Peptides derived from non-ribosomal peptide synthetases (NRPSs) have been a bountiful source for drug discovery. The modular nature of NRPS presents a great opportunity to reprogram the enzymatic assembly line to produce novel peptides with desirable properties. Bacterial NRPS engineering have demonstrated serval successful strategies. These include the exchange unit condensation domain (XUC) concept developed by Bozhüyük et al. in 2019¹. It involves in splitting and fusing NRPSs within the condensation domains. Another strategy is adenylation domain substitution described by Calcott et al. in 2020 ². This approach demonstrated that by substituting the domain and the inter domain linker can increase the efficiency of generating functional NRPSs. However, engineering fungal NRPS has been less fruitful.

Here, we share the insights of our progress in reprogramming fungal modular NRPSs. Taken inspiration from the established enginerring methods described above. We created functional chimeric NRPSs by recombining tri-modular NRPSs. We have also characterised the peptide products by NMR. Despite the highly similar function, the evolution of fungal NRPSs have diverged from bacterial NRPSs. We are hoping to gain understanding in phylogeny and structural biology of fungal NRPSs, which can further inform us how to efficiently engineer fungal NRPSs for novel bioactive peptides.

1. Bozhüyük, K. A. J. et al. Modification and de novo design of non-ribosomal peptide synthetases using specific assembly points within condensation domains. Nat. Chem. 11, 653–661 (2019).
2. Calcott, M. J., Owen, J. G. & Ackerley, D. F. Efficient rational modification of non-ribosomal peptides by adenylation domain substitution. Nat. Commun. 11, (2020).