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Abstract
Transcriptional factor-based biosensors have been widely utilized for inducible control of gene expression in metabolic engineering and relative applications in pathway regulation and high-throughput screening. Two major ways to expand the sensor pool are engineering existing biosensors and exploring unknown transcriptional factors. In this study, we characterized and engineered a phenolic acid-responsive regulator PadR from Bacillus Amyloliquefaciens (BaPadR). The BaPadR sensor system possessed a unique inducer preference and showed a high output intensity compared with other commonly used inducible biosensors. The DNA binding region of BaPadR was engineered to enhance the dynamic range of the biosensor system. By promoter truncation and the construction of hybrid promoters, the DNA binding sequence of BaPadR was identified. To further explore the tunability of the sensor system, base substitutions were performed on the BaPadR binding region of the PpadC promoter. This novel biosensor system can serve as a valuable tool in future synthetic biology applications.