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Streptococcus sanguinis  is an early colonizer of tooth surfaces and a key player in plaque biofilm development. However, the mechanism of biofilm formation of  S. sanguinis  is still unclear. Here, we showed that deletion of a transcription factor,  brpL , promotes cell aggregation and biofilm formation in  S. sanguinis  SK36. Glucan, a polysaccharide synthesized from sucrose, was over-produced and aggregated in the biofilm of Δ brpL , which was necessary for better biofilm formation ability of Δ brpL . Quantitative RT-PCR demonstrated that  gtfP  was significantly up-regulated in Δ brpL , which increased the productions of water-insoluble and water-soluble glucans. The Δ brpL Δ gtfP  double mutant decreased biofilm formation ability of Δ brpL  to a level similar like that of Δ gtfP . Interestingly, the biofilm of Δ brpL  had an increased tolerance to ampicillin treatment, which might be due to better biofilm formation ability through the mechanisms of cellular and glucan aggregation. RNA sequencing and quantitative RT-PCR revealed the modulation of a group of genes in Δ brpL  was mediated by activating the expression of  ciaR , another  gtfP -related biofilm formation regulator. Double deletion of  brpL  and  ciaR  decreased biofilm formation ability to the phenotype of a Δ ciaR  mutant. Additionally, RNA sequencing elucidated a broad range of genes, related to carbohydrate metabolism and uptake, were activated in Δ brpL . SSA_0222, a gene involved in the phosphotransferase system, was dramatically up-regulated in Δ brpL  and essential for  S. sanguinis  survival under our experimental conditions. In summary,  brpL  modulates glucan production, cell aggregation and biofilm formation by regulating the expression of  ciaR  in  S. sanguinis  SK36.

A Novel Regulator Modulates Glucan Production, Cell Aggregation and Biofilm Formation in Streptococcus sanguinis SK36