Mining public genomic databases for novel lanthipeptides: a case study on <i>Bacillus amyloliquefaciens</i>

The increasing prevalence of drug-resistant bacteria has prompted the search for alternative approaches to address bacterial infections. Lanthipeptides, which are small molecule peptides, are biosynthesized and post-translationally modified by ribosomes in gram-positive bacteria. These compounds possess distinctive target recognition properties, making them highly potential candidates for combating antimicrobial resistance. Consequently, they hold promising potential as innovative substitutes for traditional antibiotics. We conducted a comprehensive analysis of 65 publicly available Bacillus amyloliquefaciens genomes’ data to explore the potential for lanthipeptide synthesis in this species. We annotated 21 putative precursor peptide genes for these novel lanthipeptides. Through heterologous expression experiments, we successfully obtained a novel lanthipeptide named amyA3. We accomplished a novel co-transformation strategy by introducing the synthetic enzyme obtained from B. amyloliquefaciens WS-8 into the E. coli BL21 (DE3) strain along with the heterologous precursor peptide amyA3. Through mass spectrometry analysis, we confirmed the successful modification of amyA3 by the synthetic enzyme derived from B. amyloliquefaciens WS-8, which represents a significant breakthrough in this research field. The identification and characterization of the lanthipeptide amyA3 along with the elucidation of its biosynthetic pathway provide novel perspectives into the discovery of naturally derived products and the intricate processes of translation and post-translational modifications.