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Research ArticleArticle

Glucuronide Production by Whole-Cell Biotransformation Using Genetically Engineered Fission Yeast Schizosaccharomyces pombe

Călin-Aurel Drăgan, Daniela Buchheit, Daniel Bischoff, Thomas Ebner and Matthias Bureik
Drug Metabolism and Disposition March 2010, 38 (3) 509-515; DOI: https://doi.org/10.1124/dmd.109.030965
Călin-Aurel Drăgan
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Daniela Buchheit
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Daniel Bischoff
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Thomas Ebner
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Matthias Bureik
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Abstract

Drug metabolites generated by UDP glycosyltransferases (UGTs) are needed for drug development and toxicity studies, especially in the context of safety testing of metabolites during drug development. Because chemical metabolite synthesis can be arduous, various biological approaches have been developed; however, no whole-cell biotransformation with recombinant microbes that express human UGTs was yet achieved. In this study we expressed human UDP glucose-6-dehydrogenase together with several human or rat UGT isoforms in the fission yeast Schizosaccharomyces pombe and generated strains that catalyze the whole-cell glucuronidation of standard substrates. Moreover, we established two methods to obtain stable isotope-labeled glucuronide metabolites: the first uses a labeled aglycon, whereas the second uses 13C6-glucose as a metabolic precursor of isotope-labeled UDP-glucuronic acid and yields a 6-fold labeled glucuronide. The system described here should lead to a significant facilitation in the production of both labeled and unlabeled drug glucuronides for industry and academia.

Footnotes

  • This work is part of the following patent application: Dragan C-A, Bureik M, and Buchheit D (2008) Drug metabolism. European patent application EP 08 164 826.3.

  • Article, publication date, and citation information can be found at http://dmd.aspetjournals.org.

    doi:10.1124/dmd.109.030965.

  • ABBREVIATIONS:

    P450
    cytochrome P450
    UGT
    UDP glycosyltransferase
    UDP-GA
    UDP glucuronic acid
    4MU
    4-methylumbelliferone
    T
    testosterone
    TG
    testosterone glucuronide
    4MUG
    4-methylumbelliferone-β-D-glucuronide
    HPLC
    high-performance liquid chromatography
    UGDH
    UDP glucose-6-dehydrogenase
    EMM
    Edinburgh minimal medium
    LC/MS
    liquid chromatography/mass spectrometry
    FTMS
    Fourier transform mass spectrometry
    EIC
    extracted ion currents.

    • Received October 29, 2009.
    • Accepted December 11, 2009.
  • Copyright © 2010 by The American Society for Pharmacology and Experimental Therapeutics
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Drug Metabolism and Disposition: 38 (3)
Drug Metabolism and Disposition
Vol. 38, Issue 3
1 Mar 2010
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Research ArticleArticle

Glucuronide Production by Whole-Cell Biotransformation Using Genetically Engineered Fission Yeast Schizosaccharomyces pombe

Călin-Aurel Drăgan, Daniela Buchheit, Daniel Bischoff, Thomas Ebner and Matthias Bureik
Drug Metabolism and Disposition March 1, 2010, 38 (3) 509-515; DOI: https://doi.org/10.1124/dmd.109.030965

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Research ArticleArticle

Glucuronide Production by Whole-Cell Biotransformation Using Genetically Engineered Fission Yeast Schizosaccharomyces pombe

Călin-Aurel Drăgan, Daniela Buchheit, Daniel Bischoff, Thomas Ebner and Matthias Bureik
Drug Metabolism and Disposition March 1, 2010, 38 (3) 509-515; DOI: https://doi.org/10.1124/dmd.109.030965
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