Abstract
Glucoamylase (SGA) was purified approximately 250-fold from sporulating Saccharomyces cerevisiae cells. The partially purified enzyme was active against glycogen, starch, maltotriose and maltose. It exhibited maximum catalytic activity against glycogen at pH 5.5. The enzyme appears to be glycosylated, because it bound to lentil-lectin Sepharose. SGA was expressed in vegetatively growing cells under the control of the GAL1 promoter, and the cellular location of the enzymatic activity determined by fractionation techniques. SGA was preferentially recovered in fractions which were enriched for the vacuolar hydrolases, carboxypeptidase Y and alpha-mannosidase.
Publication types
-
Research Support, U.S. Gov't, Non-P.H.S.
-
Research Support, U.S. Gov't, P.H.S.
MeSH terms
-
Carbohydrate Conformation
-
Cloning, Molecular
-
Electrophoresis, Polyacrylamide Gel
-
Gene Expression Regulation
-
Glucan 1,4-alpha-Glucosidase / genetics
-
Glucan 1,4-alpha-Glucosidase / isolation & purification
-
Glucan 1,4-alpha-Glucosidase / metabolism*
-
Glycogen / metabolism
-
Glycosylation
-
Hydrogen-Ion Concentration
-
Lectins / metabolism
-
Maltose / metabolism
-
Molecular Weight
-
Plant Lectins*
-
Plasmids
-
Saccharomyces cerevisiae / enzymology*
-
Saccharomyces cerevisiae / genetics
-
Saccharomyces cerevisiae / physiology
-
Spores, Fungal
-
Starch / metabolism
-
Substrate Specificity
-
Transformation, Genetic
-
Trisaccharides / metabolism
-
Vacuoles / enzymology
Substances
-
Lectins
-
Plant Lectins
-
Trisaccharides
-
lentil lectin
-
maltotriose
-
Maltose
-
Starch
-
Glycogen
-
Glucan 1,4-alpha-Glucosidase