title: Evaluation of immobilized metal affinity chromatography for purification of penicillin acylase.
abstract: The aim of this work was to test immobilized metal affinity chromatography (IMAC) for the purification of penicillin acylase. After evaluation of different metals, Cu2+ was selected. Different samples were tested: pure penicillin acylase, industrial clarified feedstock and crude extract. After comparing two eluents, NH4Cl and imidazole, it appeared that although both gave good results for recovery and activity, NH4Cl was a more selective eluent with a higher fold purification than imidazole (4.64 versus 2.04). Moreover, we shown that a multistep gradient of NH4Cl, greatly increased the degree of purification (12.36) compared with the one-step process as control (4.64). In addition, good recovery was obtained (97-100%).
authors.affiliation: Ecole Supérieure de Technologie des Biomolécules de Bordeaux (ESTBB), UMR 5544, Université Victor Segalen Bordeaux 2, France.
authors.forename: V
authors.initials: V
authors.lastname: Fitton
authors.forename: X
authors.initials: X
authors.lastname: Santarelli
mesh_terms: D000821:Animal Feed / Q000032:analysis; D002413:Cations, Divalent; D002846:Chromatography, Affinity / Q000379:methods*; D003300:Copper; D004926:Escherichia coli; D008670:Metals; D009532:Nickel; D010399:Penicillin Amidase / Q000302:isolation & purification*; D011485:Protein Binding; D015032:Zinc
chemical_list: D002413:Cations, Divalent; D008670:Metals; D003300:Copper; D009532:Nickel; D010399:Penicillin Amidase; D015032:Zinc