Improved chromatographic purification of pea seedlings diamine oxidase.

M Wimmerová, Z Glatz, O Janiczek, L Macholán
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引用次数: 4

Abstract

An improved and simplified purification procedure has been developed for the isolation of diamine oxidase from pea seedlings (DAO EC 1.4.3.6). It involves ammonium sulphate precipitation, hydrophobic interaction chromatography, ion-exchange chromatography and size-exclusion chromatography. The homogeneity of the final enzyme preparations and molecular weight were determined by size-exclusion chromatography and by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate (SDS PAGE). The isoelectric point of 7.35 +/- 0.05 was determined by chromatofocusing and by polyacrylamide gel isoelectric focusing.

豌豆幼苗二胺氧化酶的改进色谱纯化。
建立了一种改进和简化的分离豌豆幼苗二胺氧化酶的纯化方法(DAO EC 1.4.3.6)。它包括硫酸铵沉淀,疏水相互作用层析,离子交换层析和尺寸排除层析。在十二烷基硫酸钠(SDS PAGE)的存在下,采用粒径排除色谱法和聚丙烯酰胺凝胶电泳法测定最终酶制剂的均匀性和分子量。采用色谱聚焦法和聚丙烯酰胺凝胶等电聚焦法测定等电点为7.35 +/- 0.05。
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