How to detect the enzyme naringinase

The enzyme naringinase can hydrolyze many kinds of polysaccharide, enzyme naringinase is often used in food processing industry. The process of enzymatic hydrolysis of naringinase reported determination method can not detect and monitor food glucoside. The substrate and product determination of naringin hydrolysis of naringin, arbutin and Yang Meigan glucoside by using liquid chromatography. The chemical reaction and study of naringinase in the application process and determine its activity. Analysis of the chromatographic conditions: SymmetryC18 reversed-phase column. The detection wavelength of 280rim, the column temperature of 35 DEG C, injection 20 L, flow rate of 0.4mL / min; the mobile phase of methanol water (A), (B) and acetonitrile (C), solvent gradient elution procedure is: O ~ 7min (V, MB / C V / V / =62 / 12 / 26); 7 9min, A / B / C (V / V / V) =15 / 35 / 50; 9 ~ 15min, A / B / C (V / V / V) =15 / 35 / 50; 15-17min, A / B / C (v v / V / =62 / 12 / 26); 17 ~ 20min, A / B / C (V, V, V, =62 / 12, 26). This method can effectively separate the naringin, Yang Meigan, arbutin and salicin as substrate and its hydrolysis products; Determination of the relative standard deviation was less than 1% of naringin and naringenin by using this method, the recovery rate in 100.4%. 105.2%, 94.5%-117.3%. In 50oC, pH4.0, Michaelis constant Km of naringinase hydrolysis of naringin was 0.10mmoUL, the maximum reaction rate of Vmax was 256U / Mg, Kat / s 1487. The HPLC method can detect naringinase on various glycoside hydrolysis process, enzyme activity and enzyme hydrolysis kinetics can accurate determination of naringin hydrolysis of naringin, plays an important role in the further application of naringinase in food.