Purification of ACV synthetase fromStreptomyces clavuligerus

Summary By a combination of protamine sulfate treatment, ammonium sulfate fractionation, gel filtration and hydrophobic interaction chromatography, an active -(L--aminoadipyl)-L-cysteinyl-D-valine (ACV) synthetase from the prokaryoteStreptomyces clavuligerus was purified 135-fold to give a single major protein band on SDS-PAGE. Its size appears to be approximately 360 kDa which is very similar to that of the enzyme from the eukaryote,Cephalosporium acremonium.