Nonlinear Biosynthetic Assembly of Alpiniamide by a Hybrid /-AT PKS-NRPS.

Alpiniamide A is a linear polyketide produced by endophytic bacteria. Despite its relatively simple chemical structure suggestive of a linear assembly line biosynthetic construction involving a hybrid polyketide synthase-nonribosomal peptide synthetase enzymatic protein machine, we report an unexpected nonlinear synthesis of this bacterial natural product. Using a combination of genomics, heterologous expression, mutagenesis, isotope-labeling, and chain terminator experiments, we propose that alpiniamide A is assembled in two halves and then ligated into the mature molecule. We show that each polyketide half is constructed using orthogonal biosynthetic strategies, employing either - or -acyl transferase mechanisms, thus prompting an alternative proposal for the operation of this PKS-NRPS.