Direct comparison between ion-exchange chromatography and aqueous two-phase processes for the partial purification of penicillin acylase produced by E. coli

摘要

A direct comparison of a chromatography and an aqueous two-phase system (ATPS) processes for the partial purification of penicillin acylase (PA) produced by a recombinant strain of E. coli, was performed. An established chromatography process for the recovery of PA was selected as a model system and characterised for comparison with a developed ATPS prototype process. PEG-phosphate systems were selected for the recovery of PA over PEG-citrate systems, since higher enzyme recovery and increased purity was obtained. ATPS proved to be suitable to process highly concentrated disrupted extract (35%, w/w) and maintain a high top phase enzyme recovery. In the direct comparison of the processes, the superiority of the ATPS approach was highlighted since a reduction of the number of unit operations from 7 to 4 was achieved. An outline economic analysis based on the cost of separation agent of the processes favour the ATPS process. in which a gross cost reduction of 37% (from