Iterative chain elongation by a pikromycin monomodular polyketide synthase.
The unique ability of the pikromycin polyketide synthase (Pik PKS) to generate 12- and 14-membered ring macrolactones presents an opportunity to explore the fundamental processes of polyketide synthesis, specifically, the mechanistic details of the chain extension process. We have overexpressed and purified PikAIII and PikAIV and demonstrated the ability of these proteins to generate triketide lactone products using (14)C-methylmalonyl-CoA as the sole substrate. Monomodular PikAIII generates TKL (1) when reacted alone, and synthesizes TKL (2) upon reaction in combination with PikAIV. Product formation remains dependent on the enzymatic decarboxylation of methylmalonyl-CoA and transfer of the acyl chain within the enzyme rather than acylation by propionyl-CoA from spontaneous decarboxylation. We propose that synthesis of TKL (1) by PikAIII involves iterative assembly of the triketide chain within a PikAIII homodimer analogous to the nonmodular type I PKS systems.
Beck
Brian J
BJ
Department of Microbiology, University of Minnesota, Minneapolis, Minnesota, USA.
Aldrich
Courtney C
CC
Fecik
Robert A
RA
Reynolds
Kevin A
KA
Sherman
David H
DH
D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D007783:Lactones / Q000378:metabolism; D018942:Macrolides; D009097:Multienzyme Complexes / Q000737:chemistry / Q000378:metabolism*; D013302:Streptomyces / Q000201:enzymology
D000900:Anti-Bacterial Agents; D007783:Lactones; D018942:Macrolides; D009097:Multienzyme Complexes; C003145:picromycin