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

1996 Fiscal Year Final Research Report Summary

• Project/Area Number: 07660176
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: 食品科学・栄養科学
• Research Institution: Nagaoka College of Technology
• Principal Investigator: SEKIZAWA Tsuneo Department of lndustrial Chemistry Nagaoka College of Technology Professor, 工業化学科, 教授 (90042746)
• Co-Investigator(Kenkyū-buntansha): SUGAWARA Masayoshi Department of Materials Engineering Nagaoka College of Technology Associcte Prof, 物質工学科, 助教授 (30259840)
• Project Period (FY): 1995 – 1996
• Keywords: intestinal bacteria / glucuronidase / activity assay / electrophoresis / food substance

Research Abstract

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 feces
By 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.