Evaluation of immobilized metal affinity chromatography for purification of penicillin acylase

The aim of this work was to test immobilized metal affinity chromatography (TMAC) for the purification of penicillin acylase. After evaluation of diff erent metals, Cu2+ was selected. Different samples were tested: pure penici llin acylase, industrial clarified feedstock and crude extract. After compa ring two eluents, NH,CI and imidazole, it appeared that although both gave good results for recovery and activity, NH,CI was a more selective eluent w ith 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%). (C) 2001 Elsevier Sci ence B.V. All rights reserved.