猪肝半乳糖激酶的纯化及性质
Purification and properties of galactokinase from pig liver.
作者信息
Ballard F J
出版信息
Biochem J. 1966 Jan;98(1):347-52. doi: 10.1042/bj0980347.
Abstract
- Galactokinase has been purified from the liver of young pigs by high-speed centrifugation, chromatography on Sephadex G-100 and DEAE-cellulose, and ammonium sulphate fractionation. 2. The enzyme preparation has a specific activity of 10-18mumoles of galactose phosphorylated/mg. of protein/min. at 37 degrees and has been purified 400-fold from the liver supernatant. 3. Purified liver galactokinase has Michaelis constants of 1x10(-4)-3x10(-4)m for galactose and 2x10(-4)m for ATP-Mg(2+), and the enzyme reaction produces equimolar amounts of galactose 1-phosphate and ADP. 4. Galactokinase phosphorylates 2-deoxygalactose and galactosamine in addition to galactose, has a pH optimum of 7.8, a Q(10) of 2, and is stimulated by cysteine and other thiols. 5. With the exception of substrate specificity, the properties of liver galactokinase are similar to galactokinase purified from yeast and Escherichia coli.
摘要
- 已通过高速离心、在葡聚糖凝胶G - 100和二乙氨基乙基纤维素上进行层析以及硫酸铵分级分离，从幼猪肝脏中纯化出半乳糖激酶。2. 该酶制剂在37℃时的比活性为每毫克蛋白质每分钟使10 - 18微摩尔半乳糖磷酸化，并且已从肝脏上清液中纯化了400倍。3. 纯化的肝脏半乳糖激酶对半乳糖的米氏常数为1×10(-4) - 3×10(-4)摩尔，对ATP - Mg(2+)的米氏常数为2×10(-4)摩尔，并且酶反应产生等摩尔量的1 - 磷酸半乳糖和ADP。4. 半乳糖激酶除了能使半乳糖磷酸化外，还能使2 - 脱氧半乳糖和半乳糖胺磷酸化，最适pH为7.8，温度系数Q(10)为2，并且受半胱氨酸和其他硫醇的刺激。5. 除底物特异性外，肝脏半乳糖激酶的性质与从酵母和大肠杆菌中纯化的半乳糖激酶相似。
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Purification and properties of galactokinase from pig liver.猪肝半乳糖激酶的纯化及性质
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