Pueraria candollei var. mirifica (PM) has a significant beneficial effect on postmenopausal symptoms associated with estrogen deficiency. However, the estrogenic activity and intestinal absorption of isoflavonoid glycosides derived from PM, such as daidzin and genistin, are significantly lower than those of their aglycones. To enhance the estrogenic activity of the PM extract, we developed β-glucosidase and its immobilized form to increase the PM aglycone content (daidzein and genistein). The enzyme immobilization was done by alginate beads, and the resulting β-glucosidase alginate beads have a diameter of about 0.20 cm. Response surface methodology (RSM) was used to optimize certain parameters, such as the pH, temperature, and ethanol concentration. The optimal conditions of β-glucosidase for daidzein and genistein production were pH of 4.8-4.9, a temperature in the range 46.3-49.1 °C, and ethanol concentration of 10.0-11.0%. The ANOVA results indicated that the design experiment involving free and immobilized β-glucosidase was the best fit by quadratic models, which had adjusted R 2 values between 0.8625 and 0.9318. Immobilized β-glucosidase can be reused up to nine times and maintained efficacy of greater than 90%. Treatment of the PM extract with β-glucosidase increased the estrogenic activity of the PM extract by 8.71- to 23.2-fold compared to that of the untreated extract. Thus, β-glucosidase has a high potential for enhancing the estrogenic activity of PM constituents, and it can be applied on an industrial scale to increase the utility of these natural products.
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