Mechanistic Insights into the Antibiofilm Activity of Simvastatin and Lovastatin against <i>Bacillus subtilis</i>

Statins have been reported for diverse pleiotropic activities, including antimicrobial and antibiofilm. However, due to the limited understanding of their mode of action, none of the statins have gained approval for antimicrobial or antibiofilm applications. In a recent drug repurposing study, we observed that two statins (<i>i.e</i>., Simvastatin and Lovastatin) interact stably with TasA_(28-261), the principal extracellular matrix protein of <i>Bacillus subtilis</i>, and also induce inhibition of biofilm formation. Nevertheless, the underlying mechanism remained elusive. In the present study, we examined the impact of these statins on the physiological activity of TasA_(28-261), specifically its interaction with TapA_(33-253) and aggregation into the amyloid-like structure using purified recombinant TasA_(28-261) and TapA_(33-253) in amyloid detection-specific <i>in vitro</i> assays (<i>i.e</i>., CR binding and ThT staining assays). Results revealed that both statins interfered with amyloid formation by the TasA_(28-261)-TapA_(33-253) complex, while neither statin inhibited amyloid formation by lysozyme, a model amyloid-forming protein. Moreover, neither statin significantly altered the expressions of terminal regulatory genes (<i>viz</i>, <i>sinR</i>, <i>sinI</i>) and terminal effector genes (<i>viz</i>, <i>tasA</i>, <i>tapA</i>, and <i>bslA</i>) involved in biofilm formation by <i>B. subtilis</i>. While the intricate interplay between Simvastatin and Lovastatin with the diverse molecular constituents of <i>B. subtilis</i> biofilm remains to be elucidated conclusively, the findings obtained during the present study suggest that the underlying mechanism for Simvastatin- and Lovastatin-mediated inhibition of <i>B. subtilis</i> biofilm formation is manifested by interfering with the aggregation and amyloid formation by TasA_(28-261)-TapA_(33-253). These results represent one of the first experimental evidence for the underlying mechanism of antibiofilm activity of statins and offer valuable directions for future research to harness statins as antibiofilm therapeutics.