One-step purification and characterization of an intracellular beta-glucosidase from Metschnikowia pulcherrima.
A collection of 60 non-Saccharomyces yeasts isolated from grape musts in Uruguayan vineyards was screened for beta-glucosidase activity and Metschnikowia pulcherrima was the best source of this enzyme activity. Its major beta-glucosidase was successfully purified to homogeneity by ion-exchange chromatography on amino-agarose gel. The enzyme exhibited an optimum catalytic activity at 50 degrees C and pH 4.5 and was active against (1 --> 4)-beta and (1 --> 2)-beta glycosidic linkages. In spite of preserving 100% of its activity and stability in the presence of 12% (v/v) ethanol and 5 g glucose/l, the enzyme was unstable below pH 4. We characterized the beta-glucosidase from M. pulcherrima with a view to its potential applications in wine-making.
González-Pombo
Paula
P
Departamento de Biociencias, Facultad de Química, Cátedra de Bioquímica, Gral Flores 2124, CC1157 Montevideo, Uruguay.
Pérez
Gabriel
G
Carrau
Francisco
F
Guisán
José Manuel
JM
Batista-Viera
Francisco
F
Brena
Beatriz M
BM
D000327:Adsorption / Q000187:drug effects; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D042541:Intracellular Space / Q000201:enzymology*; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008670:Metals / Q000494:pharmacology; D004718:Saccharomycetales / Q000187:drug effects / Q000201:enzymology*; D012965:Sodium Chloride / Q000494:pharmacology; D013379:Substrate Specificity / Q000187:drug effects; D001617:beta-Glucosidase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism*
D007527:Isoenzymes; D008670:Metals; D012965:Sodium Chloride; D001617:beta-Glucosidase