title: Direct electrochemistry of cytochrome c immobilized on gold electrode surface via Zr(IV) ion glue and its activity for ascorbic acid.
abstract: Direct electrochemistry of cytochrome c (Cyt c) is achieved via Zr(IV) ion as an immobilization matrix to interface Cyt c on gold surface via thiol self-assembled monolayers. Steps of surface modification and electrocatalytic activity of the immobilized Cyt c are followed by voltammetry, impedance spectroscopy, chronoampetrometry, and attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy. The results indicate that the native structure of Cyt c is conserved during the immobilization process. The immobilization method is rather simple, effective and free of immobilizing activators and reagents. Direct electron transfer rate constant and surface coverage of the immobilized Cyt c are found as 8.62(±1.98) s(-1) and 1.15(±0.38)×10(-11)molcm(-2), respectively. Bioactivity studies of the immobilized Cyt c toward oxidation of the ascorbic acid (AA) substrate show a linear response, from 10.0μM to 1.30mM AA, with a detection limit of 5.0(±1.8) μM AA and mean relative standard deviations varied from 13.7% to 3.7% for n=4 at each point. A value of 1.6(±0.8) mM AA is found for the Michaelis-Menten constant of Au-MPA-Zr(IV)-Cyt c toward AA for the first time. The tightly immobilized Cyt c maintains its bioactivity for more than 32days storage at 4°C.
authors.affiliation: Department of Chemistry, University of Isfahan, Isfahan 81746-73441, I.R. Iran. Electronic address: rkarimi@sci.ui.ac.ir.
authors.forename: Reza Karimi
authors.initials: RK
authors.lastname: Shervedani
authors.affiliation: Department of Chemistry, University of Isfahan, Isfahan 81746-73441, I.R. Iran.
authors.forename: Marzieh Samiei
authors.initials: MS
authors.lastname: Foroushani
mesh_terms: D001205:Ascorbic Acid / Q000032:analysis* / Q000737:chemistry; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods; D002384:Catalysis; D002712:Chlorides / Q000737:chemistry*; D045304:Cytochromes c / Q000737:chemistry*; D055664:Electrochemical Techniques; D006046:Gold / Q000737:chemistry*; D006861:Hydrogen Peroxide / Q000737:chemistry; D055767:Immobilized Proteins / Q000737:chemistry*; D016014:Linear Models; D010084:Oxidation-Reduction; D017550:Spectroscopy, Fourier Transform Infrared; D013438:Sulfhydryl Compounds / Q000737:chemistry; D013499:Surface Properties; D015040:Zirconium / Q000737:chemistry*
chemical_list: D002712:Chlorides; D055767:Immobilized Proteins; D013438:Sulfhydryl Compounds; D006046:Gold; D045304:Cytochromes c; D006861:Hydrogen Peroxide; D015040:Zirconium; D001205:Ascorbic Acid; C429984:zirconium chloride
keywords: Ascorbic acid; Cytochrome c; Direct electrochemistry; Impedance spectroscopy; Zr(IV) ion