Investigation of antipyrine containing monolithic column and its use for purification of penicillin amidohydrolase

In this study, development of poly(ethyleneglycoldimethacrylate-co-methacrloyl amidoantipyrine) [poly(EGDMA-co-MAAP)] monolithic affinitysorbent and its usefulness for penicillin amidohydrolase (penicillin acylase) (E.C3.5.1.11) adsorption were investigated. In the first step, 4-aminoantipyrine wasreacted with methacryloylchloride to produce methacrloyl amidoantipyrine(MAAP) monomer. The characterization of synthesized structure was evaluatedby FTIR and NMR. Then, monolithic affinity sorbent was obtained by thepolymerization of MAAP and EGDMA comonomers.In the second stage, the effect of initial concentration, pH, temperature,ionic strength and flow rate on the adsorption of penicillin amidohydrolase fromaqueous solution were studied.In the final step, adsorption and desorption studies using crude extracts ofpenicillin amidohydrolase taken from Penicillium Chrysogenum NRRL 807 andPenicillium Purpurogenum 455 microorganisms were investigated in continuoussystem.Keywords: Penicillin Amidohydrolase, Affinity Chromatography, Methacrloylamidoantipyrine (MAAP), Enzyme Purification, Bioligand