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茶树EGCG-O-甲基转移酶的纯化及酶学性质

作  者:
吕海鹏;张悦;费冬梅;林智
单  位:
国家茶产业工程技术研究中心
关键词:
茶树;EGCG-O-甲基转移酶;纯化;性质
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).
单  位:
National Engineering Research Center for Tea Processing,Tea Research Institute,Chinese Academy of Agricultural Sciences,Hangzhou 310008,China
关键词:
tea plant%EGCG-O-transferase%purification%characterization
摘  要:
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.

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