Antimycins
are a family of natural products generated from a hybrid
nonribosomal peptide synthetase (NRPS)-polyketide synthase (PKS) assembly
line. Although they possess an array of useful biological activities,
their structural complexity makes chemical synthesis challenging,
and their biosynthesis has thus far been dependent on slow-growing
source organisms. Here, we reconstituted the biosynthesis of antimycins
in <i>Escherichia coli</i>, a versatile host that is robust
and easy to manipulate genetically. Along with <i>Streptomyces</i> genetic studies, the heterologous expression of different combinations
of <i>ant</i> genes enabled us to systematically confirm
the functions of the modification enzymes, AntHIJKL and AntO, in the
biosynthesis of the 3-formamidosalicylate pharmacophore of antimycins.
Our <i>E. coli</i>-based antimycin production system can
not only be used to engineer the increased production of these bioactive
compounds, but it also paves the way for the facile generation of
novel and diverse antimycin analogues through combinatorial biosynthesis