, F. Miura, H. Takahashi, Y. Fijita, N.
  • Journal Title: DNA Res. 17 Pages: 393-406
  • Peer Reviewed
• [Journal Article] Regio-and stereospecificity of filipin hydroxylation sites revealed by crystal structure of cytochrome P450 105P1 and 105D6 from Streptomyces avermitilis. (2010)
  • Author(s): Lin, X. Fushinobu, S. Takamatsu, S. Wakagi, T, Ikeda, H. Shoun, H.
  • Journal Title: J.Biol.Chem. 285 Pages: 16844-16853
  • Peer Reviewed
• [Journal Article] Genome mining in Streptomyces avermitilis : Cloning and characterization of SAV_76, the synthase for a new sesquiterpene. avermitilol. (2010)
  • Author(s): Chou, W.K.W. Fanizza, I. Uchiyama, T. Komatsu, M. Ikeda, H. Cane, D.E.
  • Journal Title: J.Am.Chem.Soc. 132 Pages: 8850-8851
  • Peer Reviewed
• [Journal Article] Genome-minimized Streptomyces host for the heterologous expression of secondary metabolism. (2010)
  • Author(s): Komatsu, M. Uchiyama, Takuma, Cane, D.E. Omura, S. Ikeda, H.
  • Journal Title: Proc.Natl.Acad.Sci.USA. 107 Pages: 2646-2651
  • Peer Reviewed
• [Journal Article] Nocardamin production by Streptomyces avermitilis. (2009)
  • Author(s): Ueki, M. Suzuki, R. Takamatsu, S. Takagi, H. Uramoto, M. Ikeda, H. Osada, H.
  • Journal Title: Actinomycetol 23 Pages: 34-39
  • Peer Reviewed
• [Journal Article] Genomic basis for natural product biosynthetic diversity in the actinomycetes. (2009)
  • Author(s): Nett, M. Ikeda, H. Moore, B.S.
  • Journal Title: Nat.Prod.Rep. 26 Pages: 1362-1384
  • Peer Reviewed
• [Journal Article] Genome mining in Streptomyces avermitilis : A biochemical Baeyer-Villiger reaction and discovery of a new branch of the pentalenolactone family tree. (2009)
  • Author(s): Jiang, J. Tetzlaff, C.N. Takamatsu, S. Iwatsuki, M. Komatsu, M. Ikeda, H. Cane, D.E.
  • Journal Title: Biochemistry 48 Pages: 6431-6440
  • Peer Reviewed
• [Journal Article] The site-specific recombination system of actinophage TG1. (2009)
  • Author(s): Morita, K. Yamamoto, T. Fusada, N. Komatsu, M. Ikeda, H. Hirano, N. Takahashi, H.
  • Journal Title: FEMS Microbiol.Lett. 297 Pages: 234-240
  • Peer Reviewed
• [Journal Article] In vitro characterization of the site-specific recombination system based on actinophage TG1 integrase. (2009)
  • Author(s): Morita, K. Yamamoto, T. Fusada, N. Komatsu, M. Ikeda, H. Hirano, N. Takahashi, H.
  • Journal Title: Mol.Genet.Genome. 282 Pages: 607-616
  • Peer Reviewed
• [Journal Article] Antibiotic overproduction by rpsL and rsmG mutants of various actinomycetes. (2009)
  • Author(s): Tanaka, Y. Komatsu, M. Okamoto, S. Tokuyama, S. Kaji, A. Ikeda, H. Ochi, K.
  • Journal Title: Appl.Env.Microbiol. 75 Pages: 4919-4922
  • Peer Reviewed
• [Journal Article] Characterization of a regulatory gene, aveR, for the biosynthesis of avermectin in Streptomyces avermitilis. (2009)
  • Author(s): Kitani, S. Ikeda, H. Sakamoto, T. Noguchi, S. Nihira, T.
  • Journal Title: Appl.Microbiol Biotechnol. 82 Pages: 1089-1096
  • Peer Reviewed
• [Journal Article] Crystal structures of cytochrome P450 105P1 from Streptomyces avermitilis : conformational flexibility and histidine ligand state. (2009)
  • Author(s): Lin, X. Fushinobu S. Ikeda H. Wakagi T. Shoun H.
  • Journal Title: J.Bacteriaol. 191 Pages: 1211-1219
  • Peer Reviewed
• [Journal Article] Identification and functional analysis of genes controlling biosynthesis of 2-methylisoborneol. (2008)
  • Author(s): Komatsu, M. Tsuda, M. Omura, S. Oikawa, H. Ikeda, H.
  • Journal Title: Proc.Natl.Acad.Sci.USA. 105 Pages: 7422-7427
  • Peer Reviewed
• [Journal Article] Organization of the biosynthetic gene cluster for the polyketide antitumor macrolide, Pladienolide, in Streptomyces platensis Mer-11107. (2008)
  • Author(s): Machida, K. Arisawa, A. Takeda, S. Tsuchida, T. Aritoku, Y. Yoshida, M. Ikeda, H.
  • Journal Title: Biosci.Biotechnol.Biochem. 72 Pages: 2946-2952
  • Peer Reviewed
• [Presentation] 物質生産のためのStreptomyces avermitilisのゲノム改変 (2010)
  • Author(s): 池田治生
  • Organizer: 2010年度日本放線菌学会大会
  • Place of Presentation: 東京
  • Year and Date: 2010-09-02
• [Presentation] Optimized Streptomyces host for the heterologous expression of secondary metabolism. (2010)
  • Author(s): Ikeda H., Komatsu M., Uchiyama T.
  • Organizer: 11^<th> International Symposium on the Genetics of Industrial Microorganisms.
  • Place of Presentation: Melbourone, Australia
  • Year and Date: 2010-06-29
• [Presentation] Waking up bacterial sleeping genes: a new method to elicit bacterial capabilities. (2010)
  • Author(s): Ochi K., Tanaka Y., Komatsu M., Hosaka H., Ikeda H.
  • Organizer: 11^<th> International Symposium on the Genetics of Industrial Microorganisms.
  • Place of Presentation: Melbourone, Australia
  • Year and Date: 2010-06-29
• [Presentation] Characterization of relA mutant of industrial microorganism Streptomyces avermitilis. (2010)
  • Author(s): Komatsu M., Tanaka Y., Ochi K., Ikeda H.
  • Organizer: 11^<th> International Symposium on the Genetics of Industrial Microorganisms.
  • Place of Presentation: Melbourone, Australia
  • Year and Date: 2010-06-29
• [Remarks] ホームページ
  • URL: http://avermitilis.ls.kitasato-u.ac.jp/