Part:BBa_K5439006

FRET-based system for the detection of ibuprofen

FRET-based sensor system for the detection of ibuprofen that consists of long-chain fatty acid CoA ligase from Sphingomonas spp.(BBa_K5439005),an enzyme that catalyzes the conversion of ibuprofen into isobutylcatechol, flanked by two fluorescent proteins: ECFP(BBa_K1159302)as energy donor and mVenus(BBa_K1907000)as an energy acceptor (Gibson, D. G.et al., 2009).

Usage and Biology

FRET (Fluorescence Resonance Energy Transfer) is often used in the design of biosensors as it allows for the specific and sensitive detection of biomolecules in a highly specific manner with high sensitivity, without the need to induce a change in the biomolecule. The fluorescence of the acceptor molecule is activated only when both the donor fluorophore and the acceptor molecule are in proximity. This means that any changes in their surrounding environment that affect the distance between them will also impact the fluorescence of the molecule. This mechanism of action enables the detection of changes in the environment, even if they are subtle, without the need to genetically modify the molecule. FRET is a non-radiative process, which means it does not produce any ionizing radiation. This makes this type of biosensor safer to use and handle compared to others. Additionally, they are very sensitive and versatile biosensors, allowing them to detect the presence of a wide variety of biomolecules, as well as changes in the environment. They can detect protein-protein interactions, monitor changes in pH, measure enzyme activity, among others.

[1]. Gibson, D. G., Young, L., Chuang, R.-Y., Venter, J. C., Hutchison, C. A., & Smith, H. O. (2009). Enzymatic assembly of DNA molecules up to several hundred kilobases. Nature Methods, 6(5), 343–345. https://doi.org/10.1038/nmeth.1318