The gene cluster for the biosynthesis of the heterocyclic quinone antibiotic saframycin Mx1 of DM504/15 was inactivated and tagged by Tn5 insertions. The tagged genes were cloned in and used to select overlapping cosmid clones spanning 58 kb of the genome. Gene disruption experiments defined a ≥ 18 kb contiguous DNA region involved in saframycin biosynthesis. Sequencing of part of this region revealed a large ORF containing two 600-amino-acid domains with similarity to peptide synthetase amino-acid-activating sequences, suggesting that saframycin Mx1 is synthesized by a nonribosomal multienzyme complex, similar to other bioactive peptides.


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