Purification and characterization of a broad specificity beta-glucosidase from sheep liver.

1. A soluble beta-glucosidase from sheep liver has been isolated and purified 114-fold by conventional enzyme fractionation procedures. The specific activity of the purified enzyme was 5910 mU/mg of protein. 2. The enzyme has a broad specificity and hydrolyzes the p-nitrophenyl derivatives of beta-D-glucose, beta-D-galactose, beta-D-fucose, beta-D-xylose and alpha-L-arabinose. The best Vmax/Km value corresponds to the beta-glucosidase activity. 3. The enzyme has a pH optimum between 4.5-5.5 for all the activities, and mol. wt 95,000. 4. A variety of chemicals was tested as possible activators or inhibitors. 5. The enzyme is strongly inhibited by aldono 1-5 lactones and DMDP. 6. The kinetic evidences suggest a substrate activation model and the existence of two active sites (a "gluco-fuco" site and a "galacto" site). 7. The activation energies were calculated from beta-glucosidase and beta-galactosidase activities.