title: In situ formation of H2O2 for P450 peroxygenases.
abstract: An in situ H2O2 generation approach to promote P450 peroxygenases catalysis was developed through the use of the nicotinamide cofactor analogue 1-benzyl-1,4-dihydronicotinamide (BNAH) and flavin mononucleotide (FMN). Final productivity could be enhanced due to higher enzyme stability at low H2O2 concentrations. The H2O2 generation represented the rate-limiting step, however it could be easily controlled by varying both FMN and BNAH concentrations. Further characterization can result in an optimized ratio of FMN/BNAH/O2/biocatalyst enabling high reaction rates while minimizing H2O2-related inactivation of the enzyme.
authors.affiliation: Department of Biotechnology, Delft University of Technology, Julianalaan 136, 2628 BL Delft, The Netherlands.
authors.forename: Caroline E
authors.initials: CE
authors.lastname: Paul
authors.affiliation: Department of Biotechnology, Delft University of Technology, Julianalaan 136, 2628 BL Delft, The Netherlands.
authors.forename: Ekaterina
authors.initials: E
authors.lastname: Churakova
authors.affiliation: Department of Biotechnology, Delft University of Technology, Julianalaan 136, 2628 BL Delft, The Netherlands.
authors.forename: Elmer
authors.initials: E
authors.lastname: Maurits
authors.affiliation: Institute of Biochemistry, Heinrich-Heine-Universität Düsseldorf, Universitätsstraße 1, 40225 Düsseldorf, Germany.
authors.forename: Marco
authors.initials: M
authors.lastname: Girhard
authors.affiliation: Institute of Biochemistry, Heinrich-Heine-Universität Düsseldorf, Universitätsstraße 1, 40225 Düsseldorf, Germany. Electronic address: vlada.urlacher@hhu.de.
authors.forename: Vlada B
authors.initials: VB
authors.lastname: Urlacher
authors.affiliation: Department of Biotechnology, Delft University of Technology, Julianalaan 136, 2628 BL Delft, The Netherlands. Electronic address: f.hollmann@tudelft.nl.
authors.forename: Frank
authors.initials: F
authors.lastname: Hollmann
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology; D046969:Clostridium acetobutylicum / Q000201:enzymology; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000378:metabolism*; D004138:Dinitrocresols / Q000737:chemistry / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D015394:Molecular Structure
chemical_list: D004138:Dinitrocresols; C024132:4,6-dinitro-o-cresol; D003577:Cytochrome P-450 Enzyme System; D006861:Hydrogen Peroxide
keywords: Hydrogen peroxide shunt; In situ H(2)O(2) generation; Oxyfunctionalization; P450 peroxygenases; mNADHs