Impacts of Ultrasonic Treatment for Black Soybean Okara Culture Medium Containing Choline Chloride on the β-Glucosidase Activity of BCRC 10357.

The effects of ultrasonic treatment for the culture medium of solid black soybean okara with choline chloride (ChCl) on the survival and β-glucosidase activity of BCRC 10357 (-BCRC10357) were investigated. A mixture of 3% dried black soybean okara in de Man-Rogosa-Sharpe (/) was used as the Oka medium. With ultrasonic treatment (40 kHz/300 W) of the Oka medium at 60 °C for 3 h before inoculation, the β-glucosidase activity of -BCRC10357 at 12 h and 24 h of incubation amounted to 13.35 and 15.50 U/mL, respectively, which was significantly larger than that (12.58 U/mL at 12 h and 2.86 U/mL at 24 h) without ultrasonic treatment of the medium. This indicated that ultrasonic treatment could cause the microstructure of the solid black soybean okara to be broken, facilitating the transport of ingredients and -BCRC10357 into the internal structure of the okara for utilization. For the effect of ChCl (1, 3, or 5%) added to the Oka medium (/) with ultrasonic treatment before inoculation, using 1% ChCl in the Oka medium could stimulate the best response of -BCRC10357 with the highest β-glucosidase activity of 19.47 U/mL in 12 h of incubation, showing that -BCRC10357 had a positive response when confronting the extra ChCl that acted as an osmoprotectant and nano-crowder in the extracellular environment. Furthermore, the Oka medium containing 1% ChCl with ultrasonic treatment led to higher β-glucosidase activity of -BCRC10357 than that without ultrasonic treatment, demonstrating that the ultrasonic treatment could enhance the contact of ChCl and -BCRC10357 to regulate the physiological behavior for the release of enzymes. In addition, the analysis of the isoflavone content and antioxidant activity of the fermented product revealed that the addition of 1% ChCl in the Oka medium with ultrasonic treatment before inoculation allowed a higher enhancement ratio for the biotransformation of isoflavone glycosides to their aglycones, with a slight enhancement in the antioxidant activity at 24 h of fermentation. This study developed a methodology by combining ultrasonic treatment with a limited amount of ChCl to allow the culture medium to acclimate -BCRC10357 and release high levels of β-glucosidase, and this approach has the potential to be used in the fermentation of okara-related products as nutritional supplements in foods.