Purification and characterization of trimming glucosidase I from Saccharomyces cerevisiae
Authors:
- Ernst BauseEBErnst Bause
- Rainer ErkensRERainer Erkens
- Jürgen SchwedenJSJürgen Schweden
- Lothar JaenickeLJLothar Jaenicke
- 6 October 1986
- journal article
- Published by Wiley in FEBS Letters
- Vol. 206 (2) , 208-212
Abstract
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.Keywords
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