Discovery,\nBiosynthesis, and Characterization of Rodencin,\na Two-Component Lanthipeptide, Harboring d‑Amino Acids\nIntroduced by the Unusual Dehydrogenase RodJ<sub>A</sub>

Lanthipeptides, a group of ribosomally synthesized and\npost-translationally\nmodified peptides (RiPPs), exhibit diverse structures and bioactivities.\nTheir biosynthetic enzymes serve as valuable tools for peptide bioengineering.\nHere, we report a class II lanthipeptide biosynthetic gene cluster\nin a <i>Bacillus</i> strain, driving the biosynthesis of\na two-component lanthipeptide, termed rodencin, featured by the presence\nof two different d-amino acids, i.e., d-Ala and d-Abu. Rodencin displays synergistic antimicrobial activity\nagainst food-borne pathogens such as <i>Bacillus cereus</i>, <i>Staphylococcus aureus</i>, and <i>Listeria monocytogenes</i>. The α-peptide of rodencin contains one d-Ala and\nthe β-peptide features both d-Ala and d-Abu.\nThese are installed by dehydratases RodM1 and RodM2 and dehydrogenase\nRodJ<sub>A</sub>, the activities of which were successfully reconstituted\nusing a dedicated <i>E. coli</i> expression system. To illustrate\nthe unusual d-Abu incorporation potential of the enzymes,\nanalogous to the d-amino acid-containing β peptide\nof lacticin 3147, was successfully produced with the rodencin heterologous\nexpression system, by employing RodM2 and the dehydrogenase RodJ<sub>A</sub>.