Publication de 6 numéros par an
ISSN Imprimer: 1045-4403
ISSN En ligne: 2162-6502
Indexed in
Recombinant Laccase: II. Medical Biosensor
RÉSUMÉ
Langmuir-Blodgett (LB) technology was used to build a high-sensitivity enzyme-based biosensor for medical purposes. Recombinant fungal laccase from Rigidoporous lignosus, as previously described, was used to catalyze a widely used antidepressant in a micromolar range, namely, clomipramine. The topological properties of the laccase thin film were characterized via LB π-A isotherm and AFM (mean roughness 8.22 nm, compressibility coefficient 37.5 m/N). The sensitivity of the biosensor was investigated via UV spectroscopy, and linearity was found in the absorbance peak shift at 400 nm at drug concentration varying up to 20 uM. The enzyme kinetics was subsequently investigated with potentiometric and amperometric measurements, and we found electronic transfer of at least 1 electron, ks 0.57 s−1, diffusion coefficient 3 × 10−6 cm2/s, Kcat t 6825.92 min−1, KM 4.1 uM, Kcat /KM 2.8 × 107 mol−1 s−1, sensitivity of 440 nA/uM, maximum velocity 1706.48 nA/s, and response time less than 5 s. The amperometric and potentiometric measurements were repeated after a month, confirming the stability of the biosensor.
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Spera Rosanna, Festa Fernanda, Bragazzi Nicola L., Pechkova Eugenia, LaBaer Joshua, Nicolini Claudio, Conductometric Monitoring of Protein–Protein Interactions, Journal of Proteome Research, 12, 12, 2013. Crossref