Efficient enzymatic preparation of Esculentoside B following condition optimization by response surface methodology

This study aims to provide a practical method for highly efficient preparation of EsB using esculentoside A (EsA, phytolaccagenin 3-O-β-D-glucopyranosyl (1→4)-β-D-xylopyranoside) as the starting material. β-D-glucosidase from snailase was used to catalyze the formation of EsB, and the product was then purified and fully characterized by both HRMS and NMR. To prepare EsB in a more cost-effective way, response surface methodology (RSM) was used to explore the potential effects of the reaction conditions (such as reaction temperature, pH, enzyme load, and reaction time) on the conversion rates of EsA. The highest EsB yield of 0.66mg/ml was obtained experimentally under optimized conditions as follows: temperature 48.28°C, pH 6.4, enzyme load 4.43%, and reaction time 2.73h. This result agreed well with the predicted yield of 0.68mg/ml by RSM. The enzymatic kinetics of this biotransformation was characterized at the optimum pH and temperature. The S 50 value was evaluated as 167.4μM, while the V max value was 345.6 nmol/min/mg. In summary, this study provided a mild and practical method for the highly efficient preparation of EsB from EsA, which held great promise for large scale production of EsB. Graphical abstract
Source: Journal of Molecular Catalysis B: Enzymatic - Category: Biochemistry Source Type: research