Characterization of the maduropeptin biosynthetic gene cluster from Actinomadura madurae ATCC 39144 supporting a unifying paradigm for enediyne biosynthesis.
The biosynthetic gene cluster for the enediyne antitumor antibiotic maduropeptin (MDP) from Actinomadura madurae ATCC 39144 was cloned and sequenced. Cloning of the mdp gene cluster was confirmed by heterologous complementation of enediyne polyketide synthase (PKS) mutants from the C-1027 producer Streptomyces globisporus and the neocarzinostatin producer Streptomyces carzinostaticus using the MDP enediyne PKS and associated genes. Furthermore, MDP was produced, and its apoprotein was isolated and N-terminal sequenced; the encoding gene, mdpA, was found to reside within the cluster. The biosynthesis of MDP is highlighted by two iterative type I PKSs--the enediyne PKS and a 6-methylsalicylic acid PKS; generation of (S)-3-(2-chloro-3-hydroxy-4-methoxyphenyl)-3-hydroxypropionic acid derived from L-alpha-tyrosine; a unique type of enediyne apoprotein; and a convergent biosynthetic approach to the final MDP chromophore. The results demonstrate a platform for engineering new enediynes by combinatorial biosynthesis and establish a unified paradigm for the biosynthesis of enediyne polyketides.
Van Lanen
Steven G
SG
Division of Pharmaceutical Sciences, University of Wisconsin National Cooperative Drug Discovery Group, University of Wisconsin--Madison, Madison, Wisconsin 53705, USA.
Oh
Tae-Jin
TJ
Liu
Wen
W
Wendt-Pienkowski
Evelyn
E
Shen
Ben
B
D000192:Actinomycetales / Q000235:genetics* / Q000378:metabolism*; D000606:Amino Sugars / Q000096:biosynthesis / Q000737:chemistry; D001059:Apoproteins / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D053281:Enediynes / Q000737:chemistry* / Q000378:metabolism*; D015394:Molecular Structure; D005810:Multigene Family / Q000235:genetics*; D016366:Open Reading Frames / Q000235:genetics; D010636:Phenols; D011422:Propionates / Q000737:chemistry / Q000378:metabolism
C491266:3-hydroxyphenylpropionic acid; D000606:Amino Sugars; D001059:Apoproteins; D001426:Bacterial Proteins; D053281:Enediynes; D010636:Phenols; D011422:Propionates; C509891:maduropeptin enediyne