title: Mimicking enzymatic effects of cytochrome P450 by an efficient biosensor for in vitro detection of DNA damage.
abstract: A novel biosensor for detecting DNA damage induced by benzo(a)pyrene (BP) and its metabolite was presented in this work. The nafion-solubilized single wall carbon nanotubes-ionic liquid (SWCNTs-NA-IL) composite film was prepared and then horseradish peroxidase (HRP) and double-stranded DNA were alternately assembled on the composite film by the layer-by-layer method. The biosensor was characterized by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), differential pulse voltammetry (DPV), scanning electron microscopy (SEM) and computational methods. UV-vis spectrophotometry was also used to investigate DNA damage induced by BP and its metabolites in solution. The DNA biosensor was treated separately in BP, hydrogen peroxide (H2O2) and in their mixture, respectively. The EIS analysis showed a decrease in the charge transfer resistance at the DNA/HRP/SWCNTs-NA-IL/GCE incubated in a mixture of HRP and H2O2, because HRP in the presence of H2O2 could mimic enzymatic effects of cytochrome P450 (CYP450) to metabolize BP which could cause significant DNA damage and the exposed DNA bases reduced the electrostatic repulsion of the negatively charged redox probe and leads to Faradaic impedance changes. Finally, a novel biosensor for BP determination was developed and this method provided an indirect, and quantitative estimation of DNA damage in vitro.
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemistry, Razi University, Kermanshah 671496734, Iran; Laboratorio de Desarrollo Analítico y Quimiometría (LADAQ), Cátedra de Química Analítica I, Universidad Nacional del Litoral, Ciudad Universitaria, CC 242 (S3000ZAA), Santa Fe, Argentina; Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran.
authors.forename: Ali R
authors.initials: AR
authors.lastname: Jalalvand
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemistry, Razi University, Kermanshah 671496734, Iran. Electronic address: mbgholivand2013@gmail.com.
authors.forename: Mohammad-Bagher
authors.initials: MB
authors.lastname: Gholivand
authors.affiliation: Laboratorio de Desarrollo Analítico y Quimiometría (LADAQ), Cátedra de Química Analítica I, Universidad Nacional del Litoral, Ciudad Universitaria, CC 242 (S3000ZAA), Santa Fe, Argentina.
authors.forename: Hector C
authors.initials: HC
authors.lastname: Goicoechea
authors.affiliation: Quality & Technology, Department of Food Science, Faculty of Science, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Thomas
authors.initials: T
authors.lastname: Skov
authors.affiliation: Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran.
authors.forename: Kamran
authors.initials: K
authors.lastname: Mansouri
mesh_terms: D001564:Benzo(a)pyrene / Q000302:isolation & purification* / Q000633:toxicity; D015374:Biosensing Techniques; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry*; D004247:DNA / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D004249:DNA Damage / Q000187:drug effects; D058266:Dielectric Spectroscopy; D006735:Horseradish Peroxidase / Q000737:chemistry; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry; D008855:Microscopy, Electron, Scanning; D037742:Nanotubes, Carbon / Q000737:chemistry; D010084:Oxidation-Reduction
chemical_list: D037742:Nanotubes, Carbon; D001564:Benzo(a)pyrene; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase
keywords: Benzo(a)pyrene; Biosensor; DNA damage