Lewendon A, Coggins J R
Biochem J. 1983 Jul 1;213(1):187-91. doi: 10.1042/bj2130187.
A procedure for the purification of 5-enolpyruvylshikimate 3-phosphate synthase from Escherichia coli is described. Homogeneous enzyme of specific activity 17.7 units/mg was obtained in 22% yield. The key purification step involves substrate elution of the enzyme from a cellulose phosphate column. The subunit Mr was estimated to be 49 000 by polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate. The native Mr was estimated to be 55 000 by gel filtration, indicating that the enzyme is monomeric.
本文描述了一种从大肠杆菌中纯化5-烯醇丙酮酸莽草酸-3-磷酸合酶的方法。以22%的产率获得了比活性为17.7单位/毫克的纯酶。关键的纯化步骤包括从磷酸纤维素柱上用底物洗脱该酶。在十二烷基硫酸钠存在下,通过聚丙烯酰胺凝胶电泳估计亚基分子量为49000。通过凝胶过滤估计天然分子量为55000,表明该酶是单体。