Purification and Characterization of Polyphenol Oxidase from Rhizome of Vallisneriasp inulosa
Guangfeng Wei, Meiqing Li, Shaowei Yu, XU Wen-juan, Jinling He
Abstract
Guangfeng Wei, Meiqing Li, Shaowei Yu, XU Wen-juan, Jinling He
Abstract
刺苦草根状茎PPO粗提液经过30%~80%(NH4)2SO4盐析、DEAE Sephadex A-50、SephadexG-75柱层析后,得到纯化的多酚氧化酶,纯化倍数为10.66倍,回收率为21.25%。纯化后的酶液经SDS-PAGE电泳确定为单一条带,相对分子质量约为33。对其特性研究表明,以邻苯二酚为底物,最适pH值为6.0,最适温度为50℃,Km=0.0344mol/L,90℃热处理10min可完全钝化PPO的活性,抗坏血酸、NaHSO3、草酸、焦磷酸钠、柠檬酸、L-苹果酸、β-环状糊精能有效抑制PPO的酶活性。
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刺苦草根状茎PPO粗提液经过30%~80%(NH4)2SO4盐析、DEAE Sephadex A-50、SephadexG-75柱层析后,得到纯化的多酚氧化酶,纯化倍数为10.66倍,回收率为21.25%。纯化后的酶液经SDS-PAGE电泳确定为单一条带,相对分子质量约为33。对其特性研究表明,以邻苯二酚为底物,最适pH值为6.0,最适温度为50℃,Km=0.0344mol/L,90℃热处理10min可完全钝化PPO的活性,抗坏血酸、NaHSO3、草酸、焦磷酸钠、柠檬酸、L-苹果酸、β-环状糊精能有效抑制PPO的酶活性。
Key concepts: Polyphenol oxidase, Rhizome, Polyphenol, Chemistry, Characterization (materials science), Food science, Botany, Biochemistry