Inactivation of the novel phenylalanine ammonia lyase gene encP, whose product is a key component in the biosynthetic pathway to benzoyl-coenzyme A (CoA) in the bacterium Streptomyces maritimus, resulted in the loss of production of the benzoate-primed polyketides enterocin and wailupemycin G. A series of cinnamate and benzoate derivatives were administered to the DeltaencP mutant, resulting in the formation of novel analogues bearing p-fluorobenzoate, 2- and 3-thiophenecarboxylate, and cyclohex-1-enecarboxylate residues. Given that the benzoate:CoA ligase EncN was evaluated to have broad in vitro substrate specificity towards aryl acids, the strict starter unit specificity observed in vivo indicates that the enterocin type II polyketide synthase (PKS) exerts selective control over the choice of starter units. This study represents the first mutasynthesis experiments with iterative type II PKSs.