Staphylococcus aureus is a clinically relevant bacterium in the order Bacillales that is frequently found in human-associated microbiomes. While pathogenic S. aureus strains can cause many diseases, often leveraging its ability to form biofilms and acquire antibiotic resistance, nonpathogenic colonizers are also prevalently identified. However, studying and engineering the physiology of S. aureus remains challenging due to limited availability of characterized genetic parts. Among the bacterial phylum Firmicutes and order Bacillales, Bacillus subtilis is a well-established Gram-positive model that has multiple characterized genetic toolboxes available. In our earlier work, we established a genetic toolset for S. aureus and identified high transferability of constitutive promoters from B. subtilis to S. . Here, we leveraged this toolset to further investigate the transferability and functionality of a set of synthetic sensors for four different homoserine lactones (HSLs) sensors that were previously engineered for B. subtilis in earlier work from our group. These HSL sensors use allosteric LuxR-family regulators (RpaR, LuxR, CinR and RhlR) from various. Assaying these sensors in S. aureus, we found that the HSL sensors remain functional in S. aureus with up to 80-fold dynamic range for the p-coumaroyl-HSL and N-butyryl-HSL sensors. These findings demonstrate that genetic tools and biosensors for B. subtilis might be a source for expanding toolboxes of genetic parts and sensors for S. aureus and potentially other tough to engineer bacterial relatives.
