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A biosynthetic pathway to aromatic amines that uses glycyl-tRNA as nitrogen donor

Abstract

Aromatic amines in nature are typically installed with Glu or Gln as the nitrogen donor. Here we report a pathway that features glycyl-tRNA instead. During the biosynthesis of pyrroloiminoquinone-type natural products such as ammosamides, peptide-aminoacyl tRNA ligases append amino acids to the C-terminus of a ribosomally synthesized peptide. First, \({\mathrm{Amm}}{{{\mathrm{B}}}}_{{{\mathrm{C}}}}^{{{{\mathrm{Trp}}}}}\) adds Trp in a Trp-tRNA-dependent reaction and the flavoprotein AmmC1 then carries out three hydroxylations of the indole ring of Trp. After oxidation to the corresponding ortho-hydroxy para-quinone, \({\mathrm{Amm}}{{{\mathrm{B}}}}_{{{\mathrm{D}}}}^{{{{\mathrm{Gly}}}}}\) attaches Gly to the indole ring in a Gly-tRNA dependent fashion. Subsequent decarboxylation and hydrolysis results in an amino-substituted indole. Similar transformations are catalysed by orthologous enzymes from Bacillus halodurans. This pathway features three previously unknown biochemical processes using a ribosomally synthesized peptide as scaffold for non-ribosomal peptide extension and chemical modification to generate an amino acid-derived natural product.

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Fig. 1: Structures of pyrroloiminoquinone-derived natural products and activity of PEARL enzymes.
Fig. 2: Post-translational modifications during ammosamide biosynthesis.
Fig. 3: Conversion of Trp to aminoquinone.
Fig. 4: Known pathways to aromatic amines.

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Data availability

The raw data associated with the spectra in Figs. 2 and 3, and Supplementary Figs. 35, 7, 8, 1015, 1721 and 2331 were deposited at Mendeley (van der Donk, Wilfred (2021), “PEARL 2021”, Mendeley Data, V1, https://doi.org/10.17632/mk3ttnbt5t.1).

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Acknowledgements

This work was supported by the National Institutes of Health (R37 GM058822 to W.v.d.D., T32 GM070421 to P.N.D, F32 GM105297 to R.S., F32 GM129944 to C.P.T., and R01 GM085770 to B.S.M.). We thank M. A. Funk and K.-K. A. Wang for initial attempts to activate the bha cluster and D. A. Berthold for help in purifying GlyQS.

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P.N.D., H.L., R.A.S., C.P.T., and W.A.v.d.D. designed the study. P.N.D., H.L., R.A.S., C.P.T. and X.Z. performed all experiments. L.Z. acquired and interpreted the NMR data. B.S.M. provided reagents and helpful discussions; P.N.D. and W.A.v.d.D. wrote the manuscript.

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Correspondence to Wilfred A. van der Donk.

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Peer review information Nature Chemistry thanks Seokhee Kim, A. James Link and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary Figs. 1–34, Methods and Tables 1–5.

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Daniels, P.N., Lee, H., Splain, R.A. et al. A biosynthetic pathway to aromatic amines that uses glycyl-tRNA as nitrogen donor. Nat. Chem. 14, 71–77 (2022). https://doi.org/10.1038/s41557-021-00802-2

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