title: A Peroxygenase from Chaetomium globosum Catalyzes the Selective Oxygenation of Testosterone.
abstract: Unspecific peroxygenases (UPO, EC 1.11.2.1) secreted by fungi open an efficient way to selectively oxyfunctionalize diverse organic substrates, including less-activated hydrocarbons, by transferring peroxide-borne oxygen. We investigated a cell-free approach to incorporate epoxy and hydroxyl functionalities directly into the bulky molecule testosterone by a novel unspecific peroxygenase (UPO) that is produced by the ascomycetous fungus Chaetomium globosum in a complex medium rich in carbon and nitrogen. Purification by fast protein liquid chromatography revealed two enzyme fractions with the same molecular mass (36 kDa) and with specific activity of 4.4 to 12 U mg-1. Although the well-known UPOs of Agrocybe aegerita (AaeUPO) and Marasmius rotula (MroUPO) failed to convert testosterone in a comparative study, the UPO of C. globosum (CglUPO) accepted testosterone as substrate and converted it with total turnover number (TTN) of up to 7000 into two oxygenated products: the 4,5-epoxide of testosterone in β-configuration and 16α-hydroxytestosterone. The reaction performed on a 100 mg scale resulted in the formation of about 90 % of the epoxide and 10 % of the hydroxylation product, both of which could be isolated with purities above 96 %. Thus, CglUPO is a promising biocatalyst for the oxyfunctionalization of bulky steroids and it will be a useful tool for the synthesis of pharmaceutically relevant steroidal molecules.
authors.affiliation: Faculty of Environment and Natural Sciences, Brandenburg University of Technology Cottbus-Senftenberg, Universitätsplatz 1, 01968, Senftenberg, Germany.
authors.forename: Jan
authors.initials: J
authors.lastname: Kiebist
authors.affiliation: Faculty of Environment and Natural Sciences, Brandenburg University of Technology Cottbus-Senftenberg, Universitätsplatz 1, 01968, Senftenberg, Germany.
authors.forename: Kai-Uwe
authors.initials: KU
authors.lastname: Schmidtke
authors.affiliation: Faculty of Environment and Natural Sciences, Brandenburg University of Technology Cottbus-Senftenberg, Universitätsplatz 1, 01968, Senftenberg, Germany.
authors.forename: Jörg
authors.initials: J
authors.lastname: Zimmermann
authors.affiliation: Department of Bio- and Environmental Sciences, TU Dresden, International Institute Zittau, Markt 23, 02763, Zittau, Germany.
authors.forename: Harald
authors.initials: H
authors.lastname: Kellner
authors.affiliation: Department of Molecular Systems Biology, Helmholtz-Centre for Environmental Research, UFZ, Permoserstrasse 15, 04318, Leipzig, Germany.
authors.forename: Nico
authors.initials: N
authors.lastname: Jehmlich
authors.affiliation: Department of Bio- and Environmental Sciences, TU Dresden, International Institute Zittau, Markt 23, 02763, Zittau, Germany.
authors.forename: René
authors.initials: R
authors.lastname: Ullrich
authors.affiliation: JenaBios GmbH, Löbstedter Strasse 80, 07749, Jena, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Zänder
authors.affiliation: Department of Bio- and Environmental Sciences, TU Dresden, International Institute Zittau, Markt 23, 02763, Zittau, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Hofrichter
authors.affiliation: Faculty of Environment and Natural Sciences, Brandenburg University of Technology Cottbus-Senftenberg, Universitätsplatz 1, 01968, Senftenberg, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Scheibner
mesh_terms: D000595:Amino Acid Sequence; D002384:Catalysis / Q000187:drug effects; D002597:Chaetomium / Q000201:enzymology*; D002626:Chemistry, Pharmaceutical; D002851:Chromatography, High Pressure Liquid; D006899:Mixed Function Oxygenases / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D010100:Oxygen / Q000378:metabolism*; D013739:Testosterone / Q000378:metabolism*
chemical_list: D013739:Testosterone; D006899:Mixed Function Oxygenases; C022333:peroxygenase; D010100:Oxygen
keywords: epoxidation; hydroxylation; oxyfunctionalization; peroxidase; steroid