Transformation of 5-ene steroids by the fungus Aspergillus tamarii KITA: mixed molecular fate in lactonization and hydroxylation pathways with identification of a putative 3beta-hydroxy-steroid dehydrogenase/Delta5-Delta4 isomerase pathway.
The fungus Aspergillus tamarii metabolizes progesterone to testololactone in high yield through a sequential four step enzymatic pathway which, has demonstrated flexibility in handling a range of steroidal probes. These substrates have revealed that subtle changes in the molecular structure of the steroid lead to significant changes in route of metabolism. It was therefore of interest to determine the metabolism of a range of 5-ene containing steroidal substrates. Remarkably the primary route of 5-ene steroid metabolism involved a 3beta-hydroxy-steroid dehydrogenase/Delta(5)-Delta(4) isomerase (3beta-HSD/isomerase) enzyme(s), generating 3-one-4-ene functionality and identified for the first time in a fungus with the ability to handle both dehydroepiansdrosterone (DHEA) as well as C-17 side-chain containing compounds such as pregnenolone and 3beta-hydroxy-16alpha,17alpha-epoxypregn-5-en-20-one. Uniquely in all the steroids tested, 3beta-HSD/isomerase activity only occurred following lactonization of the steroidal ring-D. Presence of C-7 allylic hydroxylation, in either epimeric form, inhibited 3beta-HSD/isomerase activity and of the substrates tested, was only observed with DHEA and its 13alpha-methyl analogue. In contrast to previous studies of fungi with 3beta-HSD/isomerase activity DHEA could also enter a minor hydroxylation pathway. Pregnenolone and 3beta-hydroxy-16alpha,17alpha-epoxypregn-5-en-20-one were metabolized solely through the putative 3beta-HSD/isomerase pathway, indicating that a 17beta-methyl ketone functionality inhibits allylic oxidation at C-7. The presence of the 3beta-HSD/isomerase in A. tamarii and the transformation results obtained in this study highlight an important potential role that fungi may have in the generation of environmental androgens.
Hunter
A Christy
AC
Molecular Targeting and Polymer Toxicology Group, School of Pharmacy, University of Brighton, Cockcroft Building, Lewes Road, Brighton, East Sussex BN2 4GJ, UK. c.hunter@bton.ac.uk
Coyle
Emma
E
Morse
Fiona
F
Dedi
Cinzia
C
Dodd
Howard T
HT
Koussoroplis
Salome-Juliette
SJ
D001230:Aspergillus / Q000201:enzymology*; D018360:Crystallography, X-Ray; D003687:Dehydroepiandrosterone / Q000737:chemistry / Q000378:metabolism; D006900:Hydroxylation; D009097:Multienzyme Complexes / Q000378:metabolism*; D011284:Pregnenolone / Q000737:chemistry / Q000378:metabolism*; D011375:Progesterone Reductase / Q000378:metabolism*; D015398:Signal Transduction; D013251:Steroid Isomerases / Q000378:metabolism*
C021425:3 beta-hydroxysteroid oxidoreductase-delta(5) 3-ketosteroid isomerase; D009097:Multienzyme Complexes; D003687:Dehydroepiandrosterone; D011284:Pregnenolone; D011375:Progesterone Reductase; D013251:Steroid Isomerases