An efficient three steps preparative purification of penicillin acylase from Escherichia coli cells.

A new and efficient safe system for the purification of the penicillin acylase from Escherichia coli G271 is presented. It was found that after a selective precipitation with ammnonium sulphate, followed by two chromatographic steps (anion exchange followed by adsorption on hydroxyapatite support), the enzyme was enriched 98 times with a 100% activity recovery. An original way has also been used to study the chromatographic separation of the protein mixture in three major categories on DEAE resin, by an analysis of the concentrations of the different species in the breakthrough curve obtained from a complete saturation of the column.