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Nonribosomal biosynthesis of backbone-modified peptides.
Journal article

Nonribosomal biosynthesis of backbone-modified peptides.

  • Niquille DL Laboratory of Organic Chemistry, ETH Zurich, 8093 Zurich, Switzerland.
  • Hansen DA Laboratory of Organic Chemistry, ETH Zurich, 8093 Zurich, Switzerland.
  • Mori T Laboratory of Organic Chemistry, ETH Zurich, 8093 Zurich, Switzerland.
  • Fercher D Laboratory of Organic Chemistry, ETH Zurich, 8093 Zurich, Switzerland.
  • Kries H Laboratory of Organic Chemistry, ETH Zurich, 8093 Zurich, Switzerland.
  • Hilvert D Laboratory of Organic Chemistry, ETH Zurich, 8093 Zurich, Switzerland.
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  • 2018-02-21
Published in:
  • Nature chemistry. - 2018
Biocatalysis
Molecular Structure
Peptide Biosynthesis
Peptide Synthases
Peptides
Protein Engineering
Ribosomes
English 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.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://sonar.ch/global/documents/40871
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