title: Direct and mediated electrochemical response of the cytochrome P450 106A2 from Bacillus megaterium ATCC 13368.
abstract: CYP106A2 is one of only a few known steroid hydroxylases of bacterial origin, which might be interesting for biotechnological applications. Despite the enzyme having been studied for more than 30 years, its physiological function remains elusive. To date, there have been no reports of the redox potential of CYP106A2, which was supposed to be unusually low for a cytochrome P450. In this work we show that cyclic voltammetry is not only suitable to determine the redox potential of challenging proteins such as CYP106A2, measured at -128 mV vs. NHE, but also to study molecular interactions of the enzyme with different interaction partners via the respective electrochemical responses. The effect of small ligands, such as carbon monoxide and cyanide, was observed on the cyclic voltammograms of CYP106A2. Furthermore, we found that Tween 80 caused a positive shift of the redox potential of immobilised CYP106A2 indicative for water expulsion from the haem environment. Moreover, electron transfer mediation phenomena with biological redox partners (e.g. ferredoxins) were studied. Finally, the influence of two different kinds of substrates on the electrochemical response of CYP106A2 was assessed, aligning observations from spectral and electrochemical studies.
authors.affiliation: Laboratoire de Chimie-Physique, UMR 8000 CNRS, Bâtiment 350, Faculté des Sciences d'Orsay, Université Paris-Sud 11, Orsay, France.
authors.forename: Hélène
authors.initials: H
authors.lastname: Colas
authors.forename: Kerstin M
authors.initials: KM
authors.lastname: Ewen
authors.forename: Frank
authors.initials: F
authors.lastname: Hannemann
authors.forename: Nikitas
authors.initials: N
authors.lastname: Bistolas
authors.forename: Ulla
authors.initials: U
authors.lastname: Wollenberger
authors.forename: Rita
authors.initials: R
authors.lastname: Bernhardt
authors.forename: Pedro
authors.initials: P
authors.lastname: de Oliveira
mesh_terms: D001410:Bacillus megaterium / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D002248:Carbon Monoxide / Q000737:chemistry; D003486:Cyanides / Q000737:chemistry; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry*; D055664:Electrochemical Techniques; D004566:Electrodes; D004579:Electron Transport; D005288:Ferredoxins / Q000737:chemistry*; D006418:Heme / Q000737:chemistry*; D008024:Ligands; D010084:Oxidation-Reduction; D011136:Polysorbates / Q000737:chemistry; D014867:Water / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; D003486:Cyanides; D005288:Ferredoxins; D008024:Ligands; D011136:Polysorbates; D014867:Water; D006418:Heme; D002248:Carbon Monoxide; D003577:Cytochrome P-450 Enzyme System