食品科学 ›› 2013, Vol. 34 ›› Issue (9): 194-197.doi: 10.7506/spkx1002-6630-201309040

• 生物工程 • 上一篇    下一篇

茶树EGCG-O-甲基转移酶的纯化及酶学性质

吕海鹏,张 悦,费冬梅,林 智*   

  1. 中国农业科学院茶叶研究所,国家茶产业工程技术研究中心,浙江 杭州 310008
  • 收稿日期:2012-07-03 修回日期:2013-03-15 出版日期:2013-05-15 发布日期:2013-05-07
  • 通讯作者: 林 智 E-mail:lvhaipeng@caas.net.cn
  • 基金资助:

    中央级公益性科研院所基本科研业务费专项(2012zl053);国家自然科学基金项目(30972404);
    国家现代农业产业技术体系建设专项(CARS-23)

Purification and Characterization of Recombinant EGCG-O-transferase from Tea Plant

LÜ Hai-peng,ZHANG Yue,FEI Dong-mei,LIN Zhi*   

  1. National Engineering Research Center for Tea Processing, Tea Research Institute, Chinese Academy of Agricultural Sciences,
    Hangzhou 310008, China
  • Received:2012-07-03 Revised:2013-03-15 Online:2013-05-15 Published:2013-05-07
  • Contact: LIN Zhi E-mail:lvhaipeng@caas.net.cn

摘要:

对重组茶树EGCG-O-甲基转移酶进行分离纯化,并对其酶学性质进行研究。结果表明:经HisTMTrap层析纯化后,获得了纯度超过95%的重组融合蛋白,纯化倍数为22.51倍,酶活回收率为34.54%,酶比活力达到0.0186U/mg;酶分子质量约为27.6kD;该酶的最适反应温度为35℃、最适pH7.5;以表没食子儿茶素没食子酸酯(EGCG)作为底物,酶学动力学常数Km为0.100mmol/L,Vmax为7.485mg/(L•min)。

关键词: 茶树, EGCG-O-甲基转移酶, 纯化, 性质

Abstract:

Recombinant EGCG-O-transferase from tea plant was expressed in E.coli. The enzyme was then purified
from cell lysate using HisTMtrap affinity column and its enzymatic properties were investigated. After purification, a
pure protein with purity over 95%, recovery of 34.54% and specific enzyme activity of 0.0186 U/mg was obtained.
Further analysis showed that the recombinant protein had a molecular mass of 27.6 kD. It exhibited the highest
activity under the conditions of pH 7.5 and 35 ℃. Kinetic studies showed that Km and Vmax of EGCG-O-transferase
using EGCG as substrate were 0.100 mmol/L and 7.485 mg/(L·min), respectively.

Key words: tea plant, EGCG-O-transferase, purification, characterization

中图分类号: