Studying trans-acting enzymes that target carrier protein-bound amino acids during nonribosomal peptide synthesis

Anja Greule, Louise K. Charkoudian, Max J. Cryle

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

1 Citation (Scopus)


Nonribosomal peptide biosynthesis is a complex enzymatic assembly responsible for producing a great diversity of bioactive peptide natural products. Due to the recurring arrangement of catalytic domains within these machineries, great interest has been shown in reengineering these pathways to produce novel, designer peptide products. However, in order to realize such ambitions, it is first necessary to develop a comprehensive understanding of the selectivity, mechanisms, and structure of these complex enzymes, which in turn requires significant in vitro experiments. Within nonribosomal biosynthesis, some modifications are performed by enzymatic domains that are not linked to the main nonribosomal peptide synthetase but rather act in trans: these systems offer great potential for redesign, but in turn require detailed study. In this chapter, we present an overview of in vitro experiments that can be used to characterize examples of such trans-interacting enzymes from nonribosomal peptide biosynthesis: Cytochrome P450 monooxygenases and flavin-dependent halogenases.

Original languageEnglish
Title of host publicationMetabolons and Supramolecular Enzyme Assemblies
EditorsClaudia Schmidt-Dannert, Maureen B. Quin
PublisherAcademic Press
Number of pages42
ISBN (Print)9780128170748
Publication statusPublished - 1 Jan 2019

Publication series

NameMethods in Enzymology
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988


  • Carrier protein
  • Cytochrome P450
  • Halogenase
  • Nonribosomal peptide synthetase
  • trans-interacting enzyme

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