Article | Published:

Amino-group carrier-protein-mediated secondary metabolite biosynthesis in Streptomyces

Nature Chemical Biology volume 12, pages 967972 (2016) | Download Citation

Abstract

Amino-group carrier proteins (AmCPs) mediate the biosynthesis of lysine and arginine in some bacteria and archaea. Here we demonstrate that an uncharacterized AmCP-mediated biosynthetic system functions to biosynthesize the previously uncharacterized and nonproteinogenic amino acid (2S,6R)-diamino-(5R,7)-dihydroxy-heptanoic acid (DADH) in Streptomyces sp. SANK 60404. DADH is incorporated into a novel peptide metabolite, vazabitide A, featuring an azabicyclo-ring structure, by nonribosomal peptide synthetases and successive modification enzymes in this bacterium. As the AmCP-mediated machinery for DADH biosynthesis is widely distributed in bacteria, further analysis of uncharacterized AmCP-containing gene clusters will lead to the discovery of novel bioactive compounds and novel biosynthetic enzymes.

  • Compound

    (2S,6R)-diamino-(5R,7)-dihydroxyheptanoic acid

  • Compound

    2-(2S-amino-3-methylbutanamido)-2-(4-hydroxy-1-azabicyclo[3.1.0]hexan-2-yl)acetic acid

  • Compound

    L-glutamic acid

  • Compound

    N-Vzb22-L-glutamate

  • Compound

    N-Vzb22-L-glutamate 5-phosphate

  • Compound

    N-Vzb22-L-glutamate 5-semialdehyde

  • Compound

    N-Vzb22-(2S,6R)-diamino-(5R,7)-dihydroxyheptanoic acid

  • Compound

    N-Vzb22-(2S)-amino-(5R,7)-dihydroxy-6-oxoheptanoic acid

  • Compound

    2-(2S-amino-3-methylbutanamido)-2-(4,5-dihydroxypiperidin-2-yl)acetic acid

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Acknowledgements

This work was supported in part by the Japan Society for the Promotion of Science (KAKENHI grant no. 24228001 to M.N.) and the Japan Foundation for Applied Enzymology (M.N.). We thank M. Tanaka (Daiichi Sankyo, Tokyo, Japan) for kindly providing Streptomyces sp. SANK 60404, and M. Kobayasi (Tsukuba University, Tsukuba, Japan) for providing pHS19. srcts-NRK cells were a gift from Y. Uehara (Iwate Medical University, Morioka, Japan). We thank H. Aono, F. Ito, M. Tanaka, and K. Yamamoto for evaluating cytotoxicity and antimicrobial activities, and Y. Kamide for evaluating regulatory activities in plants.

Author information

Author notes

    • Fumihito Hasebe
    •  & Tsutomu Fujimura

    Present addresses: School of Food and Nutritional Sciences, University of Shizuoka, Shizuoka, Japan (F.H.); Department of Medicine, Tohoku Medical and Pharmaceutical University, Sendai, Japan (T.F.).

Affiliations

  1. Biotechnology Research Center, The University of Tokyo, Tokyo, Japan.

    • Fumihito Hasebe
    • , Kenichi Matsuda
    • , Taro Shiraishi
    • , Takeo Tomita
    • , Tomohisa Kuzuyama
    •  & Makoto Nishiyama
  2. Chemical Biology Research Group, RIKEN Center for Sustainable Resource Science, Wako, Japan.

    • Yushi Futamura
    •  & Hiroyuki Osada
  3. Gene Discovery Research Group, RIKEN Center for Sustainable Resource Science, Wako, Japan.

    • Takeshi Nakano
  4. Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Kawaguchi, Japan.

    • Takeshi Nakano
  5. Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan.

    • Ken Ishigami
  6. Division of Biochemical Analysis, Central Laboratory of Medical Sciences, Juntendo University School of Medicine, Tokyo, Japan.

    • Hikari Taka
    • , Reiko Mineki
    •  & Tsutomu Fujimura

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Contributions

Research was planned and supervised by T.T., T.K., and M.N.; biochemical and genetic experiments were performed by F.H., K.M., and T.S.; cytotoxicity and antimicrobial tests were performed by Y.F. and H.O.; Arabidopsis growth experiments were performed by T.N. and H.O.; experiments using the modified Mosher's method and the acetonide derivative of 1 were performed by F.H. and K.I.; MALDI-TOF-MS/MS was performed by H.T., R.M., and T.F.; and the manuscript was written by F.H., Y.F., T.K., and M.N.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Makoto Nishiyama.

Supplementary information

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  1. 1.

    Supplementary Text and Figures

    Supplementary Results, Supplementary Tables 1–3 and Supplementary Figures 1–12

  2. 2.

    Supplementary Note

    Synthetic and isolation procedures, and NMR data.

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DOI

https://doi.org/10.1038/nchembio.2181

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