Cyanobactin biosynthesis requires proteolytic cleavage of both N-terminal and C-terminal portions of the precursor peptide. The defining proteins are thus an N-terminal protease, referred to as the A protein, and a C-terminal protease, referred to as the G protein. The G protein is also responsible for macrocyclization.
For cyanobactins, the precursor peptide is referred to as the E peptide. Minimally, the E peptide requires a leader peptide region, a core (structural) region, and both N-terminal and C-terminal protease recognition sequences. In contrast to most RiPPs, for which a single precursor peptide encodes a single natural product via a lone core peptide, cyanobactin E peptides can contain multiple core regions; multiple E peptides can even be present in a single gene cluster.
Many cyanobactins also undergo heterocyclization by a heterocyclase (referred to as the D protein), installing oxazoline or thiazoline moieties from Ser/Thr/Cys residues prior to the action of the A and G proteases. The heterocyclase is an ATP-dependent YcaO homologue that behaves biochemically in the same manner as YcaO-domain cyclodehydratases in thiopeptide and linear azol(in)e-containing peptide (LAP) biosynthesis (described above).
A common modification is prenylation of hydroxyl groups by an F protein prenyltransferase. Oxidation of azoline heterocycles to azoles can also be accomplished by an oxidase domain located on the G protein. Unusual for ribosomal peptides, cyanobactins can include D-amino acids; these can occur adjacent to azole or azoline residues. The functions of some proteins found commonly in cyanobactin biosynthetic gene clusters, the B and C proteins, are unknown.