N‐Methyl‐N‐(5‐carboxypentyl)‐1‐deoxynojirimycin, a new affinity ligand for the purification of trimming glucosidase I

Abstract

The paper describes the synthesis of a new type of affinity resin containing N-methyl-N(5-carboxypentyl)-1-deoxynojirimycin as the ligand attached to AH-Sepharose 4B, which allows the purification of trimming glucosidase 1 from a detergent extract of pig liver crude microsomes in one step and with high yield. The structure of the affinity ligand was designed on the basis of the observation that N, N-dialkylated derivatives of 1-deoxynojirimycin do strongly inhibit trimming glucosidase I, but not nonspecific aryl-a-glucosidases, including glucosidase II. The specific binding of glucosidase I eliminates the need of additional purification steps with their associated losses which were required with the previously synthesized N-5(-carboxypentyl)-AH-Sepharose 4B resin in order to achieve a homogenous enzyme preparation.