Small cyclic dipeptide produced by Lactobacillus rhamnosus with anti-biofilm properties against Streptococcus mutans biofilm

The human oral cavity harbors many bacterial species collectively termed the oral microbiome and is integral for maintaining oral health. Dysbiosis of oral microbiota leads to common oral diseases, including dental caries, gingivitis, and periodontitis. <i>Streptococcus mutans</i> is the primary causative agent of dental caries. Studies have explored the use of probiotic <i>Lactobacillus</i> spp. to mitigate <i>S. mutans</i> biofilms. In the present study, we have tested the use of <i>Lactobacillus rhamnosus</i> extracts/metabolites for anti-biofilm properties. A small organic compound/metabolite was isolated from the cell-free supernatant of <i>L. rhamnosus</i>, and this metabolite resulted in a dose-dependent inhibition of <i>S. mutans</i> biofilms. Confocal microscopy revealed that the thickness of <i>S. mutans</i> biofilms was severely reduced upon metabolite treatment. With the help of FTIR spectra and mass spectrometry analysis, the molecular formula (C₁₁H₁₉O₂N₂) was deduced. The inhibitor compound was further identified as a small cyclic peptide, cyclo (-L-Leu-L-Pro). Our data also revealed that isolated metabolite impedes <i>S. mutans</i> biofilms by modulating gene expression of several essential genes involved in biofilm establishment.