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Abstract: An extracellular β;-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 β;-glucosidase showed optimal activity at pH 5.0 and 65 °C and was very stable at 50 °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-β;-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·h) for daidzein, 0.72 mmol/(L·h) for genistein, and 0.19 mmol/(L·h) for glycitein. This is the first report on the application of A. terreus β;-glucosidase for converting isoflavone glycosides to their aglycones in soybean products.

土曲霉来源的β-葡萄糖苷酶的酶学特性及其对大豆异黄酮的水解

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[39]List of electronic supplementary materials

[40]Fig. S1 Morphology of A. terrues culture mycelium