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Lookup NU author(s): Dr James StachORCiD,
Dr Paul Race
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© 2019 Elsevier Inc. 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 I modular polyketide synthases (PKSs). These systems possess a distinctive assembly line-like architecture, comprising a series of linearly arranged, multidomain extension modules, housed in sequence within giant polypeptide chains. Due to their inherently modular structures, PKSs represent attractive targets for reengineering, enabling access to functionally optimized “nonnatural” natural products. In this chapter we describe methods for the molecular cloning, recombinant over-expression, and purification of intact PKS modules and multimodular PKS polypeptides. The usefulness of these methods is demonstrated by applying them to the study of the abyssomicin C PKS, a > 1 MDa multimodular synthase responsible for the biosynthesis of a polyketide antimicrobial lead compound.
Author(s): Maschio L, Parnell AE, Lees NR, Willis CL, Schaffitzel C, Stach JEM, Race PR
Publication type: Book Chapter
Publication status: Published
Book Title: Methods in Enzymology
Online publication date: 04/02/2019
Acceptance date: 02/04/2018
Publisher: Academic Press