Substrate controlled divergence in polyketide synthase catalysis.
Biochemical characterization of polyketide synthases (PKSs) has relied on synthetic substrates functionalized as electrophilic esters to acylate the enzyme and initiate the catalytic cycle. In these efforts, N-acetylcysteamine thioesters have typically been employed for in vitro studies of full PKS modules as well as excised domains. However, substrate engineering approaches to control the catalytic cycle of a full PKS module harboring multiple domains remain underexplored. This study examines a series of alternatively activated native hexaketide substrates on the catalytic outcome of PikAIV, the sixth and final module of the pikromycin (Pik) pathway. We demonstrate the ability to control product formation with greater than 10:1 selectivity for either full module catalysis, leading to a 14-membered macrolactone, or direct cyclization to a 12-membered ring. This outcome was achieved through modifying the type of hexaketide ester employed, demonstrating the utility of substrate engineering in PKS functional studies and biocatalysis.
Douglas A
DA
Hansen
Aaron A
AA
Koch
David H
DH
Sherman
D018942:Macrolides / Q000737:chemistry / Q000378:metabolism; D015394:Molecular Structure; D048630:Polyketide Synthases / Q000235:genetics / Q000378:metabolism*; D061065:Polyketides / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity
D018942:Macrolides; D061065:Polyketides; D048630:Polyketide Synthases; C003145:picromycin