β-Glucuronidase Activities of Feces

β-Glucuronidase Activities of Feces

To determine β-glucuronidase activities, we mixed 20 μL of the fecal suspension with 180 μL of the reaction mixture containing 1 mM p-nitrophenyl-β-D-glucuronide (FUJIFILM Wako), 50 mM HEPES-HCl (pH 7.4), and 37.2 mM 2-mercaptoethanol (24). We incubated the mixture at 37°C and measured its optical density (OD) at 405 nm, every 5 min between 10- and 30-min incubation times. We calculated the average OD change per min.

Preparation of Fecal suspension

We used fresh feces. Add 20 times the weight of the feces in water and mix thoroughly until completely suspended. Then, let it stand for about 5 minutes to remove debris. Use this suspension for the measurement.