食品科学2017，Vol.38Issue(8)：17-23,7.DOI:10.7506/spkx1002-6630-201708004
鲜切荸荠类黄酮3'-羟化酶分离纯化及其动力学性质
Isolation, Purification and Characterization of Flavonoid 3'-Hydroxylase from Fresh-Cut Chinese Wate Chestnut
摘要
Abstract
Crude flavonoid 3'-hydroxylase (F3'H) from fresh-cut Chinese water chestnut was extracted by Tris-HC1 buffer (pH 7.5),and then isolated and purified successively by ammonium sulfate precipitation,dialysis,DEAE-cellulose ionexchange column chromatography and Sephardi G-100 gel filtration chromatography.After purification,the enzyme was finally purified 14.01 folds with a specific activity of 478.49 U/mg and a protein yield of 6.38％.The purified enzyme showed a single protein band with a molecular mass of about 53.09 kD as estimated via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).Further enzymatic characterization showed that the purified enzyme activity attained its maximal value at 30 ℃ and pH 7.5,and had a Km and Vmax of 1.08 mmol/L and 416.67 U/(min·mL),respectively,using naringein as the substrate.The flavonoid 3'-hydroxylase activity was slightly inhibited by Ca2+ and citric acid and strongly inhibited by Na+.However,Fe2+,Mg2+,NADPH and ascorbic acid strongly activated its activity.关键词
鲜切荸荠/类黄酮3'-羟化酶/分离纯化/动力学性质Key words
fresh-cut Chinese water chestnut/flavonoid 3'-hydroxylase/isolation and purification/enzymatic characterization分类
轻工纺织引用本文复制引用
何凤平,潘永贵,张伟敏..鲜切荸荠类黄酮3'-羟化酶分离纯化及其动力学性质[J].食品科学,2017,38(8):17-23,7.基金项目
国家自然科学基金地区科学基金项目(31360414) （31360414）
