A Novel Biosensor for Ferrous Iron Developed via CoBiSe: A Computational Method for Rapid Biosensor Design
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Abstract
High Resolution Image Download MS PowerPoint Slide Genetically encoded biosensors enable the monitoring of metabolite dynamics in living organisms. We present CoBiSe, a co mputational bi o se nsor design approach using Constraint Network Analysis to identify optimal insertion sites for reporter modules in molecular recognition elements (MREs). Applied to the iron-binding protein DtxR from Corynebacterium glutamicum, CoBiSe identified a flexible connective loop (residues 138–150) for inserting the reporter module, resulting in IronSenseR, a novel ratiometric biosensor for ferrous iron (Fe 2+ ). IronSenseR demonstrates high specificity for Fe 2+ with dissociation constants of 1.78 ± 0.03 (FeSO 4 ) and 2.90 ± 0.12 μM (FeCl 2 ), while showing no binding to Fe 3+ and other divalent cations. In vivo assessment in Escherichia coli, Pseudomonas putida, and Corynebacterium glutamicum confirmed IronSenseR’s capability to detect changes in the intracellular iron pool. The creation of IronSenseR underlines that by reducing search space and eliminating labor-intensive screening, CoBiSe streamlines biosensor development and enables precise creation of next-generation biosensors for diverse metabolites.
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Cites
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“This vector was linearized by PCR to be used for the Gibson assembly.”
InfluentialMechanism of metal ion activation of the diphtheria toxin repressor DtxR
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- Authors
- Papadopoulos, A., Anlauf, M. T., Reiners, J., Paik, S.-H., Krüger, A., Lückel, B., Bott, M., Drepper, T., Frunzke, J., Gohlke, H., Weidtkamp-Peters, S., Smits, S. H. J., & Gertzen, C. G. W.
- Affiliations
- Heinrich Heine University Düsseldorf (DE)
- Institute of Molecular Enzyme Technology, Faculty of Mathematics and Natural Sciences, Heinrich Heine University Düsseldorf, Forschungszentrum Jülich, 52428 Jülich, Germany
- Forschungszentrum Jülich (DE)
- Center for Advanced Imaging, Faculty of Mathematics and Natural Sciences
- Corresponding author
- Smits, Sander H. J.
- Gertzen, Christoph G. W.
- ORCID
- Venue
- ACS Sensors
- Publisher
- American Chemical Society (ACS)
- Published
- 2026-01-11
- Volume
- 11
- Issue
- 1
- Pages
- 119-135
- ISSN
- 2379-3694 (ISSN-L)
- PMC
- PMC12836349
- OpenAlex
- W7121003110
- Cited by
- 5
- References
- 3
- Citations by year
- 2026: 5
- FWCI
- 20.02
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- ACS Sensors (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- Funders
- Deutsche Forschungsgemeinschaft (284074525, 417919780, 45809666, 462231789, INST 208/761-1 FUGG)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › bioluminescence and chemiluminescence research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gene Regulatory Network Analysis
- Keywords
- genetically encoded biosensors, DtxR, iron-binding proteins, Corynebacterium glutamicum, ratiometric biosensor, molecular recognition elements
- MeSH terms
- Bacterial Proteins, Escherichia coli, Ferrous Compounds, Iron, Biosensing Techniques, Pseudomonas putida, Iron-Binding Proteins, Corynebacterium glutamicum
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