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Synthetic Chain Terminators Off‐Load Intermediates from a Type I Polyketide Synthase
Author(s) -
Tosin Manuela,
Betancor Lorena,
Stephens Elaine,
Ariel Li W. M.,
Spencer Jonathan B.,
Leadlay Peter F.
Publication year - 2010
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200900772
Subject(s) - polyketide , polyketide synthase , atp synthase , chemistry , drug discovery , stereochemistry , biosynthesis , natural product , biochemistry , biocatalysis , combinatorial chemistry , enzyme , reaction mechanism , catalysis
Modular biocatalysis is responsible for the generation of countless bioactive products and its mining remains a major focus for drug discovery purposes. One of the enduring hurdles is the isolation of biosynthetic intermediates in a readily‐analysed form. We prepared a series of nonhydrolysable pantetheine and N ‐acetyl cysteamine mimics of the natural (methyl)malonyl extender units recruited for polyketide formation. Using these analogues as competitive substrates, we were able to trap and off‐load diketide and triketide species directly from an in vitro reconstituted type I polyketide synthase, the 6‐deoxyerythronolide B synthase 3 (DEBS3). The putative intermediates, which were extracted in organic solvent and characterised by LC‐HR‐ESI‐MS, are the first of their kind and prove that small‐molecule chain terminators can be used as convenient probes of the biosynthetic process.