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二氢嘧啶酶产生菌Pseudomonasputida 980 1经过超声波粉碎及分级盐析 ,DEAE 纤维素和羟基磷灰石柱层析 ,SephadexG 2 0 0凝胶过滤得到纯化 2 49倍的酶液。SDS PAGE显示为单带 ,单体分子量为 380 0 0 ,SephadexG 2 0 0凝胶过滤测得分子量为 15 6 0 0 0。二氢嘧啶酶对苯海因的km 为 2 .39× 10 -2 mol/L ,其最适 pH和最适温度分别是 8.5～ 8.7和 34℃～ 35℃。 1mmol/L的EDTA和 β 巯基乙醇对酶活力有强烈的抑制作用。
The dihydropyrimidinase-producing bacteria Pseudomonas putida 980 1 was sonicated and graded by salting-out, DEAE cellulose and hydroxyapatite column chromatography, and gel filtration was performed on Sephadex G 20 0 gel to obtain a purified enzyme solution of 494 times. SDS PAGE showed a single band, the molecular weight of 380 0 0, SephadexG 2 0 0 gel filtration measured molecular weight of 15 6 0 0 0. The dihydropyrimidinase to diphenhydramine was 2.39 × 10 -2 mol / L, and the optimum pH and optimum temperature were 8.5 ~ 8.7 and 34 ℃ ~ 35 ℃, respectively. 1mmol / L EDTA and β-mercaptoethanol had a strong inhibitory effect on the enzyme activity.