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紫色色杆菌苯丙氨酸羟化酶的异源表达及重组酶学性质研究

, PP. 152-158

Keywords: 苯丙氨酸羟化酶,紫色色杆菌,表达,纯化,酶学性质

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Abstract:

来源于紫色色杆菌(Chromobacteriumviolaceum)的苯丙氨酸羟化酶结构简单,性质更接近于人的苯丙氨酸羟化酶,具有潜在的医药应用价值。从紫色色杆菌基因组中克隆得到苯丙氨酸羟化酶基因pah。构建重组表达载体pET24a-pah,并在EscherichiacoliBL21(DE3)中实现高效表达。离子层析纯化后,重组蛋白比酶活高达503.2U/mg。酶学性质研究显示,该重组酶的最适温度为40℃左右,50℃时PAH的半衰期为15min;最适pH在7.5左右,在pH6-8范围内较稳定。37℃,pH7.5条件下,Km值为1.5mmol/L,Vmax为0.5mmol/min,kcat为5.05/s,催化效率kcat/Km为3.37L/mmol·s。

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