Difference between revisions of "Part:BBa K5439006"
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<partinfo>BBa_K5439006 short</partinfo>
 
<partinfo>BBa_K5439006 short</partinfo>
  
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FRET-based sensor system for the detection of cadmium and other heavy metals that consists of long-chain fatty acid CoA ligase from Sphingomonas spp.[https://parts.igem.org/Part:BBa_K5439005 (BBa_K5439005)],an enzyme that catalyzes the conversion of ibuprofen into isobutylcatechol, flanked by two fluorescent proteins: ECFP[https://parts.igem.org/Part:BBa_K1159302 (BBa_K1159302)]as energy donor and mVenus[https://parts.igem.org/Part:BBa_K1907000 (BBa_K1907000)]as an energy acceptor.  
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FRET-based sensor system for the detection of ibuprofen that consists of long-chain fatty acid CoA ligase from Sphingomonas spp.[https://parts.igem.org/Part:BBa_K5439005 (BBa_K5439005)],an enzyme that catalyzes the conversion of ibuprofen into isobutylcatechol, flanked by two fluorescent proteins: ECFP[https://parts.igem.org/Part:BBa_K1159302 (BBa_K1159302)]as energy donor and mVenus[https://parts.igem.org/Part:BBa_K1907000 (BBa_K1907000)]as an energy acceptor (Gibson, D. G.<i>et al.</i>, 2009).  
  
 
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=Usage and Biology=
 
=Usage and Biology=
 
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Ibuprofen is an anti-inflammatory treatment drug widely used in the world that can be bought without any necessary prescription. This makes ibuprofen a drug that everyone can consume easily, bringing problems because its disposal makes it an emerging contaminant in water bodies (Jan-Roblero, J., & Cruz-Maya, J. A.,2023). Due to this emerging contaminant, new ways to find effective treatments that can remove ibuprofen from water are needed; one particular approach has been studied which involves using different organisms that can degrade ibuprofen by metabolic pathways. An example of it is Sphingomonas Ibu-2; an organism that has been grown in an environment rich in ibuprofen. The described organism has the ability to metabolize ibuprofen to isobutylcatechol due to the adaptation, which one particular gene is in charge of this degradation which is IpfF (Murdoch, R. W., & Hay, A. G.,2013).
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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.
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The enzyme chosen for the generation of the biopart was CoA ligase (EC:6.2.1.3) as a detector for the presence of ibuprofen. The enzyme catalyzes the conversion of ibuprofen into isobutylcatechol
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[1]. Jan-Roblero, J., & Cruz-Maya, J. A. (2023). Ibuprofen: Toxicology and Biodegradation of an Emerging Contaminant. Molecules (Basel, Switzerland), 28(5), 2097. https://doi.org/10.3390/molecules28052097
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[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
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[2]. Murdoch, R. W., & Hay, A. G. (2013). Genetic and chemical characterization of ibuprofen degradation by Sphingomonas Ibu-2. Microbiology (Reading, England), 159(Pt 3), 621–632. https://doi.org/10.1099/mic.0.062273-0
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Revision as of 00:15, 1 October 2024


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