In this work, we purified and characterized a newly identified lantibiotic (salivaricin D) from Streptococcus salivarius 5M6c. Salivaricin D is a 34-amino-acid-residue peptide (3,467.55 Da); the locus of the gene encoding this peptide is a 16.5-kb DNA segment which contains genes encoding the precursor of two lantibiotics, two modification enzymes (dehydratase and cyclase), an ABC transporter, a serine-like protease, immunity proteins (lipoprotein and ABC transporters), a response regulator, and a sensor histidine kinase. The immunity gene (salI) was heterologously expressed in a sensitive indicator and provided significant protection against salivaricin D, confirming its immunity function. Salivaricin D is a naturally trypsin-resistant lantibiotic that is similar to nisin-like lantibiotics. It is a relatively broad-spectrum bacteriocin that inhibits members of many genera of Gram-positive bacteria, including the important human pathogens Streptococcus pyogenes and Streptococcus pneumoniae. Thus, Streptococcus salivarius 5M6c may be a potential biological agent for the control of oronasopharynx-colonizing streptococcal pathogens or may be used as a probiotic bacterium.
Streptococcus salivarius is a member of the lactic acid bacteria (LAB) that forms part of the normal flora of the oral cavity, throat, and upper respiratory tract (23,28,40,44). It has also been observed in the nasopharynx and intestinal tract and isolated from the human feces (25,35,44) and breast milk of healthy women (1, 46). S. salivarius strains produce a number of bacteriocins, most of which are lantibiotics (22,39,(52)(53)(54).Lantibiotics are small, heat-stable, ribosomally synthesized, posttranslationally modified antimicrobial peptides (bacteriocins) produced by Gram-positive bacteria (3). The lantibiotics, unlike other bacteriocins, are characterized by containing the thioether amino acids lanthionine (Lan) and 3-methyllanthionine (MeLan) and the modified amino acids didehydroalanine (Dha) and didehydrobutyrine (Dhb) (55). Lantibiotics are initially synthesized as inactive linear prepeptides that undergo subsequent extensive modifications to be biologically active. The modifications involve mainly dehydration of serine and threonine residues, forming the didehydro amino acids Dha and Dhb, respectively, which react with the nearby C-terminally located cysteine residues (seen among linear lantibiotics) to form a thioether linkage, which results in the formation of Lan and MeLan, respectively. Finally, the modified peptide is exported and cleaved from its leader in order to be active.According to a very recent classification system, lantibiotics and lantipeptides (class Ia), consist of four subclasses (38). Subclass I lantibiotics are modified by two different enzymes, LanB enzyme (dehydratase) and LanC enzyme (cyclase), exported by LanT, and their leader peptides are removed by the LanP enzyme. Subclass II lantibiotics are modified by a single enzyme (LanM) which has both dehydratase and cyclase activity and is exported by LanT, which als...