title: The Escherichia coli glucuronylsynthase promoted synthesis of steroid glucuronides: improved practicality and broader scope.
abstract: A library of steroid glucuronides was prepared using the glucuronylsynthase derived from Escherichia coliβ-glucuronidase, followed by purification using solid-phase extraction. A representative range of steroid substrates were screened for synthesis on the milligram scale under optimised conditions with conversions dependent on steroid substitution and stereochemistry. Epiandrosterone (3β-hydroxy-5α-androstan-17-one) provided the highest conversion of 90% (84% isolated yield). The previously unreported glucuronide conjugates of methandriol (17α-methylandrost-5-ene-3β,17β-diol), cholest-5-ene-3β,25-diol and the designer steroid trenazone (17β-hydroxyestra-4,9-dien-3-one) were prepared on a multi-milligram scale suitable for characterisation by (1)H and (13)C NMR spectroscopy. The glucuronide conjugate of d5-etiocholanolone (2,2,3,4,4-d5-3α-hydroxy-5β-androstan-17-one), a target developed by the World Anti-Doping Agency as a certified reference material, was also prepared on a milligram scale. The improved E. coli glucuronylsynthase method provides for the rapid synthesis and purification of steroid glucuronides on a scale suitable for a range of analytical applications.
authors.affiliation: Research School of Chemistry, Australian National University, Canberra, ACT 0200, Australia. malcolm.mcleod@anu.edu.au.
authors.forename: Paul
authors.initials: P
authors.lastname: Ma
authors.forename: Nicholas
authors.initials: N
authors.lastname: Kanizaj
authors.forename: Shu-Ann
authors.initials: SA
authors.lastname: Chan
authors.forename: David L
authors.initials: DL
authors.lastname: Ollis
authors.forename: Malcolm D
authors.initials: MD
authors.lastname: McLeod
mesh_terms: D066241:Carbon-13 Magnetic Resonance Spectroscopy; D004926:Escherichia coli / Q000201:enzymology*; D005966:Glucuronidase / Q000378:metabolism*; D020719:Glucuronides / Q000096:biosynthesis* / Q000737:chemistry; D066244:Proton Magnetic Resonance Spectroscopy; D013256:Steroids / Q000096:biosynthesis* / Q000737:chemistry
chemical_list: D020719:Glucuronides; D013256:Steroids; D005966:Glucuronidase