Purification and Characterization of an Ethanol-Tolerant β-Glucosidase from Sporidiobolus pararoseus and Its Potential for Hydrolysis of Wine Aroma Precursors

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作者
Milla Alves Baffi
Natália Martin
Thaise Mariá Tobal
Ana Lúcia Ferrarezi
João Henrique Ghilardi Lago
Maurício Boscolo
Eleni Gomes
Roberto Da-Silva
机构
[1] Uberlândia Federal University (UFU),Agricultural Sciences Institute (ICIAG)
[2] São Paulo State University (UNESP),Laboratory of Biochemistry and Applied Microbiology
[3] São Paulo Federal University (UNIFESP),undefined
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Yeasts; β-Glucosidase; Purification; Wine; Terpenes;
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摘要
An extracellular ethanol-tolerant β-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-βgl-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-βgl-P has optimal activity at pH 4.0 and at 55 °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-βgl-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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页码:1681 / 1691
页数:10
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