Purification and characterization of trimming glucosidase I from Saccharomyces cerevisiae

FEBS Letters

Schweden, Jürgen¹  Jaenicke, Lothar¹  Erkens, Rainer¹  Bause, Ernst¹ 

[1] Institut für Biochemie, Zülpicher Str. 47, 5000 Köln 1, FRG

关键词: Trimming enzyme;    Glucosidase I;    Enzyme purification;    (S. cerevisiae);    PMSF;    phenylmethylsulfonyl fluoride;    SDS-PAGE;    SDS-polyacrylamide gel electrophoresis;    dNM;    1-deoxynojirimycin;    CP-dNM;    N-5-carboxypentyl-1-deoxynojirimycin;    Nph-;    p-nitrophenyl-;   

DOI  :  10.1016/0014-5793(86)80982-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Glucosidase I was purified about 1900-fold from yeast microsomal preparations by DEAE-Sephacel chromatography, affinity chromatography on AH-Sepharose 4B-linked N-5-carboxypentyl-1-deoxynojirimycin and Con A-Sepharose chromatography. The enzyme is a glycoprotein with a subunit molecular mass of 95 kDa. Its reaction has a pH optimum close to 6.8 and does not require metal ions. Purified glucosidase I hydrolyses the distal α1,2-linked glucose residue from the Glc₃-Man₉-GlcNAc₂ chain of its natural substrate, but is not active against Glc₂-Man₉-GlcNAc₂ and aryl-α-glucosides. Like glucosidase I from calf liver, the yeast enzyme is strongly inhibited by 1-deoxynojirimycin (dNM), N-methyl-dNM and N-5-carboxypentyl-dNM with K _i values of 16, 0.3 and 3μM, respectively.

【 授权许可】

Unknown