title: P450 BM3-Catalyzed Regio- and Stereoselective Hydroxylation Aiming at the Synthesis of Phthalides and Isocoumarins.
abstract: Cytochrome P450 BM3 monooxygenases are able to catalyze the regio- and stereoselective oxygenation of a broad range of substrates, with promising potential for synthetic applications. To study the suitability of P450 BM3 variants for stereoselective benzylic hydroxylation of 2-alkylated benzoic acid esters, the biotransformation of methyl 2-ethylbenzoate, resulting in both enantiomeric forms of 3-methylphthalide, was investigated. In the case of methyl 2-propylbenzoate as a substrate the regioselectivity of the reaction was shifted towards β-hydroxylation, resulting in the synthesis of enantioenriched R- and S-configured 3-methylisochroman-1-one. The potential of P450 BM3 variants for regio- and stereoselective synthesis of phthalides and isocoumarins offers a new route to a class of compounds that are valuable synthons for a variety of natural compounds.
authors.affiliation: Institut für Bioorganische Chemie der Heinrich-Heine-Universität Düsseldorf, Forschungszentrum Jülich, Stetternicher Forst, Gebäude 15.8, 52426, Jülich, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Holec
authors.affiliation: Institut für Bio- und Geowissenschaften (IBG-1: Biotechnologie), Forschungszentrum Jülich, 52426, Jülich, Germany.
authors.forename: Ute
authors.initials: U
authors.lastname: Hartrampf
authors.affiliation: Institut für Bioorganische Chemie der Heinrich-Heine-Universität Düsseldorf, Forschungszentrum Jülich, Stetternicher Forst, Gebäude 15.8, 52426, Jülich, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Neufeld
authors.affiliation: Institut für Bioorganische Chemie der Heinrich-Heine-Universität Düsseldorf, Forschungszentrum Jülich, Stetternicher Forst, Gebäude 15.8, 52426, Jülich, Germany.|Institut für Bio- und Geowissenschaften (IBG-1: Biotechnologie), Forschungszentrum Jülich, 52426, Jülich, Germany.
authors.forename: Jörg
authors.identifier: 0000-0002-9819-889X
authors.initials: J
authors.lastname: Pietruszka
mesh_terms: D019943:Amino Acid Substitution; D001410:Bacillus megaterium; D001426:Bacterial Proteins / Q000737:chemistry*; D001572:Benzofurans / Q000138:chemical synthesis*; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry*; D006900:Hydroxylation; D049934:Isocoumarins / Q000138:chemical synthesis*; D007783:Lactones / Q000138:chemical synthesis*; D009251:NADPH-Ferrihemoprotein Reductase / Q000737:chemistry*; D015202:Protein Engineering; D013237:Stereoisomerism
chemical_list: D001426:Bacterial Proteins; D001572:Benzofurans; D049934:Isocoumarins; D007783:Lactones; D003577:Cytochrome P-450 Enzyme System; D009251:NADPH-Ferrihemoprotein Reductase; C052423:flavocytochrome P450 BM3 monoxygenases
keywords: C−H activation; P450 enzymes; benzylic hydroxylation; biotransformations; protein engineering