The purification of 5‐enolpyruvylshikimate 3‐phosphate synthase from an overproducing strain of Escherichia coli
The purification of 5‐enolpyruvylshikimate 3‐phosphate synthase from an overproducing strain of Escherichia coli
复制标题
从大肠杆菌过量生产菌株中纯化 5-烯醇丙酮莽草酸 3-磷酸合酶
DOI：
10.1016/0014-5793(84)80027-7
复制
发表时间：
1984
期刊：
影响因子：
3.5
通讯作者：
J. Coggins
中科院分区：
文献类型：
--
作者：
K. Duncan;A. Lewendon;J. Coggins
从λ转导噬菌体λpserC中克隆了编码5-烯醇丙酮基莽草酸3-磷酸合成酶(EPSP合成酶)的大肠杆菌aroA基因。该基因位于4.7kb的PstI DNA片段上，插入到多拷贝载体pAT153中，转化大肠杆菌后，EPSP合成酶的产量提高了100倍。设计了一种纯化毫克量均一酶的简单方法。所产生的酶与从未转化的E分离出来的酶没有区别。Coli.
TheEscherichia coli aroAgene which codes for the enzyme 5‐enolpyruvylshikimate 3‐phosphate synthase (EPSP synthase) has been cloned from the λ‐transducing bacteriophage λpserC. The gene has been located on a 4.7 kilobase pairPstI DNA fragment which has been inserted into the multiple copy plasmid pAT153.E. colicells transformed with this recombinant plasmid overproduce EPSP synthase 100‐fold. A simple method for the purification of homogeneous enzyme in milligram quantities has been devised. The resulting enzyme is indistinguishable from enzyme isolated from untransformedE. coli.