Discovery, functional characterization, and structural studies of the NRPS-independent siderophore synthetases

To adapt to low-iron environments, many bacteria produce siderophores, low molecular weight iron chelators that are secreted into the environment where they bind ferric iron. The production of siderophore uptake systems then allows retrieval of the iron-complexed siderophore into the cell, where the metal ion can be used for structural and catalytic roles in many proteins. While many siderophores are produced by the activity of a family of large modular nonribosomal peptide synthetase (NRPS) enzymes, a second class of siderophores are produced by an alternate pathway. These so-called NRPS-independent siderophores (NIS) are biosynthesized through a shared catalytic step that is performed by an NIS synthetase. These enzymes catalyze the formation of an amide linkage between a carboxylate and an amine or, more rarely, form an ester with a hydroxyl substrate. Here we describe the discovery and biochemical studies of diverse NIS synthetases from different siderophore pathways to provide insight into their substrate specificity and catalytic mechanism. The structures of a small number of family members are additionally described that correlates the functional work with the enzyme structure. While the field has come a long way since it was described as a “long-overlooked” family in 2009, there remains much to discover in this large and important enzyme family.