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bioRxiv · 10.1101/2022.10.10.511241

Elucidating the molecular programming of a nonlinear nonribosomal peptide synthetase responsible for fungal siderophore biosynthesis.

Jenner, M.·Hai, Y.·Nguyen, H. H.·Passmore, M.·Skyrud, W.·Kim, J.·Garg, N. K.·Zhang, W.·Ogorzalek Loo, R. R.·Tang, Y.
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Abstract

Siderophores belonging to the ferrichrome family are essential for the viability of fungal species and play a key role for virulence of numerous pathogenic fungi. Despite their biological significance, our understanding of how these iron-chelating cyclic hexapeptides are assembled by non-ribosomal peptide synthetase (NRPS) assembly lines remains poorly understood, primarily due to the nonlinearity exhibited by the domain architecture. Herein, we report the biochemical characterization of the SidC NRPS, responsible for construction of the intracellular siderophore ferricrocin. In vitro reconstitution of purified SidC revealed its ability to produce ferricrocin and its structural variant, ferrichrome. Application of intact protein mass spectrometry uncovered several non-canonical events during peptidyl siderophore biosynthesis, including inter-modular loading of amino acid substrates and an adenylation domain capable of poly-amide bond formation. This work expands the scope of NRPS programming, allows biosynthetic assignment of ferrichrome NRPSs, and sets the stage for reprogramming towards novel hydroxamate scaffolds.

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Jenner, M., Hai, Y., Nguyen, H. H., Passmore, M., Skyrud, W., Kim, J., Garg, N. K., Zhang, W., Ogorzalek Loo, R. R., Tang, Y.. 2022-10-10. Elucidating the molecular programming of a nonlinear nonribosomal peptide synthetase responsible for fungal siderophore biosynthesis.. https://doi.org/10.1101/2022.10.10.511241

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