Purification and Characterization of a Naringinase from <italic>Cryptococcus albidus</italic>.

Naringinase which was extracted from the fermented broth of <italic>Cryptococcus albidus</italic> was purified about 42-folds with yield 0.7% by sulfate fractionation and chromatography on Toyopearl HW-60, Fractogel DEAE-650-s, and Sepharose 6B columns. Molecular weight of protein determined by gel filtration and SDS-PAGE was 50 kDa. Naringinase of <italic>C. albidus</italic> includes high content of the dicarbonic and hydrophobic amino acids. Enzyme contains also carbohydrate component, represented by mannose, galactose, rhamnose, ribose, arabinose, xylose, and glucose. The enzyme was optimally active at pH 5.0 and 60 °C. Naringinase was found to exhibit specificity towards <italic>p</italic>-nitrophenyl-α-L-rhamnose, <italic>p</italic>-nitrophenyl-β-D-glucose, naringin, and neohesperidin. Its <italic>K</italic>_m towards naringin was 0.77 mM and the <italic>V</italic>_max was 36 U/mg. Naringinase was inhibited by high concentrations of reaction product—L-rhamnose. Enzyme revealed stability to 20% ethanol and 500 mM glucose in the reaction mixture that makes it possible to forecast its practical use in the food industry in the production of juices and wines. [ABSTRACT FROM AUTHOR]