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

FEBS Letters

Schweden, Jürgen¹  Gross, Achim¹  Bause, Ernst¹ 

[1] Institut für Physiologische Chemie der Universität, Nußallee 11.53 Bonn I, Germany

关键词: Trimming glucosidase I;    Pig liver;    Affinity purification;    1-Deoxinojirimycin derivative;    dNM;    1-deoxynorjirimycin;    CP-;    N-(5-carboxypentyl)-;    Nph-;    nitrophenyl;    AH-Sepharose 4B;    aminohexyl-Sepharose 4B;   

DOI  :  10.1016/0014-5793(91)80108-F

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown