Purification and Characterization of Glycyrrhetic Acid Mono-Glucuronide beta-D-glucuronidase in Eubacterium Sp. GLH

Biol Pharm Bull. 1999 Jan;22(1):80-2. doi: 10.1248/bpb.22.80.

Abstract

Glycyrrhetic acid mono-glucuronide (GAMG), 1-(18beta-glycyrrhet-3-yl)-beta-D-glucopyranuroic acid, was hydrolyzed to glycyrrhetic acid (GA) by GAMG beta-D-glucuronidase in Eubacterium sp. GLH from human intestinal bacteria. The enzyme had an optimum pH of 5.0 and was purified from a crude extract by Butyl Toyopearl 650 S, Toyopearl HW-55 S, Hydroxyapatite and DEAE-Toyopearl 650 M column chromatography. The purified enzyme showed a specific activity of 495 nmol/min/mg protein and a single band on Coomassie brilliant blue staining and a molecular weight of about 43 kDa on sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. The apparent molecular weight was 49.5 kDa, as estimated by Toyopearl HW-55 S column chromatography. Also, the enzyme seemed to have a sulfhydryl group(s) in its active site with a Km value of 77 x 10(-3) M.

MeSH terms

  • Chromatography, Gel
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Stability
  • Eubacterium / enzymology*
  • Eubacterium / isolation & purification
  • Glucuronidase / chemistry
  • Glucuronidase / isolation & purification*
  • Glucuronidase / metabolism
  • Glycyrrhetinic Acid / analogs & derivatives*
  • Glycyrrhetinic Acid / metabolism
  • Humans
  • Molecular Weight

Substances

  • glycyrrhetyl 3-monoglucuronide
  • glycyrrhetic acid mono-glucuronide beta-glucuronidase
  • glycyrrhizin beta-D-glucuronidase
  • Glucuronidase
  • Glycyrrhetinic Acid