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

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.

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Figure 1: Putative biosynthetic pathways of vazabitides through DADH.
Figure 2: Identification of DADH.
Figure 3: DADH biosynthesis in vitro using [1,2-13C2]glucose and the proposed DADH biosynthetic pathway.
Figure 4: Production and structure of vazabitides.

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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.

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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.

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Correspondence to Makoto Nishiyama.

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The authors declare no competing financial interests.

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Supplementary Text and Figures

Supplementary Results, Supplementary Tables 1–3 and Supplementary Figures 1–12 (PDF 1770 kb)

Supplementary Note

Synthetic and isolation procedures, and NMR data. (PDF 8763 kb)

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Hasebe, F., Matsuda, K., Shiraishi, T. et al. Amino-group carrier-protein-mediated secondary metabolite biosynthesis in Streptomyces. Nat Chem Biol 12, 967–972 (2016). https://doi.org/10.1038/nchembio.2181

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