利用青霉素-琼脂糖凝胶柱从大肠杆菌B中纯化D-丙氨酸羧肽酶
Purification of D-alanine carboxypeptidase from Escherichia coli B on a penicillin-Sepharose column.
作者信息
Gorecki M, Bar-Eli A, Burstein Y, Patchornik A, Chain E B
出版信息
Biochem J. 1975 Apr;147(1):131-7. doi: 10.1042/bj1470131.
Abstract
- A soluble D-alanine carboxypeptidase from Escherichia coli strain B was purified on a p-aminobenzylpenicillin-Sepharose column. This one-step chromatography followed by an (NH4)2SO4 precipitation yielded an enzyme purified 1200-fold and some of its properties are reported. 2. The pure D-alanine carboxypeptidase was devoid of D-alanine carboxypeptidase II activity and migrated as a single protein band on analytical disc gel electrophoresis. 3. Triton X-100 in the purification procedure is an absolute requirement for obtaining a stable enzyme. 4. The enzymic activity of D-alanine carboxypeptidase was greatly affected in solution of high salt concentrations and varied somewhat with the nature of the cation tested.
摘要
- 从大肠杆菌B菌株中纯化出一种可溶性D-丙氨酸羧肽酶，该酶在对氨基苄青霉素-琼脂糖柱上进行纯化。经这一步层析，再进行硫酸铵沉淀，得到了纯化1200倍的酶，并报道了其一些性质。2. 纯化的D-丙氨酸羧肽酶没有D-丙氨酸羧肽酶II活性，在分析盘状凝胶电泳上迁移为单一蛋白条带。3. 纯化过程中的吐温X-100是获得稳定酶的绝对必要条件。4. D-丙氨酸羧肽酶的酶活性在高盐浓度溶液中受到极大影响，并且随所测试阳离子的性质略有变化。
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