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Cloning, Expression and Purification of Intact Polyketide Synthase Modules

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

Original languageEnglish
Title of host publication[forthcoming Methods in Enzymology volume]
Publisher or commissioning bodyElsevier Inc.
Chapter4
Pages63-82
Number of pages20
Volume617
DOIs
DateAccepted/In press - 31 Oct 2018

Publication series

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

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.

    Research areas

  • Polyketide synthase, protein expression, enzymology, protein structure, natural products

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