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Портал Begell Электронная Бибилиотека e-Книги Журналы Справочники и Сборники статей Коллекции
Critical Reviews™ in Eukaryotic Gene Expression
Импакт фактор: 1.841 5-летний Импакт фактор: 1.927 SJR: 0.649 SNIP: 0.516 CiteScore™: 1.96

ISSN Печать: 1045-4403
ISSN Онлайн: 2162-6502

Выпуски:
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Critical Reviews™ in Eukaryotic Gene Expression

DOI: 10.1615/CritRevEukarGeneExpr.v22.i3.30
pages 197-203

Recombinant Laccase: II. Medical Biosensor

Nicola Luigi Bragazzi
Nanoworld Institute Fondazione EL.B.A Nicolini (FEN), Pradalunga, Largo Redaelli 7, Bergamo 24100, Italy; Biophysics and Nanobiotechnology Laboratories (BNL), Department of Experimental Medicine (DIMES), University of Genoa, Via Antonio Pastore 3, Genoa
Eugenia Pechkova
Nanoworld Institute Fondazione ELBA Nicolini, Pradalunga, Bergamo 24100, Italy ; Biophysics and Nanobiotechnology Laboratories, Department of Experimental Medicine, University of Genova, Genoa 16121-16167, Italy; European Synchrotron Radiation Facility
Dora Scudieri
Biophysics and Nanobiotechnology Laboratories, Department of Experimental Medicine, University of Genova, Genoa 16121-16167, Italy
Tercio Bezerra Correia Terencio
Biophysics and Nanobiotechnology Laboratories, Department of Experimental Medicine, University of Genova, Genoa 16121-16167, Italy
Manuela Adami
Nanoworld Institute Fondazione ELBA Nicolini, Pradalunga, Bergamo 24100, Italy
Claudio Nicolini
Nanoworld Institute Fondazione ELBA Nicolini, Pradalunga, Bergamo 24100, Italy; Biophysics and Nanobiotechnology Laboratories, Department of Experimental Medicine, University of Genova, Genoa 16121-16167, Italy; European Synchrotron Radiation Facility

Краткое описание

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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