Premium
Purification and characterization of trimming glucosidase I from Saccharomyces cerevisiae
Author(s) -
Bause Ernst,
Erkens Rainer,
Schweden Jürgen,
Jaenicke Lothar
Publication year - 1986
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(86)80982-6
Subject(s) - chemistry , glucosidases , yeast , chromatography , enzyme , sepharose , affinity chromatography , saccharomyces cerevisiae , biochemistry , glycoside hydrolase , molecular mass , residue (chemistry) , glycoprotein , substrate (aquarium) , biology , ecology
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 Glc 3 ‐Man 9 ‐GlcNAc 2 chain of its natural substrate, but is not active against Glc 2 ‐Man 9 ‐GlcNAc 2 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.