Department of Agronomy and Range Science and Department of Veterinary Microbiology, University of California, Davis, California 95616.
Plant Physiol. 1970 Aug;46(2):204-7. doi: 10.1104/pp.46.2.204.
A rapid procedure was developed for purifying ribulose 1,5-diphosphate carboxylase from barley leaves. After (NH(4))(2)SO(4) fractionation, the unique sedimentation properties of the enzyme were exploited to effect a single step purification to 90% homogeneity. High speed centrifugation pelleted the enzyme with complete recovery of activity. Residual impurities were then removed by diethylaminoethyl cellulose chromatography and density gradient centrifugation. The purified protein exhibited size heterogeneity due to polymerization. The polymerization products were enzymatically active aggregates of ribulose 1,5-diphosphate carboxylase and were precipitated by an antibody specific for the enzyme.
建立了从大麦叶片中快速纯化核酮糖 1,5-二磷酸羧化酶的方法。经（NH4）2SO4分级沉淀后，利用该酶独特的沉降特性进行一步纯化，可达到 90%的均一性。高速离心可使酶完全回收并保持活性。然后通过二乙基氨基乙基纤维素层析和密度梯度离心去除残留杂质。纯化的蛋白由于聚合而表现出大小不均一性。聚合产物是核酮糖 1,5-二磷酸羧化酶的酶活性聚合体，可被针对该酶的抗体沉淀。
