title: Polycyclic Ketone Monooxygenase from the Thermophilic Fungus Thermothelomyces thermophila: A Structurally Distinct Biocatalyst for Bulky Substrates.
abstract: Regio- and stereoselective Baeyer-Villiger oxidations are difficult to achieve by classical chemical means, particularly when large, functionalized molecules are to be converted. Biocatalysis using flavin-containing Baeyer-Villiger monooxygenases (BVMOs) is a well-established tool to address these challenges, but known BVMOs have shortcomings in either stability or substrate selectivity. We characterized a novel BVMO from the thermophilic fungus Thermothelomyces thermophila, determined its three-dimensional structure, and demonstrated its use as a promising biocatalyst. This fungal enzyme displays excellent enantioselectivity, acts on various ketones, and is particularly active on polycyclic molecules. Most notably we observed that the enzyme can perform oxidations on both the A and D ring when converting steroids. These functional properties can be linked to unique structural features, which identify enzymes acting on bulky substrates as a distinct subgroup of the BVMO class.
authors.affiliation: Molecular Enzymology Group, University of Groningen , Nijenborgh 4, 9747AG, Groningen, The Netherlands.
authors.forename: Maximilian J L J
authors.initials: MJ
authors.lastname: Fürst
authors.affiliation: Department of Biology and Biotechnology, University of Pavia , Via Ferrata 1, 27100, Pavia, Italy.
authors.forename: Simone
authors.initials: S
authors.lastname: Savino
authors.affiliation: Molecular Enzymology Group, University of Groningen , Nijenborgh 4, 9747AG, Groningen, The Netherlands.
authors.forename: Hanna M
authors.initials: HM
authors.lastname: Dudek
authors.affiliation: Department of Biology and Biotechnology, University of Pavia , Via Ferrata 1, 27100, Pavia, Italy.
authors.forename: J Rúben
authors.initials: JR
authors.lastname: Gómez Castellanos
authors.affiliation: Stratingh Institute for Chemistry, University of Groningen , Nijenborgh 4, 9747AG, Groningen, The Netherlands.
authors.forename: Cora
authors.initials: C
authors.lastname: Gutiérrez de Souza
authors.affiliation: Department of Biology and Biotechnology, University of Pavia , Via Ferrata 1, 27100, Pavia, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Rovida
authors.affiliation: Molecular Enzymology Group, University of Groningen , Nijenborgh 4, 9747AG, Groningen, The Netherlands.
authors.forename: Marco W
authors.identifier: 0000-0001-6346-5014
authors.initials: MW
authors.lastname: Fraaije
authors.affiliation: Department of Biology and Biotechnology, University of Pavia , Via Ferrata 1, 27100, Pavia, Italy.
authors.forename: Andrea
authors.identifier: 0000-0002-9523-7128
authors.initials: A
authors.lastname: Mattevi
mesh_terms: D003500:Cyclization; D005658:Fungi / Q000201:enzymology*; D007659:Ketones / Q000737:chemistry*; D006899:Mixed Function Oxygenases / Q000737:chemistry*; D013237:Stereoisomerism
chemical_list: D007659:Ketones; D006899:Mixed Function Oxygenases