Abstract
Malate is a key intermediate in the citric acid cycle, an enzymatic cascade that is central to cellular energy metabolism and that has been applied to make biofuel cells. To enable real-time sensing of malate levels, we have engineered a genetically encoded, protein-based fluorescent biosensor called Malon specifically responsive to malate by performing structure-based mutagenesis of the Cache-binding domain of the Citron GFP-based biosensor. Malon demonstrates high specificity and fluorescence activation in response to malate and has been applied to monitor enzymatic reactions in vitro. Furthermore, we successfully incorporated Malon into redox polymer hydrogels and bacterial cells, enabling analysis of malate levels in these materials and living systems. These results show the potential for fluorescent biosensors in enzymatic cascade monitoring within biomaterials and present Malon as a novel tool for bioelectronic devices.
Recommended Citation
N. J. Ricks et al., "Development of Malate Biosensor-Containing Hydrogels and Living Cell-Based Sensors," International Journal of Molecular Sciences, vol. 25, no. 20, article no. 11098, MDPI, Oct 2024.
The definitive version is available at https://doi.org/10.3390/ijms252011098
Department(s)
Chemistry
Publication Status
Open Access
Keywords and Phrases
bioelectrode; Cache; cell-based biosensor; DcuS
International Standard Serial Number (ISSN)
1422-0067; 1661-6596
Document Type
Article - Journal
Document Version
Final Version
File Type
text
Language(s)
English
Rights
© 2024 The Authors, All rights reserved.
Creative Commons Licensing
This work is licensed under a Creative Commons Attribution 4.0 License.
Publication Date
01 Oct 2024
PubMed ID
39456881
Comments
Office of Naval Research, Grant N000142112188