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Two sepharose-bound 1-deoxynojirimycin N-alkyl derivatives, N-(9-carboxynonyl)- and N-(l l-carboxyundecyl)-deoxynojirimycin, were used for the affinity purification of acid ß-glucosidase (ß-Glc) from normal and type-1 Ashkenazi Jewish Gaucher disease (AJGD) sources. The capacities of these nondegradable inhibitor supports were 0.5 and 0.75 mg of normal ß-Glc/ml of settled gel, respectively. The purified normal enzyme (14- 18% yield) had a specific activity of 1.6 X 10^6 nmol/h/mg protein and was homogeneous as evidenced by a single protein species of M(r) = 67,000 on sodium dodecylsulfate-polyacrylamide gel electrophoresis and reverse phase high-performance liquid chromatography (HPLC). Microsequencing demonstrated a single N terminus, and the sequence of the first 22 N-terminal amino acids was colinear with that predicted from the ß-Glc cDNA. Amino acid composition analyses of ß-Glc revealed a high content (35%) of hydrophobic amino acids. The N-decyl-deoxynojirimycin support facilitated the purification of the residual enzyme from type-1 AJGD spleen to about 7,500-fold in four steps with a yield of about 11 %. These new affinity supports provided improved stability, capacity and/or specificity compared to other affinity or HPLC methods for purifying this lysosomal glycosidase.

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