Cloning, Expression and Purification of Intact Polyketide Synthase Modules

Laurence Maschio, Alice Parnell, Nick R Lees, Chris Willis, Christiane Berger-Schaffitzel, James E M Stach, Paul Race

Research output: Chapter in Book/Report/Conference proceedingChapter in a book

Abstract

Polyketides are a structurally and functionally diverse family of bioactive natural products that have proven to be a rich source of pharmaceutical and agrochemical lead compounds. Many polyketides are biosynthesized by large multifunctional megaenzymes termed type 1 modular polyketide synthases (PKSs). These systems possess a distinctive assembly line-like architecture, comprising a series of linearly arranged, multi-domain extension modules, housed in sequence within giant polypeptide chains. Due to their inherently modular structures, PKSs represent attractive targets for re-engineering, enabling access to functionally optimized ‘non-natural’ natural products. In this chapter we describe methods for the molecular cloning, recombinant over-expression, and purification of intact PKS modules and multi-modular PKS polypeptides. The usefulness of these methods is demonstrated by applying them to the study of the abyssomicin C PKS, a >1 MDa multi-modular synthase responsible for the biosynthesis of a polyketide antimicrobial lead compound.
Original languageEnglish
Title of host publication[forthcoming Methods in Enzymology volume]
PublisherElsevier Inc.
Chapter4
Pages63-82
Number of pages20
Volume617
DOIs
Publication statusAccepted/In press - 31 Oct 2018

Publication series

NameMethods in Enzymology
PublisherAcademic Press
ISSN (Print)0076-6879

Structured keywords

  • BrisSynBio
  • Bristol BioDesign Institute
  • BCS and TECS CDTs

Keywords

  • Polyketide synthase
  • protein expression
  • enzymology
  • protein structure
  • natural products
  • Synthetic biology

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  • Cite this

    Maschio, L., Parnell, A., Lees, N. R., Willis, C., Berger-Schaffitzel, C., Stach, J. E. M., & Race, P. (Accepted/In press). Cloning, Expression and Purification of Intact Polyketide Synthase Modules. In [forthcoming Methods in Enzymology volume] (Vol. 617, pp. 63-82). (Methods in Enzymology). Elsevier Inc.. https://doi.org/10.1016/bs.mie.2018.12.018