Akao T
Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University, Sugitani, Japan.
Biol Pharm Bull. 2000 Jan;23(1):6-11. doi: 10.1248/bpb.23.6.
Eubacterium sp. GLH with Ruminococcus sp. PO1-3 and Clostridium innocuum ES24-06 possessing enzymes involved in the metabolism of glycyrrhizin (GL) was cultured in GAM medium with and without 1.0 mM GL or 1.0 mM glycyrrhetic acid (GA). GL (1.0 mM) enhanced 3alpha-hydroxyglycyrrhetinate (3alpha-hydroxyGA) dehydrogenase activity, GA (1.0 mm) suppressed 3alpha-hydroxyGA dehydrogenase activity, GL beta-D-glucuronidase activity and the mixed bacterial growth, and GL and GA showed almost no change in a lower level of 3beta-hydroxysteroid dehydrogenase (3beta-HSD) activity during 5 d of culture. GL (1.0 mM) and GA (1.0 mM) were metabolized to a small amount of GA and a negligible amount of 3-oxo-glycyrrhetic acid (3-oxo-GA) and 3alpha-hydroxyGA, and to a negligible amount of 3-oxo-GA, respectively, by these mixed bacteria. These amounts coincided with those of metabolites produced from 1.0 mM GL and 1.0 mM GA added to these mixed bacteria after 24 h culture. Whole bacteria and sonicated bacteria derived from the collection of these mixed bacteria reached a maximal stage and metabolized GL to a relatively large amount of GA and 3-oxo-GA, and a negligible amount of 3alpha-hydroxyGA and GA to a small amount of 3-oxo-GA and 3alpha-hydroxyGA within 180 min. GL beta-D-glucuronidase with 3beta-HSD and 3alpha-hydroxyGA dehydrogenase partially purified from each bacterium was converted GL to 3alpha-hydroxyGA, producing metabolites of about 60% after 10 min of incubation. These mixed bacteria possessed high enzyme activities could produce the metabolites of GL in under one hour under conditions.
将含有参与甘草酸(GL)代谢的酶的真杆菌属GLH与瘤胃球菌属PO1 - 3和无害梭菌ES24 - 06在添加和不添加1.0 mM GL或1.0 mM甘草次酸(GA)的GAM培养基中培养。1.0 mM的GL增强了3α - 羟基甘草次酸(3α - 羟基GA)脱氢酶的活性,1.0 mM的GA抑制了3α - 羟基GA脱氢酶的活性、GL β - D - 葡萄糖醛酸酶的活性以及混合细菌的生长,并且在培养5天期间,GL和GA在较低水平的3β - 羟基类固醇脱氢酶(3β - HSD)活性方面几乎没有变化。1.0 mM的GL和1.0 mM的GA分别被这些混合细菌代谢为少量的GA和可忽略量的3 - 氧代甘草次酸(3 - 氧代 - GA)和3α - 羟基GA,以及可忽略量的3 - 氧代 - GA。这些量与在24小时培养后添加到这些混合细菌中的1.0 mM GL和1.0 mM GA产生的代谢物量一致。源自这些混合细菌的完整细菌和超声破碎细菌在180分钟内达到最大阶段,并将GL代谢为相对大量的GA和3 - 氧代 - GA,以及可忽略量的3α - 羟基GA,将GA代谢为少量的3 - 氧代 - GA和3α - 羟基GA。从每种细菌中部分纯化的具有3β - HSD和3α - 羟基GA脱氢酶的GL β - D - 葡萄糖醛酸酶在孵育10分钟后将GL转化为3α - 羟基GA,产生约60%的代谢物。这些具有高酶活性的混合细菌在该条件下可在一小时内产生GL的代谢物。