Glucuronide production by whole-cell biotransformation using genetically engineered fission yeast Schizosaccharomyces pombe.
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 (13)C(6)-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.
Drăgan
Călin-Aurel
CA
PomBioTech GmbH, Saarbrücken, Germany.
Buchheit
Daniela
D
Bischoff
Daniel
D
Ebner
Thomas
T
Bureik
Matthias
M
D000818:Animals; D001711:Biotransformation; D002851:Chromatography, High Pressure Liquid; D005818:Genetic Engineering / Q000379:methods*; D020719:Glucuronides / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D014453:Glucuronosyltransferase / Q000235:genetics / Q000378:metabolism; D016695:Glycosyltransferases / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007553:Isotope Labeling / Q000379:methods; D015394:Molecular Structure; D051381:Rats; D012568:Schizosaccharomyces / Q000235:genetics* / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization; D017550:Spectroscopy, Fourier Transform Infrared; D014170:Transformation, Genetic; D014533:Uridine Diphosphate Glucose Dehydrogenase / Q000235:genetics / Q000378:metabolism
D020719:Glucuronides; D014533:Uridine Diphosphate Glucose Dehydrogenase; D016695:Glycosyltransferases; D014453:Glucuronosyltransferase