摘要
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。
Dihydropyrimidinase is purified from \%Pseudomonas putida\% 9801,by a procedure including ammonium sulfate fractionation, column chromatography on DEAE-cellulose and hydroxylapatite,and gel filtration on Sephadex G-200.The enzyme is homogeneous according to the criteria of SDS-PAGE and gel filtration. The molecular weight of the enzyme is determined to be 156000 by Sephadex G-200 gel filtration. Its optimum pH and temperature are 8.5~8.7 and 34℃~35℃ respectively.
出处
《中国药科大学学报》
CAS
CSCD
北大核心
2000年第5期389-392,共4页
Journal of China Pharmaceutical University
基金
江苏省应用基础研究计划项目!BJ970 80
关键词
二氢嘧啶酶
苯海因
分离
纯化
Dihydropyrimidinase
Sephadex G-200 gel filtration
DEAE-cellulose
Hydroxylapatite column chromatography
\%Pseudomonas putida 9801\%
Isolation
Purification
