Preliminary studies on continuous recovery and purification of the penicillin acylase under pseudo-affinity conditions using phenyl-Sepharose gel

A continuous system for the recovery and purification of the penicillin acylase from crude extracts by recycling phenyl-Sepharose gel through three agitated vessels with disc filters of stainless steel is presented. The penicillin acylase present in the crude extract was absorbed into the phenyl-Sepharose gel under pseudo-affinity conditions (16% w/v of ammonium sulphate). After gel washing under the same conditions in the second vessel, enzyme desorption was performed using the same salt but at a lower concentration (6% w/v) in the third vessel. The preliminary studies reported here occurred without experimental difficulties, even at a gel concentration as high as 40% (v/v). The recovery of the penicillin acylase was achieved with high yield (74%), but a low purification factor of 2.4 was obtained owing to the use of a crude extract with low specific activity.