title: Substrate Hunting for the Myxobacterial CYP260A1 Revealed New 1α-Hydroxylated Products from C-19 Steroids.
abstract: Cytochromes P450 catalyze a variety of synthetically useful reactions. However, it is difficult to determine their physiological or artificial functions when a plethora of orphan P450 systems are present in a genome. CYP260A1 from Sorangium cellulosum So ce56 is a new member among the 21 available P450s in the strain. To identify putative substrates for CYP260A1 we used high-throughput screening of a compound library (ca. 17,000 ligands). Structural analogues of the type I hits were searched for biotechnologically relevant compounds, and this led us to select C-19 steroids as potential substrates. We identified efficient surrogate redox partners for CYP260A1, and an Escherichia coli-based whole-cell biocatalyst system was developed to convert testosterone, androstenedione, and their derivatives methyltestosterone and 11-oxoandrostenedione. A detailed (1) H and (13) C NMR characterization of the product(s) from C-19 steroids revealed that CYP260A1 is the very first 1α-steroid hydroxylase.
authors.affiliation: Universität des Saarlandes, Biochemie, Campus B2.2, 66123, Saarbrücken, Germany.
authors.forename: Yogan
authors.initials: Y
authors.lastname: Khatri
authors.affiliation: Universität des Saarlandes, Biochemie, Campus B2.2, 66123, Saarbrücken, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Ringle
authors.affiliation: Forschungsinstitut für Molekulare Pharmakologie, Robert-Rössle-Strasse 10, 13125, Berlin, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Lisurek
authors.affiliation: Forschungsinstitut für Molekulare Pharmakologie, Robert-Rössle-Strasse 10, 13125, Berlin, Germany.
authors.forename: Jens Peter
authors.initials: JP
authors.lastname: von Kries
authors.affiliation: Universität des Saarlandes, Pharmazeutische Biologie, Campus C2.2, 66123, Saarbrücken, Germany.
authors.forename: Josef
authors.initials: J
authors.lastname: Zapp
authors.affiliation: Universität des Saarlandes, Biochemie, Campus B2.2, 66123, Saarbrücken, Germany. ritabern@mx.uni-saarland.de.
authors.forename: Rita
authors.initials: R
authors.lastname: Bernhardt
mesh_terms: D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology; D059014:High-Throughput Nucleotide Sequencing; D006900:Hydroxylation; D009231:Myxococcales / Q000201:enzymology*; D013256:Steroids / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D013256:Steroids; D003577:Cytochrome P-450 Enzyme System
keywords: Sorangium cellulosum; cytochromes P450; high-throughput screening; hydroxylation; steroids