Effect of Food Substances on beta-glucuronidase Activity of lntestinal Bacteria.

食品物质对肠道细菌β-葡萄糖醛酸酶活性的影响。

• 批准号: 07660176
• 负责人: SEKIZAWA Tsuneo
• 金额: $ 1.09万
• 依托单位: Nagaoka College of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07660176/
• 关键词: intestinal bacteria; glucuronidase; activity assay; electrophoresis; food substance

This study was conducted to clarify the effect of food substances on beta-glucuronidase metabolism of intestinal microflora by using the electrophoresis and the activity staining enzyme method. By using together with above methods, the measuring assay for the activity of each producing bacteria, respectively was established. This method was applied to the rat cecal content and human fecesBy supplemented spices to rats, the activity of cecal beta-glucronidase activity was increased significantly. In tha rat cecum, we detected only one of beta-glucuronidase active band on the PAGE gel, and this band was in accord with that established in Esherichia coli beta-glucuronidase. This increase was resulted from activity increase of beta-glucuronidase of E.coli. Changes of total activity in cecum were detected as the enzyme band concentrations of E.coli. From the experimental results of the total number of E.coli did not varied, It may be reasonable to conclude that the increases of the activities depend on the increases of production rate of beta-glucronidase from E.coli.We detected two active bands on PAGE gel in healthy human feces. One was major in accord with that established in E.coli beta-glucuronidase, and another miner was in accord with Bacteroides fragilis enzyme. In human feces, major activity of beta-glucuronidase was E.coli orgin.The problem in this study is the low detectable limit activity staining enzyme method with the substrate as 6-brom-2 naphtyl-beta-D-glucuronide or alpha-naphtol-AS-Bl beta-glucuronide. Therefore, this method are inadequate to detect low activity of the isozyme.

本研究采用电泳和活性染色酶法，研究了食物物质对肠道菌群β -葡糖苷酶代谢的影响。结合上述方法，分别建立了各产菌活性的测定方法。将该方法应用于大鼠盲肠内容物和人粪便中，通过添加香料，大鼠盲肠β -葡萄糖苷酶活性显著提高。在该大鼠盲肠中，PAGE凝胶只检测到一条β -葡糖醛酸酶活性条带，该条带与大肠杆菌β -葡糖醛酸酶的活性条带一致。这种增加是由于大肠杆菌β -葡萄糖醛酸酶活性增加所致。以大肠杆菌酶带浓度检测盲肠总活性的变化。从实验结果来看，大肠杆菌的总数没有变化，可以合理地推断，活性的增加与大肠杆菌产生β -葡萄糖苷酶的速度的增加有关。我们在健康人粪便中检测到PAGE凝胶上的两个活性条带。其中一种主要与大肠杆菌β -葡萄糖醛酸酶一致，另一种与脆弱拟杆菌酶一致。在人类粪便中，β -葡糖醛酸酶的主要活性是大肠杆菌。本研究的问题是用底物为6-溴-2萘基- β - d -葡糖苷或α -萘基- as - bl -葡糖苷的低检测限活性染色酶法。因此，这种方法不适用于检测低活性的同工酶。