The isolation by ligand affinity chromatography of a novel form of alpha-L-fucosidase from almond.
An alpha-fucosidase has been extracted from almond meal and purified 163,000-fold to apparent homogeneity using a novel affinity ligand, N-(5-carboxy-1-pentyl)-1,5-dideoxy-1,5-imino-L-fucitol, coupled to Affi-Gel 102. Substrate specificity studies demonstrate that the enzyme hydrolyzes the alpha-fucosidic linkages in Gal(beta 1----3)(Fuc(alpha 1----4]GlcNAc(beta 1----3)Gal(beta 1----4)Glc and Gal(beta 1----4)(Fuc(alpha 1----3]GlcNAc(beta 1----3)Gal(beta 1----4)Glc at similar rates but is unable to hydrolyze Fuc(alpha 1----2)Gal, Fuc(alpha 1----6)GlcNAc, or the synthetic substrate, p-nitrophenyl alpha-L-fucopyranoside. Hence, the enzyme closely resembles an alpha-fucosidase I isolated previously from a commercial preparation of partially purified almond beta-glucosidase (Ogata-Arakawa, M., Muramatsu, T., and Kobata, A. (1977) Arch. Biochem. Biophys. 181, 353-358). However, native and subunit relative molecular masses of 106,000 and 54,000 respectively, different charge and hydrophobicity properties, and the absence of stimulation by NaCl clearly distinguish this enzyme, designated alpha-fucosidase III, from other almond alpha-fucosidases reported previously.
Department of Biochemistry, G.D. Searle & Company, University of Oxford, United Kingdom.
P
P
Scudder
D C
DC
Neville
T D
TD
Butters
G W
GW
Fleet
R A
RA
Dwek
T W
TW
Rademacher
G S
GS
Jacob
D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002846:Chromatography, Affinity / Q000379:methods; D002850:Chromatography, Gel / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D005260:Female; D006801:Humans; D007700:Kinetics; D008024:Ligands; D008895:Milk, Human / Q000032:analysis; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000302:isolation & purification; D011247:Pregnancy; D012639:Seeds / Q000201:enzymology*; D005644:alpha-L-Fucosidase / Q000302:isolation & purification* / Q000378:metabolism
D008024:Ligands; D009844:Oligosaccharides; D005644:alpha-L-Fucosidase