title: Direct electron transfer of heme- and molybdopterin cofactor-containing chicken liver sulfite oxidase on alkanethiol-modified gold electrodes.
abstract: Direct heterogeneous electron transfer (ET) of sulfite oxidase (SOx), a heme- and molybdopterin cofactor-containing intermembrane enzyme, was studied on alkanethiol-modified Au electrodes both with SOx entrapped between the modified Au electrode and a permselective membrane and with SOx adsorbed at the electrode surface, in the absence of any membrane. SOx in direct electronic communication with the electrode surface gave a quasi-reversible electrochemical signal with a midpoint potential of--120 mV vs Ag/AgCl corresponding to the redox transformations of the heme domain of SOx and with a heterogeneous ET constant in the order of 15 s(-1). The efficiency of the bioelectrocatalytic 2e- oxidation of sulfite catalyzed by SOx in direct ET exchange with the electrode was shown to depend essentially on the nature of the alkanethiol layer. Adsorption and orientation of SOx on an 11-mercapto-1-undecanol (MuD-OH) self-assembled monolayer, i.e., terminally functionalized with OH groups, provided efficient catalytic oxidation of sulfite, contrary to nonfunctionalized alkanethiols, e.g., 1-decanethiol, or alkanethiol layers terminally functionalized with NH2 groups. Comparative studies with short-chain alkanethiols, e.g., cysteamine and 2-mercaptoethanol, revealed an evidently different mode of adsorption of SOx on these layers, onto which SOx was not catalytically active. Coadsorption of MuD-OH and 11-mercapto-1-undecanamine improved the surface properties of the SAM, resulting in a higher surface coverage with bioelectrocatalytically active SOx but not in an increased apparent catalytic rate constant, kcat, ranging in the order of 18-24 s(-1) at pH 7.4. The achieved efficiency of SOx bioelectrocatalysis in direct ET reaction between the modified electrode and the enzyme approached the rates characteristic for the catalysis mediated by cytochrome c, the natural redox partner of SOx, thus implying the retention of the biological function of SOx under the heterogeneous electrode reaction conditions. Results obtained enable the development of a third-generation biosensor for sulfite monitoring.
authors.affiliation: Department of Analytical Chemistry, Lund University, PO Box 124, SE 221 00 Lund, Sweden. elena.ferapontova@analykem.lu.se
authors.forename: Elena E
authors.initials: EE
authors.lastname: Ferapontova
authors.forename: Tautgirdas
authors.initials: T
authors.lastname: Ruzgas
authors.forename: Lo
authors.initials: L
authors.lastname: Gorton
mesh_terms: D000818:Animals; D002645:Chickens; D003067:Coenzymes; D004566:Electrodes; D004579:Electron Transport; D004800:Enzymes, Immobilized / Q000737:chemistry*; D006046:Gold; D006418:Heme / Q000737:chemistry*; D008099:Liver / Q000201:enzymology*; D008667:Metalloproteins / Q000737:chemistry*; D000090006:Molybdenum Cofactors; D050862:Oxidoreductases Acting on Sulfur Group Donors / Q000737:chemistry*; D011621:Pteridines / Q000737:chemistry*; D013438:Sulfhydryl Compounds / Q000737:chemistry*
chemical_list: D003067:Coenzymes; D004800:Enzymes, Immobilized; D006046:Gold; D006418:Heme; D008667:Metalloproteins; D000090006:Molybdenum Cofactors; D050862:Oxidoreductases Acting on Sulfur Group Donors; D011621:Pteridines; D013438:Sulfhydryl Compounds; C022261:molybdopterin