Evolution-inspired engineering of nonribosomal peptide synthetases

Science. 2024 Mar 22;383(6689):eadg4320. doi: 10.1126/science.adg4320. Epub 2024 Mar 22.

Abstract

Many clinically used drugs are derived from or inspired by bacterial natural products that often are produced through nonribosomal peptide synthetases (NRPSs), megasynthetases that activate and join individual amino acids in an assembly line fashion. In this work, we describe a detailed phylogenetic analysis of several bacterial NRPSs that led to the identification of yet undescribed recombination sites within the thiolation (T) domain that can be used for NRPS engineering. We then developed an evolution-inspired "eXchange Unit between T domains" (XUT) approach, which allows the assembly of NRPS fragments over a broad range of GC contents, protein similarities, and extender unit specificities, as demonstrated for the specific production of a proteasome inhibitor designed and assembled from five different NRPS fragments.

MeSH terms

  • Amino Acid Sequence / genetics
  • Bacterial Proteins* / chemistry
  • Bacterial Proteins* / classification
  • Bacterial Proteins* / genetics
  • Evolution, Molecular*
  • Peptide Synthases* / chemistry
  • Peptide Synthases* / classification
  • Peptide Synthases* / genetics
  • Phylogeny
  • Protein Engineering*
  • Sequence Analysis, Protein

Substances

  • non-ribosomal peptide synthase
  • Peptide Synthases
  • Bacterial Proteins