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The purification of 5‐enolpyruvylshikimate 3‐phosphate synthase from an overproducing strain of Escherichia coli
Author(s) -
Duncan Kenneth,
Lewendon Ann,
Coggins John R.
Publication year - 1984
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(84)80027-7
Subject(s) - aroa , escherichia coli , microbiology and biotechnology , plasmid , biology , atp synthase , enzyme , recombinant dna , psti , bacteriophage , gene , enterobacteriaceae , biochemistry
The Escherichia coli aroA gene which codes for the enzyme 5‐enolpyruvylshikimate 3‐phosphate synthase (EPSP synthase) has been cloned from the λ‐transducing bacteriophage λp serC . The gene has been located on a 4.7 kilobase pair Pst I DNA fragment which has been inserted into the multiple copy plasmid pAT153. E. coli cells 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 untransformed E. coli .