Heterologous expression and structural characterisation of a pyrazinone natural product assembly line

Chembiochem. 2012 Nov 5;13(16):2408-15. doi: 10.1002/cbic.201200340. Epub 2012 Oct 15.

Abstract

Through a number of strategies nonribosomal peptide assembly lines give rise to a metabolic diversity not possible by ribosomal synthesis. One distinction within nonribosomal assembly is that products are elaborated on an enzyme-tethered substrate, and their release is enzyme catalysed. Reductive release by NAD(P)H-dependent catalysts is one observed nonribosomal termination and release strategy. Here we probed the selectivity of a terminal reductase domain by using a full-length heterologously expressed nonribosomal peptide synthetase for the dipeptide aureusimine and were able to generate 17 new analogues. Further, we generated an X-ray structure of aureusimine terminal reductase to gain insight into the structural details associated with this enzymatic domain.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Products / chemistry*
  • Biological Products / metabolism*
  • Crystallography, X-Ray
  • Escherichia coli / chemistry
  • Escherichia coli / metabolism
  • Models, Molecular
  • Molecular Structure
  • Oxidoreductases / chemistry
  • Oxidoreductases / metabolism
  • Peptide Synthases / chemistry
  • Peptide Synthases / metabolism
  • Pyrazines / chemistry*
  • Pyrazines / metabolism*

Substances

  • Biological Products
  • Pyrazines
  • Oxidoreductases
  • Peptide Synthases
  • non-ribosomal peptide synthase