| Title |
Maleylpyruvic acid-inducible gene expression system and its application for the development of gentisic acid biosensors / |
| Authors |
Kutraite, Ingrida ; Augustiniene, Ernesta ; Malys, Naglis |
| DOI |
10.1021/acs.analchem.4c03906 |
| Full Text |
|
| Is Part of |
Analytical chemistry.. Washington, DC : American Chemical Society. 2024, vol. 96, iss. 47, p. 18727-18735.. ISSN 0003-2700. eISSN 1520-6882 |
| Keywords [eng] |
anatomy ; bacteria ; biotechnology ; genetics ; sensors |
| Abstract [eng] |
Gentisic acid is a secondary plant metabolite, known for its health benefits, not only widely used as a supplement but also implicated as a potential biomarker for cancer-associated metabolism alterations. To advance bioproduction and detection of this compound or its derivatives, cell-based approaches have become of interest in recent years. However, the lack of tools for high-throughput gentisic acid monitoring and compound-metabolizing organism screening limits the progress in this area. Here, we analyzed the gene cluster responsible for gentisic acid metabolism in Cupriavidus necator H16. The transcriptional regulator GtdR-based inducible gene expression system CnGtdR/PgtdA was elucidated, showing that it was activated when C. necator cells were subjected to gentisic acid. Subsequently, a 3- maleylpyruvic acid was identified as a primary inducer for this inducible system. Furthermore, genes gtdA and gtdT, encoding for gentisate 1,2-dioxygenase and MFS transporter, were shown to be essential for inducible system activation in the presence of gentisic acid with GtdA enabling conversion of this phenolic acid into the inducer. The CnGtdRAT/PgtdA-based inducible system was employed to develop a whole-cell biosensor for the intracellular and extracellular detection of gentisic acid. The potential of the 3-maleylpyruvic acid-inducible system was demonstrated by its application in metabolic pathway research, detection of highly unstable 3-maleylpyruvic acid, and development of biosensors for the intracellular or extracellular determination of gentisic acid. In addition, the utility of the biosensor was emphasized by its application for detection of gentisic acid as a potential biomarker for cancer in urine samples. |
| Published |
Washington, DC : American Chemical Society |
| Type |
Journal article |
| Language |
English |
| Publication date |
2024 |
| CC license |
|
