title: Transformation of some 3alpha-substituted steroids by Aspergillus tamarii KITA reveals stereochemical restriction of steroid binding orientation in the minor hydroxylation pathway.
abstract: Aspergillus tamarii contains an endogenous lactonization pathway which can transform progesterone to testololactone in high yield through a sequential four step enzymatic pathway. In this pathway testosterone is formed which primarily undergoes oxidation of the C-17beta-alcohol to a C-17 ketone but, can also enter a minor hydroxylation pathway where 11beta-hydroxytestosterone is produced. It was recently demonstrated that this hydroxylase could monohydroxylate 3beta-hydroxy substituted saturated steroidal lactones in all four possible binding orientations (normal, reverse, inverted normal, inverted reverse) on rings B and C of the steroid nucleus. It was therefore of interest to determine the fate of a series of 3alpha-substituted steroidal analogues to determine stereochemical effect on transformation. Hydroxylation on the central rings was found to be restricted to the 11beta-position (normal binding), indicating that the 3alpha-stereochemistry removes freedom of binding orientation within the hydroxylase. The only other hydroxylation observed was at the 1beta-position. Interestingly the presence of this functional group did not prevent lactonization of the C-17 ketone. In contrast the presence of the 11beta-hydroxyl completely inhibited Baeyer-Villiger oxidation, a result which again demonstrates that single functional groups can exert significant control over metabolic handling of steroids in this organism. This may also explain why lactonization of 11beta-hydroxytestosterone does not occur. Lactonization of the C-17 ketone was not significantly affected by the 3alpha-alcohol with significant yields achieved (53%). Interestingly a time course experiment demonstrated that the presence of the 3alpha-acetate inhibited the Baeyer-Villiger monooxygenase with its activity being observed 24h later than non-acetate containing analogues. Apart from oxidative transformations observed a minor reductive pathway was revealed with the C-17 ketone being reduced to a C-17beta-alcohol for the first time in this organism.
authors.affiliation: Molecular Targeting and Polymer Toxicology Group, School of Pharmacy, University of Brighton, East Sussex BN2 4GJ, UK. c.hunter@bton.ac.uk <c.hunter@bton.ac.uk>
authors.forename: A
authors.initials: A
authors.lastname: Christy Hunter
authors.forename: Hedda
authors.initials: H
authors.lastname: Khuenl-Brady
authors.forename: Patrice
authors.initials: P
authors.lastname: Barrett
authors.forename: Howard T
authors.initials: HT
authors.lastname: Dodd
authors.forename: Cinzia
authors.initials: C
authors.lastname: Dedi
mesh_terms: D000732:Androstanols / Q000032:analysis / Q000378:metabolism*; D001230:Aspergillus / Q000201:enzymology*; D055162:Biocatalysis; D001711:Biotransformation / Q000502:physiology; D006900:Hydroxylation; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D010084:Oxidation-Reduction; D013055:Spectrophotometry, Infrared; D013237:Stereoisomerism; D013252:Steroid 11-beta-Hydroxylase / Q000378:metabolism; D013250:Steroid Hydroxylases / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D000732:Androstanols; D013250:Steroid Hydroxylases; D013252:Steroid 11-beta-Hydroxylase