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To combat antibiotic resistance of  Enterococcus faecalis , a better understanding of the molecular mechanisms, particularly of antibiotic detection, signal transduction and gene regulation is needed. Because molecular studies in this bacterium can be challenging, we aimed at exploiting the genetically highly tractable Gram-positive model organism  Bacillus subtilis  as a heterologous host. Two fundamentally different regulators of  E. faecalis  resistance against cell wall antibiotics, the bacitracin sensor BcrR and the vancomycin-sensing two-component system VanS B -VanR B , were produced in  B. subtilis  and their functions were monitored using target promoters fused to reporter genes ( lacZ  and  luxABCDE ). The bacitracin resistance system BcrR-BcrAB of  E. faecalis  was fully functional in  B. subtilis , both regarding regulation of  bcrAB  expression and resistance mediated by the transporter BcrAB. Removal of intrinsic bacitracin resistance of  B. subtilis  increased the sensitivity of the system. The  lacZ  and  luxABCDE  reporters were found to both offer sensitive detection of promoter induction on solid media, which is useful for screening of large mutant libraries. The VanS B -VanR B  system displayed a gradual dose-response behaviour to vancomycin, but only when produced at low levels in the cell. Taken together, our data show that  B. subtilis  is a well-suited host for the molecular characterization of regulatory systems controlling resistance against cell wall active compounds in  E. faecalis . Importantly,  B. subtilis  facilitates the careful adjustment of expression levels and genetic background required for full functionality of the introduced regulators.

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Bacillus subtilis as a Platform for Molecular Characterisation of Regulatory Mechanisms of Enterococcus faecalis Resistance against Cell Wall Antibiotics