Purification and functional studies of a potent modified quorum-sensing peptide and a two-peptide bacteriocin in Streptococcus mutans.

Bacteria use quorum-sensing signals or autoinducers to communicate. The signals in Gram-positive bacteria are often peptides activated by proteolytic removal of an N-terminal leader sequence. While investigating stimulation of antimicrobial peptide production by the Streptococcus mutans synthetic competence stimulating peptide signal (21-CSP), we found a peptide similar to the 21-CSP, but lacking the three C-terminal amino acid residues (18-CSP). The 18-CSP was more potent in inducing competence, biofilm formation, and antimicrobial activity than the 21-CSP. Our results indicate that cleavage of the three C-terminal residues occurred post export, and was not regulated by the CSP-signalling system. Deletion of comD encoding the CSP receptor abolished the competence and biofilm responses to the 21-CSP and the 18-CSP, suggesting that signal transduction via the ComD receptor is involved in the responses to both CSPs. In S. mutans GS5, beside the 18-CSP we also purified to homogeneity a two-peptide bacteriocin which production was stimulated by the 18-CSP and the 21-CSP. Partial sequence of the two-peptide bacteriocin revealed the product of the smbAB genes recently described. We found that the peptide SmbB was slightly different from the deduced sequence, and confirmed the prediction that both peptides constituting SmbAB bacteriocin are post-translationally modified. SmbAB exhibited antimicrobial activity against 11 species of streptococci, Enterococcus faecalis and Staphylocococcus epidermidis. Taken together, the findings support the involvement of the CSP response in bacteriocin production by streptococci and suggest a novel strategy to potentiate autoinducer activity.