A novel biosensor for ferrous iron developed via CoBiSe: A computational method for rapid biosensor design
Creators
- 1. Center for Structural Studies
- 2. Center for Advanced Imaging
- 3. Institute of Molecular Enzyme Technology
- 4. Institute of Bio- and Geosciences
- 5. Institute for Pharmaceutical and Medicinal Chemistry
- 6. Center for Structural Studies & Institute for Biochemistry
- 7. Center for Structural Studies & Institute for Pharmaceutical and Medicinal Chemistry
Description
Genetically encoded biosensors enable monitoring of metabolite dynamics in living organisms. We present CoBiSe, a computational 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 (Fe²⁺). IronSenseR demonstrates high specificity for Fe²⁺ with dissociation constants of 1.78 ± 0.03 µM (FeSO₄) and 2.90 ± 0.12 µM (FeCl₂), while showing no binding to Fe³⁺ 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.