Eubacterium sp. GLH possessing glycyrrhizin (GL) and glycyrrhetic acid mono-glucuronide (GAMG) beta-D-glucuronidases, Ruminococcus sp. PO1-3 possessing GL and GAMG beta-D-glucuronidases and 3beta-hydroxysteroid dehydrogenase and these mixed bacteria were cultured in GAM medium with and without GL, GAMG or both. GL added to Eubacterium sp. GLH accelerated the peaks of enhanced GL beta-D-glucuronidase activity and suppressed GAMG beta-D-glucuronidase activity, and GAMG delayed the peaks of the enhanced growth with GL and GAMG beta-D-glucuronidase activities. GL added to Ruminococcus sp. PO1-3 enhanced gradually the growth with GL and GAMG beta-D-glucuronidase activities, and GAMG enhanced slowly GL beta-D-glucuronidase activity and rapidly the growth with GAMG beta-D-glucuronidase activity. The metabolite glycyrrhetic acid (GA) was produced by Eubacterium sp. GLH and Ruminococcus sp. PO1-3 in larger amounts and faster from GAMG than from GL. GL (1.0 mM) and 1.0 mM GAMG added to these mixed bacteria enhanced the growth with GL and GAMG beta-D-glucuronidase activities and were metabolized almost completely to GA in culture of 2 d and 1 d, respectively. It was found that the metabolism of GAMG was faster than that of GL. GL with GAMG added to mixed Eubacterium sp. GLH and Ruminococcus sp. PO1-3 cultured for 0 and 1 d led to a lower level of these enzyme activities and the consumption of GAMG more quickly, not GL. Low GAMG beta-D-glucuronidase had the ability to hydrolyze GAMG well.