Human liver contains hydrolytic activity toward 3β-glucosido-chenodeoxycholic acid. This β-glucosidase activity, localized predominantly in the microsomal fraction, was optimally active in the presence of divalent metal ions close to pH 5.0 and was inhibited by EDTA. Kinetic parameters and other catalytic properties of hydrolytic activity towards 3β-glucosido-chenodeoxycholic acid from human liver microsomes are described.