MEDIUM OPTIMIZATION FOR ISOFLAVONE AGLYCONES PRODUCTION FROM FERMENTED DEFATTED SOYBEAN FLOUR BY DISPLAY OF Β-GLUCOSIDASE FROM BACILLUS LICHENIFORMIS NRC24 AND EVALUATION OF THEIR ANTIVIRAL AND ANTITUMOR ACTIVITIES

Abstract

The submerged fermentation parameters of defatted soybean flour with Bacillus licheniformis NRC24 were optimized using the one-factor at-a-time method and Box-Behnken design to maximize the production of β-glucosidase and transformation of glucosidic isoflavones (daidzin and genistin) into their aglycones. One-factor-at-a-time optimization studies revealed that 80 g/L of defatted soybean flour , fermentation time of 16 h, temperature at 37°C, initial pH value of 7, inoculum volume of 5% (6x10⁷ CFU/mL), and medium volume of 50 mL in 250 mL Erlenmeyer flask with shaking conditions at 100 rpm were the most suitable conditions for aglycones production. The addition of cellobiose (1 percent w/v) to the soy flour medium resulted in a significantly increase in total aglycone. Further optimization using the Box-Behnken design resulted in a 1.11-fold increase in daidzein and genistein. Antiviral activity was observed against the (H5N1) influenza, herpes simplex type 1, and rota virus. The MTT cytotoxicity assay of the extract using MDCK, Vero, and MA104 cell lines revealed that fermented soybean flour reduced cytotoxicity more than the unfermented one. However, it showed higher cytotoxicity against HCT-116 colon cancer cells, MCF-7 breast and PC3 prostate cancer cells than the unfermented soy extract.