Deconjugation Kinetics of Glucuronidated Phase II Flavonoid Metabolites by beta-glucuronidase from Neutrophils

R. Bartholomé, G. Haenen, P.C.H. Hollman, A. Bast, P.C. Dagnelie, D. Roos, J. Keijer, P.A. Kroon, P.W. Needs, I.C.W. Arts

Research output: Contribution to journalArticleAcademic

Abstract

Flavonoids are inactivated by phase II metabolism and occur in the body as glucuronides. Mammalian beta-glucuronidase released from neutrophils at inflammatory sites may be able to deconjugate and thus activate flavonoid glucuronides. We have studied deconjugation kinetics and pH optimum for four sources of beta-glucuronidase (human neutrophil, human recombinant, myeloid PLB-985 cells, Helix pomatia) with five flavonoid glucuronides (quercetin-3-glucuronide, quercetin-3'-glucuronide, quercetin-4'-glucuronide, quercetin-7-glucuronide, 3'-methylquercetin-3-glucuronide), 4-methylumbelliferyl-beta-D-glucuronide, and paranitrophenol-glucuronide. All substrate-enzyme combinations tested exhibited first order kinetics. The optimum pH for hydrolysis was between 3.5-5, with appreciable hydrolysis activities up to pH 5.5. At pH 4, the Km ranged 44-fold from 22 mu M for quercetin-4'-glucuronide with Helix pomatia beta-glucuronidase, to 981 mu M for para-nitrophenol-glucuronide with recombinant beta-glucuronidase. V-max (range: 0.735-24.012 mmol.min(-1).unit(-1) [1 unit is defined as the release of 1 mu M 4-methylumbelliferyl-beta-D-glucuronide per min]) and the reaction rate constants at low substrate concentrations (k) (range: 0.002-0.062 min(-1). (unit/L)(-1) were similar for all substrates-enzyme combinations tested. In conclusion, we show that beta-glucuronidase from four different sources, including human neutrophils, is able to deconjugate flavonoid glucuronides and non-flavonoid substrates at fairly similar kinetic rates. At inflammatory sites in vivo the pH, neutrophil and flavonoid glucuronide concentrations seem favorable for deconjugation. However, it remains to be confirmed whether this is actually the case
Original languageUndefined/Unknown
Pages (from-to)379-387
JournalDrug Metabolism and Pharmacokinetics
Volume25
Issue number4
DOIs
Publication statusPublished - 2010

Cite this