Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Purification and Characterization of Two Novel $\beta$-D-Glucuronidases Converting Glycyrrhizin to 18$\beta$-Glycyrrhetinic Acid-3-O-$\beta$-D-Glucuronide from Streptococcus LJ-22

  • PARK HYE-YOUNG (Department of Food and Nutrition, Kyung Hee University) ;
  • KIM NA-YOUNG (Department of Food and Nutrition, Kyung Hee University) ;
  • HAN MYUNG JOO (Department of Food and Nutrition, Kyung Hee University) ;
  • BAE EUN-AH (College of Pharmacy, Kyung Hee University) ;
  • KIM DONG-HYUN (College of Pharmacy, Kyung Hee University)
  • Published : 2005.08.01
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Abstract

Two novel $\beta$-glucuronidases, which metabolize glycyrrhizin (GL) to 18$\beta$-glycyrrhetinic acid-3-O-$\beta$-D-glucuronide (GAMG), were purified from Streptococcus LJ-22 isolated from human intestinal microflora. $\beta$-Glucuronidases I and II were purified to apparent homogeneity, using a combination of ammonium sulfate fractionation, butyl toyopearl, Q-Sepharose, hydroxyapatite Ultrogel, and GL-attached Sepharose column chromatographies, with the final specific activities of 137 and 190 nmole/min/mg, respectively. The molecular sizes of both $\beta$-glucuronidases were found to be 140 kDa by gel filtration, and they consisted of two identical subunits (M.W. 67 kDa by SDS-PAGE). $\beta$-Glucuronidases I and II showed optimal activity at pH 7.0 and pH 6.5, respectively. Both purified enzymes were potently inhibited by $Cu^{2+}$ and PCMS, and had maximum activity on glycyrrhizin, but did not hydrolyze p-nitrophenyl-$\beta$-glucuronides, baicalin, or GAMG These findings suggest that the biochemical properties and substrate specificities of these enzymes are different from those of the previously purified $\beta$-glucuronidases. This is the first reported purification of sugar (not aglycone)-recognizing $\beta$-glucuronidases from intestinal bacteria.

Keywords

References

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