Nonribosomal biosynthesis of backbone-modified peptides

Nat Chem. 2018 Mar;10(3):282-287. doi: 10.1038/nchem.2891. Epub 2017 Nov 20.


Biosynthetic modification of nonribosomal peptide backbones represents a potentially powerful strategy to modulate the structure and properties of an important class of therapeutics. Using a high-throughput assay for catalytic activity, we show here that an L-Phe-specific module of an archetypal nonribosomal peptide synthetase can be reprogrammed to accept and process the backbone-modified amino acid (S)-β-Phe with near-native specificity and efficiency. A co-crystal structure with a non-hydrolysable aminoacyl-AMP analogue reveals the origins of the 40,000-fold α/β-specificity switch, illuminating subtle but precise remodelling of the active site. When the engineered catalyst was paired with downstream module(s), (S)-β-Phe-containing peptides were produced at preparative scale in vitro (~1 mmol) and high titres in vivo (~100 mg l-1), highlighting the potential of biosynthetic pathway engineering for the construction of novel nonribosomal β-frameworks.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biocatalysis
  • Molecular Structure
  • Peptide Biosynthesis*
  • Peptide Synthases / metabolism*
  • Peptides / chemistry*
  • Peptides / metabolism*
  • Protein Engineering
  • Ribosomes


  • Peptides
  • Peptide Synthases
  • non-ribosomal peptide synthase