Deconstruction of iterative multidomain polyketide synthase function.
PksA, which initiates biosynthesis of the environmental carcinogen aflatoxin B1, is one of the multidomain iterative polyketide synthases (IPKSs), a large, poorly understood family of biosynthetic enzymes. We found that dissection of PksA and its reconstitution from selected sets of domains allows the accumulation and characterization of advanced octaketide intermediates bound to the enzyme, permitting the reactions controlled by individual catalytic domains to be identified. A product template (PT) domain unites with the ketosynthase and thioesterase in this IPKS system to assemble precisely seven malonyl-derived building blocks to a hexanoyl starter unit and mediate a specific cyclization cascade. Because the PT domain is common among nonreducing IPKSs, these mechanistic features should prove to be general for IPKS-catalyzed production of aromatic polyketides.
Crawford
Jason M
JM
Department of Chemistry, Johns Hopkins University, Baltimore, MD21218, USA.
Thomas
Paul M
PM
Scheerer
Jonathan R
JR
Vagstad
Anna L
AL
Kelleher
Neil L
NL
Townsend
Craig A
CA
D000213:Acyl Carrier Protein / Q000737:chemistry; D016604:Aflatoxin B1 / Q000096:biosynthesis*; D000465:Algorithms; D000880:Anthraquinones / Q000378:metabolism; D001230:Aspergillus / Q000201:enzymology*; D020134:Catalytic Domain; D019295:Computational Biology; D003500:Cyclization; D013058:Mass Spectrometry; D010084:Oxidation-Reduction; D048630:Polyketide Synthases / Q000737:chemistry* / Q000378:metabolism*; D017434:Protein Structure, Tertiary
D000213:Acyl Carrier Protein; D000880:Anthraquinones; D048630:Polyketide Synthases; D016604:Aflatoxin B1; C002754:norsolorinic acid