野生荞麦籽粒中芦丁降解酶的分离纯化

Purification of Rutin Degrading Enzyme from Wild Buckwheat Kernels

从野生荞麦种金荞麦籽粒中提取出芦丁降解酶,具有将芦丁降解为槲皮素的作用。该粗酶液用葡聚糖凝胶柱G-100和DEAE阴离子交换柱分离纯化后,最终酶的比活性由59.2 U/mg提高到671.4 U/mg,纯化倍数和回收率分别为11.3倍和3.5%。所得酶液经SDS-PAGE检测酶分子量大小为60 kDa左右。对酶液反应条件进行研究,发现酶的最适温度为40℃,最适pH值为5.0。在工业生产上可借助芦丁降解酶以酶解法取代酸解法生产槲皮素。

The rutin degrading enzyme which was extracted from seeds of Fagopyrum cymosum,one of wild buckwheat species,could effectively degrade rutin into quercetin.The rutin-degrading enzyme was purified with Sephadex G-100 and DEAE cellulose-32 anion exchange column.The enzyme was purified to 11.3-fold with 3.5% recovery,and the specific activity increased from 59.2 U/mg to 671.4 U/mg.SDS-PAGE analysis revealed a single protein band corresponding to a molecular mass of 60 KDa.Maximum enzyme activity was obtained at 40 ℃ and pH 5.0.Based on the results in this study,it can be concluded that the rutin-degrading enzyme has great potential to replace the classical acid-hydrolytic method to produce quercetin in industrial application.