四川农业大学学报 ›› 2011, Vol. 29 ›› Issue (03): 391-396.doi: 10.3969/j.issn.1000-2650.2011.03.017

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野生荞麦籽粒中芦丁降解酶的分离纯化

唐宇1,2, 邵继荣1, 罗庆林1, 郑亚迪1, 孙俊秀2   

  1. 1. 四川农业大学生命科学与理学院, 四川 雅安 625014;
    2. 四川烹饪高等专科学校, 成都 610100
  • 收稿日期:2011-06-20 出版日期:2011-09-30 发布日期:2017-04-12
  • 通讯作者: 邵继荣(E-mail:shaojr007@163.com) E-mail:shaojr007@163.com
  • 基金资助:
    荞麦野生种质资源的保护和利用研究[四川省科技厅重点攻关课题(04NG001-015)];苦荞异黄酮代谢途径关键酶基因的克隆[四川省科技厅重点攻关课题(04NG001-015,2006Z08-012)]

Purification of Rutin Degrading Enzyme from Wild Buckwheat Kernels

TANG Yu1,2, SHAO Ji-rong1, LUO Qing-lin1, ZHENG Ya-di1, SUN Jun-xiu2   

  1. 1. College of Biology and Science, Sichuan Agricultural University, Yaan 625014, Sichuan, China;
    2. Sichuan Higher Institute of Cuisine, Chengdu 610100, China
  • Received:2011-06-20 Online:2011-09-30 Published:2017-04-12

摘要: 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。

关键词: 野生荞麦, 芦丁降解酶, 槲皮素, 芦丁

Abstract: 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.

Key words: wild buckwheat, rutin degrading enzyme, quercetin, rutin

中图分类号: 

  • Q599