title: Properties of immobilized fig alpha-galactosidase and effect on ceramide-3 content of plasma from patients with Fabry's disease.
abstract: The possibility of lowering the level of ceramide-3 (galactosyl-alpha(1 leads to 4)-galactosyl-beta(1 leads to 4)-glucosyl-beta(1 leads to 1)-ceramide) in the plasma of patients with Fabry's disease was investigated. An immobilized alpha-galactosidase (alpha-D-galactoside galactohydrolase, EC 3.2.1.22) was prepared by coupling purified fig alpha-galactosidase to Sepharose 4B. The pH optimum for the hydrolysis of the artificial substrate p-nitro-phenyl-alpha-D-galactopyranoside was shifted by approx. 0.5--1.0 pH unit to higher pH values upon coupling of the enzyme to Sepharose 4B. The immobilized enzyme was more stable than the native enzyme to incubation at 60 degrees C. The immobilized enzyme was able to hydrolyse ceramide-3 either at pH 4.5 or at pH 7.4 in an artificial system in which sodium taurocholate was used to solubilize the substrate. In contrast, when the immobilized enzyme was incubated with normal plasma or plasma from a patient with Fabry's disease, in which elevated levels of ceramide-3 occur, no hydrolysis of the glycosphingo-lipid could be detected. The results suggest that lowering of level of ceramide-3 in plasma from patients with Fabry's disease by enzymic means is not feasible.
mesh_terms: D004355:Drug Stability; D004800:Enzymes, Immobilized; D000795:Fabry Disease / Q000097:blood* / Q000188:drug therapy; D005696:Galactosidases / Q000378:metabolism*; D006028:Glycosphingolipids / Q000097:blood*; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D010944:Plants / Q000201:enzymology; D000519:alpha-Galactosidase / Q000378:metabolism* / Q000627:therapeutic use
chemical_list: D004800:Enzymes, Immobilized; D006028:Glycosphingolipids; D005696:Galactosidases; D000519:alpha-Galactosidase