Purification and Characterization of an Ethanol-Tolerant beta-Glucosidase from Sporidiobolus pararoseus and Its Potential for Hydrolysis of Wine Aroma Precursors
Baffi, Milla Alves
Tobal, Thaise Maria
Ferrarezi, Ana Lucia
Ghilardi Lago, Joao Henrique [UNIFESP]
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An extracellular ethanol-tolerant beta-glucosidase from Sporidiobolus pararoseus was purified to homogeneity and characterized, and its potential use for the enhancement of wine aroma was investigated. the crude enzymatic extract was purified in four steps (concentration, dialysis, ultrafiltration, and chromatography) with a yield of around 40 % for total activity. the purified enzyme (designated Sp-beta gl-P) showed a specific activity of approximately 20.0 U/mg, an estimated molecular mass of 63 kDa after sodium dodecyl sulfate polyacrylamide gel electrophoresis, and isoelectric point of 5.0 by isoelectric focusing. Sp-beta gl-P has optimal activity at pH 4.0 and at 55 A degrees C. It was stable in a broad pH range at low temperatures and it was tolerant to ethanol and glucose, indicating suitable properties for winemaking. the hydrolysis of glycosidic terpenes was analyzed by adding Sp-beta gl-P directly to the wines. the released terpene compounds were evaluated by gas chromatography/mass spectrometry. the enzymatic treatment significantly increased the amount of free terpenes, suggesting that this enzyme could potentially be applicable in wine aroma improvement.
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