Localization, purification, and characterization of a novel β‐glucosidase from Hanseniaspora uvarum Yun268

Abstract β‐Glucosidase is a key enzyme that hydrolyzes nonvolatile glycosylated precursors of aroma compounds and enhances the organoleptic quality of wines. In this study, a novel β‐glucosidase from Hanseniaspora uvarum Yun268 was localized, purified, and characterized. Results indicated that β‐glucosidase activity was mainly distributed within the cells. After purification via ammonium sulfate precipitation combined with chromatography, β‐glucosidase specific activity increased 8.36 times, and the activity recovery was 56.90%. The enzyme had a molecular mass of 74.22 kDa. It has a Michaelis constant ( K m ) of 0.65 mmol/L, and a maximum velocity ( V max ) of 5.1 nmol/min under optimum conditions; and K m of 0.94 mmol/L, and V max of 2.8 nmol/min under typical winemaking conditions. It exhibited the highest activity at 50°C and pH 5.0 and was stable at a temperature range of 20–80°C and pH range of 3.0–8.0. The enzyme has good tolerance to Fe 3+ , especially maintaining 93.68% of its activity with 10 mmol/L of Fe 3+ . Ethanol (<20%) and glucose (<150 g/L) inhibited its activity only slightly. Therefore, β‐glucosidase from H. uvarum Yun268 has excellent biochemical properties and a good application potential in winemaking. Practical Application Winemaking is a biotechnological process in which exogenous β‐glucosidase is used to overcome the deficiency of endogenous β‐glucosidase activity in grapes. By localizing, purifying, and characterizing of β‐glucosidase from Hanseniaspora uvarum Yun268, it is expected to reveal its physical and chemical characteristics to evaluate its oenological properties in winemaking. The results may provide the basis for promoting the release of varietal aroma and improving wine sensory quality in the wine industry.