title: Affinity chromatographic purification of beta-glucosidase of Candida gulliermondii.
abstract: A beta-glucosidase was isolated from Candida guilliermondii, a yeast capable of growth on cellobiose. The enzyme was partially purified by treatment with polyethyleneimine and ammonium sulfate precipitation. Further purification was achieved by affinity chromatography using a Sepharose 4B matrix to which oxidized salicin was coupled through adipic dihydrazide. The final product was a 12.5-fold purification of the crude extract with a recovery of 27% of the initial enzyme activity. Polyacrylamide disc electrophoresis of the purified enzyme gave a single band. A km of 1.25 x 10(-4)M was obtained using p-nitrophenyl beta-D-glucopyranoside as the substrate. The optimum pH for enzyme activity was 6.8. Maximum activity  was observed at temperature of 37 degrees C. Enzyme activity was completely inhibited by Hg++, Pb++, and Zn++ ions. The molecular weight of the enzyme is 48,000 as estimated by sucrose density gradient centrifugation.
mesh_terms: D002175:Candida / Q000201:enzymology* / Q000378:metabolism; D050260:Carbohydrate Metabolism; D002413:Cations, Divalent; D002414:Cations, Monovalent; D002846:Chromatography, Affinity / Q000379:methods; D005959:Glucosidases / Q000302:isolation & purification*; D007700:Kinetics; D001617:beta-Glucosidase / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D002413:Cations, Divalent; D002414:Cations, Monovalent; D005959:Glucosidases; D001617:beta-Glucosidase