Purification and characterization of β-glucosidase from Oenococcus oeni 31MBR

This study was designed to characterize a β-glucosidase from Oenococcus oeni 31MBR, a strain widely used in Chinese winemaking. An intracellular β-glucosidase (EC 3.2.1.21) was partially purified using a combination of ammonium sulfate precipitation and chromatographic methods. A single band was obtained in SDS-PAGE electrophoresis, indicating that the enzyme was highly purified and had an estimated molecular mass of 38.9 kDa. The enzyme exhibited highest activity at pH 4.5–5.0. The optimum temperature was 45 °C. Ethanol promoted the activity of this enzyme up to three times. Among the several metal ions assayed, only Mn2+ exhibited a partial promotion effect. The Km and Vmax values for p-nitrophenyl-β-d-glucopyranoside were 1.05 mmol/L and 0.957 nmol/min, respectively. Up to now, this study contains the first characterization of a native β-glucosidase purified from crude extracts of O. oeni 31MBR.