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

Abstract

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
Chapter6
Pages113-154
Number of pages42
ISBN (Print)9780128170748
DOIs
Publication statusPublished - 1 Jan 2019

Publication series

NameMethods in Enzymology
Volume617
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

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

Cite this

Greule, A., Charkoudian, L. K., & Cryle, M. J. (2019). Studying trans-acting enzymes that target carrier protein-bound amino acids during nonribosomal peptide synthesis. In C. Schmidt-Dannert, & M. B. Quin (Eds.), Metabolons and Supramolecular Enzyme Assemblies (pp. 113-154). (Methods in Enzymology; Vol. 617). Academic Press. https://doi.org/10.1016/bs.mie.2018.12.008
Greule, Anja ; Charkoudian, Louise K. ; Cryle, Max J. / Studying trans-acting enzymes that target carrier protein-bound amino acids during nonribosomal peptide synthesis. Metabolons and Supramolecular Enzyme Assemblies. editor / Claudia Schmidt-Dannert ; Maureen B. Quin. Academic Press, 2019. pp. 113-154 (Methods in Enzymology).
@inbook{e7162e60546843f1a28939f5d64ea0d7,
title = "Studying trans-acting enzymes that target carrier protein-bound amino acids during nonribosomal peptide synthesis",
abstract = "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.",
keywords = "Carrier protein, Cytochrome P450, Halogenase, Nonribosomal peptide synthetase, trans-interacting enzyme",
author = "Anja Greule and Charkoudian, {Louise K.} and Cryle, {Max J.}",
year = "2019",
month = "1",
day = "1",
doi = "10.1016/bs.mie.2018.12.008",
language = "English",
isbn = "9780128170748",
series = "Methods in Enzymology",
publisher = "Academic Press",
pages = "113--154",
editor = "Claudia Schmidt-Dannert and Quin, {Maureen B.}",
booktitle = "Metabolons and Supramolecular Enzyme Assemblies",
address = "United States",

}

Greule, A, Charkoudian, LK & Cryle, MJ 2019, Studying trans-acting enzymes that target carrier protein-bound amino acids during nonribosomal peptide synthesis. in C Schmidt-Dannert & MB Quin (eds), Metabolons and Supramolecular Enzyme Assemblies. Methods in Enzymology, vol. 617, Academic Press, pp. 113-154. https://doi.org/10.1016/bs.mie.2018.12.008

Studying trans-acting enzymes that target carrier protein-bound amino acids during nonribosomal peptide synthesis. / Greule, Anja; Charkoudian, Louise K.; Cryle, Max J.

Metabolons and Supramolecular Enzyme Assemblies. ed. / Claudia Schmidt-Dannert; Maureen B. Quin. Academic Press, 2019. p. 113-154 (Methods in Enzymology; Vol. 617).

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

TY - CHAP

T1 - Studying trans-acting enzymes that target carrier protein-bound amino acids during nonribosomal peptide synthesis

AU - Greule, Anja

AU - Charkoudian, Louise K.

AU - Cryle, Max J.

PY - 2019/1/1

Y1 - 2019/1/1

N2 - 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.

AB - 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.

KW - Carrier protein

KW - Cytochrome P450

KW - Halogenase

KW - Nonribosomal peptide synthetase

KW - trans-interacting enzyme

UR - http://www.scopus.com/inward/record.url?scp=85061277001&partnerID=8YFLogxK

U2 - 10.1016/bs.mie.2018.12.008

DO - 10.1016/bs.mie.2018.12.008

M3 - Chapter (Book)

SN - 9780128170748

T3 - Methods in Enzymology

SP - 113

EP - 154

BT - Metabolons and Supramolecular Enzyme Assemblies

A2 - Schmidt-Dannert, Claudia

A2 - Quin, Maureen B.

PB - Academic Press

ER -

Greule A, Charkoudian LK, Cryle MJ. Studying trans-acting enzymes that target carrier protein-bound amino acids during nonribosomal peptide synthesis. In Schmidt-Dannert C, Quin MB, editors, Metabolons and Supramolecular Enzyme Assemblies. Academic Press. 2019. p. 113-154. (Methods in Enzymology). https://doi.org/10.1016/bs.mie.2018.12.008