Synthetic chain terminators off-load intermediates from a type I polyketide synthase.
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.
Tosin
Manuela
M
Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge, UK. mt406@cam.ac.uk
Betancor
Lorena
L
Stephens
Elaine
E
Li
W M Ariel
WM
Spencer
Jonathan B
JB
Leadlay
Peter F
PF
D003543:Cysteamine / Q000737:chemistry / Q000378:metabolism*; D018942:Macrolides / Q000737:chemistry / Q000378:metabolism*; D008314:Malonates / Q000737:chemistry / Q000378:metabolism; D015394:Molecular Structure; D010204:Pantetheine / Q000737:chemistry / Q000378:metabolism*; D048630:Polyketide Synthases / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity
D018942:Macrolides; D008314:Malonates; D011994:Recombinant Proteins; D010204:Pantetheine; D003543:Cysteamine; D048630:Polyketide Synthases