title: Determination of the cryptic stereochemistry of the first PKS chain-extension step in ansamitocin biosynthesis by Actinosynnema pretiosum.
abstract: The biosynthesis of the antitumor antibiotic, ansamitocin, involves the assembly of a linear octaketide on the ansamitocin (asm) polyketide synthase (PKS), which is then cyclized to proansamitocin and further modified to the final product. In the first chain-extension step on the asm PKS, a stereocenter is generated which is then obliterated in a subsequent double-bond migration. The cryptic configuration at this stereocenter was determined by first synthesizing the two enantiomers of the intermediate diketide as their N-acetylcysteamine (SNAC) thioesters. These were then used to demonstrate that only the R enantiomer complements a 3-amino-5-hydroxybenzoic acid (AHBA) deficient mutant of Actinosynnema pretiosum to restore ansamitocin formation. The low efficiency of complementation by the diketide, compared to AHBA, is due to inefficient loading onto the PKS and not the inhibition of the enzyme. A presumed next chain-extension intermediate-the triketide with an unrearranged double bond-was also synthesized as its SNAC ester, but did not complement the AHBA(-) mutant.
authors.affiliation: Department of Chemistry, University of Washington, Seattle, WA 98195-1700, USA.
authors.forename: Takaaki
authors.initials: T
authors.lastname: Kubota
authors.forename: Marco
authors.initials: M
authors.lastname: Brünjes
authors.forename: Thomas
authors.initials: T
authors.lastname: Frenzel
authors.forename: Jun
authors.initials: J
authors.lastname: Xu
authors.forename: Andreas
authors.initials: A
authors.lastname: Kirschning
authors.forename: Heinz G
authors.initials: HG
authors.lastname: Floss
mesh_terms: D000192:Actinomycetales / Q000235:genetics / Q000378:metabolism*; D008453:Maytansine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D015394:Molecular Structure; D005810:Multigene Family; D048630:Polyketide Synthases / Q000378:metabolism*; D013237:Stereoisomerism
chemical_list: D008453:Maytansine; C015682:ansamitocins; D048630:Polyketide Synthases