Influence to Biofilm Infection of Surface Protein Mediated by Sortase in Cariogenic Bacteria

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In order to examine influence to biofilm infection of surface protein mediated by the Streptococcus mutans sortase, the sortase gene (srtA) of S.mutans 109c was analyzed and a srtA-deficient mutant of S.mutans 109c was constructed. Surface proteins mediated by the sortase were then identified by western blot analysis and the formation of plaque biofilm by the srtA-deficient mutant of S.mutans 109c. The srtA gene was cloned, the complete nucleotide sequence was determined and then a srtA-deficient mutant of S.mutans 109c was constructed based on the nucleotide sequence. The gene sequence was deposited in the DNA data base of Japan (DDBJ) under the accession number AB089932. Localization change of surface proteins of both wild type 109c and srtA mutant was compared and then the sortase-mediated surface proteins were identified. The result indicated that the sortase mediated the cell-wall anchoring of at least three cariogenic surface proteins: surface protein antigen (PAc), glucan-binding protein (GbpC), dextranase (Dcx). It was also clarified that in the srtA-deficient mutant physiological properties of PAc, GbpC and Dex were lost and the formation of plaque biofilm was inhibited. These results suggested that inactivation of the srtA gene or inhibition of sortase enzyme in S.mutans was useful for the prevention of plaque biofilm formation and subsequent dental caries.