Tosin, Manuela, Betancor, Lorena, Stephens, E. (Elaine), Li, W. M. Ariel, Spencer, Jonathan B. and Leadlay, Peter F.. (2010) Synthetic chain terminators off-load intermediates from a type I polyketide synthase. Chembiochem, Vol.11 (No.4). pp. 539-546. ISSN 1439-4227

Abstract

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.

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