Use of bioconversion for the preparation of [4-14C]-labeled 7 alpha- and 7 beta-hydroxylated derivatives of dehydroepiandrosterone and epiandrosterone.
The 7 alpha- and 7 beta-hydroxylated derivatives of [4-14C]-dehydroepiandrosterone were prepared with use of the yeast-expressed human cytochrome p4507B1. Epiandrosterone (EPIA), 5 alpha-androstane-3beta,17 beta-diol, and 5 alpha-androstane-3,17-dione were obtained after incubation of [4-14C]-5 alpha-dihydrotestosterone with Escherichia coli-expressed (3beta,17 beta)-hydroxysteroid dehydrogenase from Pseudomonas testosteroni. The 7 alpha- and 7 beta-hydroxylated derivatives of [4-14C]-EPIA produced were prepared after incubation with mycelium of Rhizopus nigricans. Each labeled steroid was purified by chromatography and identified by crystallization to constant specific activity after isotopic dilution with each authentic steroid carrier. Production yields and radio-purity measurements allowed the use of such procedures for the preparation of the described radio-steroids for studies of metabolism and mode of action.
Chalbot
Sonia
S
Biotechnologie, Conservatoire National des Arts et Métiers, 2 rue Conté, 75003 Paris, France
Trap
Catherine
C
Monin
Jean Paul
JP
Morfin
Robert
R
D015113:Androstane-3,17-diol / Q000378:metabolism; D000738:Androsterone / Q000031:analogs & derivatives* / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D002790:Cholesterol 7-alpha-Hydroxylase / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003687:Dehydroepiandrosterone / Q000031:analogs & derivatives / Q000378:metabolism; D004926:Escherichia coli; D006801:Humans; D006900:Hydroxylation; D006913:Hydroxysteroid Dehydrogenases / Q000378:metabolism; D015394:Molecular Structure; D011549:Pseudomonas / Q000201:enzymology; D012233:Rhizopus
D015113:Androstane-3,17-diol; D003687:Dehydroepiandrosterone; D000738:Androsterone; D006913:Hydroxysteroid Dehydrogenases; D002790:Cholesterol 7-alpha-Hydroxylase