Abstract
Here, we characterized a gene expression system induced by 3-hydroxypropionic acid (3-HP) in Pseudomonas denitrificans. The system consists of a putative LysR-type transcriptional regulator (LTTR) encoded by hpdR and a LTTR-responsive promoter that positively controls the expression of hpdH, encoding 3-HP dehydrogenase in the presence of 3-HP. In the hpdH-responsive promoter region, two operators exhibiting dyad symmetry and designated O1 and O2 and centered at the −73 and −30 positions, respectively, were located upstream of the hpdH transcription start site. When either O1, O2, or both regions were mutated, the inducibility by the HpdR-3-HP complex was significantly reduced or completely removed, indicating that both sites are required for transcriptional activation. The HpdR protein and its operator sites on hpdH were highly specific for each other, and did not engage in cross-talk with another similar 3-HP-inducible system that is also present in P. denitrificans. This 3-HP-inducible promoter system should be useful in develo** biosensors for 3-HP, and/or dynamic gene expression systems for metabolic engineering purposes.
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Acknowledgments
This study was financially supported by a grant from the C1 Gas Refinery Program (NRF-2017M3D3A1A01036927-2) through the National Research Foundation of Korea (NRF) funded by the Ministry of Science and ICT. Trinh Thi Nguyen was financially supported by the BK21 Plus Program of Pusan National University.
The authors declare no conflict of interest.
Neither ethical approval nor informed consent was required for this study.
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Nguyen, T.T., Nguyen, N.H., Kim, Y. et al. In vivo Characterization of the Inducible Promoter System of 3-hydroxypropionic Dehydrogenase in Pseudomonas denitrificans. Biotechnol Bioproc E 26, 612–620 (2021). https://doi.org/10.1007/s12257-020-0291-3
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DOI: https://doi.org/10.1007/s12257-020-0291-3