title: The oxygenating constituent of 3,6-diketocamphane monooxygenase from the CAM plasmid of Pseudomonas putida: the first crystal structure of a type II Baeyer-Villiger monooxygenase.
abstract: The three-dimensional structures of the native enzyme and the FMN complex of the overexpressed form of the oxygenating component of the type II Baeyer-Villiger 3,6-diketocamphane monooxygenase have been determined to 1.9 Å resolution. The structure of this dimeric FMN-dependent enzyme, which is encoded on the large CAM plasmid of Pseudomonas putida, has been solved by a combination of multiple anomalous dispersion from a bromine crystal soak and molecular replacement using a bacterial luciferase model. The orientation of the isoalloxazine ring of the FMN cofactor in the active site of this TIM-barrel fold enzyme differs significantly from that previously observed in enzymes of the bacterial luciferase-like superfamily. The Ala77 residue is in a cis conformation and forms a β-bulge at the C-terminus of β-strand 3, which is a feature observed in many proteins of this superfamily.
authors.affiliation: The Henry Wellcome Building for Biocatalysis, Biosciences, College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, England.
authors.forename: Michail N
authors.initials: MN
authors.lastname: Isupov
authors.affiliation: The Henry Wellcome Building for Biocatalysis, Biosciences, College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, England.
authors.forename: Ewald
authors.initials: E
authors.lastname: Schröder
authors.affiliation: The Henry Wellcome Building for Biocatalysis, Biosciences, College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, England.
authors.forename: Robert P
authors.initials: RP
authors.lastname: Gibson
authors.affiliation: The Henry Wellcome Building for Biocatalysis, Biosciences, College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, England.
authors.forename: Jean
authors.initials: J
authors.lastname: Beecher
authors.affiliation: The Henry Wellcome Building for Biocatalysis, Biosciences, College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, England.
authors.forename: Giuliana
authors.initials: G
authors.lastname: Donadio
authors.affiliation: The Henry Wellcome Building for Biocatalysis, Biosciences, College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, England.
authors.forename: Vahid
authors.initials: V
authors.lastname: Saneei
authors.affiliation: The Henry Wellcome Building for Biocatalysis, Biosciences, College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, England.
authors.forename: Stephlina A
authors.initials: SA
authors.lastname: Dcunha
authors.affiliation: The Henry Wellcome Building for Biocatalysis, Biosciences, College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, England.
authors.forename: Emma J
authors.initials: EJ
authors.lastname: McGhie
authors.affiliation: The Henry Wellcome Building for Biocatalysis, Biosciences, College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, England.
authors.forename: Christopher
authors.initials: C
authors.lastname: Sayer
authors.affiliation: The Henry Wellcome Building for Biocatalysis, Biosciences, College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, England.
authors.forename: Colin F
authors.initials: CF
authors.lastname: Davenport
authors.affiliation: Biotechnology Research Institute, National Research Council Canada, 6100 Royalmount Avenue, Montreal, QC H4P 2R2, Canada.
authors.forename: Peter C
authors.initials: PC
authors.lastname: Lau
authors.affiliation: Department of Biotechnology, Faculty of Engineering, Kansai University, Japan.
authors.forename: Yoshie
authors.initials: Y
authors.lastname: Hasegawa
authors.affiliation: Department of Biotechnology, Faculty of Engineering, Kansai University, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Iwaki
authors.affiliation: Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Strasse 4, 17487 Greifswald, Germany.
authors.forename: Maria
authors.initials: M
authors.lastname: Kadow
authors.affiliation: Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Strasse 4, 17487 Greifswald, Germany.
authors.forename: Kathleen
authors.initials: K
authors.lastname: Balke
authors.affiliation: Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Strasse 4, 17487 Greifswald, Germany.
authors.forename: Uwe T
authors.initials: UT
authors.lastname: Bornscheuer
authors.affiliation: European Molecular Biology Laboratory (EMBL), Hamburg Outstation, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Gleb
authors.initials: G
authors.lastname: Bourenkov
authors.affiliation: The Henry Wellcome Building for Biocatalysis, Biosciences, College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, England.
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Littlechild
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D038181:FMN Reductase / Q000378:metabolism; D005486:Flavin Mononucleotide / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010105:Oxygenases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011487:Protein Conformation; D017510:Protein Folding; D016958:Pseudomonas putida / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment
chemical_list: D001426:Bacterial Proteins; D005486:Flavin Mononucleotide; D010105:Oxygenases; C115882:3,6-diketocamphane monooxygenase; D038181:FMN Reductase
keywords: FMN-dependent monooxygenase; industrial biocatalysis; protein structure