Pectinase and naringinase were investigated for improving the production of pummelo juice by increasing the juice yield and eliminating juice bitterness. Compared to a control, the enzymatic treatment significantly (p<0.05) increased the juice yield, soluble pectin and total soluble solid (TSS) contents, and the clarity, while decreasing the concentrations of the bitter chemicals naringin, limonin, and nomilin. A combined processing treatment of peeling and enzymatic hydrolyses using 5U/g of pectinase and 0.4 U/g of naringinase at 50oC for 60 min resulted in a juice yield of 42.3%, a TSS content of 11.4oBx, a titrable acidity (TA) of 0.96%, a vitamin C concentration of 38.4 mg/100 mL, and concentrations of naringin, limonin, and nomilin of 42.4 μg/mL, 33.5 μg/mL, and 13.3 μg/mL, respectively. The enzymatic method was effective and practical for production of high quantity pummelo juice.

An HPLC method that can separate naringin, prunin, and naringenin was used to help accurately measure the activities of naringinase and its subunits (α-l-rhamnosidase and β-d-glucosidase). The activities of the naringinase and β-d-glucosidase were determined through an indirect calculation of the naringenin concentration to avoid interference from its poor solubility. The measured enzymatic activities of the naringinase complex, α-l-rhamnosidase, and β-d-glucosidase were the as same as their theoretical activities when the substrates’ (i.e., naringin or prunin) concentrations were 200 μg/mL, and the enzyme concentrations were within the range of 0.06–0.43, 0.067–0.53, and 0.15–1.13 U/mL, respectively. The β-d-glucosidase had a much higher Vmax than either naringinase or α-l-rhamnosidase, implying the hydrolysis of naringin to prunin was the limiting step of the enzyme reaction. The reliability of the method was finally validated through the repeatability test, indicating its feasibility for the determinations of the naringinase complex.