CYP21-catalyzed production of the long-term urinary metandienone metabolite 17β-hydroxymethyl-17α-methyl-18-norandrosta-1,4,13-trien-3-one: a contribution to the fight against doping
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Andy Zöllner
Abstract
Anabolic-androgenic steroids are some of the most frequently misused drugs in human sports. Recently, a previously unknown urinary metabolite of metandienone, 17β-hydroxymethyl-17α-methyl-18-norandrosta-1,4,13-trien-3-one (20OH-NorMD), was discovered via LC-MS/MS and GC-MS. This metabolite was reported to be detected in urine samples up to 19 days after administration of metandienone. However, so far it was not possible to obtain purified reference material of this metabolite and to confirm its structure via NMR. Eleven recombinant strains of the fission yeast Schizosaccharomyces pombe that express different human hepatic or steroidogenic cytochrome P450 enzymes were screened for production of this metabolite in a whole-cell biotransformation reaction. 17,17-Dimethyl-18-norandrosta-1,4,13-trien-3-one, chemically derived from metandienone, was used as substrate for the bioconversion, because it could be converted to the final product in a single hydroxylation step. The obtained results demonstrate that CYP21 and to a lesser extent also CYP3A4 expressing strains can catalyze this steroid hydroxylation. Subsequent 5 l-scale fermentation resulted in the production and purification of 10 mg of metabolite and its unequivocal structure determination via NMR. The synthesis of this urinary metandienone metabolite via S. pombe-based whole-cell biotransformation now allows its use as a reference substance in doping control assays.
©2010 by Walter de Gruyter Berlin New York
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Artikel in diesem Heft
- REVIEW
- Hexose-6-phosphate dehydrogenase in the endoplasmic reticulum
- GENES AND NUCLEIC ACIDS
- Biochemical characterization of human Ecdysoneless reveals a role in transcriptional regulation
- PROTEIN STRUCTURE AND FUNCTION
- Bovine β-lactoglobulin acts as an acid-resistant drug carrier by exploiting its diverse binding regions
- Structural studies of the phosphatidylinositol 3-kinase (PI3K) SH3 domain in complex with a peptide ligand: role of the anchor residue in ligand binding
- A fluorescence correlation spectroscopy study of ligand interaction with cytokinin-specific binding protein from mung bean
- The oxygen-independent coproporphyrinogen III oxidase HemN utilizes harderoporphyrinogen as a reaction intermediate during conversion of coproporphyrinogen III to protoporphyrinogen IX
- MEMBRANES, LIPIDS, GLYCOBIOLOGY
- Phytosphingosine kills Candida albicans by disrupting its cell membrane
- CELL BIOLOGY AND SIGNALING
- Dexamethasone-dependent versus -independent markers of epithelial to mesenchymal transition in primary hepatocytes
- PROTEOLYSIS
- Potential role of multiple members of the kallikrein-related peptidase family of serine proteases in activating latent TGFβ1 in semen
- Binding and activation of the human plasma kinin-forming system on the cell walls of Candida albicans and Candida tropicalis
- A novel matrix metalloprotease-like enzyme (karilysin) of the periodontal pathogen Tannerella forsythia ATCC 43037
- NOVEL TECHNIQUES
- CYP21-catalyzed production of the long-term urinary metandienone metabolite 17β-hydroxymethyl-17α-methyl-18-norandrosta-1,4,13-trien-3-one: a contribution to the fight against doping
