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PARTIAL PURIFICATION AND CHARACTERIZATION OF NEUTRAL TREHALASE FROM COMMERCIAL BAKER'S YEAST, SACCHAROMYCES CEREVISIAE
Author(s) -
YARAR SANIYE,
HAMAMCI HALUK,
BAKIR UFUR
Publication year - 2000
Publication title -
journal of food biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.507
H-Index - 47
eISSN - 1745-4514
pISSN - 0145-8884
DOI - 10.1111/j.1745-4514.2000.tb00714.x
Subject(s) - trehalase , chromatography , chemistry , yeast , column chromatography , fractionation , ion chromatography , saccharomyces cerevisiae , ammonium sulfate , biochemistry , enzyme
The neutral trehalase of a commercial baker's yeast (S. cerevisiae) strain has been partially purified using ammonium sidfate fractionation and DEAE‐cellulose column chromatography techniques. Trehalase was precipitated between 35–50% ammonium sulfate saturation and approximately 5–8 fold purification was achieved. The yeast cAMP‐dependent protein kinase was also precipitated in the same fraction and these two proteins were separated by DEAE‐cellulose column chromatography. Trehalase became totally inactive after ion exchange chromatography, “cryptic trehalase” (tre‐c), but was later activated with the addition of partially purified protem kinase together with cAMP and ATP. A 215 fold purification was obtained after DEAE‐ceUulose column chromatography. One mM EDTA caused complete inhibition of the enzyme in crude extract, however the inhibition levels in ammonium sulfate and DEAE‐cellulose fractions were 73.5% and 50%, respectively. Optimal pH range and temperature of the enzyme were determined as pH 6–6.8 and 30C, respectively. The kinetic parameters, K m and V max , were estimated as 11.78 mM trehalose and 12.47 μmole glucose/min‐mg protein, respectively .