title: Recombinant cytochrome p450 immobilization for biosensor applications.
abstract: As a result of the very attractive pleiotropic properties of the heme-enzymes, three P450 cytochrome isoforms (P4501A2, P4502B4, P450SCC) have been utilized to identify a general optimal procedure to biodevice assembly for sensing a wide range of organic substances. The Langmuir-Blodgett films appears to yield the best stable working conditions as shown by UV-vis spectrophotometry, nanogravimetry, circular dichroism, and electrochemical characterization, to identify the ordered nanostructures of P450 cytochromes optimal for clozapine, styrene, and cholesterol sensing. Only in the presence of low purity grade protein, as in the case of P4501A2, a gel-matrix was needed to warrant the optimal clozapine sensing. By the combination of proper immobilization, transducer and nanostructured mutants of high-grade stable and selective P450-based sensors appear capable to detect the interaction with a wide range of organic substrates such as fatty acids, drugs, and toxic compounds.
authors.affiliation: Nanoworld Institute and Biophysics Division, University of Genova, Corso Europa, 30, 16132 Genova, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Paternolli
authors.forename: Mirco
authors.initials: M
authors.lastname: Antonini
authors.forename: Paola
authors.initials: P
authors.lastname: Ghisellini
authors.forename: Claudio
authors.initials: C
authors.lastname: Nicolini
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D002784:Cholesterol / Q000032:analysis; D003024:Clozapine / Q000032:analysis; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008855:Microscopy, Electron, Scanning; D015394:Molecular Structure; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013057:Spectrum Analysis
chemical_list: D004800:Enzymes, Immobilized; D011994:Recombinant Proteins; D003577:Cytochrome P-450 Enzyme System; D002784:Cholesterol; D003024:Clozapine