New support material for penicillin g acylase purification

Penicillin G acylase (EC 3.5.1.11) has been used in the production ofsemi-synthetic penicillins in industrial processes. Purification of penicillin acylaseenzyme enables the production of semi-synthetic penicillins and cephalosporins.In the present study, bioaffinity chromatography adsorbent has prepared for thepurification of penicillin acylase in the form of reusable cryogel which has macropores. These characteristics have enabled cryogels to be used in nano-micro scalerange without any diffusion problem. In this study, firstly, methacryloylamidoantipyrine (MAAP) monomer has synthesized reacting 4-aminoantipyrinewith methacryloyl chloride and the obtained structure has characterized by FTIR.Then, poly(HEMA-co-MAAP) cryogel column has synthesized by thepolymerization of MAAP and 2-hydoxyethylmethacrylate (HEMA) comonomers.The specific surface area and swelling value of prepared cryogels have found tobe 25.8 m2g-1 and 1463.97 %, respectively. In the second stage of the study, theadsorption of penicillin acylase has investigated from aqueous solutions andnatural sources on cryogel columns containing MAAP. The maximum adsorptioncapacity of poly(HEMA-co-MAAP) cryogels has calculated as 190.2 mgg-1 (pH5.0 and 2h). The purification of Penicillin acylase from natural sources hasachieved by fast protein liquid chromatography (FPLC). Penicillin acylasepurification factor from the extract of Penicillium chrysogenum NRRL 1951,Penicillium crysogenum 807 and Penicillium purpurogenum 455 has found to be17.89, 54.9 and 10.0, respectively. Desorption of penicillin acylase from cryogelshas achieved by using 1 M NaOH. It has been determined that there was nosignificant decrease in adsorption capacity of cryogels after 6 adsorptiondesorptioncycles. As a result, poly(HEMA-co-MAAP) cryogels have a suitable