title: Properties of partially purified goat kidney beta-D-mannosidase.
abstract: Lysosomal beta-D-mannosidase (EC 3.2.1.25) was purified 6900-fold from normal goat kidney by serial Concanavalin A-Sepharose 4B and Red A dye ligand affinity chromatography, followed by anion exchange and molecular sieve high performance liquid chromatography. The relative molecular mass of the enzyme was estimated by molecular sieving to be 79,000 +/- 3000. The apparent Km for the synthetic substrate, 4-methylumbelliferyl-beta-D-mannopyranoside, was 2.3-2.8 mM and the sharp, unimodal pH optimum was 5.5. Enzyme activity was inhibited by Hg2+, Zn2+, Ag+, Co2+ and the thiol reactive agent N-ethylmaleimide. The mannose derivatives p-nitrophenyl-beta-D- thiomannopyranoside and p-aminophenyl-beta-D-thiomannopyranoside inhibited enzyme activity and may be of use as immobilized ligands in future attempts to purify beta-D-mannosidase by specific affinity chromatography.
authors.affiliation: Department of Paediatrics, Hospital for Sick Children, Toronto, Ontario, Canada.
authors.forename: R D
authors.initials: RD
authors.lastname: Pearce
authors.forename: J W
authors.initials: JW
authors.lastname: Callahan
authors.forename: A
authors.initials: A
authors.lastname: Novak
authors.forename: P B
authors.initials: PB
authors.lastname: Little
authors.forename: J T
authors.initials: JT
authors.lastname: Clarke
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid; D006041:Goats / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007668:Kidney / Q000201:enzymology*; D007700:Kinetics; D008361:Mannosidases / Q000032:analysis* / Q000302:isolation & purification; D008970:Molecular Weight; D044902:beta-Mannosidase
chemical_list: D008361:Mannosidases; D044902:beta-Mannosidase