title: Scrutiny of electrochemically-driven electrocatalysis of C-19 steroid 1α-hydroxylase (CYP260A1) from Sorangium cellulosum So ce56.
abstract: Direct electrochemistry and bioelectrocatalysis of a newly discovered C-19 steroid 1α-hydroxylase (CYP260A1) from the myxobacterium Sorangium cellulosum So ce56 were investigated. CYP260A1 was immobilized on screen-printed graphite electrodes (SPE) modified with gold nanoparticles, stabilized by didodecyldimethylammonium bromide (SPE/DDAB/Au). Cyclic voltammograms in argon-saturated substrate free 0.1 M potassium phosphate buffer, pH 7.4, and in enzyme-substrate complex with androstenedione demonstrated a redox processes with a single redox couple of E(0') of -299 ± 16 mV and -297.5 ± 21 mV (vs. Ag/AgCl), respectively. CYP260A1 exhibited an electrocatalytic activity detected by an increase of the reduction current in the presence of dissolved oxygen and upon addition of the substrate (androstenedione) in the air-saturated buffer. The catalytic current of the enzyme correlated with substrate concentration in the electrochemical system and this dependence can be described by electrochemical Michaelis-Menten model. The products of CYP260A1-depended electrolysis at controlled working electrode potential of androstenedione were analyzed by mass-spectrometry. MS analysis revealed a mono-hydroxylated product of CYP260A1-dependent electrocatalytic reaction towards androstenedione.
authors: Institute of Biomedical Chemistry, Pogodinskaya Street, 10, Moscow 119121, Russia; Pirogov Russian National Research Medical University, Ostrovitianov Street, 1, Moscow 117997, Russia.
authors: Alexey V
authors: AV
authors: Kuzikov
authors: Institute of Biomedical Chemistry, Pogodinskaya Street, 10, Moscow 119121, Russia; Pirogov Russian National Research Medical University, Ostrovitianov Street, 1, Moscow 117997, Russia.
authors: Rami A
authors: RA
authors: Masamrekh
authors: Institute of Biochemistry, Saarland University, Saarbruecken 66123, Germany.
authors: Yogan
authors: Y
authors: Khatri
authors: Institute of Biomedical Chemistry, Pogodinskaya Street, 10, Moscow 119121, Russia.
authors: Maria G
authors: MG
authors: Zavialova
authors: Institute of Biochemistry, Saarland University, Saarbruecken 66123, Germany.
authors: Rita
authors: R
authors: Bernhardt
authors: Institute of Biomedical Chemistry, Pogodinskaya Street, 10, Moscow 119121, Russia; Pirogov Russian National Research Medical University, Ostrovitianov Street, 1, Moscow 117997, Russia.
authors: Alexander I
authors: AI
authors: Archakov
authors: Institute of Biomedical Chemistry, Pogodinskaya Street, 10, Moscow 119121, Russia; Pirogov Russian National Research Medical University, Ostrovitianov Street, 1, Moscow 117997, Russia. Electronic address: viktoria.shumyantseva@ibmc.msk.ru.
authors: Victoria V
authors: VV
authors: Shumyantseva
mesh_terms: D000738:Androsterone / Q000032:analysis*; D001426:Bacterial Proteins / Q000737:chemistry*; D002384:Catalysis; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry*; D055664:Electrochemical Techniques; D004800:Enzymes, Immobilized / Q000737:chemistry*; D006046:Gold / Q000737:chemistry; D006108:Graphite / Q000737:chemistry; D053768:Metal Nanoparticles / Q000737:chemistry; D009231:Myxococcales / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; D004800:Enzymes, Immobilized; D006046:Gold; D006108:Graphite; D003577:Cytochrome P-450 Enzyme System; D000738:Androsterone
keywords: Analytical bioelectrochemistry; C-19 steroid 1α-hydroxylase; Direct electron transfer; Electrocatalysis; Enzyme electrodes; Sorangium cellulosum So ce56