title: Whole-cell biotransformation with recombinant cytochrome P450 for the selective oxidation of Grundmann's ketone.
abstract: 25-Hydroxy-Grundmann's ketone is a key building block in the chemical synthesis of vitamin D3 and its derivatives through convergent routes. Generally, the chemical synthesis of this compound involves tedious procedures and results in a mixture of several products. Recently, the selective hydroxylation of Grundmann's ketone at position C25 by cytochrome P450 (CYP) 154E1 from Thermobifida fusca YX was described. In this study a recombinant whole-cell biocatalyst was developed and applied for hydroxylation of Grundmann's ketone. Biotransformation was performed by Escherichia coli cells expressing CYP154E1 along with two redox partner systems, Pdx/PdR and YkuN/FdR. The system comprising CYP154E1/Pdx/PdR showed the highest production of 25-hydroxy-Grundmann's ketone and resulted in 1.1mM (300mgL(-1)) product concentration.
authors.affiliation: Institute of Biochemistry, Heinrich-Heine University Düsseldorf, Universitätsstrasse 1, 40225 Düsseldorf, Germany; Departamento de Química Orgánica e Inorgánica, Instituto Universitario de Biotecnología de Asturias, Universidad de Oviedo, 33006 Oviedo (Asturias), Spain.
authors.forename: Alba
authors.initials: A
authors.lastname: Hernández-Martín
authors.affiliation: Institute of Biochemistry, Heinrich-Heine University Düsseldorf, Universitätsstrasse 1, 40225 Düsseldorf, Germany.
authors.forename: Clemens J
authors.initials: CJ
authors.lastname: von Bühler
authors.affiliation: Institute of Biochemistry, Heinrich-Heine University Düsseldorf, Universitätsstrasse 1, 40225 Düsseldorf, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Tieves
authors.affiliation: Departamento de Química Orgánica e Inorgánica, Instituto Universitario de Biotecnología de Asturias, Universidad de Oviedo, 33006 Oviedo (Asturias), Spain.
authors.forename: Susana
authors.initials: S
authors.lastname: Fernández
authors.affiliation: Departamento de Química Orgánica e Inorgánica, Instituto Universitario de Biotecnología de Asturias, Universidad de Oviedo, 33006 Oviedo (Asturias), Spain.
authors.forename: Miguel
authors.initials: M
authors.lastname: Ferrero
authors.affiliation: Institute of Biochemistry, Heinrich-Heine University Düsseldorf, Universitätsstrasse 1, 40225 Düsseldorf, Germany. Electronic address: vlada.urlacher@uni-duesseldorf.de.
authors.forename: Vlada B
authors.initials: VB
authors.lastname: Urlacher
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology* / Q000378:metabolism; D055162:Biocatalysis; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004926:Escherichia coli / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism; D007659:Ketones / Q000737:chemistry / Q000378:metabolism*; D008968:Molecular Conformation; D010084:Oxidation-Reduction; D016958:Pseudomonas putida / Q000201:enzymology* / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D007659:Ketones; D011994:Recombinant Proteins; D003577:Cytochrome P-450 Enzyme System
keywords: Cytochrome P450; Grundmann’s ketone; Hydroxylation; Vitamin D(3); Whole-cell biocatalysis