Purification and characterization of acid phosphatase from yellow lupin (Lupinus luteus) seeds
Purification and characterization of acid phosphatase from yellow lupin (Lupinus luteus) seeds
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DOI：
10.1016/s0167-4838(97)00055-1
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发表时间：
1997-08-15
期刊：
影响因子：
--
通讯作者：
Morawiecka, B
中科院分区：
文献类型：
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作者：
Olczak, M;Watorek, W;Morawiecka, B
黄羽扇豆(Lupinus Luteus)种子经硫酸铵分级沉淀、亲和层析、阳离子交换层析、凝胶过滤或反相高效液相色谱分离纯化得到均一的酸性磷酸酶(EC 3.1.3.2)。该酶是一个含有50kD和44kD亚基的二聚体，含有7.3%的碳水化合物，形成至少四条寡糖链，酶的最适pH为5.4。对硝基苯磷酸的表观K-m=0.28 mM，V-max=1780IU/mg蛋白质，纯化的磷酸酶在任何天然或合成底物中具有最高的比活力，该酶具有广泛的特异性，但环核苷酸、焦磷酸或植酸盐不被切割。钼酸盐、氟化物和磷酸盐对其有抑制作用。在EDTA、邻菲咯啉和酒石酸盐的存在下，酶的活性没有变化。(C)1997年爱思唯尔科学公司。
Acid phosphatase (EC 3.1.3.2) from yellow lupin (Lupinus luteus) seeds was purified to homogeneity by ammonium sulphate fractionation, affinity chromatography, cation-exchange chromatography, gel filtration or reverse-phase HPLC. The enzyme is a dimer with the 50 kD and 44 kD subunits and contains 7.3% of carbohydrate, forming at least four oligosaccharide chains, The optimum pH for the enzyme is 5.4. The apparent K-m for p-nitrophenyl phosphate was estimated to be 0.28 mM and V-max = 1780 IU/mg of protein, The purified phosphatase has the highest specific activities reported for any plant acid phosphatases measured for any native or synthetic substrate, The enzyme has broad specificity; however, cyclic nucleotides, pyrophosphate or phytate are not cleaved. It is inhibited by molybdate, fluoride and phosphate. There is no change in the enzyme activity in the presence of EDTA, phenanthroline and tartrate. (C) 1997 Elsevier Science B.V.