Characterization of β-glucosidase from Aspergillus terreus and its application in the hydrolysis of soybean isoflavones

An extracellular beta-glucosidase produced by Aspergillus terreus was identified, purified, characterized and was tested for the hydrolysis of soybean isoflavone. Matrix-assisted laser desorption/ionization with tandem time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF MS) revealed the protein to be a member of the glycosyl hydrolase family 3 with an apparent molecular mass of about 120 kDa. The purified beta-glucosidase showed optimal activity at pH 5.0 and 65 A degrees C and was very stable at 50 A degrees C. Moreover, the enzyme exhibited good stability over pH 3.0-8.0 and possessed high tolerance towards pepsin and trypsin. The kinetic parameters K (m) (apparent Michaelis-Menten constant) and V (max) (maximal reaction velocity) for p-nitrophenyl-beta-d-glucopyranoside (pNPG) were 1.73 mmol/L and 42.37 U/mg, respectively. The K (m) and V (max) for cellobiose were 4.11 mmol/L and 5.7 U/mg, respectively. The enzyme efficiently converted isoflavone glycosides to aglycones, with a hydrolysis rate of 95.8% for daidzin, 86.7% for genistin, and 72.1% for glycitin. Meanwhile, the productivities were 1.14 mmol/(L center dot h) for daidzein, 0.72 mmol/(L center dot h) for genistein, and 0.19 mmol/(L center dot h) for glycitein. This is the first report on the application of A. terreus beta-glucosidase for converting isoflavone glycosides to their aglycones in soybean products.